Open Collections

UBC Theses and Dissertations

UBC Theses Logo

UBC Theses and Dissertations

Studies on early development and endogenous gibberellins in red alder (Alnus rubra Bong.) Elliott, Dave Michael 1980

Your browser doesn't seem to have a PDF viewer, please download the PDF to view this item.

Notice for Google Chrome users:
If you are having trouble viewing or searching the PDF with Google Chrome, please download it here instead.

Item Metadata

Download

Media
831-UBC_1981_A1 E44.pdf [ 8.65MB ]
Metadata
JSON: 831-1.0095116.json
JSON-LD: 831-1.0095116-ld.json
RDF/XML (Pretty): 831-1.0095116-rdf.xml
RDF/JSON: 831-1.0095116-rdf.json
Turtle: 831-1.0095116-turtle.txt
N-Triples: 831-1.0095116-rdf-ntriples.txt
Original Record: 831-1.0095116-source.json
Full Text
831-1.0095116-fulltext.txt
Citation
831-1.0095116.ris

Full Text

STUDIES ON EARLY DEVELOPMENT AND ENDOGENOUS GIBBERELLINS IN RED ALDER (ALNUS RUBRA BONG.) B . S c , ( F o r . ) H o n s . , A u s t r a l i a n N a t i o n a l U n i v e r s i t y , 1971 A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY i n THE FACULTY OF GRADUATE STUDIES ( F a c u l t y o f F o r e s t r y ) We a c c e p t t h i s t h e s i s as c o n f o r m i n g t o the r e q u i r e d s t a n d a THE UNIVERSITY OF BRITISH COLUMBIA SEPTEMBER 1980 (c)Dave M i c h a e l E l l i o t t by DAVE MICHAEL ELLIOTT In p r e s e n t i n g t h i s t h e s i s in p a r t i a l f u l f i l m e n t o f the r e q u i r e m e n t s f o r an advanced degree at the U n i v e r s i t y o f B r i t i s h C o l u m b i a , I a g r e e that the L i b r a r y s h a l l make i t f r e e l y a v a i l a b l e f o r r e f e r e n c e and s t u d y . I f u r t h e r agree t h a t p e r m i s s i o n f o r e x t e n s i v e c o p y i n g o f t h i s t h e s i s f o r s c h o l a r l y p u r p o s e s may be g r a n t e d by the Head o f my Department o r by h i s r e p r e s e n t a t i v e s . It i s u n d e r s t o o d tha t c o p y i n g o r p u b l i c a t i o n o f t h i s t h e s i s f o r f i n a n c i a l g a i n s h a l l not be a l l o w e d w i t h o u t my w r i t t e n p e r m i s s i o n . Department o f F o r e s t r y  The U n i v e r s i t y o f B r i t i s h Co lumbia V ancouver 8, Canada Date December 1 9 8 0 i i ABSTRACT The t h e s i s i s p r e s e n t e d i n f i v e p a r t s . In the f i r s t , our p r e s e n t knowledge o f red a l d e r {Alnus rubra Bong . ) i s summar ized and shows a l a c k o f s p e c i f i c knowledge about i t s seed and r e p r o d u c t i v e b i o l o g y . The aims o f the f i r s t s t u d y were (1) t o d e t e r m i n e the o c c u r r e n c e and ( i f p r e s e n t ) the n a t u r e o f seed do rmancy ; (2) t o d e t e r m i n e whe the r seed v i g o u r was r e l a t e d t o g e o g r a p h i c l o c a t i o n o f d i f f e r e n t p r o v e n a n c e s ; and (3) t o examine the i n f l u e n c e o f s t r a t i f i c a t i o n and p r e s o a k i n g on seed g e r m i n a t i o n . G e r m i n a t i o n t e s t s were c a r r i e d out on s t r a t i f i e d , p r e s o a k e d and u n t r e a t e d seeds c o l l e c t e d f rom s e v e r a l l o c a t i o n s t h r o u g h o u t the s p e c i e s ' r a n g e . W i t h one e x c e p t i o n ( seed f rom Haney , B. C. c o l l e c t e d i n 1976) , dormancy was no t a s i g n i f i c a n t f a c t o r i n p r e v e n t i n g g e r m i n a t i o n o f u n t r e a t e d s e e d s . G e r m i n a t i o n o f t he se e x c e p t i o n a l seeds was improved by c o l d s t r a t i f i c a t i o n f o r e i g h t days and t o a l e s s e r e x t e n t , by p r e -s o a k i n g i n t ap w a t e r f o r 2h h o u r s . Seed c o l l e c t e d f rom the same a r e a b e f o r e and a f t e r t h e s e t e s t s showed no dormancy . G e r m i n a t i o n p e r c e n t was r e l a t i v e l y h i g h f o r t r e a t e d and u n t r e a t e d seeds of o t h e r p rovenances and showed few d i f f e r e n c e s between p r o v e n a n c e s . The re were more p ronounced d i f f e r e n c e s i n seed v i g o u r , as measured by g e r m i n a t i o n v a l u e , between p r o v e n a n c e s . The re appea red t o be no d i r e c t r e l a t i o n s h i p between the d i f f e r e n c e s o b s e r v e d and g e o g r a p h i c f a c t o r s , as found i n many w ide r a n g i n g s p e c i e s n a t i v e t o w e s t e r n N o r t h A m e r i c a . The aims o f the second s t u d y were (1) t o examine the e f f e c t s o f f e r t i l i z a t i o n and d i f f e r e n t s o i l t y p e s on r o o t and n o d u l e d e v e l o p m e n t , h e i g h t g r o w t h , d r y w e i g h t a c c u m u l a t i o n and r o o t - s h o o t d r y w e i g h t r a t i o o f s e e d l i n g s o f red a l d e r f rom d i f f e r e n t l o c a t i o n s i n i t s n a t u r a l d i s t r i b u t i o n ; and (2) t o r e l a t e o b s e r v e d i i i d i f f e r e n c e s between p rovenances t o t h e i r g e o g r a p h i c l o c a t i o n . E a r l y s e e d l i n g deve lopment i n f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s front s e v e r a l s o u r c e s and grown i n d i f f e r e n t c o m b i n a t i o n s o f sand and a loamy s o i l , was e x a m i n e d . Root deve lopment was i n f l u e n c e d by the t ype o f growth medium and the n u t r i e n t s t a t u s o f t he same. N o d u l a t i o n , howeve r , was l i t t l e a f f e c t e d by t h e s e f a c t o r s . Nodu le deve lopment was more e x t e n s i v e on upper r oo t b r anches than on lower o n e s . F e r t i l i z a t i o n i n c r e a s e d h e i g h t and d r y w e i g h t , and lowered r o o t - s h o o t d r y w e i g h t r a t i o o f s e e d l i n g s . H e i g h t g rowth and d r y w e i g h t a c c u m u l a t i o n were g r e a t e r , and r o o t - s h o o t r a t i o was l ower i n loam and i n a loam-sand m i x t u r e (1:1 w/w) than i n s and . H e i g h t g rowth d i d no t d i f f e r between p r o v e n a n c e s . There were d i f f e r e n c e s i n d r y w e i g h t and r o o t - s h o o t r a t i o between p r o v e n a n c e s but t he se d i f f e r e n c e s were no t c o r r e l a t e d w i t h any g e o g r a p h i c f a c t o r . In the l a s t s t u d y , an a t t e m p t was made t o i d e n t i f y endogenous g i b b e r e l l i n s e x t r a c t e d f rom q u i e s c e n t v e g e t a t i v e , male and fema le buds o f red a l d e r . F o l l o w i n g f r a c t i o n a t i o n o f the e x t r a c t s by s i l i c a g e l p a r t i t i o n c h r o m a t o g r a p h y , p o s i t i v e g rowth r e s p o n s e s were o b t a i n e d i n the dwar f r i c e b i o a s s a y of s e v e r a l o f the f r a c t i o n s . In g a s - l i q u i d c h r o m a t o g r a p h y , v a r i o u s f r a c t i o n s p roduced peaks w i t h r e t e n t i o n t i m e s s i m i l a r t o t hose o f s t a n d a r d g i b b e r e l l i n s . A l t h o u g h some q u a l i t a t i v e i n d i c a t i o n s were n o t e d , i t was not p o s s i b l e t o c o n f i r m the p r e s e n c e and i d e n t i t y o f g i b b e r e l l i n s u s i n g combined g a s - 1 i q u i d chromatography-mass s p e c t r o m e t r y . Inadequate p u r i f i c a t i o n o f e x t r a c t s and low c o n c e n t r a t i o n s o f endogenous g i b b e r e l l i n s i n the e x t r a c t s were thought t o be m a i n l y r e s p o n s i b l e i v for th i s fa i1ure . The impact and impl icat ions a r i s i n g from the resul ts of these studies and -poss ib le future d i r e c t i o n s for research are discussed in the f i n a l chapter . TABLE OF CONTENTS . ...Page CHAPTER I THE IMPORTANCE OF RED ALDER (Ainus rubra Bong. ) IN THE PACIF IC NORTHWEST. I n t r o d u c t i o n 1 Economic Impor tance o f Red A l d e r 5 E c o l o g i c a l Impor tance o f Red A l d e r 8 I n f l u e n c e o f Red A l d e r on S o i l P r o p e r t i e s 9 P o t e n t i a l o f Red A l d e r i n C o n t r o l l i n g Root Rot D i s e a s e s 13 Management I m p l i c a t i o n s 17 CHAPTER II GERMINATION OF RED ALDER SEED FROM DIFFERENT LOCATIONS. I n t r o d u c t i o n 20 M a t e r i a l s and methods 21 F i r s t S tudy 21 Second S tudy 2k R e s u l t s and O b s e r v a t i o n s 15 F i r s t S tudy 15 T r e a t m e n t E f f e c t s on I n d i v i d u a l Seed C o l l e c t i o n s . . 36 Haney P rovenance 36 Ki t s a u l t. P ro venance 38 Nanaimo Lakes P rovenance 38 Idaho P rovenance 38 Cedar C r e e k P rovenance k2 P o i n t Reyes P rovenance k2 v i Page E f f e c t s o f I n d i v i d u a l T r e a t m e n t s on A l l P r o v e n a n c e s . . k5 T rea tmen t E f f e c t s on A l l P rovenances kS Second S tudy 55 D i s c u s s i o n 59 F i r s t S tudy 59 Second S tudy 66 G e n e t i c C o n s i d e r a t i o n s 67 C o n c l u s i o n s . . . 68 CHAPTER III THE IMPORTANCE OF FERTIL ITY AND PHYSICAL CHARACTERISTICS OF SOIL IN EARLY SEEDLING DEVELOPMENT IN RED ALDER. I n t r o d u c t i o n 69 M a t e r i a l s and Methods 70 R e s u l t s and O b s e r v a t i o n s Ik P rovenance Response t o I n d i v i d u a l T r ea tmen t s . . 78 P rovenance Response t o F e r t i l i z a t i o n i n Each Growth Med i um 8 l Response o f I n d i v i d u a l P rovenances t o a l l T r e a t m e n t s . . . Sk K i t s a u l t P rovenance Sk C r e s c e n t C i t y P rovenance 86 M y r t l e C reek P rovenance 89 Vancouve r P rovenance 91 C e n t r a l i a P rovenance 91 L a y t o n v i l l e P rovenance 91 C o t t a g e Grove P rovenance • • • 95 Nanaimo Lakes P rovenance 95 CHAPTER IV CHAPTER V V I I Page T o t a l A n a l y s e s 95 He i g h t 9 5 Dry w e i g h t 39 Root-s h o o t R a t i o . . . . . . 1 0 1 D i s c u s s i o n 'O** C o n c l u s i o n s ''' IDENTIFICATION OF ENDOGENOUS GIBBERELLINS IN BUDS OF RED ALDER. I nt r o d u c t i on • • • '13 M a t e r i a l s and Methods E x t r a c t i o n ''5 F r a c t i o n a t i o n ''*> B i oassay '' 7 G a s - L i q u i d Chromatography ''8 Combined G a s - L i q u i d chromatography and Mass-s p e c t o m e t r y ' ' 9 R e s u l t s . . • 1 2 0 D i s c u s s i o n '30 C o n c l u s i o n s '32 GENERAL DISCUSSION 135 LITERATURE CITED 139 v i i i L IST OF TABLES T a b l e Page 2 . 1 . G e o g r a p h i c l o c a t i o n o f p r o v e n a n c e s used i n the f i r s t g e r m i n a t i o n s t u d y and d a t e s o f seed c o l l e c t i o n 22 2 . 2 . G e o g r a p h i c l o c a t i o n o f p r o v e n a n c e s used i n the second g e r m i n a t i o n s t u d y and d a t e s o f seed c o l l e c t i o n 22 2 . 3 . Mean g e r m i n a t i o n p e r c e n t a g e s and p e r c e n t a g e s o f v i a b l e s e e d s , n o n - v i a b l e seeds and abnorma l g e r m i n a n t s i n each o f the seed c o l l e c t i o n s used i n the f i r s t g e r m i n a t i o n s t u d y . . 26 2 . 4 . G e r m i n a t i o n d a t a f o r s t r a t i f i e d , p r e s o a k e d and u n t r e a t e d seeds o f the Haney p rovenance 36 2 . 5 . G e r m i n a t i o n d a t a f o r s t r a t i f i e d , p r e s o a k e d and u n t r e a t e d seeds o f the K i t s a u l t p ro venance 39 2 . 6 . G e r m i n a t i o n d a t a f o r s t r a t i f i e d , p r e s o a k e d and u n t r e a t e d seeds o f the Nanaimo Lakes p rovenance 40 2 . 7 . G e r m i n a t i o n d a t a f o r s t r a t i f i e d , p r e s o a k e d and u n t r e a t e d seeds o f the Idaho p rovenance 41 2 . 8 . G e r m i n a t i o n d a t a f o r s t r a t i f i e d , p r e s o a k e d and u n t r e a t e d seeds o f the Cedar C reek (Wash ing ton ) p rovenance 43 2 . 9 . G e r m i n a t i o n d a t a f o r s t r a t i f i e d , p r e s o a k e d and u n t r e a t e d seeds o f the P o i n t Reyes ( C a l i f o r n i a ) p rovenance 44 2 .10 . Summary o f a n a l y s i s o f g e r m i n a t i o n p e r c e n t a g e s f o r s t r a t i f i e d , p r e s o a k e d and u n t r e a t e d seeds o f a l l p r o v e n a n c e s t e s t e d i n the f i r s t g e r m i n a t i o n s t udy 46 2 . 1 1 . Summary o f a n a l y s i s o f g e r m i n a t i o n v a l u e s f o r s t r a t i f i e d , p r e s o a k e d and u n t r e a t e d seeds o f a l l p rovenances t e s t e d i n the f i r s t g e r m i n a t i o n s t u d y 47 2 . 1 2 . Summary o f a n a l y s i s o f c o l l e c t i v e g e r m i n a t i o n p e r c e n t a g e s and g e r m i n a t i o n v a l u e s f o r a l l p rovenances and t r e a t m e n t s i n the f i r s t g e r m i n a t i o n s t u d y 52 2 . 1 3 . Summary o f a n a l y s i s o f g e r m i n a t i o n d a t a f o r p r e s o a k e d seeds o f a l l p r o v e n a n c e s t e s t e d i n the second g e r m i n a t i o n s t u d y . . 58 3 . 1 . C o m p o s i t i o n o f the c o m m e r c i a l f e r t i l i z e r " H i - s o l " 73 3 . 2 . Summary o f d a t a c o l l e c t e d f o r n o d u l e f o r m a t i o n i n f e r t i l i z e d (F) and u n f e r t i l i z e d (NF) s e e d l i n g s f rom K i t s a u l t , M y r t l e C r e e k , Vancouve r and C r e s c e n t C i t y grown i n the d i f f e r e n t c o m b i n a t i o n s o f c o a r s e sand (SD) and loam (LM) . . . 77 i x T a b l e Page 3 - 3 - Summary o f a n a l y s i s o f d a t a f o r h e i g h t (mm.) o f s e e d l i n g s o f t he d i f f e r e n t p rovenances i n i n d i v i d u a l t r e a t m e n t s 79 3 . 4 . Summary o f a n a l y s i s o f d a t a f o r d r y w e i g h t (mg.) o f s e e d l i n g s o f t he d i f f e r e n t p rovenances i n i n d i v i d u a l t r e a t m e n t s 79 3 -5 - Summary o f a n a l y s i s o f d a t a f o r r o o t - s h o o t r a t i o o f s e e d l i n g s o f the d i f f e r e n t p r o v e n a n c e s i n i n d i v i d u a l t r e a t m e n t s 80 3 . 6 . Summary o f a n a l y s i s o f d a t a f o r h e i g h t (mm.) o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f the d i f f e r e n t p rovenances i n each g rowth medium 82 3 .7 - Summary o f a n a l y s i s o f d a t a f o r d r y w e i g h t (mg.) o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f the d i f f e r e n t p rovenances i n each g rowth medium 82 3 . 8 . Summary o f a n a l y s i s o f d a t a f o r r o o t - s h o o t r a t i o o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f the d i f f e r e n t p rovenances i n each g rowth medium 83 3 .9 - Summary o f a n a l y s i s o f d a t a f o r h e i g h t (mm.), d r y w e i g h t (mg.) and r o o t - s h o o t r a t i o o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f the K i t s a u l t p rovenance grown in s a n d , loam-sand and loam . . . 87 3 . 1 0 . Summary o f a n a l y s i s o f d a t a f o r h e i g h t (mm.), d r y w e i g h t (mg.) and r o o t - s h o o t r a t i o o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f t he C r e s c e n t C i t y p rovenance grown i n s a n d , loam-sand and loam 88 3 . 1 1 . Summary o f a n a l y s i s o f d a t a f o r h e i g h t (mm.), d r y w e i g h t (mg.) and r o o t - s h o o t r a t i o o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f the M y r t l e C reek p rovenance grown i n s a n d , loam-sand and loam 90 3 .12 . Summary o f a n a l y s i s o f d a t a f o r h e i g h t (mm.), d r y w e i g h t (mg.) and r o o t - s h o o t r a t i o o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f the Vancouve r p rovenance grown in s a n d , loam-sand and loam . . . 92 3 .13- Summary o f a n a l y s i s o f d a t a f o r h e i g h t (mm.), d r y w e i g h t (mg.) and r o o t - s h o o t r a t i o o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f the C e n t r a l i a p rovenance grown in s a n d , loam-sand and loam . . . 3 .1A. Summary o f a n a l y s i s o f d a t a f o r h e i g h t (mm.), d r y w e i g h t (mg.) and r o o t - s h o o t r a t i o o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f the L a y t o n v i l l e p rovenance grown in s a n d , loam-sand and loam . . 3h X T a b l e Page 3 . 1 5 . Summary o f a n a l y s i s o f d a t a f o r h e i g h t (mm.), d r y w e i g h t (mg.) and r o o t - s h o o t r a t i o o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f t he C o t t a g e Grove p rovenance grown i n s a n d , loam-sand and loam 96 3 . 1 6 . Summary o f a n a l y s i s o f d a t a f o r h e i g h t (mm.), d r y w e i g h t (mg.) and r o o t - s h o o t r a t i o o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f the Nanaimo Lakes p rovenance grown i n s a n d , loam-sand and loam 96 3 .17 - Summary o f a n a l y s i s o f d a t a f o r h e i g h t (mm.) o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f the d i f f e r e n t p rovenances grown i n s a n d , loam-sand and loam 98 3 . 1 8 . Summary o f a n a l y s i s o f d a t a f o r d r y w e i g h t (mg.) o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f the d i f f e r e n t p rovenances grown i n s a n d , loam-sand and loam 100 3 . 1 9 . Summary o f a n a l y s i s o f d a t a f o r r o o t - s h o o t r a t i o o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f the d i f f e r e n t p rovenances grown i n s a n d , loam-sand and loam 102 4 . 1 . F r a c t i o n s o f e x t r a c t s f rom v e g e t a t i v e , male and f e m a l e buds w h i c h showed a p o s i t i v e g r o w t h r e s p o n s e i n the dwar f r i c e b i o a s s a y and w i t h GLC peaks whose r e t e n t i o n t imes were s i m i l a r t o t h a t o f s t a n d a r d GA^ 124 4 . 2 . F r a c t i o n s o f e x t r a c t s f rom v e g e t a t i v e , male and f ema l e buds w h i c h showed a p o s i t i v e g r o w t h r e s p o n s e i n the dwar f r i c e b i o a s s a y and w i t h GLC peaks whose r e t e n t i o n t imes were s i m i l a r t o t h a t o f GA^ 124 4 . 3 . F r a c t i o n s o f e x t r a c t s f rom v e g e t a t i v e , male and f e m a l e buds w h i c h showed a p o s i t i v e g r o w t h r e s p o n s e i n the dwar f r i c e b i o a s s a y and w i t h GLC peaks whose r e t e n t i o n t imes were s i m i l a r t o t h a t o f GA-, 125 4 . 4 . F r a c t i o n s o f e x t r a c t s f r om v e g e t a t i v e , male and f ema le buds w h i c h showed a p o s i t i v e g rowth r e s p o n s e i n the dwar f r i c e b i o a s s a y and w i t h GLC peaks whose r e t e n t i o n t imes were s i m i l a r t o t h a t o f GA g 125 4 .5 - F r a c t i o n s o f e x t r a c t s f rom v e g e t a t i v e , male and f ema le buds w h i c h showed a p o s i t i v e g rowth r e s p o n s e i n the dwar f r i c e b i o a s s a y and w i t h GLC peaks whose r e t e n t i o n t imes were s i m i l a r t o t h a t o f GA 126 4 . 6 . P r e s e n c e (+) o r absence (-) o f GLC peaks w i t h s i m i l a r r e t e n t i o n t imes as t h o s e o f s t a n d a r d GAs , f rom f r a c t i o n s i n w h i c h the s t a n d a r d s were e x p e c t e d 127 x i Figure LIST OF FIGURES Page 1 .1 . Natural range of Alnus rubra Bong, (taken from Plank, ( 1 9 7 0 ) • 2 2 . 1 . Course of germination for s t r a t i f i e d , presoaked and untreated seeds of the Haney provenance 36 2 . 2 . Course of germination for s t r a t i f i e d , presoaked and untreated seeds of the Kitsault provenance 39 2 . 3 - Course of germination for s t r a t i f i e d , presoaked and untreated seeds of the Nanaimo Lakes provenance 40 l.k. Course of germination for s t r a t i f i e d , presoaked and untreated seeds of the Idaho provenance 41 2 . 5 . Course of germination for s t r a t i f i e d , presoaked and untreated seeds of the Cedar Creek (Washington) provenance A 3 2 . 6 . Course of germination for s t r a t i f i e d , presoaked and untreated seeds of the Point Reyes (California) provenance kk 2 . 7 . Course of germination for s t r a t i f i e d seeds from each source tested in the f i r s t germination study kS 2 . 8 . Course of germination for presoaked seeds from each source tested in the f i r s t germination study 50 2 . 9 . Course of germination for untreated seeds from each source tested in the f i r s t germination study 51 2 . 1 0 . Mean germination percent for s t r a t i f i e d , presoaked and untreated seeds of each provenance tested in the f i r s t germination study 5^ 2 . 1 1 . Mean germination value for s t r a t i f i e d , presoaked and untreated seeds of each provenance tested in the f i r s t germination study 5** 2 . 1 2 . Course of germination for s t r a t i f i e d , presoaked and untreated seeds of a l l provenances tested in the f i r s t germination study 56 xi i Course of germination for treated and untreated seeds of each co l l e c t i o n tested in the f i r s t germination study Course of germination for presoaked seeds from each source tested in the second germination study Root-shoot ratios of seedlings of the different provenances grown in loam-sand (A) and loam (B) . . Mean height of f e r t i l i z e d and u n f e r t i l i z e d seedlings of the Kitsault provenance grown in sand, loam-sand and loam Mean root-shoot ra t i o of f e r t i l i z e d and u n f e r t i l i z e d seedlings of the Crescent City provenance grown in sand, loam-sand and loam Mean root-shoot ra t i o of f e r t i l i z e d and u n f e r t i l i z e d seedlings of the Myrtle Creek provenance grown in sand, loam-sand and loam Mean height of f e r t i l i z e d and u n f e r t i l i z e d seedlings of the Vancouver provenance grown in sand, loam-sand and loam Mean dry weight of f e r t i l i z e d and u n f e r t i l i z e d seedlings of the Vancouver provenance grown in sand, loam-sand and loam. . Mean height of f e r t i l i z e d and u n f e r t i l i z e d seedlings of the Nanaimo Lakes provenance grown in sand, loam-sand and loam Mean root-shoot ratio of f e r t i l i z e d and u n f e r t i l i z e d seedlings of the Nanaimo Lakes provenance grown in sand, loam-sand and loam Mean height of f e r t i l i z e d and u n f e r t i l i z e d seedlings of a l l provenances grown in sand, loam-sand and loam Mean dry weight of f e r t i l i z e d and u n f e r t i l i z e d seedlings of a l l provenances grown in sand, loam-sand and loam . . . . Mean root-shoot ratio of f e r t i l i z e d and u n f e r t i l i z e d seedlings of a l l provenances grown in sand, loam-sand and loam Mean root-shoot r a t i o of f e r t i l i z e d and u n f e r t i l i z e d seedlings of each provenance grown in a l l growth media . . . x i i i F i g u r e Page 4 . 1 . Dwarf r i c e b i o a s s a y s o f f r a c t i o n s c o l l e c t e d d u r i n g s i l i c a g e l p a r t i t i o n chromatography o f e x t r a c t s from v e g e t a t i v e buds 121 4 . 2 . Dwarf r i c e b i o a s s a y s o f f r a c t i o n s c o l l e c t e d d u r i n g s i l i c a g e l p a r t i t i o n chromatography o f e x t r a c t s from male r e p r o d u c t i v e buds 122 4 . 3 - Dwarf r i c e b i o a s s a y s o f f r a c t i o n s c o l l e c t e d d u r i n g s i l i c a g e l p a r t i t i o n chromatography o f e x t r a c t s from female r e p r o d u c t i v e buds . 123 4 . 4 . Mass spe c t r u m o f the TMSi e i t h e r o f GA^ 127 4.5. Mass s p e c t r u m o f scan a t peak whose r e t e n t i o n time was s i m i l a r t o t h a t o f the TMSi e i t h e r o f GA_ ( F r a c t i o n No. 15-Repl i c a t i o n 3 o f femal e bud e x t r a c t ) 128 4 . 6 . Mass spectrum o f scan a t peak whose r e t e n t i o n time was s i m i l a r t o t h a t o f the TMSi e i t h e r o f GA, ( F r a c t i o n No. 15-Repl i c a t i o n 3 o f female bud e x t r a c t ) 129 x i v L IST OF PLATES P l a t e Page 2.1. Funga l decay i n embryo o f seed f rom the Haney ( l ) c o l l e c t i o n a t the end o f i n i t i a l t e s t f o r the f i r s t g e r m i n a t i o n s t u d y (X20) 27 2 . 2 . Funga l decay i n embryo o f seed f rom the Haney ( l ) c o l l e c t i o n a t t he end o f i n i t i a l t e s t f o r the f i r s t g e r m i n a t i o n s t u d y (X20) 27 2 . 3 . Embryo o f seed f rom the Haney ( l ) c o l l e c t i o n w i t h p a r t o f d e c a y i n g s e c t i o n removed w i t h a n e e d l e (X20) 28 2 . 4 . Abnormal g e r m i n a n t f rom the K i t s a u l t c o l l e c t i o n w i t h d o u b l e r a d i c l e s (R) (X15) 2 8 2.5. Abnormal ge rminant f rom the K i t s a u l t c o l l e c t i o n w i t h p r e -emergent c o t y l e d o n s (X15) • • 29 2 . 6 . Abnormal g e r m i n a n t f r om the K i t s a u l t c o l l e c t i o n showing the unusua l emergence o f the h y p o c o t y l (H) (X20) 31 2 . 7 . Abnormal g e r m i n a n t f rom the K i t s a u l t c o l l e c t i o n w i t h d o u b l e embryos and p re-emergen t c o t y l e d o n s (X1 5) 32 2 . 8 . Abnormal g e r m i n a n t f rom the K i t s a u l t c o l l e c t i o n showing r a d i c l e (R) g r o w i n g a round i n n e r edge o f p e r i c a r p w h i l e h y p o c o t y l (H) emerges a t m i c r o p y l a r end o f seed (X20) 33 2 . 9 . Abnormal g e r m i n a n t f rom the K i t s a u l t c o l l e c t i o n showing the l o o p - l i k e emergence o f h y p o c o t y l (H) a t m i c r o p y l a r end o f the seed (X20) 33 2 . 1 0 . Abnormal g e r m i n a n t f rom t h e K i t s a u l t c o l l e c t i o n show ing the emergence o f h y p o c o t y l (H) a l o n g the edge o f the t e s t a r a t h e r than a t t he m i c r o p y l a r end o f seed (X20) 34 2 . 1 1 . Abnormal g e r m i n a n t f rom the K i t s a u l t c o l l e c t i o n showing r a d i c l e (R) a p p a r e n t l y g r o w i n g n o r m a l l y a f t e r e a r l i e r r e s t r i c t i o n was overcome (X20) 34 2 . 1 2 . Embryo o f seed f rom the K i t s a u l t c o l l e c t i o n showing s w e l l i n g (SW) a t r a d i c l e end a p p a r e n t l y formed where p h y s i c a l r e s t r i c t i o n was e n c o u n t e r e d 35 2.13 Abnormal g e r m i n a n t f rom the K i t s a u l t c o l l e c t i o n showing p r e -emerg ing c o t y l e d o n s (C) c aused by g rowth o f h y p o c o t y l w i t h i n the t e s t a (X15) 35 XV Plate Page 3.1. U n f e r t i l i z e d (UF) and f e r t i l i z e d (F) seedlings of the Laytonville provenance grown in sand (SD) , loam-sand (SS) and loam (SL) 75 3 .2 . U n f e r t i l i z e d (-FERT) and f e r t i l i z e d (+FERT) seedlings of the Kitsault provenance grown in sand (SD), loam-sand (SS) and loam (SL) 75 xv i ACKNOWLEDGEMENTS The research towards this thesis was supported by operating grant #A5397 to Dr. I.E.P. Taylor from the National Research Council of Canada. I gratefully acknowledge the bursaries provided by the National Research Council of Canada, and by the Faculties of Forestry and Graduate Studies of the University of B r i t i s h Columbia. I also wish to acknowledge the assistance and advice of the many people who contributed towards the thesis. In par t i c u l a r , gratitude is expressed for the support and positive c r i t i c i s m provided by my supervisors, Dr. I.E.P. Taylor and Dr. A. Kozak, and for the guidance and helpful comments of my committee members. Special thanks are extended to Mr. P. Saunders of the University of B r i t i s h Columbia's Research Forest for his assistance in seed c o l l e c t i o n ; to Professor F.D. Johnson of the University of Idaho and to Dr. W.I. Stein of the U.S.D.A. Forestry Sciences Laboratory, Oregon for the provision of seeds from Idaho and Cedar Creek, Washington respectively; to Dr. D.C. Aldridge, Imperial Chemical Industries Ltd., Cheshire, England for providing samples of gibberellins; and to Dr. R.P. Pharis of the University of Calgary for his advice on the extraction of gibberellins from plant materials. F i n a l l y , I wish to thank Dr. G. Eigendorf of the Department of Chemistry, and Dr. F.S. Abbott and Mr. R. Burton of the Faculty of Pharmaceutical Sciences, University of B r i t i s h Columbia, for .their help with the combined gas-liquid chromatography-mass spectrometry phase of these studies. xvi i INTRODUCTION The economic development of a tree species requires an understanding of it s s i l v i c a l characteristics as wel1 as relevant aspects of i t s basic botany. Since the pioneer days, forestry in western North America has been dominated by coniferous species with the naturally occurring hardwoods l i k e red alder (Alnus rubra Bong.), bigleaf maple (Acer macrophyllum Pursh) and black cottonwood (Populus trichocarpa Torr. and Gray), playing only a minor role to that of the conifers (Tarrant, 1978). Present circumstances are that supplies of old growth conifers are rapidly being depleted and regeneration has not yet provided adequate replacements to maintain the level of forestry a c t i v i t y that was enjoyed for many years (Tarrant, 1978). In addition, environmental and economic restraints are c u r t a i l i n g the harvesting of large volumes of timber that are not eas i l y accessible. The dominance of conifers in the forest industry has resulted in hardwood species being largely under-used. Red alder has long been viewed as a weed species mainly because of i t s abundance, i t s rapid growth and suppression of the more desirable conifer regeneration, and i t s generally poor tree quality. It possesses a number of attributes which would enable i t to f i l l part of the void in wood fib r e that is anticipated when the available supply of old growth softwoods is exhausted. At the same time, i t can provide on a continual basis, a favourable s o i l environment for the regeneration of the much favoured conifers. Red alder's nitrogen f i x i n g xv i i i a b i l i t y can be exploited to enrich a variety of s o i l s . Its a b i l i t y to colonize various types of bared ground make i t useful for improving conditions on sites which are unsuitable for the growth of other commer-c i a l l y important species. In addition to these ecological attributes from which connifers can benefit, technological advances have made red alder an increasingly important pulping species in the P a c i f i c Northwest today, and more and more foresters are beginning to recognize i t s wood as one of high quali ty. Attitudes towards red alder are changing in a more positive d i r e c t i o n , and attention is now being directed towards i t s intensive management. To make wise use of this species, the establishment of a strong information base dealing with a l l aspects of i t s growth is needed. There is surprisingly l i t t l e r e l i a b l e information available for red alder compared with that for species l i k e Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco). Institutions in Oregon and Washington States (United States of America) have become major places of research and have begun to accumulate relevant information, but there s t i l l is a substantial lack of sp e c i f i c knowledge about the biology of red alder. This is p a r t i c u l a r l y important in light of the fact that improvement in tree quality through breeding programs w i l l be a major objective for the intensive management of red alder. The selection of individual trees with desirable t r a i t s , controlled cross-pollination between these trees, and the co l l e c t i o n of seeds thus produced are major f i r s t steps in a tree breeding program. These are followed by cul t i v a t i o n of seedlings and their establishment in the f i e l d . Thus, in the f i r s t instance, information on the biology of red alder seed and seedlings w i l l be most valuable. A xf'x sound knowledge of seed production and s u r v i v a l , and of early development of seedlings under different environmental conditions, becomes central to tree breeding programs for this or any other species. I n i t i a l l y , seed quality w i l l rely on wild grown stock but an understanding of i t s re-productive growth is also necessary. The y i e l d of sound seed in alder {Alnus) collections can be very e r r a t i c and often, collections contain a large percentage of empty seeds (McVean, 1955 ; Schalin, 1968; Schopmeyer, 197*0. Seed production re l i e s on the f e r t i l i z a t i o n process and f e r t i l i z a t i o n depends on the production of viable pollen and "healthy" ovules. Work by Pharis (1976a and 1976b) has shown that in conifers, s p e c i f i c gibberellins (GAs) are associated with the development of maleness and femaleness and this knowledge is being applied in nursery programs to produce and harvest sustained yields of seed. It is possible that a similar situation exists in hardwoods such as red alder but the s p e c i f i c GAs may not be the same as those in conifers. A long term study of red alder reproductive growth and hormonal control, w i l l assist in improving pollen and ovule y i e l d , and subsequently, the y i e l d of viable seeds. The application of synthetic GAs to improve vegetative growth and to accelerate and increase the production of reproductive structures is being increasingly used in agriculture and horticulture and may become more important in forestry in the near future. It may not be practical to employ this technique on a large scale but i t certainly can be used in seed orchards where good yields of viable seed can then be anticipated more regularly. XX Red alder is a widely distributed species occurring along the western coast of North America with a few disjunct populations in Idaho. Several west coast species display great genetic v a r i a b i l i t y and differences between provenances can usually be related to environmental factors in their respective locations (Wright, 1976). Hence, studies on such species, where possible, should include some indication as to whether the findings can be applied to the species as a whole, or whether one should expect variations between different provenances. If there are variations between provenances, then relationships between them and environmental parameters should be investigated. The work in this thesis was undertaken to provide some of the information on seed biology presently not available in the l i t e r a t u r e . It was necessary to establish r e l a t i v e l y simple information but I believe that i t is extremely unsound to rely on the extension of work on other species to plan management programs for a particular species. For example, the sp e c i f i c GAs applied to the different reproductive structures of conifers to increase seed y i e l d w i l l not necessarily produce the same results i f they were applied to corresponding structures in red alder. Also, results from work on other alders in Europe may not necessarily apply to red alder. The thesis consists of four parts followed by a general discussion. The f i r s t chapter is essenti a l l y a l i t e r a t u r e review which was undertaken because of the lack of a recent review focussing on the value of red alder. It summarizes the present knowledge of the species. It reveals a lack of sp e c i f i c knowledge about i t s biology, knowledge which is important for the development of tree breeding programs and intensive management. xx i Seed germination behaviour and early seedling development under varying s o i l conditions are r e l a t i v e l y poorly understood. These are reported in the second and third chapters respectively. Growth parameters and nodule formation which may be early signs of good performance, are measured and related to provenance. The i n i t i a l hypothesis in the project on seed germination was that red alder seeds show increasing incidence of dormancy as i t s di s t r i b u t i o n is followed from i t s southern li m i t s northwards. Seeds of many wide ranging west coast species commonly display varying degrees of dormancy and vigour, and one might expect red alder seeds to behave s i m i l a r l y . There is limited information in the l i t e r a t u r e on the occurrence of dormancy in seeds of this species throughout i t s range, and as such, one does not know i f dormancy e x i s t s , what type may occur, nor the manner in which i t can be overcome i f encountered. Information on the effects of pretreatment on germination rate is also lacking. The germination studies on seeds from provenances in the southern two-thirds of i t s range, should f i l l some of the gap in our knowledge of the species' seed biology. The i n i t i a l hypothesis in the seedling development study was that s o i l and f e r t i l i z a t i o n would not affect growth and nodule formation s i g n i f i c a n t l y . The li t e r a t u r e gives the impression that red alder seedlings can grow on almost any type of ground providing s o i l moisture is adequate. Also, there is a wide range of figures reported for the amount of growth that takes place in the f i r s t year of development. Most of these measurements were xxi i made on naturally growing seedlings often without consideration of the effects of environmental variations. The effects of individual environ-mental factors on early development have not been examined extensively. Soil conditions are very important in the establishment of seedlings and are commonly overlooked p a r t i c u l a r l y in species l i k e red alder whose nitrogen f i x i n g a b i l i t y enriches the s o i l on which i t is growing. The study reported in Chapter III was done mainly to provide some information on how sensitive the development of red alder seedlings are to variations in s o i l texture and f e r t i l i t y . A f i e l d t r i a l was i n i t i a t e d in parallel with these experiments but poor seedling survival forced me to abandon this project. The GA project was exploratory. Different provenances were not studied. The technical and physiological complexity of hormonal chemistry make i t important to establish the chemistry using a single source of material. Thus a l l material was taken from trees close to the laboratory. The study reported in Chapter IV is the i n i t i a l stage of a long term study. It is becoming clear that various combinations of GAs are characteristic of different developmental processes and correspondingly, are present in different parts of the tree at different times. In the long term, i t is intended to characterize the differences in GAs present in vegetative, male and female buds at various stages of their development, and to study the metabolism of GAs during the passage from vegetative development, through sexual reproduction, to seed germination and seedling development. Thus the extraction, isolation and characterization of GAs present in quiescent buds was thought an appropriate starting point in this extensive study. CHAPTER I THE IMPORTANCE OF RED ALDER (ALNUS RUBRA BONG.) IN THE PACIFIC NORTHWEST Introduction Red alder {Alnus rubra Bong.) is the largest tree of i t s genus and commercially, the most important hardwood species in the P a c i f i c Northwest (Oregon, Washington, B r i t i s h Columbia and Alaska). It is essentially a coastal species (Figure 1) whose natural d i s t r i b u t i o n is in western North America between latitudes 60° north (Yakutat Bay, Alaska) and 3^° north (Santa Barbara, C a l i f o r n i a ) . There are some isolated inland populations in Idaho (Johnson, 1968a, 1968b). It occurs in pure stands or mixed with either coniferous species such as Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco), western red cedar {Thuja plicata Donn.), western hemlock (rsuga beterophylla (Raf.) Sarg.), grand f i r (Abies grandis (Dougl.) Lindl.) and Sitka spruce (Picea sitchensis (Bong.) C a m ) , or other hardwood species eg. bigleaf maple (Acer macrophyllum Pursh), vine maple (Acer circinatum Pursh) and black cottonwood (Populus trichocarpa Torr. and Gray). The species thrives in areas having abundant s o i l moisture, and deep, well drained, r i c h , a l l u v i a l s o i l s . It attains tree size on coarser s o i l s provided moisture is adequate: thus red alder is commonly found along streams, Figure 1. Natural range of Alnus rubra Bong, (taken from Plank, 1971) . - 3 -benches and on moist h i l l s i d e s . Growth is however, poor in hot or continu-ously cool environments regardless of moisture regime (Hoyer et al., 1978). Though the larger pure stands of red alder are confined to wet sites along the coast and below an elevation of 800 feet (Rymer, 1951), trees are found up to an elevation of 2500 feet and generally less than 100 miles from the P a c i f i c coast (Worthington, 1965; Worthington et al., 1962; Plank, 1971). Red alder has the characteristics of a pioneer species vi.z. the p r o l i f i c production of small, winged seeds which are easily disseminated by wind; rapid germination on logged, burned or otherwise bared ground under favourable environmental conditions; aggressive early growth; the a b i l i t y to improve s o i l conditions for the occupation of the s i t e by succeeding vegetation; and a rel a t i v e l y short natural l i f e . In addition, i t occurs naturally in an area where forest f i r e s are frequent during the warmer periods of the year, and where the commercially more valuable and desirable conifers are abundant and logged regularly. As a result, there are extensive areas in the P a c i f i c Northwest which are occupied by dense stands of red alder of varying ages. These are naturally regenerated after f i r e s and logging operations. In such situations, an understorey with slower growing and more tolerant conifers is commonly present and these come to dominate the s i t e after the short-lived red alder dies (Johnson et al., 1926; Rymer, 1951). Red alder outgrows coastal conifers during early development, probably because of i t s foliage characteristics and i t s r e l a t i v e l y high photosynthetic rate at high light _ 4 _ i n t e n s i t i e s (Krueger and Ruth, 1968 and 1969) - However, growth of coniferous species l i k e Douglas-fir w i l l surpass that of red alder which is older than 25 to 30 years (Nielson, 1977; M i l l e r and Murray, 1 9 7 8 ) . Red alder is r e l a t i v e l y f i r e resistant. Some foresters consider this attribute makes i t an effective nurse crop for developing conifers (Johnson, 1 9 1 7 ) . However, i t s competitive edge over seedlings of coniferous species (Berntsen, 1961 a) leads others to view the species as less valuable. The importance of red alder as a forest type was stressed as early as 1917 (Johnson, 1 9 1 7 ) , but the species has also been c l a s s i f i e d as a "weed tree" and as an "un-desirable woody plant" (Tschirley, 1956; Leonard, 1961 ; Stewart, 1972a and b). There has been extensive research to find suitable methods for i t s control and for i t s eradication in favour of the more valuable coniferous species (e.g. Ruth and Berntsen, 1956; Rediske and Staebler, 1962; Finnis, 1964; Hetherington, 1964 ; Stewart, 1 9 7 4 ) . In B r i t i s h Columbia, red alder was not recorded by timber cruisers u n t i l 1930 (Rymer, 1951) and, although i t has been used as a lumber species in this province since at least 1910 , the f i r s t s i g n i f i c a n t volume of alder lumber was not produced un t i l 1938 (Nielson, 1 9 7 7 ) . Its potential importance was not recognized in the western United States of America un t i l r e l a t i v e l y recently (Clark, 1 9 5 7 ) . The control or eradication of red alder may be necessary under certain circumstances and management objectives. The negative attitude of many foresters towards this species as being a weed tree arose from i t s widespread occurrence, i t s suppression of coniferous regeneration on cleared ground, the - 5 -l a c k o f p r o p e r r e c o g n i t i o n o f i t s v a l u e both from an e c o l o g i c a l and u t i l i z a -t i o n a l v i e w p o i n t , and t h e narrow t h i n k i n g o f t h o s e f o r e s t e r s c h a r a c t e r i z e d by C l a r k (1957) as b e i n g " a d d i c t e d t o a o n e - s p e c i e s D o u g l a s - f i r regime". R e c e n t l y , however, t h e r e has been a more f a v o u r a b l e a t t i t u d e towards r e d a l d e r both as a c o m m e r c i a l l y i m p o r t a n t s p e c i e s and as an a m e l i o r a t o r o f f o r e s t ecosystems i n the P a c i f i c N o r t h w e s t . Economic Importance o f Red A l d e r The a c c e l e r a t i n g demand f o r red a l d e r w h i c h has s t i m u l a t e d i n t e r e s t i n i t s management and i t s u t i l i z a t i o n was prompted by s e v e r a l f a c t o r s i n c l u d i n g t e c h n o l o g i c a l advances i n i t s p u l p i n g , a g r o w i n g r e c o g n i t i o n o f i t s wood as one o f h i g h q u a l i t y and v a l u e , and improved market c o n d i t i o n s on the west c o a s t f a v o u r i n g l o c a l l y grown hardwoods ( W o r t h i n g t o n e t al., 1962). T a r r a n t (1978) c i t e d two more reasons f o r t h e c h a n g i n g a t t i t u d e towards r e d a l d e r viz. (1) t h e need f o r more e f f i c i e n t use o f t h e s h r i n k i n g s u p p l y o f o l d growth s o f t w o o d s and the need t o f i n d a l t e r n a t e s o u r c e s o f wood f i b r e t o r e p l a c e them and (2) t h e r e c o g n i t i o n o f red a l d e r , a n i t r o g e n f i x i n g s p e c i e s , as a v i a b l e a l t e r n a t i v e t o e x p e n s i v e f e r t i l i z e r s f o r t h e maintenance and improvement o f s o i l f e r t i l i t y . S e v e r a l a u t h o r s ( J o h n s o n , et al. 3 1926; Rymer, 1951; C l a r k , 1957; W o r t h i n g t o n et al., 1962; N i e l s o n , 1977; T a r r a n t 1978) have d i s c u s s e d t h e p r o p e r t i e s and t r e n d s i n the u t i l i z a t i o n o f red a l d e r wood, h a r v e s t i n g methods, m a n u f a c t u r i n g p r o c e s s e s , p o t e n t i a l uses and consequent management i m p l i c a t i o n s . Economic and m a r k e t i n g c o n s i d e r a t i o n s r e l e v a n t t o t h e management o f t h i s s p e c i e s a r e - 6 -contained in publications by Berntsen (196 l a ) , Yoho et al., (1969a and b) , Hildenbrand ( 1972) , Manning ( 1975 ) , Briggs et al., (1978) and Feddern (1978) . The potential of red alder as an energy source was considered by Evans (1974) and Smith (1978) . Additional information with respect to the various uses of it s wood and treatment procedures in the manufacture of i t s products are included in reports by Espenas (1951 and 1971) , Quick ( 1957 ) , Kozlik (1962, 1967 and 1978) Schroeder and Hansen ( 1968) , Chow and Pickles ( 1971) , Chow (1972) , Hillstrom (197*0, Atterbury ( 1978) , Hrutfiord ( 1978 ) , Maloney (1978) and Remington (1978) . The structure of red alder wood was described by Johnson et a l . , (1926), Ha 11 (1952) and Panshin and De Zeeuw (1964) . In e a r l i e r times, i t was used only for fuelwood and for smoking meat. Today, i t s main uses are as pulpwood and lumber for the manufacture of furniture. Its wood properties compare favourably with those of other commercially important hardwoods in North America (Clark, 1957; Leney et al., 1978) but there is s t i l l only slow progress towards the realization of i t s f u l l potential. It seems that we require s t i l l more changes in attitudes about the species (Behm, 1978; L i t t l e , 1978) . Red alder may have substantial potential as a means of meeting future demands for various wood products. The production of adequate quantities of high quality timber w i l l depend on the si1vicu1tura1 and management practices adopted by the various forestry agencies. Such practices must be based upon a cle°ar understanding of i t s growth characteristics. This is always a requirement for the proper management of any species, but i t is important in - 7 -t h i s c a se because some o f i t s g rowth c h a r a c t e r i s t i c s have r a i s e d doub t s as t o i t s s u i t a b i l i t y f o r i n t e n s i v e management. The a l t e r n a t i v e o f c h a n n e l l i n g more e f f o r t i n t o management o f t he more v a l u a b l e c o n i f e r s rema ins an a t t r a c t i v e p r o p o s i t i o n t o many f o r e s t e r s i n the P a c i f i c N o r t h w e s t . P r e s e n t e v i d e n c e s u g g e s t s t h a t a s h o r t r o t a t i o n i s bes t f o r i n t e n s i v e management (Rymer, 1951 ; S m i t h , 1972; Sm i th and De B e l l , 1 9 7 ^ ; Z a v i t k o w s k i and S t e v e n s , 1972; De B e l l et al., 1 9 7 8 ) . The l a c k o f s t a b i l i t y and c o n t i n u i t y i n p r o d u c t i o n and ma rke t s n e c e s s a r y f o r e f f i c i e n t u t i l i z a t i o n was n o t e d by Haddock (19^9) and War rack ( 1 9 ^ 9 ) . I t i s a p p a r e n t t h a t a major cause o f t h i s s i t u a t i o n as f a r as lumber p r o d u c t i o n i s c o n c e r n e d , i s the i n a d e q u a t e s u p p l y o f h i g h q u a l i t y l o g s . The need f o r a p p r o p r i a t e c u l t u r a l p r a c t i c e s t o improve l o g q u a l i t y i s a p p r e c i a t e d by f o r e s t e r s i n t e r e s t e d i n the c o n t i n u e d use o f red a l d e r . A l t h o u g h decay i s i n s i g n i f i c a n t , sweep and c r o o k a r e v e r y p r e v a l e n t N i e l s o n , 1 9 7 7 ) . It was b e l i e v e d t h a t the p rob l ems e n c o u n t e r e d i n f e l l i n g t r e e s and saw ing l o g s a r o s e f rom the p r e s e n c e o f t e n s i o n wood ( C l a r k , 1957) • However , Leney et al.,(1978) found t h a t t e n s i o n wood was not p r e v a l e n t and t h a t g rowth s t r e s s e s were the more l i k e l y cause o f the p r o b l e m s . Improv ing l o g q u a l i t y by r e d u c i n g the amount o f s p i r a l g r a i n i n the o u t e r c o r e s appea r s p o s s i b l e i f g rowth r a t e i s c o n t r o l l e d (Kennedy and E l l i o t t , 1 9 5 7 ) . The o c c u r r e n c e o f e p i c o r m i c b r anches on red a l d e r t r u n k s i s a n o t h e r i m p o r t a n t f a c t o r t h a t r educes saw log q u a l i t y . B e r n t s e n (1961b) showed t h a t , p r u n i n g was e f f e c t i v e i n r e d u c i n g the i n c i d e n c e o f k n o t s i n 43 y e a r o l d t r e e s examined - 8 -22 y e a r s a f t e r t r e a t m e n t but t h a t t h e s e g a i n s i n wood q u a l i t y were o f f s e t by the development o f s i n g l e o r c l u s t e r e d e p i c o r m i c branches where n a t u r a l o r a r t i f i c i a l p r u n i n g took p l a c e . These d e f e c t s do not appear t o pose s e r i o u s problems i n the p u l p i n g i n d u s t r y and red a l d e r has been s u g g e s t e d as the b e s t p r o s p e c t f o r whole t r e e c h i p p i n g i n t h e P a c i f i c Northwest ( H r u t f i o r d , 1978). Very few red a l d e r t r e e s produce t r u n k s s u i t a b l e f o r v e neer (Bene, 1950) and because o f t h e k n o t t i n e s s o f l o g s , a l a r g e amount o f red a l d e r v e neer i s o f i n f e r i o r q u a l i t y (McGuane, 1978). Thus, i t seems t h a t a t l e a s t i n t h e s h o r t term, the wood o f red a l d e r has a l i m i t e d number o f uses and t h a t i t s p r o d u c t s cannot compete s u c c e s s f u l l y w i t h t h e h i g h q u a l i t y ones from o t h e r s p e c i e s growing i n t h e r e g i o n . In th e f u t u r e however, i t seems l i k e l y t h a t changes i n a t t i t u d e s and demands, t e c h n o l o g i c a l a d v a n c e s , and/or a s h o r t a g e o f wood from o t h e r s p e c i e s t h a t p r e s e n t l y s u p p o r t t h e t h r i v i n g f o r e s t i n d u s t r y i n the P a c i f i c N o r t h w e s t , a l l w i l l c o n s p i r e t o f a v o u r i n t e n s i v e management o f red a l d e r . The i d e a l o b j e c t i v e o f t h i s management would i n v o l v e a r o t a t i o n and s t a n d d e n s i t y w h i c h w i l l reduce t h e p r o d u c t i o n o f e p i c o r m i c branches and g a i n the most increment and s t r a i g h t e s t stems (Warrack, 19^9). T r e e improvement programmes (De B e l l and W i l s o n , 1978; S t e t t l e r , 1978) and p r o p e r s p a c i n g and t h i n n i n g regimes ( S m i t h , 1978) s h o u l d improve t r e e s i z e . E c o l o g i c a l Importance o f Red A l d e r S y m b i o t i c n i t r o g e n f i x a t i o n i n r o o t n o d u l e s o f non-leguminous angiosperms has - 9 -been e x t e n s i v e l y r e v i e w e d ( A l l e n and A l l e n , 1958; Bond, 1958, 1959, 1963, 1967 and 197A; S t e w a r t , 1966; B e c k i n g , 1970 and 1975). Members o f s e v e r a l f a m i l i e s and genera i n c l u d i n g t h e genus Alnus, a r e known t o form s y m b i o t i c a s s o c i a t i o n s w i t h a number o f m i c r o b e s . T h i s a t t r i b u t e o f red a l d e r and i t s r e s i s t a n c e t o the c o n i f e r r o o t r o t d i s e a s e , Phellinus (Porta) weirii (Murr.) G i l b . a r e r e s p o n s i b l e f o r i t s r e c o g n i t i o n as a p o t e n t i a l l y i m p o r t a n t s p e c i e s f o r i m p r o v i n g t h e s o i l e n v i r o n m e n t i n a r e a s where i t grows. I n f l u e n c e o f Red A l d e r on S o i l P r o p e r t i e s N i t r o g e n i s one o f the most i m p o r t a n t c o n s t i t u e n t s o f s y n t h e t i c f e r t i l i z e r s w h i c h a r e a p p l i e d t o n u t r i e n t d e f i c i e n t f o r e s t and a g r i c u l t u r a l s o i l s t o i n c r e a s e p r o d u c t i v i t y . However, w i t h the d w i n d l i n g s u p p l i e s and i n c r e a s i n g c o s t s o f f o s s i l f u e l r e q u i r e d t o produce such f e r t i l i z e r s , a t t e n t i o n i s now be i n g f o c u s s e d on i n c r e a s i n g b i o l o g i c a l n i t r o g e n f i x a t i o n as a s u b s t i t u t e t o improve s o i l f e r t i l i t y (Evans and B a r b e r , 1977)-T a r r a n t and Trappe (1971) and T a r r a n t (1972) d i s c u s s e d s e v e r a l s t u d i e s w h i c h have showed t h a t Alnus s p e c i e s improve s o i l f e r t i l i t y by a d d i n g s i g n i f i c a n t amounts o f n i t r o g e n t o the s o i l on wh i c h t h e y a r e g r o w i n g . T h i s i n c r e a s e d s o i l f e r t i l i t y i s u s u a l l y a t t r i b u t e d , e i t h e r d i r e c t l y o r i n d i r e c t l y , t o n i t r o g e n f i x a t i o n t a k i n g p l a c e i n r o o t n o d u l e s and t o a l e s s e r e x t e n t , i n f r e e - l i v i n g s o i l m i c r o - o r g a n i s m s . Red a l d e r i s a l s o known t o improve t h e p h y s i c a l n a t u r e o f s o i l s . I t r e n d e r s - 10 -t h e s o i l o f d r y u n p r o t e c t e d s l o p e s more f r i a b l e and c r e a t e s b e t t e r m o i s t u r e c o n d i t i o n s t h r o u g h r o o t a c t i v i t y and l i t t e r a ccummulation on t h e s e s i t e s ( J o hnson, 1917). I t s l i t t e r decomposes r a p i d l y and forms a mu l l l a y e r , t h e r e b y i m p r o v i n g s o i l s t r u c t u r e ( W o r t h i n g t o n etal.3 1962). T a r r a n t et al. (1951) compared l i t t e r f a l l and f o l i a g e n i t r o g e n c o n t e n t o f 10 major s p e c i e s o f the P a c i f i c N o r t h w e s t . They found t h a t red a l d e r added a r e l a t i v e l y l a r g e amount o f l i t t e r t o the f o r e s t f l o o r and t h a t i t s f o l i a g e had a much g r e a t e r n i t r o g e n c o n t e n t than t h e o t h e r s p e c i e s examined. S e v e r a l subsequent s t u d i e s have s u p p o r t e d t h e view t h a t red a l d e r s i g n i f i -c a n t l y i n c r e a s e s t h e n i t r o g e n c o n t e n t and improves the s t r u c t u r e o f s o i l s on whic h i t i s p r e s e n t ( F o r r i s t a l and Ge s s e l , 1955; T a r r a n t and M i l l e r , 1963; T a r r a n t et al.3 1969; B o l l e n et al.} 1967; B o l l e n and Lu, 1968; F r a n k l i n et al.3 1968; Newton et al.t 1968; Z a v i t k o w s k i and Newton, 1968 and 1971; G e s s e l and T u r n e r , 1974; C o l e et al.3 1978). A s s o c i a t e d s p e c i e s such as D o u g l a s - f i r and b l a c k cottonwood thus d e r i v e b e n e f i t s from improved s o i l c o n d i t i o n s ( T a r r a n t , 1961; B e c k i n g , 1972; M i l l e r and M u r r a y , 1978; De B e l l and Radwan, 1979). Greenhouse s t u d i e s have a l s o d e m onstrated t h e b e n e f i c i a l i n f l u e n c e o f red a l d e r on the growth o f Monterey p i n e {Pinus vadiata D. Don) s e e d l i n g s (wbllum and Youngberg, 1964). N i t r o g e n e n r i c h m e n t by red a l d e r may t a k e p l a c e t h r o u g h l i t t e r f a l l and l i t t e r d e c o m p o s i t i o n , t h r o u g h t h e a c t i v i t y o f f r e e - l i v i n g o r ganisms g r o w i n g i n t h e - 11 -s o i l beneath i t , by d i r e c t s e c r e t i o n i n t o the s o i l from n o d u l e s and l i v i n g r o o t s o r via washings from above-ground p a r t s o f t r e e s i n p r e c i p i t a t i o n r e a c h i n g t h e s o i l ( T a r r a n t and T r a p p e , 1971; T a r r a n t , 1972). E s t i m a t e s o f the amount o f n i t r o g e n added t o t h e s o i l as a consequence o f red a l d e r ' s p r e s e n c e d i f f e r , presumably because o f d i f f e r e n c e s i n s i t e c o n d i t i o n s , t r e e ages and e x p e r i m e n t a l p r o c e d u r e s used i n t h e v a r i o u s s t u d i e s . T a r r a n t (1972) i n d i c a t e d t h a t t h i s s p e c i e s enhances s o i l n i t r o g e n by an average o f a p p r o x i m a t e l y 168 k i l o g r a m s / h e c t a r e / y e a r . Z a v i t k o w s k i and Newton (1968) found t h a t i n the two t o 14 y e a r o l d s t a n d s t h e y examined, a p p r o x i m a t e l y 100 k i l o g r a m s / h e c t a r e / y e a r c o u l d be added t o t h e s o i l t h r o u g h 1 i t t e r d e c o m p o s i t i o n and t h a t t h i s c o n d i t i o n remained s t a b l e d u r i n g f u l l occupancy o f the s i t e . An a d d i t i o n a l 35 k i l o g r a m s / h e c t a r e / y e a r were p r e s e n t i n t h e above ground p a r t s o f t r e e s . These components r e p r e s e n t e d o n l y k0% o f the n i t r o g e n added t o the e c o s y s t e m ; the r e m a i n i n g 60% was thought t o come from d i r e c t s e c r e t i o n s by n o d u l e s and a c t i v e r o o t s , from f i x a t i o n by f r e e - l i v i n g m i c r o - o r g a n i s m s and from d e c o m p o s i t i o n o f n i t r o g e n r i c h dead r o o t s and n o d u l e s . L i t t e r f a l l and l i t t e r d e c o m p o s i t i o n p l a y an i m p o r t a n t p a r t i n the n i t r o g e n e n r i c h m e n t p r o c e s s and as s u c h , t h e amount o f l i t t e r a c c u m u l a t i n g on the f o r e s t f l o o r and t h e r a t e o f d e c o m p o s i t i o n a r e s i g n i f i c a n t i n d e t e r m i n i n g t h e r a t e a t w h i c h n i t r o g e n i s r e t u r n e d t o t h e s o i l . Annual l i t t e r p r o d u c t i o n i n o l d e r red a l d e r s t a n d s a p pears t o be about 5000 k i l o g r a m s / h e c t a r e ( T a r r a n t et al., 1969). The s e a s o n a l d i s t r i b u t i o n o f i t s l i t t e r f a l l f o l l o w e d a p a t t e r n t y p i c a l o f d e c i d u o u s s p e c i e s ( Z a v i t k o w s k i and Newton, 1971). B e f o r e age 10, annual l i t t e r p r o d u c t i o n was p o s i t i v e l y c o r r e l a t e d w i t h age. The next decade was one o f maximum p r o d u c t i o n and then t h e r e was a d e c l i n e i n l i t t e r p r o d u c t i o n , presumably as a r e s u l t o f canopy c l o s u r e . T h e i r s t u d y a l s o showed t h a t t h e r e was l i t t l e y e a r l y v a r i a t i o n i n p r o d u c t i o n and t h a t l i t t e r p r o d u c t i o n was not a f f e c t e d by s t a n d d e n s i t y , age, h e i g h t o r s i t e p r o d u c t i v i t y . G e s s e l and T u r n e r (1974) and T a r r a n t et al., (1969) r e p o r t e d t h a t annual v a r i a t i o n i n weather p a t t e r n s c o u l d a f f e c t l i t t e r p r o d u c t i o n . L i t t e r breakdown i n pure o r mixed red a l d e r s t a n d s o c c u r s a t a much more r a p i d r a t e than i n c o n i f e r o u s s t a n d s ( T a r r a n t and M i l l e r , 1963; T a r r a n t et al., 1969; B o l l e n et al., 1967; C o l e et al., 1978). There a r e q u a l i t a t i v e and q u a n t i t a t i v e d i f f e r e n c e s between f u n g a l p o p u l a t i o n s i n s o i l beneath red a l d e r and t h o s e i n s o i l beneath c o n i f e r s ( T a r r a n t , 1961 ; B o l l e n and W r i g h t , 1961; Wicklow et al.3 1974; Neal et al., 1968a and b ) . These d i f f e r e n c e s p r o b a b l y r e f l e c t d i f f e r e n c e s i n r a t e s and p r o c e s s e s o f n u t r i e n t t u r n o v e r under the d i f f e r e n t t y p e s o f v e g e t a t i o n . There i s no c l e a r p i c t u r e o f t h e i n f l u e n c e o f red a l d e r on t h e a v a i l a b i l i t y o f o t h e r n u t r i e n t s i n the s o i l . Data o b t a i n e d from such s t u d i e s may be i n f l u e n c e d by such f a c t o r s as s o i l a c i d i t y , l e a c h i n g and the f o r m a t i o n o f complexes ( B o l l e n et al., 1967) and t h u s , d i f f e r e n c e s i n s i t e f a c t o r s m i ght have c o n t r i b u t e d t o t h e i n c o n s i s t e n c y o f r e s u l t s . C o l e et aZ.,(l978) s t u d i e d red a l d e r and D o u g l a s - f i r ecosystems o f s i m i l a r ages and found l i t t l e d i f f e r e n c e w i t h r e s p e c t t o t o t a l c a l c i u m and p o t a s s i u m a c c u m u l a t e d i n t h e v e g e t a t i o n . They a l s o p o i n t e d o u t t h a t a l t h o u g h e l e m e n t a l c y c l i n g was much more r a p i d i n t h e - 13 -red a l d e r e c o s y s t e m than i n t h a t o f D o u g l a s - f i r , n u t r i e n t l e a c h i n g appeared t o be g r e a t e r i n t h e s o i l under the f o r m e r . That red a l d e r i s e f f e c t i v e i n i n c r e a s i n g t h e n i t r o g e n c o n t e n t o f s o i l i s w e l l documented. However, i t s h o u l d be not e d t h a t a c c u m u l a t i o n o f n i t r o g e n i n such s o i l s c e a s e s w e l l b e f o r e t r e e s a r e mature, p r o b a b l y as a r e s u l t o f the i n v e r s e r e l a t i o n s h i p between n i t r o g e n f i x a t i o n and a v a i l a b l e s o i l ni-trogen (Newton et al., 1968). I t a l s o appears t h a t the p r o p o r t i o n o f s o i l n i t r o g e n d e r i v e d from l i t t e r d e c o m p o s i t i o n i s r e l a t i v e l y s m a l l d u r i n g the f i r s t few y e a r s o f growth. T h i s i s the t i m e when a b u i l d up o f l i t t e r on the f o r e s t f l o o r o c c u r s and seems t o p r o v i d e a s u i t a b l e e n vironment f o r the p r o l i f e r a t i o n and i n c r e a s e d a c t i v i t y o f t h e s o i l m i c r o - f l o r a and s u b t e r r a n e a n fauna ( Z a v i t k o w s k i and Newton, 1971)- However, on more p r o d u c t i v e s i t e s , s o i l n i t r o g e n can be g r e a t l y i n c r e a s e d by red a l d e r growth i n as l i t t l e as f o u r y e a r s (De B e l l and Radwan, 1979)-P o t e n t i a l o f Red A l d e r i n C o n t r o l l i n g Root Rot D i s e a s e s Root r o t i s r e c o g n i z e d as a major cause o f growth l o s s e s and m o r t a l i t y i n c o n i f e r o u s f o r e s t s i n the P a c i f i c N o r t h w e s t . C o n s i d e r a b l e a t t e n t i o n has been f o c u s s e d on t h e i n t e r a c t i o n between t h e f u n g u s , Phellinus (Poria) weirii w h i c h i n f e c t s a number o f c o n i f e r s p e c i e s . Chi Ids and Shea (1967) e s t i m a t e d t h a t i t causes an annual l o s s o f 32 m i l l i o n c u b i c f e e t (about 0.91 m i l l i o n c u b i c metres) o f c o n i f e r o u s wood i n t h e west s i d e D o u g l a s - f i r r e g i o n o f Oregon and Washington a l o n e . L o s s e s a r e g r e a t e s t i n D o u g l a s - f i r , t r u e f i r s (Abies) - 14 -and w e s t e r n hemlock but r e l a t i v e l y s m a l l in p o n d e r o s a p i n e (Pinus ponderosa L a w s . ) . I n t e n s i v e management o f D o u g l a s - f i r i s r e p o r t e d t o i n c r e a s e l o s s e s in some a r e a s (Ne lson et al., 1978) . Symptoms o f the d i s e a s e and d i s t i n g u i s h i n g f e a t u r e s f rom o t h e r r o o t r o t d i s e a s e s were d e s c r i b e d by Buchanan (19**8), Hansen (1975) and Wal l i s ( 1976) . S i m i l a r l y , W a l l i s and R e y n o l d s (1965) d e s c r i b e d the i n i t i a t i o n and s p r e a d o f the fungus in D o u g l a s - f i r g rowing in c o a s t a l B r i t i s h C o l u m b i a . They a l s o found t h a t whereas D o u g l a s - f i r and w e s t e r n hemlock were q u i t e s u s c e p t i b l e t o i n f e c t i o n , w e s t e r n red c e d a r , b i g l e a f maple and red a l d e r were q u i t e r e s i s t a n t . Red a l d e r ' s r e s i s t a n c e was l a t e r c o n f i r m e d by W a l l i s (1968) . R e g u l a t i o n o f s o i l o r g a n i s m s by red a l d e r and a l t e r n a t i v e methods t o c o n t r o l the s p r e a d o f the d i s e a s e were d i s c u s s e d by T r a p p e (1972) and N e l s o n et al., (1978) , hence o n l y e v i d e n c e w h i c h l e d t o the b e l i e f t h a t red a l d e r has some p o t e n t i a l in r e s t r i c t i n g fungus s p r e a d w i l l be c o n s i d e r e d h e r e . B o l l e n et a Z . , 0 9 6 7 ) and Lu et al., (1968) s u g g e s t e d t h a t a n t i b i o t i c s p r o d u c e d by the p r e p o n d e r a n c e o f Streptomyoes s p e c i e s in s o i l under mixed red a l d e r - c o n i f e r s t a n d s were i m p o r t a n t in r e d u c i n g p o p u l a t i o n s o f f u n g i w h i c h a t t a c k e d c o n i f e r r o o t s . L i et al.,(1967) f ound t h a t u n l i k e t h e s e a n t a g o n i s t s w h i c h t h r i v e on n i t r a t e as a n i t r o g e n s o u r c e , Phellinus (Povia) w e i r i i d i d not use n i t r a t e as a s o u r c e o f n i t r o g e n nor d i d i t s c e l l f r e e e x t r a c t s c o n t a i n n i t r a t e r e d u c t a s e a c t i v i t y . S o i l s under red a l d e r a r e r i c h in n i t r a t e and i t was thought t h a t the i n a b i l i t y o f the fungus t o u t i l i z e n i t r a t e - 15 -would cause i t s growth t o be s u p p r e s s e d by a n t a g o n i s t i c o r g a n i s m s . E v i d e n c e o f t h i s s u p p r e s s i o n was o b t a i n e d by N e l s o n (1967) . In a subsequent s t u d y ( N e l s o n , 1968) , he examined growth o f the fungus i n cubes o f D o u g l a s - f i r heartwood b u r i e d f o r 18 months i n s o i l under c o n i f e r , red a l d e r and mixed c o n i f e r - r e d a l d e r s t a n d s . He c l a i m e d t h a t s u r v i v a l o f t h e fungus was g r e a t e r under c o n i f e r s than under s t a n d s c o n t a i n i n g red a l d e r but d i d not t e s t t h e s t a t i s t i c a l s i g n i f i c a n c e o f t h e s e d i f f e r e n c e s . The e f f e c t s o f lower pH o f red a l d e r s o i l s on the growth o f the fungus might have c o n t r i b u t e d t o t h e o b s e r v e d d i f f e r e n c e s ( N e l s o n et al., 1978) . I t has been s u g g e s t e d t h a t n i t r o g e n f e r t i l i z a t i o n might a l s o promote t h e growth o f m i c r o b i a l a n t a g o n i s t s and reduce t h a t o f the pathogen ( N e l s o n , 1970 and 1975a) but Wa l l i s and Reynolds (197*0 showed t h a t t h e a p p l i c a t i o n o f urea and n i t r a t e f e r t i l i z e r s t o c o n i f e r s t a n d s d i d not reduce t h e s p r e a d o f the d i s e a s e n or l o s s e s caused by i t . S i m i l a r l y , N e l s o n (1975b) found t h a t s u r v i v a l o f the fungus d i d not d i f f e r s i g n i f i c a n t l y between r e d a l d e r and c o n i f e r s t a n d s a l t h o u g h pH was s i g n i f i c a n t l y lower and n i t r a t e c o n t e n t s i g n i f i c a n t l y h i g h e r i n t h e s o i l beneath red a l d e r . Red a l d e r a p p e ars t o have some i n f l u e n c e on the s u r v i v a l o f t h e pathogen but the p r o c e s s by w h i c h t h i s i s a c h i e v e d may r e q u i r e a long t i m e p e r i o d and may i n v o l v e a c o m b i n a t i o n o f f a c t o r s ( N e l s o n et al., 1978) . I t does not appear t o be v e r y e f f e c t i v e i n p r e v e n t i n g t h e s p r e a d o f the d i s e a s e i n mixed s t a n d s . Hansen (1975) found i t t o be i n e f f e c t i v e i n p r e v e n t i n g i n i t i a l i n f e c t i o n s o f - 16 -10 t o 17 y e a r o l d D o u g l a s - f i r t r e e s i n an even aged m i x t u r e o f both s p e c i e s . S i m i l a r f i n d i n g s were r e p o r t e d by Wa l l i s (1976) who warned t h a t c a u t i o n s h o u l d be e x e r c i s e d when i n t e r p r e t i n g r e s u l t s from s t u d i e s w h i c h s u g g e s t a p o t e n t i a l r o l e f o r red a l d e r i n c o n t r o l l i n g t h e s p r e a d o f the pathogen. Red a l d e r i t s e l f i s r e s i s t a n t t o i n f e c t i o n by Phellinus (Poria) w e i r i i but t h e r e s i s t a n c e mechanism has not been d e f i n e d . P o t a s s i u m s a l t s o f l i n o l e i c a c i d , a l i p i d e x t r a c t e d from red a l d e r by K u r t h and Becker (1953), were found t o have s t r o n g i n h i b i t o r y e f f e c t s on t h i s fungus and on Fomes annosus ( F r . ) Cooke, a n o t h e r i m p o r t a n t r o o t pathogen i n the P a c i f i c N o r t h -west ( L i et al., 1970b). L i et al.s (1969) found t h a t many p h e n o l i c com-pounds i n h i b i t e d two d i f f e r e n t i s o l a t e s o f the fungus in v i t r o . S u b s e q u e n t l y , some o f t h e s e i n h i b i t o r s were found i n h y d r o l y z e d e x t r a c t s from s o i l under red a l d e r ( L i et al., 1970a) and i n s i m i l a r e x t r a c t s from i t s r o o t s ( L i et al., 1972b). E x t r a c t s from D o u g l a s - f i r r o o t s were shown t o c o n t a i n v a n i l l i c a c i d w h i c h was p r e s e n t i n t h e e x t r a c t s from r e d a l d e r , but t h i s compound by i t s e l f had l i t t l e i n h i b i t o r y e f f e c t on growth o f the pathogen. The e f f e c t o f p h e n o l i c compounds e x t r a c t e d from red a l d e r may a l s o v a r y w i t h t h e s t r a i n o f the fungus b e i n g t e s t e d (Trappe et al., 1973). D i f f e r e n c e s i n enzyme a c t i v i t y ( p h e n o l o x i d a s e , p e r o x i d a s e and n i t r a t e r e d u c t a s e a c t i v i t y ) were found between l e a v e s o f red a l d e r and t h o s e o f D o u g l a s - f i r and t h e s e were c o n s i d e r e d as a p o s s i b l e c o n t r i b u t i o n t o the f o r m e r ' s r e s i s t a n c e mechanism ( L i et al., 1968). The same w o r k e r s r e p o r t e d - 17 -s i m i l a r d i f f e r e n c e s i n enzyme a c t i v i t y between r o o t s and n o d u l e s o f red a l d e r , and r o o t s o f D o u g l a s - f i r ( L i et al.3 1972a). A l l t h e s e f a c t o r s may be i n v o l v e d i n the r e s i s t a n c e mechanism o f red a l d e r t o the p a t hogen, a mechanism w h i c h seems more complex than p r e v i o u s l y t h o u g h t . Management I m p l i c a t i o n s The most v a l u a b l e t r e e s p e c i e s i n t h e P a c i f i c Northwest i s u n d o u b t e d l y D o u g l a s - f i r . F o r e s t management i n t h i s r e g i o n has been f o c u s s e d on t h i s as w e l l as on o t h e r c o n i f e r s p e c i e s . Red a l d e r on the o t h e r hand, seems i n f e r i o r from t h e economic v i e w p o i n t . Both s p e c i e s a r e abundant i n the a r e a but i n the p a s t , the e c o l o g i c a l a t t r i b u t e s o f red a l d e r were o v e r l o o k e d because i t was seen as an a g g r e s s i v e and u n d e s i r a b l e c o m p e t i t o r o f the e c o n o m i c a l l y v a l u a b l e c o n i f e r s . The r e c e n t change i n a t t i t u d e towards red a l d e r has r e s u l t e d i n d i s c u s s i o n s on the f e a s i b i l i t y o f i n t e n s i v e management o f t h i s s p e c i e s t a k i n g i n t o c o n s i d e r a t i o n s i 1 v i c u l t u r a l systems c o m p a t i b l e w i t h t h e i n t e n s i v e management o f D o u g l a s - f i r . These systems u s u a l l y f a l l i n t o two c a t e g o r i e s : ( l ) c r o p r o t a t i o n s u s i n g t h e two as a l t e r n a t e s , and (2) a d m i x t u r e s i n w h i c h both s p e c i e s a r e grown t o g e t h e r . The main o b j e c t i v e o f t h e s e systems i s t o e x p l o i t the n i t r o g e n f i x i n g a b i l i t y o f red a l d e r t o p r o v i d e D o u g l a s - f i r w i t h a more f a v o u r a b l e growth e n v i r o n m e n t . At t h e same t i m e , red a l d e r w i l l p r o v i d e a u s e f u l t i m b e r r e s o u r c e . - 18 -At p r e s e n t c o s t s , a p p l i c a t i o n o f s y n t h e t i c f e r t i l i z e r s t o m a i n t a i n a d e q u a t e s o i l n i t r o g e n l e v e l s in D o u g l a s - f i r s t a n d s a p p e a r t o be more c o s t e f f e c t i v e than u s i n g mixed p l a n t i n g s w i t h red a l d e r . However , minor changes in c o s t s and v a l u e s c o u l d r e v e r s e t h i s s i t u a t i o n in the f u t u r e ( A t k i n s o n and H a m i l t o n , 1978; A t k i n s o n et al.3 1979). It i s l i k e l y t h a t the s t a t u s o f red a l d e r w i l l not change so l o n g as p r e s e n t c o n d i t i o n s p r e v a i l a l t h o u g h t h e r e i s a wide s c o p e f o r f u t u r e use i f the economic p a t t e r n c h a n g e s . T h e r e i s c o n s i d e r a b l e g e n e t i c v a r i a t i o n w i t h i n red a l d e r , even in e c o n o m i c a l l y impor tan t t r a i t s between i n d i v i d u a l t r e e s in a s t a n d and t h i s a u g e r s w e l l f o r t r e e improvement programmes (De B e l l and W i l s o n , 1978; S t e t t l e r , 1978). D e l i b e r a t e management s h o u l d a l s o i n c r e a s e y i e l d s o v e r t h a t wh ich i s o b t a i n e d n a t u r a l l y (De B e l l et al.3 1978). Both t h e s e f a c t o r s s h o u l d p r o v i d e more and improved l o g s than t h o s e p r e s e n t l y a v a i l a b l e . Red a l d e r i s a f a s t g rowing s p e c i e s and w i t h s h o r t r o t a t i o n s , b e n e f i t s f rom i t s h o u l d be o b t a i n e d in a r e l a t i v e l y s h o r t t ime p e r i o d . In a d d i t i o n , a l d e r s g e n e r a l l y have few n a t u r a l e n e m i e s . T h i s t o g e t h e r w i t h i t s g r e a t g e n e t i c d i v e r s i t y makes e x t e n s i v e p e s t damage u n l i k e l y (Gordon and Dawson, 1979). It i s a p i o n e e r s p e c i e s w h i c h c a n N b e used t o c o l o n i z e s i t e s l a i d b a r e by commerc ia l a c t i v i t y and u n s u i t e d f o r c o n i f e r g r o w t h . It can a l s o grow in s o i l s p h y s i c a l l y u n s u i t a b l e f o r c o n i f e r s (Minore et al.3 1969; M i n o r e , 1970; M i n o r e and S m i t h , 1971). In e i t h e r c a s e , s i t e c o n d i t i o n s s h o u l d improve - 19 -s u f f i c i e n t l y a t l e a s t a f t e r one r o t a t i o n , t o p e r m i t t h e o c c u p a t i o n o f the s i t e by o t h e r s p e c i e s . Stream bank p r o t e c t i o n i s a n o t h e r p o t e n t i a l use ( N i e l s o n , 1977). I n d i c a t i o n s a r e t h a t red a l d e r c o u l d become a v e r y u s e f u l s p e c i e s i n the P a c i f i c Northwest but r e a l i z a t i o n o f t h i s p o t e n t i a l w i l l depend on the n a t u r e and e x t e n t o f changes t h a t t a k e p l a c e i n the f o r e s t r y i n d u s t r y i n the next few y e a r s . In the meantime, c o n t i n u e d r e s e a r c h i s n e c e s s a r y t o d e t e r m i n e the r e a l e x t e n t o f i t s e c o l o g i c a l and economic p o t e n t i a l . - 20 -CHAPTER I I GERMINATION OF RED ALDER SEED FROM DIFFERENT LOCATIONS Introduct ion Red alder seeds collected at maturity can germinate with l i t t l e or no pretreatment (Schopmeyer, 197 ;^ Kenady, 1978). This is an indication that they do not enter f u l l dormancy but remain quiescent between maturity and germination which occurs when environmental conditions become favourable. The species can tolerate substantial environmental divers i t y within i t s natural range. Hence, one might expect this f l e x i b l e performance to be reflected by great genetic v a r i a b i l i t y between populations from different locations throughout i t s range. Seeds of different species and of different provenances within a species exhibit varying degrees of dormancy. Geographic location is commonly related to the degree of dormancy shown by such seeds. In temperate regions, seed dormancy ensures survival over winters, the severity of which can vary with geographic location and from year to year within a location. Seed dormancy c l a s s i f i c a t i o n s usually do not take into account the l a t t e r variations nor the different degrees of dormancy they may cause. Although geographic location appears to have a more important bearing on the degree of dormancy shown by seeds of most species, there is a need to examine more - 21 -c a r e f u l l y , the i n f l u e n c e o f annua l v a r i a t i o n s in w i n t e r c o n d i t i o n s on the e x t e n t o f dormancy p r e s e n t in seeds o f d i f f e r e n t s p e c i e s , e s p e c i a l l y t h o s e t h a t a r e r e p u t e d to show no dormancy . The p r e s e n t s t u d y was u n d e r t a k e n t o examine the g e r m i n a t i o n b e h a v i o u r o f c o l d s t r a t i f i e d , p r e s o a k e d and u n t r e a t e d red a l d e r s e e d s c o l l e c t e d f rom d i f f e r e n t p r o v e n a n c e s and to d e t e r m i n e whether any o b s e r v e d d i f f e r e n c e s were r e l a t e d t o g e o g r a p h i c l o c a t i o n o f the seed s o u r c e s . M a t e r i a l s and Methods The i n v e s t i g a t i o n was c a r r i e d o u t in two p a r t s . The f i r s t took p l a c e f rom l a t e 1976 t o e a r l y 1977, and the s e c o n d in e a r l y 1978. T a b l e s 2.1 and 2 . 2 g i v e the l a t i t u d e , l o n g i t u d e and e l e v a t i o n o f the v a r i o u s s e e d s o u r c e s t o g e t h e r w i t h the known d a t e s o f c o l l e c t i o n . F i r s t S tudy C l e a n seed was r e c e i v e d f rom Idaho and W a s h i n g t o n in e a r l y November 1976 and J a n u a r y 1977 r e s p e c t i v e l y a n d , a l t h o u g h the a c t u a l d a t e s o f c o l l e c t i o n were not o b t a i n e d , i n d i c a t i o n s were t h a t t h e s e s e e d s were c o l l e c t e d in e a r l y and l a t e f a l l o f 1976 r e s p e c t i v e l y . S t r o b i l i f rom the B r i t i s h C o l u m b i a n and C a l i f o r n i a n p r o v e n a n c e s were a i r - d r i e d a t room t e m p e r a t u r e u n t i l t h e i r s c a l e s began t o u n f o l d . Seeds were then removed by v i g o r o u s l y s h a k i n g the d r i e d s t r o b i l i i n p a p e r b a g s . Seeds f rom Cedar C r e e k , Wash ing ton and f rom the h i g h e r e l e v a t i o n a t Haney - 22 -Location Lat tude Long i tude Elevation (to the nearest 5 meters) Date of Col l e c t i o n K i t s a u l t * B.C. 55° .29'N. 129°.31'W. 0 October 5, 1976 Nanaimo Lakes, B.C. <*9° .08'N. 123°.58'W. kZ5 October 28, 1976 Haney, B.C. (I) «.9 0 .25'N. 123°.05'W. <400 October H, 1976 Haney, B.C. (II) * 9 ° .25'N. 123°.05'W. 500 October .«•, 1976 Point Reyes, C a l i f o r n i a 38° .05'N. 122°.58'W. 60 December 31, 1976 Cedar Creek,,Washington hi0 .22'N. 1210.lt7'W. 90 _* Lake Pend O r i e l l e , Idaho *8? .20'N. 116°.15'W. 750 .** * :Received January 1977 **: Received November 1976 Table 2.1 Geographic location of provenances used in the f i r s t germination study and dates of "seed c o l l e c t i o n . Locat ion Latitude Long i tude Elevation (to the nearest 5 meters) •Date of Col 1ect ion Vancouver, Washington l(5 0.38'N. 122°.'(0,W. 30 November 25, 1977 C e n t r a l i a , Washington l)6°A3'N. 122°.58'W. 30 November 20, 1977 Myrtle Creek, Oregon 1»3°.02'N. 123°.16'W. *55 November 20, 1977 Cottage Grove, Oregon I»3°.'.8'N. 123°.05'W. 1.55 November 20, 1977 L a y t o n v i l l e , C a l i f o r n i a 39°.'»2,N. 123°.28'W. 505 November 20, 1977 Crescent C i t y , C a l i f o r n i a 'tl°.'t6'N. 12^°.31'W. 30 November 20, 1977 Or 1ck, Cal1fornia iil°.Ji8'N. \2k°.05'V. 150 November 20, 1977 Table 2.2 Geographic location of provenances used in the second germination study and dates of seed c o l l e c t i o n . - 2 3 -were o b t a i n e d f rom s i n g l e t r e e s . O t h e r c o l l e c t i o n s were made f rom s e v e r a l t r e e s in e a c h l o c a t i o n . G e r m i n a t i o n t e s t s f o r e a c h p r o v e n a n c e were c a r r i e d out on a J a c o b s e n Z e p h y r g e r m i n a t o r . Each t e s t i n v o l v e d s i x samples o f 100 s e e d s e a c h . It was not p o s s i b l e t o t e s t seeds f rom a l l s o u r c e s s i m u l t a n e o u s l y . A l l t e s t s were p e r f o r m e d w i t h normal i n d o o r l i g h t s u p p l e m e n t e d by f l u o r e s c e n t l i g h t (w i th an i n t e n s i t y o f about 2 5 0 0 lux ) f rom 6 : 0 0 a . m . to 6 : 0 0 p .m. d a i l y t o g i v e a 1 2 hour day and a day t ime t e m p e r a t u r e o f 2 5 ° C . The n i g h t t e m p e r a t u r e ( 6 : 0 0 p .m. t o 6 : 0 0 a . m . ) was 1 8 ° C . Two s e r i e s o f t e s t s were c a r r i e d out f o r e a c h p r o v e n a n c e . The f i r s t was done on u n t r e a t e d seed as soon as i t a r r i v e d in the l a b o r a t o r y . T h i s s t u d y a l l o w e d e s t i m a t i o n o f seed v i a b i l i t y and r e v e a l e d any a p p a r e n t l y dormant m a t e r i a l . Unused seeds were s t o r e d a t 0 ° C . w h i l e the f i r s t t e s t s were b e i n g c o n d u c t e d . T h e r e a f t e r , t e s t s were p e r f o r m e d on t h e s e seeds w i t h o u t f u r t h e r p r e t r e a t m e n t , on seeds soaked in tap w a t e r f o r 2k h o u r s and on s i m i l a r l y soaked s e e d s w i t h s u b s e q u e n t c o l d s t r a t i f i c a t i o n a t 0 ° C . f o r e i g h t d a y s . Each t e s t l a s t e d \k days e x c e p t in the c a s e o f the i n i t i a l t e s t i n g o f s e e d s f rom the Nanaimo Lakes a r e a w h i c h was t e r m i n a t e d a f t e r the t w e l f t h d a y . - 2k -Second Study ^ A l l t e s t s In the second p a r t o f t h i s g e r m i n a t i o n s t u d y were r e s t r i c t e d by seed s h o r t a g e and were done o n l y w i t h seeds p r e s o a k e d f o r 2k h o u r s . Seeds were e x t r a c t e d f rom a i r d r i e d s t r o b i l i as d e s c r i b e d in the f i r s t s t u d y . T h e s e t e s t s were a l s o o f ]k d a y s ' d u r a t i o n and took p l a c e under the same c o n d i t i o n s as were t h o s e i n the f i r s t s t u d y . In bo th s t u d i e s , g e r m i n a t i o n was r e c o r d e d as b e i n g normal when the r a d i c l e emerged and grew to a l e n g t h e x c e e d i n g t h a t o f the t e s t a . The f i n a l g e r m i n a t i o n c o u n t was o b t a i n e d by a d d i n g t h i s f i g u r e t o the number o f p a r t i a l l y g e r m i n a t e d seeds ( t h o s e w i t h a r a d i c l e l e s s than the p r e s c r i b e d l e n g t h ) a t the end o f the t e s t . G e r m i n a t i o n c o u n t s were made a t r e g u l a r i n t e r v a l s d u r i n g the t e s t s so t h a t g e r m i n a t i o n v a l u e s ( C z a b a t o r , 1962) c o u l d be c a l c u l a t e d . Abnormal g e r m i n a n t s were a l s o r e c o r d e d but t h e s e were e x c l u d e d f rom the f i n a l g e r m i n a t i o n c o u n t . The c o n t e n t s o f u n g e r m i n a t e d seeds were examined m i c r o s c o p i c a l l y , and t h o s e c o n t a i n i n g i n t a c t embryos were c o n s i d e r e d as b e i n g v i a b l e whereas t h o s e t h a t e i t h e r l a c k e d embryos o r c o n t a i n e d embryos showing s i g n s o f f u n g a l d e c a y , were deemed n o n - v i a b l e . The number o f v i a b l e u n g e r m i n a t e d seeds was added t o the number o f g e r m i n a n t s t o o b t a i n the number o f v i a b l e seeds in e a c h sample on wh ich g e r m i n a t i o n p e r c e n t was b a s e d . C o m p a r i s o n s between p r o v e n a n c e s were made on the b a s i s o f g e r m i n a t i o n p e r c e n t and g e r m i n a t i o n v a l u e . Where n e c e s s a r y i n the s t a t i s t i c a l a n a l y s i s o f d a t a - 25 -o b t a i n e d , a r c - s i n e t r a n s f o r m a t i o n s were used ( F r e e s e , 197^ *) • R e s u l t s and O b s e r v a t i o n s  F i r s t Study T a b l e 2.3 i s a summary o f the r e s u l t s o f the i n i t i a l g e r m i n a t i o n t e s t s on u n t r e a t e d s eeds. The mean g e r m i n a t i o n p e r c e n t exceeded 32% i n a l l t e s t s e x c e p t t h o s e from t h e lower e l e v a t i o n s i t e a t Haney ( T a b l e 2 . 1 ) . . The p e r c e n t a g e o f v i a b l e seeds c o l l e c t e d ranged from 7«3% (Haney 11 c o l l e c t i o n ) t o 79.0% (Nanaimo Lakes c o l l e c t i o n ) . A l l n o n - v i a b l e seeds from Haney 11 l a c k e d embryos and, as a r e s u l t o f t h i s poor y i e l d , the provenance was e l i m i n a t e d from f u r t h e r s t u d y . Most n o n - v i a b l e seeds l a c k e d embryos and f u n g a l i n f e c t i o n was r e l a t i v e l y r a r e e x c e p t i n the case o f the Idaho provenance. A p p r o x i m a t e l y 25% o f the n o n - v i a b l e seeds from t h i s s o u r c e had embryos t h a t were a p p a r e n t l y once h e a l t h y but had d e t e r i o r a t e d due t o f u n g a l a c t i v i t y . I n f e c t i o n was u s u a l l y d e t e c t e d ( P l a t e s 2.1 and 2.2) a l t h o u g h t h e r e were t i m e s when i t was n e c e s s a r y t o p r o d the embryo w i t h a s h a r p - p o i n t e d i n s t r u m e n t t o r e v e a l t h e d e c a y i n g p o r t i o n ( P l a t e 2.3)• Abnormal g e r m i n a n t s were uncommon f o r a l l provenances a l t h o u g h t h e r e were more i n the K i t s a u l t and Cedar Creek c o l l e c t i o n s . Abnormal g e r m i n a n t s f e l l i n t o t h r e e main c a t e g o r i e s ; t h o s e w i t h d o u b l e embryos ( P l a t e 2 . 4 ) ; t h o s e w i t h c o t y l e d o n s emerging w i t h o u t r a d i c l e emergence ( P l a t e 2 . 5 ) ; and - 26 -Provenance Mean Germination % Viable Seed % Non-viable Seed % Abnorma1 Germination % Ki tsaul t 93.9 65.2 33.2 1 .6 Haney (l) 66.9 74.4 25.5 0.3 Haney (l l) 95.9 7.3 92.5 0.2 Nanaimo Lakes 99.8 79.0 20.7 0.2 1 daho 92.8 49.1 50.5 0.3 Point Reyes 94.2 17.8 81.9 0.3 Cedar Creek 100.0 23.2 75.4 1 .3 Table 2.3 Mean germination percentages and percentages of viable seeds, non-viable seeds and abnormal germinants in each of the seed collections used in the f i r s t germination study. - 27 -P l a t e 2 . 1 . Funga l decay i n embryo o f seed f rom the Haney ( I ) c o l l e c t i o n a t the end o f i n i t i a l t e s t f o r the f i r s t g e r m i n a t i o n s t udy (X20) . P l a t e 2 . 2 . Funga l decay i n embryo o f seed f rom the Haney ( l ) c o l l e c t i o n a t the end o f i n i t i a l t e s t f o r the f i r s t g e r m i n a t i o n s t u d y (X20 ) . - 28 -PI ate 2.3•• Embryo of seed from the Haney (I) c o l l e c t i o n with part of decaying section removed with a needle (X20). Plate 2.k. Abnormal germinant from the Kitsa u l t c o l l e c t i o n with double radicles (R) (XI5) . ( H - h y p o c o t y l ) . I - 29 P l a t e 2 . 5 - Abnormal germinant from the K i t s a u l t C o l l e c t i o n w i t h pre-emergent c o t y l e d o n s (X15) J - 30 -those in which hypocotyl emergency was abnormal ( P l a t e 2.6). Some were abnormal by two c r i t e r i a , e.g. double embryos and pre-emergent cotyledons (Plate 2.1). The d i f f e r e n t types of abnormality were observed in a l l provenances and, except f o r the pre-emergence of cotyledons in seeds from Idaho, no one type of abnormality appeared to be more common in a p a r t i c u l a r provenance. The cause of abnormal emergence o f hypocotyls was not c l e a r . The r a d i c l e was observed growing around the inner edge of the p e r i c a r p instead of through the micropyle (P l a t e 2.8) and as elongation of the hypocotyl continued, that part adjacent to the micropyle was forced through t h i s opening in the form of a loop ( P l a t e 2.9). There was a l s o evidence of phy s i c a l r e s t r i c t i o n at the micropylar end of the seed causing the elongating hypocotyl to emerge at some weaker point along the edge of the t e s t a (Plate 2.10). The aberrant c o n d i t i o n was oft e n temporary as once the r e s t r i c t i o n was overcome ( P l a t e 2.11), r a d i c l e growth continued normally. There was u s u a l l y a swel1ing where the apparent r e s t r i c t i o n was encountered (Plate 2.12). Growth of the hypocotyl w i t h i n the t e s t a in some cases caused the e a r l y emergence of the cotyledons through openings in the sides of the t e s t a ( P l a t e 2.13). Treatment E f f e c t s on In d i v i d u a l Seed C o l l e c t i o n s  Haney Provenance Table 2.k shows a summary of r e s u l t s from germination t e s t s on seeds from - 31 -P l a t e 2.6. Abnormal g e r m i n a n t f r o m t h e K i t s a u l t c o l l e c t i o n showing t h e u n u s u a l emergence of h y p o c o t y l (H) ( X 2 0 ) . - 32 -P l a t e 2 . 7 . Abnormal ge rminan t f r o m t h e K i t s a u l t c o l l e c t i o n w i t h d o u b l e embryos and p r e - e m e r g e n t c o t y l e d o n s ( X 1 5 ) . - 33 -2.8. Abnormal germinant from the K i t s a u l t c o l l e c t i o n showing r a d i c l e (R) growing around inner edge of pericarp while hypocotyl (H) emerges at the micropylar end of sec-d ( X 2 0 ) . 2 . 9 . Abnormal germinant from the K i t s a u l t c o l l e c t i o n showing the l o o p - l i k e emergence of hypocotyl (H) at micropylar end of seed (X20). - 34 -2.10. Abnormal germinant from the K i t s a u l t c o l l e c t i o n showing the emergence of hypocotyl (H) along the edge of t e s t a r a t h e r than at the micropylar end of seed (X20). : — " J 2,11. Abnormal germinant from the K i t s a u l t c o l l e c t i o n showing r a d i c l e (R) appparently growing normally a f t e r e a r l i e r r e s t r i c t i o n was overcome (X20). - 35 -s w P l a t e 2.12. Embryo of seed f r o m t h e K i t s a u l t c o l l e c t i o n showing s w e l l i n g (SW) a t r a d i c l e end a p p a r e n t l y formed where p h y s i c a l r e s t r i c t i o n was e n c o u n t e r e d ( X 1 5 ) . P l a t e 2.13. Abnormal g e r m i n a n t f r o m t h e K i t s a u l t c o l l e c t i o n showing p r e - e m e r g i n g c o t y l e d o n s (C) caused by g r o w t h of h y p o c o t y l w i t h i n t h e t e s t a ( X 1 5 ) . - 36 -Treatment Germination measure Stratification Presoaking No pretreatment F-value Significance Germination percent 99.4 a 67.4 b 73.3 c 145.38 • Germination value 67.03 a 47.52 b 27.91 c 167.96 • • : significant at the 5% level. Figures followed by different letters are significantly different from each other at the 5% level. Table 2.h. Germination data for s t r a t i f i e d , presoaked and untreated seeds of the Haney provenance. Germination percent Days » e i Stratified seeds -•— .a i Presoaked seeds - Untreated seeds Figure 2.1. Course of germination for s t r a t i f i e d , presoaked and untreated seeds of the Haney provenance. - 37 -t h e Haney l o c a t i o n . Mean g e r m i n a t i o n p e r c e n t and mean g e r m i n a t i o n v a l u e f o r s t r a t i f i e d seeds were s i g n i f i c a n t l y h i g h e r than t h o s e f o r p r e s o a k e d seeds w h i c h i n t u r n were s i g n i f i c a n t l y g r e a t e r than the c o r r e s p o n d i n g measurements f o r u n t r e a t e d seeds. The c o u r s e o f g e r m i n a t i o n f o r each t r e a t m e n t i s shown i n F i g u r e 2 .1 . The graph shows t h a t whereas the i n i t i a l r a t e s o f g e r m i n a t i o n f o r s t r a t i f i e d and p r e s o a k e d seeds were s i m i l a r and g r e a t e r t h a n t h a t f o r u n t r e a t e d s e e d s , a h i g h e r r a t e was m a i n t a i n e d i n the s t r a t i f i e d seeds o v e r t h e p r e s o a k e d ones a f t e r the f o u r t h day. The t r e n d f o r g e r m i n a t i o n o f u n t r e a t e d seeds was more o r l e s s s t e a d y t h r o u g h o u t the t e s t p e r i o d . At t h e end o f the t e s t s , embryos o f u n g e r m i n a t e d , v i a b l e , p r e s o a k e d and u n t r e a t e d seeds were removed, c h i l l e d a t 0°C. f o r f i v e days and r e t u r n e d t o th e same c o n d i t i o n s under w h i c h t h e g e r m i n a t i o n t e s t s were c o n d u c t e d . A f t e r f o u r d a y s , a l l but two showed n o t i c e a b l e growth w i t h h y p o c o t y l s e x c e e d i n g 1 cm. i n l e n g t h and w i t h w e l l d e v e l o p e d c o t y l e d o n s . The e x c e p t i o n s d i d have c o t y l e d o n s but t h e i r r a d i c l e ends showed f u n g a l decay w h i c h d e v e l o p e d a f t e r the c o n c l u s i o n o f t h e g e r m i n a t i o n t e s t s . The growth o f embryos a f t e r the p e r i o d o f c h i l l i n g s u g g e s t e d the p r e s e n c e o f dormant seeds i n the c o l l e c t i o n . T h i s h a v i n g been t h e c a s e , s t r a t i f i c a t i o n and, t o a l e s s e r e x t e n t , p r e s o a k i n g were e f f e c t i v e i n o v e r c o m i n g the dormancy and In i m p r o v i n g g e r m i n a t i o n o v e r t h a t o f u n t r e a t e d s e e d s . - 38 -K i t s a u l t Provenance N e i t h e r s t r a t i f i c a t i o n nor p r e s o a k i n g had a s i g n i f i c a n t e f f e c t on g e r m i n a t i o n p e r c e n t and g e r m i n a t i o n v a l u e f o r seeds o f t h i s provenance ( T a b l e 2.5). The l a t t e r showed a s l i g h t i n c r e a s e w i t h p r e s o a k i n g and s t r a t i f i c a t i o n . The o n l y a p p a r e n t e f f e c t o f t h e s e t r e a t m e n t s was t o i n c r e a s e the r a t e o f g e r m i n a t i o n i n the e a r l y s t a g e s o f t h e t e s t p e r i o d ( F i g u r e 2.2) but t h i s was not s u f f i c i e n t l y g r e a t t o b r i n g about s i g n i f i c a n t d i f f e r e n c e s i n germina-t i o n v a l u e . Nanaimo Lakes Provenance G e r m i n a t i o n p e r c e n t was s i m i l a r f o r s t r a t i f i e d , p r e s o a k e d and u n t r e a t e d seeds o f the Nanaimo Lakes p r o v e n a n c e , whereas g e r m i n a t i o n v a l u e d e c r e a s e d s i g n i f i c a n t l y from s t r a t i f i e d t h r o u g h p r e s o a k e d t o u n t r e a t e d seeds ( T a b l e 2.6). D i f f e r e n c e s i n t h e r a t e o f g e r m i n a t i o n brought about by t r e a t m e n t s ( F i g u r e 2.3) were a p p a r e n t l y r e s p o n s i b l e f o r the s i g n i f i c a n t d i f f e r e n c e s i n g e r m i n a t i o n v a l u e d e s p i t e n o n - s i g n i f i c a n t d i f f e r e n c e s i n g e r m i n a t i o n p e r c e n t . Idaho Provenance G e r m i n a t i o n p e r c e n t d i d not d i f f e r s i g n i f i c a n t l y between t r e a t m e n t s ( T a b l e 2.7). G e r m i n a t i o n v a l u e f o r soaked seeds was s i g n i f i c a n t l y lower than t h a t f o r s t r a t i f i e d and u n t r e a t e d seeds but they were not s i g n i f i c a n t l y d i f f e r e n t from each o t h e r ( T a b l e 2.7). F i g u r e l . h shows t h a t w h i l e the p a t t e r n o f g e r m i n a t i o n d i d not d i f f e r much between t r e a t m e n t s , the r a t e f o r p r e s o a k e d seeds was lower than t h a t o f the o t h e r seeds a l m o s t t h r o u g h o u t t h e t e s t p e r i o d . - 39 -Treatment Germination measure Stratification Presoaking No pretreatment F-value Germination percent 95.0 a 92.7 a 95.0 a 1.01 N.S. Germination value 107.12 a 104.97 a 96.03 a 1.02 N.S. N.S. : not significant at the 5* level. Figures followed by the same letter are not significantly different from each other at the 5% level. Table 2.5. Germination data for s t r a t i f i e d , presoaked and untreated seeds of the Kitsault provenance. 100 r Days » > i Stratified seeds _ . a , Presoaked seeds .— —~-—. Untreated seeds Figure 2.2. Course of germination for s t r a t i f i e d , presoaked and untreated seeds of the Kitsault provenance. - ko -Treatment Germination measure Stratification Presoaking No pretreatment F-value S i g n i f i c a n c e Germination percent 99.9 a 99.8 a 99.9 a 0.45 N.S. Germination value 149.21 a 130.41 b 113.06 c 56.15 • N .S. : not significant at the 5% level. • : significant at the 5% level. Figures followed by the same letter are not significantly different from each other at the 5% l e v e l . Table 2.6. Germination data for s t r a t i f i e d , presoaked and untreated seeds of the Nanaimo Lakes provenance. 100 Germination 50 percent Days : Stratified seeds • Presoaked seeds , Untreated seeds Figure 2.3. Course of germination for stratified, presoaked and untreated seeds of the Nanaimo Lakes provenance. - k] -Treatment Germination measure Stratification Presoaking No pretreatment F-value Significance Germination percent 94.7 a 96.2 a 96.7 a 0.67 N.S. Germination value 76.66 a 67.89 b 76.85 a 3.85 • N.S. : not significant at the 5% level. • : significant at the 5% level. Figures followed by the same letter are not significantly different from each other at the 5% level. Table 2 . 7 . Germination data for s t r a t i f i e d , presoaked and untreated seeds of the Idaho provenance. loof-Ccrmi nation percent Diys « » i Stratified seeds -• —« 1 Presoaked seeds —*— . — : Untreated seeds Figure 2.h. Course of germination for s t r a t i f i e d , presoaked and untreated seeds of the Idaho provenance. - kl -The c a l c u l a t e d g e r m i n a t i o n v a l u e f o r the i n i t i a l t e s t f o r t h i s c o l l e c t i o n was 7b.k2% wh ich was s i m i l a r t o t h o s e f o r s t r a t i f i e d and u n t r e a t e d seeds in t h i s t e s t . T h i s s i m i l a r i t y and the a p p a r e n t d e p r e s s i o n in the r a t e o f g e r m i n a t i o n c a u s e d by p r o l o n g e d s o a k i n g w i t h o u t s u b s e q u e n t s t r a t i f i c a t i o n , prompted a f u r t h e r t e s t in which s i x samples each o f 1.00 seeds were soaked f o r 36 hours and then g e r m i n a t e d under the same c o n d i t i o n s o f the o t h e r t e s t s . On ly 16 o f the 600 seeds g e r m i n a t e d s u c c e s s f u l l y and a l l o f t h e s e d i d so w i t h i n the l a s t seven days o f the 14 day t e s t p e r i o d . At the end o f the t e s t , f u n g a l a c t i v i t y had damaged a l l the unger mina ted embryos . Cedar Creek P r o v e n a n c e G e r m i n a t i o n p e r c e n t f o r the t r e a t e d and u n t r e a t e d seeds d i d not d i f f e r s i g n i f i c a n t l y but g e r m i n a t i o n v a l u e f o r s t r a t i f i e d seeds was s i g n i f i c a n t l y h i g h e r than v a l u e s f o r p r e s o a k e d and u n t r e a t e d s e e d s b o t h o f w h i c h were s i m i l a r ( T a b l e 2 . 8 ) . S t r a t i f i c a t i o n a g a i n a p p e a r e d t o have the e f f e c t o f i n c r e a s i n g the i n i t i a l r a t e o f g e r m i n a t i o n o v e r t h a t o f o t h e r s e e d s ( F i g u r e 2 . 5 ) , g i v i n g a s i g n i f i c a n t l y h i g h e r g e r m i n a t i o n v a l u e d e s p i t e s i m i l a r g e r m i n a t i o n p e r c e n t a g e s . P o i n t Reyes P r o v e n a n c e T a b l e 2 . 9 shows t h a t f o r s e e d s o f t h i s c o l l e c t i o n , o v e r a l l g e r m i n a t i o n as measured by g e r m i n a t i o n v a l u e s , was s i m i l a r in a l l t r e a t m e n t s . G e r m i n a t i o n p e r c e n t a g e s f o r the t r e a t e d s e e d s were not s i g n i f i c a n t l y d i f f e r e n t f rom e a c h o t h e r but were s i g n i f i c a n t l y lower than t h a t f o r u n t r e a t e d s e e d s . - 43 -Treatment Germination measure S t r a t i f i c a t i o n Presoaking No pretreatment F-value Significance Germination percent 100.0 a 100.0 a 99.9 a 1.00 N.S. Germination value 180.40 a 139.55 b 129.99 b 19.57 * N.S. : not s i g n i f i c a n t at the 5% l e v e l . • : s i g n i f i c a n t at the 5% l e v e l . Figures followed by the same l e t t e r are not s i g n i f i c a n t l y d i f f e r e n t from each other at the 5% l e v e l . Table 2.8. Germination data for s t r a t i f i e d , presoaked and untreated seeds of the Cedar Creek (Washington) provenance. 6"! 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Diyt « « i Strttifwd M o d s — — Prr^cf Ked s»«ds — — - i Uilrsalcd M * d t Figure 2.5. Course of germination for s t r a t i f i e d , presoaked and untreated seeds of the Cedar Creek (Washington) provenance. - kk -Germination measure Stratification Presoakinq No pretreatment F-value Significance Germination percent 94.4 a 94.2 a 99.5 b 4.52 • Germination value 51.33 a 54.32 a 58.90 a 1.41 N .S .  N . S . : not significant at the 5% level. • : significant at the 5% level. Figures followed by the same letter are not significantly different from each other at the 5% l e v e l . Table 2.9. Germination data for s t r a t i f i e d , presoaked and untreated seeds of the Point Reyes (California) provenance. Germination 5 0 cercent 0«y» : Stratified seeds • Presoaked seeds : Untreated seeds T3 14 Figure 2.6. Course of germination for s t r a t i f i e d , presoaked and untreated seeds of the Point Reyes (California) provenance. - k5 -The course of germination as represented in Figure 2 . 6 was similar for s t r a t i f i e d and presoaked seeds. Germination of these seeds was recorded from the t h i r d day up to which time the f i r s t germinants from untreated seeds were not observed. However, whereas the treated seeds maintained a more or less steady rate of germination during the f i r s t 10 days of testing, germination of untreated seeds appeared to follow two phases, the f i r s t , a slower steady rate preceding a sharp increase after day eight. Effects of Individual Treatments on a l l Provenances Results from the Cedar Creek provenance were included in the subsequent c o l l e c t i v e comparisons between seed l o t s , although seed from this source was obtained from a single tree. This was legitimate because in the analyses performed, neither seed to seed variation nor tree to tree variation was separated. A summary of the results for individual treatments is presented in Tables 2 . 1 0 and 2 . 1 1 . Except in the case of untreated and presoaked seeds from Haney, germination percent was always above 32%, an observation consistent with the results of the i n i t i a l tests which suggested some degree of dormancy within the Haney seed l o t , but l i t t l e or none in those from other sources. Seeds from Cedar Creek and Nanaimo Lakes showed similar germination percentages regardless of treatment and generally, they were superior to Treatment Germination percent F-value SignifIcance Stratification Cedar Nanaimo Creek Lakes Haney Kitsault 100.0 a 99.9 a 99.4 b 95.0 c Point Idaho Reyes 94.7 c 94.4 c 30.74 Presoaking Cedar Nanaimo Creek Lakes 100.0 a 99.8 a Point Idaho Reyes Kitsault Haney 96.2 b 94.2 bc 92.7 c 87.4 d 48.26 Haney Cedar Nanaimo Point Creek Lakes Reyes Idaho Kitsault No pretreatment 99.9 a 99.9 a 99.5 ab 96.7 bc 95.0 c 73.3 d 23.34 ON • t significant at the 5% level. Figures followed by the same letter are not significantly different from each other at the 5% level. Table 2.10. Summary of analysis of germination percentages for stratified, presoaked and untreated seeds of a l l provenances tested in the f i r s t germination study. Treatment F-value Significance Stratification Cedar Creek 180.40 a Nanaimo Lakes 149.21 b Kitsault Idaho Haney 107.12 c 76.66 d 67.03 d Point Reyes 51.33 e 152.05 Presoaklng Cedar Creek 139.55 a Nanaimo Lakes 130.41 a Kitsault 104.97 b Idaho 67.89 c Point Reyes 54.32 d Haney 47.52 d 94.81 Cedar Nanaimo Point Creek Lakes Kitsault Idaho Reyes Haney No pretreatment 129.99 a 113.06 b 96.03 c 76.85 d 58.90 e 27.91 f 95.36 * • : significant at the 5% level. Figures followed by the same letter are not significantly different from each other at the 5% level. Table 2.11. Summary of analysis of germination values for stratified, presoaked and untreated seeds of a l l provenances tested in the f i r s t germination study. - 48 -s e e d s f rom o t h e r l o c a t i o n s . In no t r e a t m e n t d i d the C a l i f o r n i a n p r o v e n a n c e d i f f e r s i g n i f i c a n t l y f rom the Idaho p r o v e n a n c e , and in no t r e a t m e n t was the Haney p r o v e n a n c e s i m i l a r to any o t h e r . In a d d i t i o n , s e e d s f rom the l a t t e r showed the lowest g e r m i n a t i o n p e r c e n t f o r p r e s o a k e d and u n t r e a t e d s e e d s . Wi th s t r a t i f i c a t i o n however , t h i s measure though s i g n i f i c a n t l y l o w e r , was c l o s e r t o t h a t o f i t s g e o g r a p h i c n e i g h b o u r s . G e r m i n a t i o n p e r c e n t f o r K i t s a u l t s e e d s was s i m i l a r t o e i t h e r t h a t o f seeds f rom P o i n t R e y e s , Idaho o r bo th d e p e n d i n g on the t r e a t m e n t . The r e s u l t s in T a b l e 2.11 show t h a t in d e s c e n d i n g o r d e r , the g r e a t e s t g e r m i n a t i o n as measured by g e r m i n a t i o n v a l u e s , came f rom the Cedar C r e e k , Nanaimo L a k e s , K i t s a u l t and Idaho p r o v e n a n c e s . W i t h o u t s t r a t i f i c a t i o n , seeds f rom P o i n t Reyes were s u p e r i o r t o t h o s e f rom Haney but the r e v e r s e was t r u e where t h i s t r e a t m e n t was a p p l i e d . G e r m i n a t i o n p e r c e n t a g e s were g e n e r a l l y much c l o s e r t o e a c h o t h e r than were g e r m i n a t i o n v a l u e s . S i n c e the l a t t e r t a k e s both g e r m i n a t i o n p e r c e n t and g e r m i n a t i o n r a t e i n t o c o n s i d e r a t i o n , i t may be c o n c l u d e d t h a t the e f f e c t o f t h e s e t r e a t m e n t s where s i g n i f i c a n t , was t o i n c r e a s e the r a t e o f g e r m i n a t i o n p a r t i c u l a r l y in the e a r l y s t a g e s o f the t e s t p e r i o d ( F i g u r e s 2 . 7 , 2 . 8 and 2 . 9 ) . T r e a t m e n t E f f e c t s on A l l P r o v e n a n c e s The s t a t i s t i c a l a n a l y s i s o f c o l l e c t i v e d a t a f o r a l l p r o v e n a n c e s and t r e a t m e n t s i s summarized in T a b l e 2 . 1 2 . T h i s shows a s i g n i f i c a n t i n t e r a c t i o n - hs -Figure 2.7- Course of germination for s t r a t i f i e d seeds from each source tested in the f i r s t germination study. Fi gure 2 . 8 . Course of germination for presoaked seeds from each source tested in the f i r s t germination study. - 51 -Figure 2.9. Course of germination for untreated seeds from each source tested in the f i r s t germination study. - 52 -Germination percent Source of variation' F-value Sign i f icance Provenances 56.17 • Treatment 6.15 • Provenance X treatment interaction 14.70 * Provenance: Cedar Creek Nanaimo Lakes Point Reyes Idaho Kitsault Haney Mean : 100.0 a 99.9 a 96.6 b 95.9 bc 94.3 c 89.6 d Treatment : Stratification Presoaking No pretreatment 98.3 a 96.6 b 96.9 b Germination value Source of variation F-value Significance Provenances 325.58 • Treatment 45.17 • Provenance X treatment interaction 9.76 * Provenance: Cedar Creek Nanaimo Lakes Kitsault Idaho Point Reyes Haney Mean : 150.00 a 130.90 b 102.70 c 73.80 d 54.85 e 47.49 f Treatment : Stratification Presoaking No pretreatment 105.30 a 90.78 b 83.79 c * : significant at the 5% level. Figures followed by the same letter are not significantly different from each other at the 5% level. Table 2.12. Summary of analysis of c o l l e c t i v e germination percentages and germination values for a l l provenances and treatments in the f i r s t germination study. - 53 -between provenances and treatments, both for germination percent and germination value. Figures 2 . 1 0 and 2 . 1 1 show the trend of these measures from provenance to provenance. Differences in germination percent between treatments are most noticeable in the Haney provenance (Figure 2 . 1 0 ) and to a lesser extent, the Point Reyes provenance. There was r e l a t i v e l y l i t t l e variation between treatments in the remaining provenances. Thus, i t appears as though the effect of treatments on the germination of seeds from Haney and the apparent depression these treatments caused in seeds from Point Reyes, were mainly responsible for the interaction for germination percent. Germination value in seeds from Haney, Nanaimo Lakes, Cedar Creek and Kitsault increased with, presoaking and further with s t r a t i f i c a t i o n but there were variations in the extent to which these increases took place, especially in those brought about by s t r a t i f i c a t i o n (Figure 2 . 1 1 ) . There was l i t t l e increase caused by this treatment in seeds from Kitsault while i t was r e l a t i v e l y large in the Cedar Creek seeds. The increase in germination value caused by s t r a t i f i c a t i o n was similar for the Nanaimo Lakes and Haney provenances. However, germination values for the Haney provenance were lower than those for the Nanaimo Lakes provenance. There was l i t t l e variation in germination value between treatments in the Point Reyes and Idaho provenances. These factors most l i k e l y led to the interaction between provenances and treatments for germination value. - 5k -too 80 Germination percent 60 70. Haney s Stratification : Presoaking No pretreatment Point Reyes Idaho Provenance Nanaimo Lakes Cedar Creek Kitsault Figure 2.10. Mean germination percent for s t r a t i f i e d , presoaked and untreated seeds of each provenance tested in the f i r s t germination study. Figure 2 . 1 1 . Mean germination value for s t r a t i f i e d , presoaked and untreated seeds of each provenance tested in the f i r s t germination study. - 55 -A l l provenances and a l l treatments yielded germination values that were s i g n i f i c a n t l y different from each other (Table 2.12). The same was not true however, when germination percentages were compared. In this case, pre-soaked and untreated seeds had similar percentages but these were s i g n i f i -cantly lower than that for s t r a t i f i e d seeds. Seeds from Cedar Creek and Nanaimo Lakes behaved s i m i l a r l y in this respect and their germination percentages were s i g n i f i c a n t l y higher than those for the other provenances. The germination percent of seeds from Haney was s i g n i f i c a n t l y lower than1 that of every other c o l l e c t i o n . Germination percent for seeds from Idaho was not s i g n i f i c a n t l y different from that of seeds from Kitsault nor Point Reyes. The l a t t e r seeds had a s i g n i f i c a n t l y higher germination percent than that of seeds from K i t s a u l t . Differences between provenances and between treatments were again more evident when germination value was used as the measure of germination rather than germination percent. The courses of germination represented in Figures 2.12 and 2.13 also suggest that differences in the rate of germination in the early part of testing contributed substantially to the differences in germination value observed. Second Study Table 2.13 summarizes the analysis of data collected in the second study. Final germination percentages were a l l above 33% and there were some sig n i f i c a n t differences in this measure between provenances. Germination Figure 2 . 1 2 . Course of germination for s t r a t i f i e d , presoaked and untreated seeds of a l l provenances tested in the f i r s t germination study. - 57 -Course o f g e r m i n a t i o n f o r t r e a t e d and u n t r e a t e d seeds o f each c o l l e c t i o n t e s t e d i n the f i r s t g e r m i n a t i o n s t u d y (where not r e c o r d e d , d a t a f o r a p a r t i c u l a r day was o b t a i n e d by e x t r a p o l a t i o n f rom f i g u r e s 1 t o 6 ) . Germination measure F-value Significance Crescent Cottage Myrtle Germination Centralis Vancouver City Grove Creek Irvine OricJc percent 99.2 a 99.4 ab 99.6 ab 100.0 be 100.0 be 100.0 be 100.0 c 2.95 • Germination value 96.54 a 107.60 a 125.96 b 134.93 be 140.00 cd 145.22 cd 151.59 d 20.63 * * i significant at the 5% level. Figures followed by the same letter are not significantly different from each other at the 5% level. Table 2.13. Summary of analysis of germination data for presoaked seeds of a l l provenances tested in the second germination study. - 59 -values between provenances were s i g n i f i c a n t l y different. Both seed lots from Washington showed similar germination values but they were s i g n i f i c a n t l y lower than germination values for the Oregon and Californian provenances. Germination values for the different Oregon collections were not s i g n i f i c a n t l y different from each other. This value for seeds from Crescent City was s i g n i f i c a n t l y lower than those for the other Californian provenances. Values for the Crescent City and Cottage Grove collections were similar but were s i g n i f i c a n t l y lower than that for the Orick seeds which in turn was not s i g n i f i c a n t l y different from those for seeds from Myrtle Creek and Laytonville. The order in which germination values increased through the various provenances was the same as that for germination percent. Differences in germination percent, though small, and differences in germination ratei as shown in Figure 2.1A, would have contributed to the observed differences in germination value. Di scuss ion  F i r s t Study Red alder is a widely distributed species and different populations are exposed to different c l i m a t i c conditions. This favours considerable geographic v a r i a b i l i t y . There is however, l i t t l e discontinuity throughout i t s range, a factor which should encourage gene exchange between populations (Wright, 1976). - 60 -Figure 2.14. Course of germination for presoaked seeds from each source tested in the second germination study. - 61 -Size, age, vigour, crown exposure, heredity and climate are a l l factors which influence the quantity and quality of seeds produced in a particular year by individual trees of a species (Kramer and Kozlowski, I 960) . Some of these factors are inter-related. It is also a common t r a i t of trees of seed bearing age to produce heavy and light crops at regular intervals (Kozlowski, 1971) . In red alder, seed production begins at about age 10 and heavy crops are produced about every four years (Worthington, 1965; Plank, 1971) . When seed was collected for these studies, there was no discrimination as to the above tree characteristics. It was not therefore surprising to find a wide variation in the percentage of viable and non-viable seeds contained in the various seed lot s . McVean (1955) found that the percentage of seeds with embryos and from single trees of black alder (Alnus glutinosa ( L . ) Gaertn.) varied from 1% to 78% in two successive years. The percentage of viable seeds of black alder and grey alder (Alnus inaana ( L . ) Moench) in collections examined by Schalin (1968) was 29.8% and 28.9% respectively. The varied figures for red alder probably r e f l e c t the variation that is, to be expected from alders as a whole. The relationship between time of col l e c t i o n of the various seed lots and the quality of seeds obtained, (especially in those cases where seed was collected after natural seed dispersal began), was not determined in this investigation. The nature of the Point Reyes co l l e c t i o n also might not have been truly representative since these seeds were produced and collected during a period when the region was experiencing an unusually prolonged drought (Californian Department of Water Resources Report, 1977) . - 62 -No attempt was made to determine the cause of extensive embryo decay in seeds from Idaho. It could have arisen as a result of a greater abundance of fungal spores present in their seed coats and most active under the conditions of testing. A l t e r n a t i v e l y , spores might have been equally prevalent in a l l collections with seeds from Idaho having a lower resistance to infection than seeds from other areas. Inhibition of radicle development by fungi while growth of the remainder of the embryo proceeded, was the main cause of the early emergence of cotyledons. This type of abnormality was more common in seeds from Idaho than in the other collections and probably resulted from the high incidence of fungal a c t i v i t y in these seeds. Polyembryony was found in this study and was reported in seeds of black and grey alders examined by Schalin (1968) . The various causes were discussed by Bhatnagar and Johri (1972) . This phenomenon may be common in alders as a whole but the contributing factors and the ways in which i t develops have not been i d e n t i f i e d . A l i k e l y cause of the unusual emergence of the hypocotyl in cases where there appeared to be a physical r e s t r i c t i o n to radicle growth at the micropylar end, was injury to the meristematic tissues or breaking of the vascular connections within the radicle (Pollock and Roos, 1972; Justice, 1972) . The subsequent regeneration of tissues would have caused the characteristic swelling observed in the affected radicles. Also, the delay - 63 -in growth due to the regeneration process would create the impression that radicle growth was being physically impeded. The cause was less clear in the other cases where hypocotyl emergence through the micropyle was accompanied by the curling and growth of the radicle around the testa. Gavrielit-Gelmond (1971) warned that the orientation of a seed in the planting medium with i t s radicle end facing upward might cause bending or curling of the hypocotyl in the early stages of germination, giving the false impression of an abnormality. This does not apply here. The physiological, mechanical or environmental factor responsible for this disorder was not evident. It should be specified here that indications obtained and conclusions drawn from the results of these studies are based solely on tests using seed collected in the areas and at the times indicated, and performed under the conditions described e a r l i e r . The c r i t e r i o n of radicle length exceeding that of the testa before germination was considered successful, was not the usual one recommended by forestry and agricultural i n s t i t u t i o n s . It was chosen in these studies for convenience in measuring and to achieve consistency throughout the tests. There were some apparently dormant seeds in the Haney 1 c o l l e c t i o n . C h i l l i n g was followed by growth of previously u n s t r a t i f i e d embryos of viable seeds that f a i l e d to germinate during testing and this indicated that this - bk -form of dormancy was physiological. Germination f a i l u r e did not appear to be a result of seed immaturity. Seeds collected from various areas throughout the Research Forest at Haney in the 1975 -76 winter showed a 3k% germination in preliminary tests carried out early in 1976. Examination of the climatic data for this location revealed no major differences in weather patterns from 1975 to 1976. Hence, i t does not appear as though differences in weather contributed to the observed differences in the extent of dormancy present in the two collections. Differences in the time of year when these collections were made (winter 1975-76 and f a l l 1976) did not appear to have any influence. Both seed lots were stored under the same low temperature conditions for f i v e and six months respectively between co l l e c t i o n and testing. It was also possible that the extent of dormancy was related to elevation. If seeds from a higher elevation, where winter temperatures would be lower, tended to develop dormancy more readily than those from the lower areas, a broad c o l l e c t i o n from a mixture of low and high elevations would be expected to contain fewer dormant seeds than one from a r e l a t i v e l y high location of A00 metres. This hypothesis was tested. Seed was collected in the f a l l of 1978 from eight trees at the Research Forest at elevations ranging from 90 to 520 metres. Untreated seed from each tree was tested separately in the same way and under the same conditions as the other tests. Almost every viable seed germinated, indicating a near total absence of dormancy. - 65 -Kenady (1978) pointed out that individual trees of red alder of similar age can produce seed with s t r i k i n g l y different patterns of germination, the causes of which are unknown. The results of these tests only indicate that whilst physiological dormancy does not appear to be widespread in seeds of red alder, i t can e x i s t . Further investigations are required to determine whether the production of dormant seeds in this species is confined to certain provenances, whether i t is periodic and occurs only in response to some environmental factor, whether i t is a genetic t r a i t of some trees or of a l l trees, and the environmental factor or factors which trigger their production. Germination value measured germination rate, germination percent and any interactions between the two. The effect of presoaking in improving germination is usually attributed to the removal of water soluble agents causing dormancy, softening the seed coat to f a c i l i t a t e radicle emergence and raising the moisture content within seeds to the c r i t i c a l level for the i n i t i a t i o n of germination more quickly than i f this moisture had to be absorbed from the germinating medium. S t r a t i f i c a t i o n is thought to remove physiological dormancy and create favourable conditions within the seed for immediate germination under favourable environmental conditions. In this investigation, dormancy was not a s i g n i f i c a n t factor in preventing germination except in seeds from Haney. Red alder seeds possess r e l a t i v e l y soft seed coats which do not normally appear to hinder radicle emergence. Thus, i t seems that these treatments increased germination rate as a result - 66 -of the favourable conditions they created within the seeds affected. Their effectiveness in removing dormancy was an added factor in the case of the Haney seeds. There seems to be no satisfactory d e f i n i t i o n of the term "seed vigour" (Bonner, 197^; Perry, 1976) . It is used here to refer to the t o t a l i t y and rate of germination as measured by germination value. The effects of s t r a t i f i c a t i o n and presoaking were most evident in the more vigorous untreated seeds, and in the Haney c o l l e c t i o n . There was no direct relationship between seed vigour and any geographic factor. Rather, i t seemed to decrease from Cedar Creek northward and southward for the more coastal areas, and inland to Idaho. The results of the tests in the f i r s t study might be interpreted as suggesting that red alder seeds are most vigorous in the central portion of i t s range, and less vigorous in the more southerly and northerly latitudes, and with increasing distance from the coast. Confirmation of this requires testing of various collections from areas along i t s l a t i t u d i n a l range and at varying distances from the coast. If the behaviour of seeds from Haney over successive years is any indication of that in other populations, meaningful studies in the future may have to involve testing of seed collected at the same locations over several successive years. Second Study In the second study, germination of presoaked seeds from provenances within a - 67 -narrower l a t i t u d i n a l range, was examined. On the basis of germination percentages obtained, dormancy was not a s i g n i f i c a n t factor among these seeds. Differences in germination value arose mainly in the rate of germination during the early part of testing (Table 2.13 and Figure 2.14). As in the f i r s t study, there appeared to be no relationship between seed vigour and geographic location of the provenances. When the more northerly provenances from Oregon and Washington are considered, there is a tendency for germination value to increase as latitude decreases. However, this trend did not continue in the Californian provenances. Collections from Orick and Laytonville were similar and more vigorous than that from Crescent City. Orick and Crescent City are closely situated and Laytonville is the most southerly and the highest of the three. Vigour of the Crescent City seeds was similar to that of those from Cottage Grove, the more northerly Oregon provenance. Genetic Considerations The absence of geographic barriers Within a species' range permits the constant exchange of pollen and seeds between segments of a population or adjoining populations which may not d i f f e r very much from each other (Wright, 1976). The reproductive characteristics that favour considerable variation within red alder were discussed by S t e t t l e r (1978). The results of these germination tests indicate that unlike many western North American species with a wide d i s t r i b u t i o n , this species does not contain d i s t i n c t - 68 -geographic races, each adapted to i t s particular environment within i t s range. Free gene exchange along i t s range has probably led to con-siderable genetic variation within individual populations and a r e l a t i v e l y small amount of variation between populations. Localized environmental factors are probably more important in bringing about different physiological adaptations than are broader geographic factors l i k e elevation and latitude. Both suggestions would explain the lack of trends with broad geographic parameters and the observed differences between closely situated provenances. Conclus ions Dormancy does not appear to be a widespread phenomenon in red alder but can be present in a number of seeds as was shown in tests with seeds from the Haney c o l l e c t i o n . Low temperature s t r a t i f i c a t i o n and presoaking of seeds remove dormancy when i t occurs and increase the rate of germination over that in the absence of pretreatment. The tests showed that there were differences in seed vigour between provenances but there was no relationship between these differences and geographic location of the provenances. There do not appear to be d i s t i n c t races of red alder related to broad geographic factors l i k e latitude and elevation. This apparent uniformity is thought to be due to considerable gene exchange between populations throughout i t s range. - 69 -CHAPTER THE IMPORTANCE OF FERTILITY AND PHYSICAL CHARACTERISTICS OF SOIL IN EARLY SEEDLING DEVELOPMENT IN RED ALDER Introduction Well drained loamy s o i l s of a l l u v i a l o r i g i n are best for early seedling development in red alder. Seedlings can grow successfully on any s o i l type 11 once s o i l moisture is not a li m i t i n g factor (Johnson et al., 1926; Worthington et al., 1962) and providing s o i l bulk densities are below 1.50 grams per cubic centimetre (Forri s t a l and Gessel, 1955; Minore et al., I 969) . They may not persist on poor to medium sites (Smith, 1974) and may suffer heavy mortality after the f i r s t year (Zavitkovski and Stevens, 1972) . Heights attained after f i r s t year's growth vary from s i x inches (152.4 millimetres) to as much as 18 inches (457.2 millimetres) under excellent conditions (Worthington, 1965; Plank, 1971) . Ruth (1968) observed an average height of 21.4 millimetres in 38 seedlings after f i r s t season's growth in prepared seedspots subjected to various levels of solar radiation whereas 100 naturally regenerated seedlings grew to an average height of 45 -8 millimetres under similar conditions. Smith (1972) measured only the largest seedlings in studies on the campus and at the Research Forest of the University of B r i t i s h Columbia. The average height of these was 4 .49 feet (1368.6 millimetres). - 70 -The variation in figures obtained probably resulted from differences in the numbers of trees measured and in s i t e factors in the areas where measurements were made. Genetic variation within the species might also have contributed to the wide range of figures. Except for a few studies e.g. those by Fo r r i s t a l and Gessel ( 1955 ) , Hughes et a l . 3 ( 1968 ) , Minore (1968 and 1970) and Minore et al. 3 ( 1969 ) , most investigations on the interaction between red alder and i t s s o i l environment have dealt with i t s a b i l i t y to improve the physical and chemical properties of the s o i l , and with the species' associated microflora. Both the physica1,nature of the s o i l and i t s f e r t i l i t y influence growth in any growing season, even though other s i t e factors may not be l i m i t i n g . Soil f e r t i l i t y is probably p a r t i c u l a r l y important in the f i r s t year of development in red alder before nodule development is extensive. This study was undertaken to determine the relative importance of the physical nature of the s o i l and i t s f e r t i l i t y in the early growth of red alder seedlings raised from seed collected from several locations within i t s range. Materials and Methods Seedling growth of two B r i t i s h Columbian and s i x American provenances used in the germination studies, was examined. These were K i t s a u l t , Nanaimo Lakes, Centralia, Vancouver, Myrtle Creek, Cottage Grove, Laytonville and - 71 -and Crescent City. Two weeks after seeds were sown, germinants were transplanted into four inch diameter pots containing either coarse sand, a loamy s o i l or a loamrsand mixture (1:1 w/w). For each provenance, one pair of seedlings was grown in each pot and each growth medium was contained in six of 18 pots. A mixture of tap water and crushed nodules from one year old seedlings was evenly applied to a l l , p o t s . Seedlings were then transferred to a growth chamber and maintained at 25°C. Fluorescent lig h t i n g with an intensity of about 3500 lux, was provided for 12 hours each day. The relative humidity fluctuated between 65% and 75% during the l i g h t period and between 70% and 80% during the dark period. Plants were watered with an automatic spray system and manually when i t was necessary. At no time were the growth media allowed to dry out or become waterlogged. The \kk pairs of plants were divided into two equal, randomly arranged sections. A 1% solution of " H i - s o l " (Green Valley F e r t i l i z e r and Chemical Co. Ltd., Surrey, B r i t i s h Columbia), the composition of which is given in Table 3 - 1 , was applied every three weeks during the growth period to one of the two sections at a rate of approximately 10 mi 11i1itres per pot whilst the other section received only tap water. Seedlings were harvested after 95 days. The height of the f i r s t order branch was recorded for each seec'ling, and root systems were separated from the s o i l p a r t i cles to which they were attached. The presence or absence of nodules - 72 -Chemical Constituent Concentration (%) Total nitrogen Available phosphoric acid Soluble potash Elemental magnesium Boron Molybdenum I ron Copper Manganese Zinc 20 .000 20 .000 20 .000 0 .056 0 .026 0.001 0 .100 0.051 0.051 0 .110 Constituent materials: Urea, diamonium phosphate, nitrate of potash, and muriate of potash. Table 3 - 1 . Composition of the commercial f e r t i l i z e r " H i - s o l " . - 73 -was recorded. Each pair of seedlings was divided into shoots (above the point of attachment of the cotyledons), roots (below the point of attachment of the cotyledons) and nodules. The dry weights of these components were measured after they were dried at 105°C. for at least 48 hours. The mean height, dry weight and root-shoot dry weight r a t i o of each seedling pair were analyzed to determine whether there were any growth differences caused by f e r t i l i z a t i o n or the different growth media, and whether seedlings of the different provenances performed d i f f e r e n t l y under the conditions described. A second study was done to examine nodule formation in sand and loam. Seedlings from K i t s a u l t , Myrtle Creek, Vancouver and Crescent City were grown in four inch diameter pots. Six seedlings of each provenance were grown in each of two pots containing coarse sand placed over loamy s o i l , and s i x were grown in each of two pots with loamy s o i l over sand, both s o i l types in about equal amounts. They were kept under the same conditions as those seedlings in the f i r s t study. They were divided into two equal sections with the same random arrangement. One section was f e r t i l i z e d with Hi-sol as described e a r l i e r . - 74 -The seedlings from each treatment were harvested after 116 days and the number of plants with nodules and the number of nodules in each s o i l section were recorded at harvest. Seedling and nodule dry weights were determined after drying at 105°C. for at least 48 hours. Results and Observations The results showed considerable variation within individual provenances regardless of the type of growth medium or whether f e r t i l i z e r was applied. An indication of this height variation is presented in Plate 3-1 which shows f e r t i l i z e d and u n f e r t i l i z e d seedlings of the Laytonville provenance. The physical nature of roots produced by seedlings of a l l provenances varied with the s o i l type (Plate 3-2). Roots formed in sand were r e l a t i v e l y thick in cross-section and very long. In the loamy s o i l , they were short and tended to be more fibrous. Roots formed in the loam-sand mixture were intermediate between those formed in the other media. Plants in the sand medium which was not f e r t i l i z e d , showed a greatly extended primary root.from which there were few short branches. Where f e r t i l i z e r was added to this medium, root development was very vigorous and roots had a red tinge. The colour of the f e r t i l i z e r was red but i t was unlikely that the colour of the roots resulted from staining by the f e r t i l i z e r since this colour was absent in the other f e r t i l i z e d media except in some nodules formed at the root c o l l a r and p a r t i a l l y exposed to the atmosphere. It is l i k e l y that the red colour observed reflected the presence of anthocyanins produced under the conditions described (Bond, 1958; Becking, 1975). - 75 -P l a t e 3.1. U n f e r t i l i z e d (UF) and f e r t i l i z e d (F) s e e d l i n g s o f t h e L a y t o n v i l l e provenance grown i n sand ( s d ) , loam-sand (ss) and loam ( s l ) . P l a t e 3.2. U n f e r t i l i z e d (-FERT) and f e r t i l i z e d (+FERT) s e e d l i n g s of t h e K i t s a u l t provenance grown i n sand ( s d ) , loam-sand (ss) and loam ( s l ) . - 76 -Nodules were formed in seedlings from a l l provenances tested. Of the 144 pairs of seedlings grown, nodule formation was i n i t i a t e d or took place on roots of at least one seedling of each of 110 pairs, 52 of which were not f e r t i l i z e d and 58 of which were f e r t i l i z e d . Without f e r t i l i z a t i o n , 75% of the seedlings pairs grown in sand, and 63% and 73% of those in loam-sand and loam respectively, had nodules. Corresponding figures for f e r t i l i z e d treatments were 83%, 92% and 62% in sand, loam-sand and loam respectively. In general, large nodules were produced by large seedlings, however, not a l l large seedlings produced nodules. Many nodules formed in the absence of f e r t i l i z a t i o n were small and the average dry weight of those formed with f e r t i l i z a t i o n was about twice that of those formed without. These observations are consistent with those from the second study. Table 3.2 shows that a l l seedlings formed nodules only in the f e r t i l i z e d Crescent City seedlings grown in the sand/loam medium. There was great variation in the sizes of nodules but large nodules were generally produced by large seedlings. Some of the smallest nodules were, however, present on roots of large seedlings. Sixty-three nodules developed of which 50 were present in the upper section of the medium and 13 in the lower. The type of s o i l did not appear to influence the location of nodules. T h i r t y - s i x nodules were present on roots grown in sand and 27 on roots grown in loam. Twenty-four nodules developed in the absence of f e r t i l i z a t i o n while 39 were formed on roots of f e r t i l i z e d seedlings. On roots of u n f e r t i l i z e d seedlings, 14 nodules were - 77 -Provenance Treatment Mean Seedling No. Seedlings No. Nodules No. Nodules Dry Weight (Mg.) With Nodules In Upper Section In Lower Section Total Dry Weight of Nodules (to Nearest 10 mg.) Kitsault SD/LM-NF LM/SD-NF SD/LM-F LM/SD-F 20. 1 65.0 520.0 650.0 10 10 30 30 Myrtle Creek SD/LM-NF LM/SD-NF SD/LM-F LM/SD-F 26.7 238.3 5*5.3 *t03.3 10 60 70 350 Vancouver SD/LM-NF LM/SD-NF SD/LM-F LM/SD-F 88.3 79.7 331.7 200.0 k 0 2 20 10 kO 20 Crescent City SD/LM-NF LM/SD-NF SD/LM-F LM/SD-F 151.7 36.7 395.0 660.1 10 10 60 10 Table 3-2 Summary of data collected for nodule formation in f e r t i l i z e d (F) and u n f e r t i l i z e d (NF) seedlings from K i t s a u l t , Myrtle Creek, Vancouver and Crescent City, grown in the different combinations of coarse sand (SD) and loam (LM). - 78 -formed in sand and 10 in loam. C o r r e s p o n d i n g f i g u r e s f o r f e r t i l i z e d s e e d l i n g s were 22 and 17 in sand and loam r e s p e c t i v e l y . T h e s e o b s e r v a t i o n s s u g g e s t t h a t e n v i r o n m e n t has l i t t l e i n f l u e n c e on n o d u l e deve lopment w h i c h seems t o be under s t r o n g g e n e t i c c o n t r o l . F e r t i l i z a t i o n may f a v o u r the deve lopment o f l a r g e n o d u l e s in many s e e d l i n g s in t h a t i t i n c r e a s e s s e e d l i n g s i z e . N o d u l e s a p p e a r t o fo rm more r e a d i l y on upper r o o t b r a n c h e s than on lower o n e s , r e g a r d l e s s o f t h e type o f s o i l medium in wh ich the r o o t s a r e g r o w i n g . P r o v e n a n c e Response t o I n d i v i d u a l T r e a t m e n t s H e i g h t growth in a l l t r e a t m e n t s d i d not d i f f e r s i g n i f i c a n t l y between p r o v e n a n c e s ( T a b l e 3-3). It was o n l y in the u n f e r t i l i z e d loam-sand medium t h a t s i g n i f i c a n t d i f f e r e n c e s between p r o v e n a n c e s were o b t a i n e d f o r d r y w e i g h t ( T a b l e 3-4). The mean dry w e i g h t o f s e e d l i n g s o f the L a y t o n v i l l e p r o v e n a n c e was s i g n i f i c a n t l y h i g h e r than t h o s e o f a l l o t h e r p r o v e n a n c e s e x c e p t Nanaimo L a k e s . Dry w e i g h t s o f s e e d l i n g s f rom C r e s c e n t C i t y and M y r t l e Creek were s i g n i f i c a n t l y lower than t h a t o f the Nanaimo Lakes s e e d l i n g s . S e e d l i n g dry w e i g h t in the C o t t a g e G r o v e , V a n c o u v e r , C e n t r a l i a and K i t s a u l t p r o v e n a n c e s were s i m i l a r t o each o t h e r and to a l l o t h e r p r o v e n a n c e s e x c e p t L a y t o n v i l l e . R o o t - s h o o t r a t i o s were not s i g n i f i c a n t l y d i f f e r e n t f rom each o t h e r in a l l f e r t i l i z e d t r e a t m e n t s and in the u n f e r t i l i z e d sand medium ( T a b l e 3 . 5 ) . In - 7 9 -Hean height (mm.) Crescent Myrtle Cottage Nanaimo Treatment City Creek Grove Vancouver Centralia Kitsault Lakes Laytonville F-value Sign ificance Sand • NF 9.2 8.8 7.7 30.3 13.8 8.3 13.0 29.3 1.31 N.S Sand • F lSl .5 169.3 188.5 158.0 119.0 156.8 186.5 96.8 1.64 N.S. Loam-sand • NF 44.0 35.2 64.3 56.7 47.7 51.5 89.3 84.3 1.50 N.S. Loan-sand • F 115.2 193.3 146.7 197.7 191.0 190.2 121.0 129.8 1.05 N.S. Loam * NF 79.0 112.0 82.8 72.7 86.2 62.0 74.5 95.8 0.82 N.S. Loam + F 178.0 209.3 115.5 107.7 129.7 103.0 147.7 148.7 1.65 N.S. N.S. : not significant at the 5% level . NF : no fer t i l izat ion F : fer t i l izat ion Table 3.3. Summary of analysis of data for height (mm.) of seedlings of the different provenances in individual treatments. Crescent Myrtle Cottage Nanaimo 'Treatment City Creek Grove Vancouver Centralia Kitsault Lakes Laytonville F-value Significance Sand • NF 16.7 18.3 13.3 58.3 25.0 11.7 25.0 71.7 1.23 N.S. Sand • F 476.7 508.3 495.0 918.3 628.3 418.3 746.7 621.7 1.73 N.S. Loam-sand • NF 105.0 153.3 216.7 238.3 246.7 276.7 478.3 528.3 3.49 .1 Loam-sand • F 315.0 868.3 603.3 1195.0 896.7 848.3 743.3 611.7 1.18 N.S. Loam * tlF 261.7 353.3 368.3 298.3 295.0 293.3 275.0 483.3 0.54 N.S. Loam • F 526.7 940.0 491.7 551.7 510.0 750.0 676.7 800.0 1.93 N.S. 1. Duncan's multiple range test (at the 5% level) : Crescent Myrtle Cottage Nanaimo Provenance : c l f c y Creek Grove Vancouver Centralia Kitsault Lakes Laytonville Mean : 105.0 153.3 216.7 238.3 246.7 -276.7 478.3 528.3 N.S. : not significant at the 5% level . * : significant at the 5% level . NF : no fer t i l izat ion F : fer t i l iza t ion Table 3.4 Summary cf analysis of data for dry weight (mg.) of seedlings of the different provenances ln individual treatments. - 80 -Mean root-shoot ratio Crescent Myrtle Cottage Nanaimo Treatment City Creek Grove Vancouver Centralis Kitsault Lakes Laytonville F-value Siqnificance Sand * NF 1.67 1.17 0.67 1.48 1.17 1.33 1.17 0.59 1.70 N.S. Sand + F 0.50 0.41 0.40 0.48 0.54 0.58 0.39 0.54 0.69 N.S. Loam-sand *• NF 0.69 0.88 0.57 0.70 0.90 0.88 0.53 0.44 3.58 .1 Loan-sand + F 0.40 0.33 0.42 0.38 0.38 0.39 0.41 0.35 0.66 N.S. Loam + NF 0.50 0.48 0.51 0.68 0.57 0.62 0.67 0.41 3.64 .2 Loan • F 0.32 0.29 0.33 0.46 0.41 0.36 0.42 0.40 0.65 N.S. 1. Duncan's multiple range test (at the 5% level) : Nanaimo Cottage Crescent Pwenance i , . . ^ , c Mean Laytonville Lakes Grove 0.44 0.53 0.57 0.69 Myrtle Vancouver Kitsault Creek Centralia 0.70 .0.88 0.88 0.90 2. Duncan's multiple range test (at the 5% level) : Myrtle Crescent Cottage Provenance : . . , , , _ , „ . . Laytonville Creek City Mean 0.41 0.48 0.50 Nanaimo Grove Centralis Kitsault Lakes Vancouver 0.51 0.57 0.62 0.67 0.68 N.S. : not significant at the 5% l evel . • : significant at the 5* level . NF : no fer t i l izat ion F : fer t i l izat ion Table 3.5. Summary of analysis of data for root-shoot ratio of seedlings of the different provenances in individual treatments. - 81 -the u n f e r t i l i z e d loam-sand medium, root-shoot ratios of seedlings from Ki t s a u l t , Myrtle Creek and Centralia were s i g n i f i c a n t l y higher than those of seedlings from Laytonville, Nanaimo Lakes and Cottage Grove. Root-shoot ratios of seedlings from Vancouver and Crescent City were similar to each other and to that of seedlings of a l l other provenances. Root-shoot ratios of seedlings from the four most northerly provenances (Centralia, K i t s a u l t , Nanaimo Lakes and Vancouver) grown in the u n f e r t i l i z e d loam medium were not s i g n i f i c a n t l y different from each other, neither were the root-shoot ratios of seedlings from the more southerly locations (Laytonville, Myrtle Creek, Crescent City and Cottage Grove). Northern seedlings tended to have higher root-shoot ratios than the southern ones. Vancouver seedlings had a s i g n i f i c a n t l y higher ratio than that of a l l the southern seedlings whereas seedlings from Centralia were similar to a l l of the southern group. The root-shoot rat i o of seedlings from Nanaimo Lakes was s i g n i f i c a n t l y higher than that of each of the southern group except Cottage Grove. The ratio for the Kitsault seedlings was s i g n i f i c a n t l y higher than that for the Laytonville seedlings. Provenance Response to F e r t i l i z a t i o n in Each Growth Medium Height and dry weight increased s i g n i f i c a n t l y with f e r t i l i z a t i o n but there were no s i g n i f i c a n t differences between provenances in either measure (Tables 3-6 and 3.7). F e r t i l i z a t i o n reduced root-shoot rat i o in each medium (Table 3.8). There were no s i g n i f i c a n t differences between provenances - 82 -Wean height (nan.) Source of variation Growth medium Sand Loam-sand Loam P-value Significance F-value Significance F-value Significance F e r t i l i z a t i o n Provenance Fer t i l iza t ion X provenance interaction 225.59 • 1.13 N.S. 2.08 N.S, 56.17 • 0.58 N.S. 1.68 N.S. 31.66 • 2.23 N.S. 0.60 N.S. Without f e r t i l i z a t i o n With f e r t i l i z a t i o n 15.1 a 153.3 b 59.1 a 160.6 b 83.1 a 147.4 b N.S. : not significant at the Si l eve l . • : significant at the 5S level . Figures not followed by the same letter are significantly different from each other at the 5% level . •.Table 3.6. Summary of analysis of data for height (mm.) of fer t i l ized and unfertilized seedlings of the different provenances in each growth medium. Mean dry weight (mg.) Source of variation Growth medium Sand Loam-sand Loam F-value Significance F-value Significance F-value Significance F e r t i l i z a t i o n Provenance F e r t i l i z a t i o n X provenance interaction 160.62 • 2.01 N.S. 1.42 N.S. 29.04 • 1.46 N.S. 1.36 N.S. 36.52 • 1.79 N.S. 0.93 N.S. Without f e r t i l i z a t i o n With f e r t i l i z a t i o n 30.0 a 601.7 b 280.4 a 760.2 b 328.5 a 655.8 b N.S. > not significant at the 5% level . • : significant at the 5% level . Figures not followed by the same letter axe significantly different from each other at the 5% level . Table 3.7. Summary of analysis of data for dry weight (mg.) of fer t i l ized and unfertilized seedlings of the different provenances in each growth medium. - 83 -Source of variation Growth medium Sand Loam-sand Loam F-value Significance F-value Significance F-value Significance Fertil ization Provenance Ferti l ization X provenance interaction 41.20 • 1.71 N.S. 1.51 N.S. 82.07 • 2.92 • 1 3.47 • 34.55 • 2 2.51 0.76 N.S. Without fer t i l izat ion With fer t i l izat ion 1.16 a 0.46 b 0.70 a 0.38 b 0.56 a 0.38 b 1. Duncan's multiple range test (at the 5% level) : Nanaimo Cottage Laytonville Lakes Grove Vancouver : 0.40 0.47 0.49 0.54 Provenance : Mean Crescent Myrtle City Creek 0.55 0.61 Kitsault Centralia 0.64 0.64 2. Duncan's multiple range test (at the 5% level) : Myrtle Crescent Cottage Provenance Mean Creek Laytonville City Grove : 0.39 0.41 0.41 0.42 Nanaimo Centralia Kitsault Lakes 0.49 0.49 0.55 Vancouver 0.57 N.S. : not significant at the 5% level . • : significant at the 5* level . Table 3.8. Summary of analysis of data for root-shoot ratio of fer t i l ized and unfertilized seedlings of the different provenances in each growth medium. - 84 -for seedlings grown in sand and there was a si g n i f i c a n t provenance x f e r t i l i z a t i o n interaction for seedlings grown in the loam-sand mixture. Figure 3•1A shows that the extent to which f e r t i l i z a t i o n reduced root-shoot rat i o in this medium, varied with provenance and this may have contributed to the observed interaction. The effect of f e r t i l i z a t i o n was greatest in seedlings from Centralia and K i t s a u l t , and smallest in those from Nanaimo Lakes and Laytonville. Both these groups were s i g n i f i c a n t l y different from each other. Seedlings of the Laytonville provenance also had a s i g n i f i c a n t l y lower root-shoot r a t i o than that of the Myrtle Creek seedlings. Seedlings from Cottage Grove, Vancouver and Crescent City were similar to each other and to a l l other seedlings. There was l i t t l e reduction in root-shoot rat i o with f e r t i l i z a t i o n for seedlings from Laytonville grown in the loam medium (Figure 3 -IB). Seedlings of the Vancouver provenance showed a s i g n i f i c a n t l y higher root-shoot rat i o than those of the Cottage Grove, Crescent City, Laytonville and Myrtle Creek provenances (Table 3 . 8 ) . Also, the root-shoot ratio for the Nanaimo Lakes seedlings was s i g n i f i c a n t l y higher than that for the Myrtle Creek seedlings. The Centralia and Kitsault provenances did not d i f f e r from each other nor any other provenance. Response of Individual Provenances to a l l Treatments Kitsault Provenance Growth medium and f e r t i l i z a t i o n s i g n i f i c a n t l y affected dry weight and root-shoot - 85 -A. Loam-sand Root-shoot r a t i o 0.9 0.8 0.7 0.6 0.5 0.4 0.3-0.8 0.7 0.6 0.5 0.4 0.3 0.2-u to >i QJ +J U -H u u rH M +> QJ U QJ >< u s u QJ fO QJ O U ID O O § > ro rH (0 M u 3 0) 4J •H • • •rl Ul rrj OJ 2 J A — A • A 4J 6 u OJ -p M -rl U U OJ r-l M -P OJ l l OJ OJ 0> ro OJ •P -P _ O M U O • A • A A A u OJ ' fe (0 c 5 •H 3 o iH > u (0 S-i +J c QJ U A A • A A A A A 3 05 -p •H g •H W (0 OJ 2 iJ A A A A A A A A A A A A •rl o •p >1 (0 •H & O -P A A A A A A A A EZ3 s B. Loam Provenance Without f e r t i l i z a t i o n With f e r t i l i z a t i o n F i gure 3.1. Root-shoot r a t i o s of seedlings of the d i f f e r e n t provenances grown i n loam-sand (A.) and loam (B.). - 86 -r a t i o , and t h e i n t e r a c t i o n between the two had a s i g n i f i c a n t e f f e c t on h e i g h t growth o f t h e K i t s a u l t s e e d l i n g s ( T a b l e 3.9)• F e r t i l i z a t i o n i n c r e a s e d d r y w e i g h t and reduced r o o t - s h o o t r a t i o as compared t o n o n - f e r t i l i z a t i o n . Dry w e i g h t i n t h e loam-sand m i x t u r e and i n loam were not s i g n i f i c a n t l y d i f f e r e n t f rom each o t h e r but were s i g n i f i c a n t l y h i g h e r than t h a t i n sand. The r o o t - s h o o t r a t i o o f s e e d l i n g s grown i n sand was s i g n i f i c a n t l y h i g h e r than t h a t o f t h o s e grown i n loam. The r o o t - s h o o t r a t i o o f t h o s e grown i n t h e loam-sand m i x t u r e was s i m i l a r t o t h a t o f the o t h e r s e e d l i n g s . F i g u r e 3.2 shows t h a t w i t h o u t f e r t i l i z a t i o n , h e i g h t growth was n o t i c e a b l y s m a l l e r i n sand than i n t h e o t h e r media. W i t h f e r t i l i z a t i o n t h e r e was a g r e a t i n c r e a s e i n h e i g h t i n the sand and loam-sand media, and a r e l a t i v e l y s m a l l i n c r e a s e i n loam. As a r e s u l t , h e i g h t o f t h o s e s e e d l i n g s grown i n sand and i n loam were s i m i l a r but were s i g n i f i c a n t l y lower than t h a t o f s e e d l i n g s grown i n t h e loam-sand m i x t u r e . H e i g h t growth o f s e e d l i n g s t h a t were f e r t i l i z e d was s i g n i f i c a n t l y g r e a t e r than t h a t o f s e e d l i n g s not f e r t i 1 i z e d . C r e s c e n t C i t y Provenance The d i f f e r e n t t y p e s o f growth medium d i d not have a s i g n i f i c a n t e f f e c t on h e i g h t o r d r y w e i g h t o f s e e d l i n g s o f t h e C r e s c e n t C i t y provenance but f e r t i l i z a t i o n s i g n i f i c a n t l y i n c r e a s e d both measures ( T a b l e 3.10). There - 87 -Source of variation Growth parameter Heiqht (mm.) Dry weight (mq.) Root-shoot ratio F-value Significance F-value Significance F-value Significance Growth medium Fertil ization Growth medium X fer t i l izat ion interaction 5.70 • 104.70 • 10.30 • 4.46 • 21.20 • 0.22 N.S. 4.73 • 16.13 • 1.30 N.S. Mean in sand Mean in loam-sand Mean in loam 82.6 a 120.8 b 82.5 a 215.0 a 562.5 b 521.7 b 0.96 a 0.64 ab 0.49 b Mean without fer t i l izat ion Mean with fer t i l izat ion 40.6 a 150.0 b 193.9 a 672.2 b 0.95 a 0.44 b N.S. : not significant at the 5* level . • : significant at the 5% level . Figures not followed by the same letter are significantly different from each other at the 5% level . Table 3.9. Summary of analysis of data for height (ran.), dry weight (mg.) and root-shoot ratio of fer t i l ized and unfertilized seedlings of the Kitsault provenance grown in sand, loam-sand and loam. 200 • 180 • 160 • 140 • 120 • Height (mm.) 100 . 80 • 60 . 40 . 20 • 0 ' 1 •— Sand Loam-sand Loam Growth-medium •- • without f e r t i l i z a t i o n — • ••- : With fe r t i l i z a t i o n Figure 3.2. Mean height of fer t i l i z e d end unfertilized seedlings of the Kitsault provenance grown in sand, loam-sand and loam. \ \ \ \ \ - 88 -Source of variation Growth parameter Heiqht (mm.) Dry weiqht (mq.) Root-shoot ratio F-value Siqnif icance F-value Significance F-value Significance Growth medium Ferti l ization Growth medium X fert i l izat ion interaction 3.83 N.S. 39.65 • 1.57 N.S. 2.10 N.S. 16.09 • 0.95 N.S. 11.69 • 20.88 • 6.81 • Mean in sand Mean in loam-sand Mean in loam 80.3 79.6 128.5 246.7 210.0 394.2 1.08 a 0.55 b 0.41 b Mean without fert i l izat ion Mean with fert i l izat ion 44.1 a 148.2 b 127.8 a 439.4 b 0.95 a 0.41 b N.S. : not significant at the 5% level . • : significant at the 5% level . Figures not followed by the same letter are significantly different from each other at the 5% level . Table 3.10. Summary of analysis of data for height (mm.), dry weight (mg.) and root-shoot ratio of fer t i l ized and unfertilized seedlings of the Crescent City provenance grown in sand,loam-sand and loam. 1.7 1.6 1.5 1.4 1.3 1.2 1.1 1 .0 0 . 9 0 . 8 0 . 7 0.6 0 . 5 0.4 0 . 3 -Root-shoot ratio Sand Loam-sand Growth medium Loam Without fer t i l i z a t i o n With fe r t i l i z a t i o n Figure 3.3. Mean root-shoot ratio of fer t i l i z e d and unfertilized seedlings of the Crescent City provenance grown in sand, loam-sand and loam. - 89 -was a l s o a s i g n i f i c a n t growth medium x f e r t i l i z a t i o n i n t e r a c t i o n on r o o t -shoot r a t i o . R oot-shoot r a t i o d e c r e a s e d w i t h f e r t i l i z a t i o n i n a l l media ( F i g u r e 3.3) but t h e r e were d i f f e r e n c e s i n t h e e x t e n t t o w h i c h t h i s d e c r e a s e took p l a c e . The r e l a t i v e l y l a r g e d e c r e a s e t h a t o c c u r r e d i n t h e sand medium as compared w i t h t h o s e i n the o t h e r media, was p r o b a b l y the main c o n t r i b u t i n g f a c t o r t o t h e s i g n i f i c a n t i n t e r a c t i o n o b s e r v e d . R o o t - s h o o t r a t i o o f s e e d l i n g s grown i n sand was s i g n i f i c a n t l y h i g h e r than t h a t o f s e e d l i n g s grown i n the o t h e r media. M y r t l e Creek Provenance F e r t i l i z a t i o n s i g n i f i c a n t l y i n c r e a s e d h e i g h t and d r y w e i g h t o f the M y r t l e Creek s e e d l i n g s o v e r t h a t a t t a i n e d w i t h o u t f e r t i l i z a t i o n ( T a b l e 3-11). H e i g h t i n the loam-sand and sand media were s i m i l a r but s i g n i f i c a n t l y lower than t h a t i n loam. A l s o , d r y w e i g h t i n sand was s i g n i f i c a n t l y lower than t h a t i n loam-sand and i n loam. The i n t e r a c t i o n between growth medium and f e r t i l i z a t i o n had a s i g n i f i c a n t e f f e c t on r o o t - s h o o t r a t i o . In a l l media, r o o t - s h o o t r a t i o was h i g h e r w i t h o u t f e r t i l i z a t i o n than w i t h f e r t i l i z a t i o n . F i g u r e 3-4 shows t h a t t h e e f f e c t o f f e r t i l i z a t i o n was r e l a t i v e l y s m a l l i n t h e loam medium as compared w i t h t h a t i n sand and t h e loam-sand m i x t u r e , and t h a t w i t h and w i t h o u t f e r t i l i z a t i o n , the r a t i o i n loam was lower than each o f t h e o t h e r two. C o n s e q u e n t l y , t h e r a t i o i n loam was s i g n i f i c a n t l y lower than t h a t i n sand and i n loam-sand. - 90 -Source of variation Growth parameter Heiqht (mm.) Dry weiqht (mq.) Root-shoot ratio F-value Siqnif icance F-value Significance F-value Significance Growth medium Ferti l ization Growth medium X fer t i l izat ion interaction 7.79 • 85.16 • 1.89 N.S. 8.38 • 59.35 • 0.71 N.S. 10.70 • 50.65 • 5.60 • Mean in sand Mean in loam-sand Mean in loam 89.1 a 114.2 a 160.7 b 263.3 a 510.8 b 646.7 b 0.79 a 0.61 a 0.39 b Mean without fer t i l izat ion Mean with fert i l izat ion 52.0 a 190.7 b 175.0 a 772.2 b 0.84 a 0.34 b N.S. : not significant at the 5% level . • : significant at the 5% level . Figures not followed by the same letter are significantly different from each other at the 5% level . Table 3.11. Summary of analysis of data for height (mm.), dry weight (mg.) and root-shoot ratio of fer t i l ized and unfertilized seedlings of the Myrtle Creek provenance grown In sand, loam-sand and loam. - 91 -Vancouver Provenance Growth medium did not have a si g n i f i c a n t effect on root-shoot ratio which was s i g n i f i c a n t l y reduced by f e r t i l i z a t i o n (Table 3-12). The interaction between growth medium and f e r t i l i z a t i o n s i g n i f i c a n t l y influenced both height and dry weight of these seedlings. F e r t i l i z a t i o n had the general effect of increasing both measures, however, as i l l u s t r a t e d in Figures 3.5 and 3.6, the extent of this increase was not similar in the different growth media. Without f e r t i l i z a t i o n , both measures were highest in the loam medium and lowest in sand but with f e r t i l i z a t i o n , they were highest in the loam-sand mixture and lowest in the loam. The result was higher mean values in the loam-sand medium and similar ones in the other two media. Only in dry weight did the loam-sand mean s i g n i f i c a n t l y d i f f e r from that of the others. Centralia Provenance F e r t i l i z a t i o n s i g n i f i c a n t l y increased height and dry weight, and reduced root-shoot ratio in seedlings from Centralia (Table 3.13). Root-shoot ra t i o was similar in loam and loam-sand. It was s i g n i f i c a n t l y lower in these media than in sand. Laytonville Provenance The only s i g n i f i c a n t effects observed in seedlings from Laytonville were the increase in height and in dry weight caused by f e r t i l i z a t i o n over those obtained without f e r t i l i z a t i o n (Table 3-14). - 92 -Source of variation Growth parameter Height (mm.) Dry wieqht (mq.) Root-shoot ratio F-value Significance F-value Significance F-value Significance Growth medium Fertil ization Growth medium X fert i l izat ion interaction 2.05 N.S. 38.27 • 4.15 • 3.01 N.S. 45.72 • 4.65 * 2.53 N.S. 8.27 • 1.85 N.S. Mean in sand Mean in loam-sand Mean in loam 94.2 a 127.2 a 90.2 a 488.3 ab 716.7 a 425.0 b 0.98 0.54 0.57 Mean without fert i l izat ion Mean with fer t i l izat ion 53.2 a 154.4 b 198.3 a 883.3 b 0.96 a 0.44 b N.S. : not significant at the 5% level . • : significant at the 5% level . Plgures not followed by the same letter are significantly different from each other at the 5% level . Table 3.12. Summary of analysis of data for height (mm.), dry weight (mg.) and root-shoot ratio of fer t i l ized and unfertilized seedlings of the Vancouver provenance grown in sand, loam-sand and loam. < - 93 -200 K 180 160 ' 140 • 120 • Height (mm.) 100 • 80 • 60 • 40 20 L 0 \ \ \ \ Sand Loam-sand Growth medium Without fe r t i l i z a t i o n With f e r t i l i z a t i o n Loam Figure 3.5. Mean heigVit of fertilized and unfertilized seedlings of the Vancouver provenance grown in sand, loam amd loam-sand. \ \ \ 1200 1100 1000 900 Dry weight 800 (mg.) 7 0 0 600 500 400 300 200 100 0 I Sand Loam-sand Loam-sand Growth medium ~% •>-: Without f e r t i l i z a t i o n -•j pj- : With fe r t i l i z a t i o n Figure 3.6. Mean dry weight of fer t i l i z e d and unfertilized seedlings of the Vancouver provenance grown in sand, loam and loam-sand. - Sk -Source of variation Growth parameter Height (mm.) Dry weight (mg.) Root-shoot ratio F-value Significance F-value Significance F-value Significance Growth medium Fertil ization Growth medium X fer t i l izat ion interaction 2.39 N.S. 21.93 • 1.96 N.S. 1.48 N.S. 16.86 • 1.34 N.S. 7.69 * 33.67 ' 3.49 N.S. Mean in sand Mean in loam-sand Mean in loam 66.4 119.3 107.9 326.7 571.7 402.5 0.85 a 0.64 b 0.49 b Mean without fert i l izat ion Mean with fer t i l izat ion 49.2 a 146.6 b 188.9 a 678.3 b 0.88 a 0.44 b N.S. : not significant at the 5% level . • : significant at the 5% level . Figures not followed by the same letter are significantly different from each other at the 5% level . Table 3.13. Summary of analysis of data for height (mm.), dry weight (mg.) and root-shoot ratio of fer t i l ized and unfertilized seedlings of the Centralia provenance grown in sand, loam-sand and loam. Source of variation Growth parameter Height (mm.) Dry weight (mg.) Root-shoot ratio F-value Significance F-value Significance F-value Significance Growth medium Ferti l ization Growth medium X fer t i l izat ion interaction 2.76 N.S. 6.69 • 0.09 N.S. 1.87 N.S. 5.95 • 1.08 N.S. 1.89 N.S. 0.41 N.S. 0.09 N.S. Mean in sand Mean in loam-sand Mean in loam 63.1 107.1 122.2 346.7 570.0 641.7 0.57 0.40 0.41 Mean without fer t i l izat ion Mean with fer t i l izat ion 69.8 a 125.1 b 361.1 a 677.8 b 0.48 0.43 N.S. : not significant at the 5% level . • : significant at the 5% level . Figures not followed by the same letter are significantly different from each other at the 5* level . Table 3.14. Summary of analysis of data for height (mm.), dry weight (mg.) and root-shoot ratio of fer t i l ized and unfertilized seedlings of the Laytonville provenance grown in sand, loam-sand and loam. - 95 -Cottage Grove Provenance F e r t i l i z a t i o n was the only treatment which had any si g n i f i c a n t effect on the growth of these seedlings (Table 3 - 1 5 ) . As compared with n o n - f e r t i l i z a t i o n , i t increased height and dry weight, and reduced root-shoot r a t i o . Nanaimo Lakes Provenance F e r t i l i z a t i o n s i g n i f i c a n t l y increased the dry weight of seedlings from the Nanaimo Lakes area over that of u n f e r t i l i z e d seedlings (Table 3 - 1 6 ) . The interaction between growth medium and f e r t i l i z a t i o n did affect both height growth and root-shoot r a t i o . F e r t i l i z a t i o n increased height and reduced root-shoot ratio but the magnitude of these changes differed in the different growth media (Figures 3-7 and 3 - 8 ) . Without f e r t i l i z a t i o n , height was lowest in sand, and greater in loam-sand than in loam. With f e r t i l i z a t i o n , seedlings were t a l l e s t in sand, and t a l l e r in loam than in loam-sand. Si m i l a r l y , with f e r t i l i z a t i o n , root-shoot ratios were a l l similar but in the absence of f e r t i l i z a t i o n , i t was r e l a t i v e l y high in sand, and lower in loam-sand than in loam. The result was a s i g n i f i c a n t l y higher root-shoot ratio in sand than that in loam. Total Analyses  Height There were no sig n i f i c a n t differences in height analysis between provenances (Table 3 . 1 7 ) . There was a si g n i f i c a n t effect on height caused by the growth medium-fertilization interaction. F e r t i l i z e d seedlings were s i g n i f i c a n t l y - 96 -Source of variation Growth parameter Height (mm.) Dry weight (mg.) Root-shoot ratio F-value Significance F-value Significance F-value Significance Growth medium Ferti l ization Growth medium X fert i l izat ion interaction 0.22 N.S. 18.05 • 1.72 N.S. 0.62 N.S. 5.45 • 0.57 N.S. 0.62 N.S. 5.95 • 0.17 N.S. Mean in sand Mean in loam-sand Mean in loam 98.1 105.5 119.2 254.2 410.0 430.0 0.53 0.49 0.42 Mean without fer t i l izat ion Mean with fer t i l izat ion 51.6 a 163.6 b 199.4 a 530.0 b 0.58 a 0.38 b N.S. : not significant at the 5% level . • : significant at the 5% level . Figures not followed by the same letter are significantly different from each other at the 5% level . Table 3.15. Summary of analysis of data for height (mm.), dry weight (mg.) and root-shoot ratio of fer t i l ized and unfertilized seedlings of the Cottage Grove provenance grown in sand, loam-sand and loam. Source of variation Growth parameter Height (mm.) Dry weight (mg.) Root-shoot ratio F-value Significance F-value Significance F-value Significance Growth medium Ferti l ization Growth medium X fer t i l izat ion interaction 0.12 N.S. 23.23 • 4.78 • 2.34 N.S. 29.26 • 2.50 N.S. 4.17 • 17.61 • 4.80 • Mean in sand Mean in loam-sand Mean in loam 99.8 a 105.2 a 111.1 a 385.8 610.8 475.8 0.78 a 0.47 b 0.55 ab Mean without fer t i l izat ion Mean with fer t i l izat ion 58.9 a 151.7 b 259.4 a 722.2 b 0.79 a 0.41 b N.S. : not significant at the 5% level . • : significant at the 5% level . Figures not followed by the same letter are significantly different from each other at the 5% level . Table 3.16. Summary of analysis of data for height (mm.), dry weight (mg.) and root-shoot ratio of fert i l ized and unfertilized seedlings of the Nanaimo Lakes provenance grown in sand, loam-sand and loam. - 97 -200 180 160 140 h 120 Height (mm.) 100 80 60 40 20 0 X Sand Loam-sand Growth medium Loam — • •-: Without fertilization - • •-; With fer t i l i z a t i o n Figure 3.7. Mean height of fer t i l i z e d and unfertilized seedlings of the Nanaimo Lakes provenance grown in sand, loam-saiid and loam. 1.2 1.1 1.0 0.9 Root-shoot 0.8 ratio 0.7 0.6 0.5 0.4 0.3. Sand Loam-sand Growth medium Without fe r t i l i z a t i o n With fer t i l i z a t i o n Loam Figure 3.8. Mean root-shoot ratio of fer t i l i z e d and unfertilized seedlings of the Nanaimo Lakes provenance grown in sand, loam-sand and loam. - 98 -Sourco of variation F-value Significance Growth medium 8.30 .1 Fertilization 231.85 .2 Provenance 0.85 N.S. Ferti l ization X growth medium interaction 10.28 • Growth medium X provenance interaction 1.43 N.S. Ferti l ization X provenance interaction 1.53 N.S. Ferti l ization X provenance X growth medium interaction 1.35 N.S. 1. Duncan's multiple range test (at the 5% level) : Growth medium : Sand Loam-sand Loam Mean : 84.2 109.9 115.3 2. Duncan's multiple range test (at the S% level) : Treatment : Without fer t i l izat ion With fer t i l izat ion Mean : 52.4 153.8 N.S. : not significant at the 5% level . • : significant at the 5% level . Table 3.17. Summary of analysis of data for height (mm.) of fert i l ized and unfertilized seedlings of the different provenances grown in sand, loam-sand and loam. 200 180 160 140 120 H e i g h t (mm.) 100 80 60 40 20 0 . Sand Loam-sand Growth medium Wi thout f e r t i l i z a t i o n Loam • - : W i t h f e r t i l i z a t i o n F i g u r e 3 . 9 . Mean h e i g h t o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f a l l p r o v e n a n c e s grown i n s a n d , loam-sand and l o a m . - 99 -t a l l e r than u n f e r t i l i z e d seedlings. As shown in Figure 3-9, the increase in height with f e r t i l i z a t i o n was smallest where the greatest height was obtained without f e r t i l i z a t i o n , and largest in sand in which seedlings were shortest in the absence of f e r t i l i z a t i o n . Height in the loam-sand mixture and in loam did not d i f f e r s i g n i f i c a n t l y from each other but were s i g n i f i c a n t l y higher than that in sand. Dry Weight There were si g n i f i c a n t differences in dry weight between provenances (Table 3-18). Seedlings of the Vancouver provenance produced s i g n i f i c a n t l y higher dry weights than those from Cottage Grove and Crescent City. Dry weights of seedlings from Laytonville, Nanaimo Lakes and Myrtle Creek were s i g n i f i c a n t l y higher than those of seedlings from Crescent City. Seedlings of the Kitsault and Centralia provenances were similar to each other and to seedlings of the other provenances. Dry weight was also affected by the fertilization-growth medium interaction. The general effect of f e r t i l i z a t i o n was to increase dry weight over that obtained without f e r t i l i z a t i o n . Figure 3.10 shows that the effect of f e r t i l i z a t i o n was not as great in loam as i t was in either sand or loam-sand. With and without f e r t i l i z a t i o n , dry weight was lowest in the sand medium. With f e r t i l i z a t i o n , i t was greater in the loam-sand medium than in loam but in the absence of f e r t i l i z a t i o n , the reverse was true. Consequently, dry weight in the l a t t e r media did not d i f f e r s i g n i f i c a n t l y but both were s i g n i f i c a n t l y higher than that of seedlings grown in sand. - 100 -Source cf variation F-value Significance Growth medium 11.43 . ! Fertil ization 147.43 .2 Provenance 2.54 Fertil ization X growth medium interaction 3.55 • Growth medium X provenance interaction 1.17 N.S. Ferti l ization X provenance interaction , 1.65 N.S. Ferti l ization X provenance X growth medium interaction 1.09 N.S. 1. Duncan's multiple range test (at the 5* level): Growth medium: Sand Loam-sand Loam Mean : 315.8 520.3 492.2 2. Duncan's multiple range test (at the 5% level) : Treatment : Without fer t i l izat ion With fer t i l izat ion Mean : 213.0 672.6 3. Duncan's multiple range test (at the 5% level) : Crescent Cottage Provenance : _ . . _ „ . . , . _ . , . City Grove Kitsault Centralia Mean : 283.6 364.7 433.1 433.6 Myrtle Nanaimo Creek Lakes 473.6 490.8 Laytonville 519.4 Vancouver 543.3 N.S. : not significant at the 5% level . • : significant at the 5% level . Table 3.18. Summary of analysis of data for dry weight (mg.) of fert i l ized and unfertilized seedlings of the different provenances grown in sand, loam-sand and loam. 800 750 700 650 600 550 500 450 Dry weight 4 0 0 (mg.) 350 300 250 200 150 100 50 0-Sand Loam-sand Growth medium Without fer t i l i z a t i o n With fe r t i l i z a t i o n Loam Table 3.10. Mean dry weight of fer t i l i z e d and unfertilized seedlings of a l l provenances grown in sand, loam-sand and loam. - 101 -Root-shoot Dry Weight Ratio There were two si g n i f i c a n t interactions in the analysis of data for root-shoot dry weight ratio (Table 3.19). These were f e r t i l i z a t i o n x growth medium, and f e r t i l i z a t i o n x provenance. F e r t i l i z e d seedlings had lower root-shoot ratios than u n f e r t i l i z e d seedlings (Figure 3.11). Root-shoot rat i o was highest in the sand, both with and without f e r t i l i z a t i o n . It was similar in f e r t i l i z e d seedlings grown in loam-sand and loam, and higher in u n f e r t i l i z e d seedlings grown in loam-sand than in similar seedlings grown in loam. The difference between the two ratios in sand was r e l a t i v e l y wide as compared with that between each of the other pairs, thus the effect of f e r t i l i z a t i o n on root-shoot ratio was not the same in a l l media. Root-shoot r a t i o in loam and in loam-sand were s i m i l a r , and s i g n i f i c a n t l y lower than that in sand. F e r t i l i z a t i o n also reduced root-shoot ratio of seedlings of a l l provenances but the effect was not the same in a l l provenances. Root-shoot rat i o in a l l f e r t i l i z e d seedlings did not vary much among provenances (Figure 3.12). This variation was r e l a t i v e l y wide in the case of u n f e r t i l i z e d seedlings. U n f e r t i l i z e d seedlings from Laytonville and Cottage Grove had comparatively low root-shoot ratios and the effect of f e r t i l i z a t i o n was not as pronounced in these seedlings as in the other provenances. Root-shoot r a t i o in the former provenances were s i g n i f i c a n t l y lower than those of the other provenances except Myrtle Creek and Nanaimo Lakes. - 102 -Source of vaxiation F-value Significance Growth medium 30.79 ,1 Fertilization 104.50 ,2 Provenance 3.05 .3 Fertilization X growth medium interaction 14.59 • Growth medium X provenance interaction 1.30 N.S. Fertilization X provenance interaction 2.63 • Fertilization X provenance X growth medium interaction 1.15 N.S. 1. Duncan's multiple range test (at the 5% level): Growth medium : Sand Loam-sand Loam Mean : 0.82 0.54 0.47 2. Duncan's multiple range test (at the 5% level): Treatment : Without fertilization With fertilization Mean : 0.80 0.41 3. Duncan's multiple range test (at the 5% level): Provenance Mean Cottage Myrtle Nanaimo Laytonville 0.46 Grove 0.48 Creek 0.59 N.S. : not significant at the 5% level. • : significant at the 5% level. Lakes 0.60 Centralia 0.66 Crescent City 0.68 Kitsault 0.69 Vancouver 0.70 Table 3•19 • Summary of a n a l y s i s of data for root-shoot r a t i o of f e r t i l i z e d and u n f e r t i l i z e d seedl ings of the d i f f e r e n t provenances grown in sand, loam-sand and loam. - 103 -1.2 0 . 5 |- , ^ 0 . 3 • • Sand Loam-sand Loam Growth medium -4 Without f e r t i l i z a t i o n m-'. With fertilization F i g u r e 3.11. Mean r o o t - s h o o t r a t i o o f f e r t i l i z e d and u n f e r t i l i z e d s e e d l i n g s o f a l l provenances grown i n sand, loam-sand and loam. 1.0 0.9 0.8 0.7 Root-shoot 0.6 ratio 0.5 0.4 0.3 0.2. U -P rH c > Q) (0 U 0 w W >i o -p <D -P c •rl U •H 3 U u rH M 4J <D > A A A A U 0) >i u s u A A A A <0 •rH rH (0 M •P C d) U A A A A •H 1 -P >i ro 0) cn ra <u JJ § o Ij u o •H W 2 »J A A A A A " - T A A A A A A A A A A Provenance Without f e r t i l i z a t i o n With f e r t i l i z a t i o n Figure 3.12. Mean root-shoot ratio of f e r t i l i z e d and unfertilized seedlings of the different provenances grown in a l l growth media. A A l - 1 0 4 -Di scuss ion Location of seed source had no effect on height growth of the seedlings examined in this study (Tables 3 . 3 , 3 . 6 , and 3 - 1 7 ) . F e r t i l i z e d seedlings, however, were s i g n i f i c a n t l y t a l l e r than u n f e r t i l i z e d seedlings, and the heights of seedlings grown in sand were s i g n i f i c a n t l y lower than that of those grown in the other two media (Table 3 . 1 7 ) . Where there was a si g n i f i c a n t f e r t i l i z a -tion x growth medium interaction, the effect of f e r t i l i z a t i o n was generally greater in sand than in loam-sand and loam. This was expected since coarse sand is a re l a t i v e l y inert material for seedling growth and without f e r t i l i z a t i o n , there would be l i t t l e available nutrients compared with those in the other media. The results collected in this study suggest that height growth of f i r s t year seedlings of red alder is more sensitive to the f e r t i l i t y than to the physical nature of the medium in which they are growing. The effects of f e r t i l i z a t i o n and type of growth medium on dry weight were much the same as those on height growth. F e r t i l i z a t i o n increased dry weight over that of u n f e r t i l i z e d seedlings (Tables 3 . 7 and 3 .18) and seedlings grown in sand were lig h t e r than those grown in loam-sand and loam (Table 3 . 1 8 ) . The effect of f e r t i l i z a t i o n was more pronounced in the presence of sand than in loam. Again, the lack of available nutrients in sand as compared with those present in loam, would have been responsible for these observations. - 105 -Unlike height growth however, dry weight of seedlings was influenced to some extent by location of seed source. As a factor, provenance did not interact with f e r t i l i z a t i o n , growth medium nor with a combination of the two (Table 3 - 1 8 ) . The analysis of data for seedlings grown in each growth medium revealed no provenance effect (Table 3 - 7 ) . S i m i l a r l y , i t was only in the case of u n f e r t i l i z e d seedlings grown in the loam-sand medium that provenance differences were obtained when individual treatments were considered (Table 3.*0. Examination of the l a t t e r data shows that there was a general tendency for seedlings from Laytonville to thrive better than other seedlings when no f e r t i l i z e r was added to the medium. Also, the response of Vancouver seedlings to f e r t i l i z a t i o n in the media containing sand, was greater than that of other seedlings. Both these factors, together with the general effects of f e r t i l i z a t i o n and type of growth medium, most l i k e l y contributed to the differences obtained (Table 3 . 1 8 ) . The physical nature of the s o i l appears in these experiments to be more important in determining dry weight of seedlings than i t is for determining height growth. However, s o i l f e r t i l i t y seems to be the more important factor for both. Provenance differences observed here might r e f l e c t differences in adaptation to localized factors in the various areas of ori g i n . Root-shoot dry weight ra t i o was lowered by f e r t i l i z a t i o n (Tables 3 .5 and 3 . 1 9 ) . This suggests that with the increased a v a i l a b i l i t y of s o i l nutrients, proportionately, - 1 0 6 -more growth occurred in the shoots than in the roots of seedlings. Root growth also was favoured by the coarser sand medium since i t was higher in this than in other media (Table 3 . 1 9 ) . Root-shoot ratios of seedlings from Laytonville and Cottage Grove did not vary as much as those of the other seedlings in the various treatments (Table 3 - 5 ) , hence the effects of f e r t i l i z a t i o n and growth medium were not as marked in these provenances. The observed provenance differences probably resulted from adaptation of seedlings from individual areas to localized s o i l factors, since there seems to be no direct relationship between geographic location of the provenances and the magnitude of the root-shoot r a t i o . Growth and development of a plant involve a strong interdependence between it s root and i t s shoot. The root depends on the shoot for photosynthates and growth regulating substances while the shoot depends on the root for water and minerals. The root also provides support for the whole plant in i t s s o i l medium. Because of this interdependence, environmental factors which affect the growth or function of one, w i l l affect the other, as well as growth of the plant as a whole. The type of root growth and the proportion of root growth to shoot growth that occur in seedlings are genetically determined in many species but both may be modified by environment (Kramer, 19^9; De Roo, 1969) . Moisture content, aeration, mineral status and temperature of the s o i l , and root competition appear to be the major factors influencing root growth and - 107 -functions d i r e c t l y (Kolsov, 197*0. Combinations of these factors may also interact to give a complex effect (Rogers and Head, 1969)- These s o i l factors a l l interact with the physical nature of the s o i l in that they influence the rate at which water, nutrients, oxygen and heat move in response to potential gradients, and may mechanically impede root extension (Eavis and Payne, 1969) . In this study, both the physical characteristics and mineral status of the rooting medium were varied. There might have been s l i g h t differences in so i l moisture content as a result of differences in porosity but these were reduced by avoiding dessication or waterlogging. Neither root competition nor s o i l temperature differences were thought to be si g n i f i c a n t factors. Aeration of the rooting medium appears to depend more on the size of pores than on i t s total pore space (Kramer, 19**9) and in the absence of other l i m i t i n g factors, increased aeration increases root development and a c t i v i t y (Kolsov, 1974) . Sand contains r e l a t i v e l y large particles and thus, pore spaces would have been large as compared with those in loam. In the mixed loam-sand medium, average pore size would have been between those of the other media. Hence, aeration would have been greater in sand than in loam-sand, which in turn would have been greater than in loam. Sand on the other hand, would have been the most i n f e r t i l e of the three media in the absence of f e r t i l i z a t i o n . Without f e r t i l i z a t i o n , aeration increased and f e r t i l i t y decreased from loam, through loam-sand to sand. The response - 108 -to s o i l f e r t i l i t y or the addition of f e r t i l i z e r s is a complex phenomenon (Rogers and Head, 1969) . It is generally accepted that with adequate moisture and aeration, the a v a i l a b i l i t y of large amounts of essential minerals especially nitrogen, increases root growth (Kramer, 1949; Kolosov, 1974) . At the same time, shoot growth is stimulated, often leading to decreased root-shoot ratios in seedlings (Kramer and Kozlowski, I960; Kozlowski, 1 9 7 0 - Increased growth (as measured by height and dry weight of seedlings) and reduced root-shoot ratios with f e r t i l i z a t i o n were observed in this study. In these f e r t i l i z e d treatments, the major-factor li m i t i n g root growth and a c t i v i t y would have been aeration. The increased root development and a c t i v i t y associated with increased s o i l aeration were observed when the physical nature of the root systems of f e r t i l i z e d seedlings grown in sand were compared with those grown in the loam-sand mixture and in loam. If the former's red colour did in fact result from the presence of anthocyanins, as speculated e a r l i e r , t h e i r formation was most l i k e l y f a c i l i t a t e d by the accumulation of carbohydrates translocated from the shoots to the roots, and by the conditions within the medium. In nutrient deficient conditions, the germinal root and i t s nodal branches develop vigorously whereas l i t t l e development and growth of lateral roots take place (Rogers and Head, 1969; Kolosov, 1974) . Nutrient deficiencies also appear to have a more pronounced effect on reducing growth in the shoot than in the root (Beadle, 1954; Kolosov, 1974) . The long unbranched nature of such roots and the r e l a t i v e l y poor shoot growth were observed in u n f e r t i l i z e d - 109 -seedlings grown in sand. Except for seedlings from Laytonville and Cottage Grove, u n f e r t i l i z e d seedlings grown in sand had r e l a t i v e l y high root-shoot ratios. They probably resulted from the greater s e n s i t i v i t y of shoot growth to nutrient deficiencies than root growth. A strong genetic influence could have prevented any modification by the environment on the root-shoot ratios of seedlings from Laytonville and Cottage Grove. The mechanisms by which growth is distributed between root and shoot were discussed by Wilson (1975) and i t appears that the control of this dis-t r i b u t i o n of growth is a complex phenomenon, s t i l l to be defined (Vaadia and Itae, 1969) . Ledig et al. (1970) stressed the importance of considering the fact that ratios involving weights of roots and shoots change as plants grow. This becomes important when comparing plants of different sizes. Thus, the reduced root-shoot ratios brought about by f e r t i l i z a t i o n could have been the result of a char a c t e r i s t i c change associated with the increase in seedling size and brought about by internal changes accompanying increased root and shoot a c t i v i t y . Excessive f e r t i l i z a t i o n can depress seedling growth through i t s effect on root growth and root a c t i v i t y (Rogers and Head, 1969) and the amount of growth depression depends on the type of f e r t i l i z e r used (Kramer and Kozlowski, I 96 0 ) . Increases in dry weight of red alder seedlings in three different media were obtained with the addition of 15 and 30 ppm. of urea and nitrate-nitrogen by Zavitkovski and Newton (1968) . Dry weight was reduced above these concentrations of both f e r t i l i z e r s . Urea-nitrogen - 110 -had a more depressing effect on both dry weight and nodulation than did nitrate-nitrogen. There were no indications of adverse effects caused by too high concentrations of the f e r t i l i z e r used in this study. The levels used were considered to be within the range in which growth is not depressed. As indicated e a r l i e r , nodulation appeared to be under strong genetic control. Nodule formation was not influenced by variations in the environmental conditions in this study, nor did i t appear to be a function of seedling size. It was intended to compare the observations in this study with the performance of seedlings from the various provenances in two f i e l d t r i a l s established at the University of B r i t i s h Columbia's Research Forest in July 1977, but because of poor seedling s u r v i v a l , these t r i a l s had to be abandoned. The differences between provenances in this investigation could not be related to geographic factors. A similar situation was encountered by De Bell and Wilson (1978) in thei r red alder provenance t r i a l s at the Cascade Head Experimental Forest in Oregon. While they were able to explain the poor performance of seedlings from the extremes of the species' range (Juneau, Alaska and Sandpoint, Idaho), reasons for the rel a t i v e performance of seedlings from B r i t i s h Columbia, Washington and Oregon were not obvious. Considerable variation within provenances was also noted by these investigators. It is possible that the amount of variation existing within individual - 111 -provenances of red alder, precludes the development of any relationship between certain t r a i t s and geographic factors when provenances within the extremes of the species' range are examined. On the other hand, the observed ^differences might represent adaptation to environmental factors not considered in experimentation (Wright, 1 9 7 6 ) . Some of the more i n f l u e n t i a l environmental factors, e.g. temperature and daylength, were not varied in this study. The great tree to tree variation that exists within red alder stands, together with i t s reproductive and growth ch a r a c t e r i s t i c s , are favourable for genetic improvement programmes ( S t e t t l e r , 1 9 7 8 ) . Present indications are that individual tree selection is most promising (De Bell and Wilson, 1 9 7 8 ) . Conclusions Height growth in red alder seedlings did not d i f f e r between the provenances examined. It was increased by f e r t i l i z a t i o n over that of u n f e r t i l i z e d seedlings, and was lower in sand than in loam-sand and loam. The effects of f e r t i l i z a t i o n and type of growth medium on dry weight were similar to those on height. F e r t i l i z a t i o n also lowered root-shoot ratios of seedlings which tended to be greater in sand than in the other media. Unlike height growth, both of these measures were affected by the location of the seed source. The differences between provenances, however, could not be related to geographic factors. - 112 -The physical nature of roots produced by seedlings was influenced by the type of growth medium and the nutrient status of the same. Nodulation, however, appeared to be under strong genetic control and was influenced l i t t l e by the environment. Nodules tended to develop more readily on upper root branches than on lower ones, regardless of the surrounding type of growth medium. There was considerable variation among seedlings of individual provenances in a l l t r a i t s measured. This characteristic of red alder populations, together with i t s reproductive and growth char a c t e r i s t i c s , are favourable for tree breeding programmes. - 113 -CHAPTER IV IDENTIFICATION OF ENDOGENOUS GIBBERELLINS IN BUDS OF RED ALDER I n t r o d u c t i o n S i n c e the i s o l a t i o n o f g i b b e r e l l i c a c i d (GA^) i n 1939, o v e r 50 a d d i t i o n a l g i b b e r e l l i n s (GAs) have been i d e n t i f i e d . The c h e m i s t r y and r e g u l a t o r y r o l e s o f t h e s e growth promoters a r e w e l l documented and were r e v i e w e d by s e v e r a l a u t h o r s i n c l u d i n g Lang ( 1 9 7 0 ) , K r i s h n a m o o r t h y (1975), P h a r i s and Kuo (1977), Hedden et al., (1978) and Hedden (1979)- P o t e n t i a l r o l e s o f GAs i n r e g u l a t i n g shoot e l o n g a t i o n , d i a m e t e r growth and crown form ( P h a r i s , 1976a), and i n m a n i p u l a t i n g f l o w e r i n g i n c o n i f e r o u s t r e e s p e c i e s ( P h a r i s , 1976b) have a l s o r e c e i v e d a t t e n t i o n . A l l t he known GAs have the same b a s i c m o l e c u l a r s t r u c t u r e but they do not show e q u a l a c t i v i t y i n the commonly used b i o a s s a y s ( C r o z i e r et al., 1970). N a t u r a l l y o c c u r r i n g GAs have been found i n s e v e r a l t r e e s p e c i e s and t h e y appear t o p l a y some r o l e i n a v a r i e t y o f p h y s i o l o g i c a l p r o c e s s e s i n c l u d i n g t he development o f , and the r e l e a s e from bud dormancy. The l a t t e r i s a s s o c i a t e d w i t h an i n c r e a s e i n growth p r o m o t e r s , a d e c r e a s e i n i n h i b i t o r s o r both (Kramer and K o z l o w s k i , 1979)- In s t u d y i n g the r o l e p l a y e d by hormones i n the c o n t r o l o f bud dormancy, two main approaches have been u s e d , v i z . ( l ) the a p p l i c a t i o n o f exogenous hormones and o b s e r v a t i o n o f subsequent changes and (2) q u a n t i t a t i v e c o r r e l a t i o n o f endogenous hormones w i t h t h e s t a t e o f dormancy. These approaches have s h o r t c o m i n g s ( S a u n d e r s , 1978) and, though they have s u g g e s t e d t h e i n v o l v e m e n t - 114 -of GAs and other hormones, i t has not yet been unequivocally demonstrated that dormancy development and release in buds are mediated through variations in hormone levels (Wareing and P h i l l i p s , 1978). Specific information on the endogenous GAs in buds is rare (Saunders, 1978). Such information w i l l be of value in understanding the role played by GAs in bud dormancy and i t s release. The id e n t i f i c a t i o n of hormones in plant extracts was d i f f i c u l t before the advent of combined gas-liquid chromatography -mass spectometry (GLC-MS). Recent advances in plant hormone research using this technique indicate that data can be obtained to provide a clearer picture of the role played by GAs in the control of bud dormancy. This study was an attempt to extract and identify endogenous GAs present in quiescent vegetative, male and female buds of red alder using bioassay, p a r t i t i o n procedures, GLC and GLC-MS. Materials and Methods Vegetative, male and female buds were collected from several red alder trees growing on the Endowment Lands of the University of B r i t i s h Columbia on January 14 and 15, 1980. These were stored in an ultra-low freezer at -70°C. In the following procedures, three samples of each bud type were used separately. The ethyl acetate and pyridine used throughout this investigation were pre-disti1 led. - 115 -E x t r a c t i o n Each sample (30 grams) o f f r o z e n m a t e r i a l was mixed w i t h 300 ml. o f 80% methanol a t -70°C. and homogenized i n a Wareing B l e n d e r . The homogenate was kept c h i l l e d ( w i t h t h e use o f d r y i c e ) and s t i r r e d f o r about 30 m i n u t e s . I t was then vacuum f i l t e r e d t h r o u g h Whatman No. 1 f i l t e r p aper. The r e s i d u e was r i n s e d t h r e e t i m e s w i t h 80% methanol a t 0°C. and vacuum f i l t e r e d . Phosphate b u f f e r ( 0 . 5 M . , pH 8.0) was added t o t h e 80% methanol e x t r a c t (30 ml. o f b u f f e r per 100 ml. o f e x t r a c t ) . The m i x t u r e was c o n c e n t r a t e d a t 35°C. t o the aqueous phase u s i n g a BL i c h l e r f l a s h e v a p o r a t o r . The aqueous phase was a d j u s t e d t o pH 9-0 w i t h IN. KOH and vacuum f i l t e r e d t h r o u g h f i l t e r paper. The f i l t r a t e was p a r t i t i o n e d t h r e e t i m e s , each t i m e w i t h an equal volume o f d i e t h y l e t h e r w h i c h was p r e s a t u r a t e d w i t h pH 9.0 phosphate b u f f e r ( 0 . 5 M . ) . The pH o f the aqueous phase a f t e r p a r t i t i o n i n g was lowered t o 3-0 w i t h IN. HC1. I t was then p a r t i t i o n e d f i v e t i m e s w i t h e t h y l a c e t a t e ( p r e s a t u r a t e d w i t h pH 9-0, 0 . 5 M . b u f f e r ) . In each p a r t i t i o n i n g , t he r a t i o o f e t h y l a c e t a t e t o t h e aqueous phase was 3"-5 by volume. The e t h y l a c e t a t e f r a c t i o n s were c o l l e c t e d , p o o l e d and s t o r e d f o r s e v e r a l hours a t -70°C. Ice formed d u r i n g s t o r a g e was f i l t e r e d , r i n s e d w i t h e t h y l a c e t a t e a t -70C and r e f i l t e r e d . The f i l t r a t e was e v a p o r a t e d a t 35°C. t o d r y n e s s . Remaining v o l a t i l e s u b s t a n c e s were removed i n a d e s s i c a t o r in vacuo. - 116 -The dried extract was redissolved, transferred to a smaller evaporator flask in a minimum of a methanol:ethyl acetate (1:1 v/v) mixture and dried on the flash evaporator. This smaller flask with the dried extract was stoppered and stored at -20°C. Fractionation S i l i c a gel p a r t i t i o n chromatography was used for fractionation. The methods used here were similar to those described by Durley et al., (1972). One column was prepared for each sample. About kO grams of Woelm S i l i c a Gel for column chromatography (70-150 mesh; p a r t i c l e size 100-200 um.) were preslurried with a mixture of ethyl acetate: hexane (1:9, v/v) saturated with 0.5 M formic acid. The slurry was poured into a 2.1 cm. I.D. glass column to a packed height of 22.5 cm. The dried extract was taken up in a minimum of a methanol:ethyl acetate (1:1 v/v) mixture, transferred onto several Whatman glass microfibre f i l t e r s , 2.1 cm. in diameter, and dried. The f i l t e r s were placed on top of the column which was gradient eluted with mixtures of ethyl acetate and hexane, both presaturated with 0.5M. formic acid. A Varigrad system with four chambers in series was used to achieve mixing of the two solvents. Chambers No. 1 to No. k contained ethyl acetate:hexane mixed by volume in the ratios of 75-0:102.5; 97-5:7.17; 127-5:30.7; and - 117 -150.0:0 respectively. Corresponding volumes in these chambers were 136.7 ml., 130.3 ml., 121.8 ml. and 115.5 ml. respectively. Twenty-six fractions were col 1ected. The f i r s t 25 of these contained 20 ml. while the remainder of the solvent volume was in the 26th. fraction. The column was then eluted with 100 ml. of methanol. A l l fractions were divided into three roughly equal parts, each of which was dried under a stream of oxygen-free nitrogen. The column eluate was dried on a rotary evaporator at 35°C. A l l the dried eluates were kept in v a c u o over KOH for at least 2h hours before being stored at ~70°C. Bioassay One part of each fraction and the column eluate were used to conduct the dwarf rice (Tan-ginbozu variety) seedling bioassay as described by Murikami (1970). Seeds were soaked in tap water at 32°C. for two days. Germinants with primary roots about 2 mm. long were selected for uniformity and transplanted in groups of 10 onto 0.9% water agar to a depth of about 5 cm. in storage dishes (100 mm. in diameter and 80 mm. in height). They were then incubated for two days at 32°C. under continuous fluorescent li g h t i n g with an intensity of about 2000 lux. Dishes were kept loosely covered to provide adequate aeration and water was added regularly to maintain a r e l a t i v e l y high humidity within the dishes. Each dried fraction was taken up in 0.05 ml. of 50% acetone. The column eluate was dissolved in a minimum volume of 50% acetone. A 1 u l . drop - 118 -of each eluate was then applied between the coleoptile and f i r s t leaf of each of 10 seedlings. Each of 10 controls received a I . ju l . drop of 50% acetone. A l l seedlings were kept under the same conditions described above for three additional days after which the length of the second leaf sheath was measured. A growth response was considered positive when the mean length was equal to or greater than 110% of that of the controls. Gas-Liquid Chromatography The second part of those fractions which showed a positive response in the bioassay was used for gas-liquid chromatography (GLC) on a Varian Aerograph 17^0 gas chromatograph (Varian Associates Ltd.). These were compared with standard GAs (GA^, GA^, GA^» and a G A / j / y mixture obtained from Dr. D.C. Aldridge, Imperial Chemical Industries Ltd., Macclesfield, Cheshire, England). The standards and dried eluates were dissolved in a minimum of dry pyridine. The s i l y l a t i n g agent BSTFA (N,0-bis-(Trimethy1silyl) Trifloroacetamide; Pierce Chemical Co.) was then added in the ratio of three parts BSTFA to one part pyridine. The reaction mixture was allowed to stand at room temperature for at least 15 minutes. Before i n j e c t i o n , s i l y l a t e d fractions were concentrated by drying under a slow stream of nitrogen. - 119 -S i l y l a t e d standards and fractions were chromatographed on glass columns 1.83 metres long and with an internal diameter of 2 mm. These were packed with 80-100 mesh Gas Chrom '0.' (Applied Science Laboratories Inc.) coated with 3% SE-33 and were preconditioned as described by Gaskin and MacMillan (1978). At i n j e c t i o n , the column temperature was held at 120°C. for two minutes after which i t was programmed to 280°C. at 10°C. per minute with a 15 minute isothermal hold at the end. The injector and detector temperatures were 250°C. and 280°C. respectively. The flow rate of the nitrogen c a r r i e r gas was 25 ml. per minute. Combined Gas-Liquid Chromatography-Mass Spectrometry The third part of those fractions with peaks whose retention times were similar to those of the standards in the GLC, were used for combined gas-liquid chromatography-mass spectrometry (GLC-MS). These fractions and standards were s i l y l a t e d as described in the preceding section and were chromatographed on a Hewlett-Packard 5700A gas chromatograph with a glass column 1.83 metres long and with an internal diameter of 2 mm. This col umn was packed with Gas Chrom 'Q' coated with 2% OV-17. The column temperature was programmed from 150°C. to 290°C. per minute with an eight minute isothermal hold at the end of the program. The gas-liquid chromatograph was connected to a Varian Mat-Ill mass spectrometer by a s l i t separator. The ionizing potential was 80 ev and the scan interval was fiv e seconds. - 120 -Results Figures 4 . 1 . , 4 . 2 . and 4 . 3 . show the results of the dwarf rice seedling bioassay for fractions of extracts from vegetative, male and female buds respectively, and the expected positions of the standard GAs based on data from Durley et al., (1972) and Pharis (unpublished data). The growth responses obtained from the different fractions were not the same in a l l replications for any one bud type although there was a general tendency for a c t i v i t y to occur in the same general areas. The biological a c t i v i t y obtained in these bioassays was not very high. Female bud fractions appeared to be less active than the vegetative and male. Those fractions with GLC peaks whose retention times were similar to those of standard GAs, are l i s t e d in Tables 4 . 1 . to 4 . 5 . The fractions in which each standard was expected, are underlined. Several peaks with varying inten s i t i e s were obtained from many fractions and this probably indicated that extracts were inadequately p u r i f i e d . Table 4 . 6 . shows the presence or absence of GLC peaks with similar retention times as those of the different standards, from fractions in which the l a t t e r were expected. There was a large degree of variation within the different replications for any one bud type and for any one standard. However, the table does show a complete absence of peaks corresponding to GA^ and GA^ from fractions.from female bud extracts and a similar absence of peaks corres-ponding to GA.- from fractions from vegetative bud extracts. - 121 -2 0 1 5 GAs I 9 l 4,7 | R E P L I C A T I O N N O . 1 GAs JL3_ 1 0 j i i i i i i i i i i i i i t i i i i t i • i i . • i 2 4 & 8 1 0 12 14 1 6 1 8 2 0 2 2 2 4 2 6 C L . 2 0 R E P L I C A T I O N N O . 2 Sl5 s: 10 "8 io 0) - 1 — I — I — I — I — I — I — I — I — I I I I I I I I I I I I I I I I I 2 4 6 8 1 0 1 2 14 1 6 1 8 2 0 22 2 4 2 6 C L R E P L I C A T I O N N O . 3 CVJ 2 0 oils c 0) a 1 0 J — i — i — i — i — i — i — i — i — i i i i i i i i I I 2 4 6 8 10 1 2 1 4 1 6 1 8 2 0 2 2 2 4 2 6 C L , F r a c t i o n N o . CL.: Column e l u a t e : 110?$ o f mean l e n g t h o f second l e a f s h e a t h o f c o n t r o l s • 1: E x p e c t e d p o s i t i o n s o f s t a n d a r d GAs F i g u r e 4.1. Dwarf r i c e b i o a s s a y o f f r a c t i o n s c o l l e c t e d d u r i n g s i l i c a g e l p a r t i t i o n chromatography o f e x t r a c t s from v e g e t a t i v e buds. G A s REPLICATION NO. 1 - 122 -G A s 13_ 2 4 6 8 10 12 14 16 18 20 22 24 26 CL. Fraction No. CL.: Column e l u a t e : 110% o f mean l e n g t h o f second l e a f s h e a t h o f c o n t r o l s i f: E x p e c t e d p o s i t i o n s o f s t a n d a r d GAs F i g u r e *K2. Dwarf r i c e b i o a s s a y o f f r a c t i o n s c o l l e c t e d d u r i n g s i l i c a g e l p a r t i t i o n chromatography o f e x t r a c t s from male "buds. 123 G A s 1 9 1 | REPLICATION NO. 1 G A s d3_ 20 15 10- -I 1 I I I I I t • • • i -1 l _ -I I L. 4 6 8 10 12 14 16 18 20 22 24 26 CL. REPLICATION NO. 2 20 -P ro £. 15 w \ 10- J 1 1 1 1 1 I I I I I I I • • . . . 4 6 8 10 12 14 16 18 20 22 24 26 CL, C CM REPLICATION NO. 3 % 20 4J 9 15 10 J — i — i — i — I _ J i i i i i i i i » • 4 6 8 10 12 14 16 18 20 22 24 26 CL. F r a c t i o n No. CL.: Column e l u a t e 110% o f mean l e n g t h o f second l e a f s h e a t h o f c o n t r o l s E x p e c t e d p o s i t i o n s o f s t a n d a r d GAs F i g u r e 4.3. Dwarf r i c e b i o a s s a y o f f r a c t i o n s c o l l e c t e d d u r i n g s i l i c a g e l p a r t i t i o n chromatography o f e x t r a c t s from female buds. - 124 -Bud Type Replication Fraction Nos.  Vegetative No. 1 1;2;4;5;12;13;16;17. No. 2 1;2;4;7;8;9;10;11;12. No. 3 5;7;9;11;13;17. Male No. 1 2;.5;13;14;15. No. 2 2;4;12;13;15;16. No. 3 2;5;11;12. Female No. 1 12;15;17;21. No. 2 U;17;20. No. 3 10;12;13;15;18;19;22. Table 4.1. Fractions of extracts from vegetative, male and female buds which showed a positive growth response in the dwarf rice bioassay and with GLC peaks whose retention times were similar to that of standard GA^ . Bud Type R e p l i c a t i o n F r a c t i o n N os. V e g e t a t i v e No. 1 1;3;A;5_;6_;7;8;9. No. 2 1;7;9;10;12;13. No. 3 2;5.;6.;7;9; 17. M a l e No. 1 5;7;8. No. 2 4;5.;7;14;21;23. No. 3 3;5;6;7. Female No. 1 12;15;17;21;22. No. 2 12;13;17;20;22. No. 3 10;12;13;19;22. Table 4.2. Fractions of extracts from vegetative, male and female buds which showed a positive growth response in the dwarf rice bioassay and with GLC peaks whose retention times were similar to that of GA... - 125 -Bud Type Replication Fraction Nos. Vegetative No. 1 1;3;4;5;J.;7;8;9. No. 2 1;7;9;10;12;13. No. 3 2;5;£;7;9. Male No. 1 5;8. No. 2 4; 5.; 7; 14; 21; 23. No. 3 5;6;7. F emale No. 1 12;15;17;21;22. No. 2 13;17;20;22. No. 3 10;12;13;19;22. Table 4.3- Fractions of extracts from vegetative, male and female buds which showed a positive growth response in the dwarf rice bioassay and with GLC peaks whose retention times were similar to that of GA.,. Bud Type Replication Fraction Nos. Vegetative No. 1 3;4;7;8. No. 2 1;2.;3;7;11;15. No. 3 1;1;7;9. Male No. 1 2;5;7. No. 2 2.; 3.; 7. No. 3 •l;7;9. Female No. 1 1;3. No. 2 1. No. 3 2. Table 4.4. Fractions of extracts from vegetative, male and female buds which showed a positive growth response in the dwarf rice bioassay and with GLC peaks whose retention times were similar to that of GA„. - 126 -Bud Type Re p l i c a t i o n F r a c t i o n No. Vegetative No. 1 9;12;13;18;19. No. 2 7;9;10;12;13. No. 3 9;13;17. Male No. 1 2;5;13;14;15; No. 2 2;4;1S;18. No. 3 2;5;19. Female No. 1 12;15;17;21. No. 2 12;13;17;20. No. 3 10;12;13;15;18;19;22. Fractions of extracts from vegetative, male and female buds which showed a positive growth response in the dwarf rice bioassay and with GLC peaks whose retention times we similar to that of GA. ; - 127 -Bud Type  Vegetative Male Female Standard Fraction No. 1 2 3 1 2 3 1 2 3 GA^ 12 + + _ _ + + + _ + 13, + _ + + + _ _ + + 14 _ _ _ + _ _ _ _ _ 15 _ _ _ + + _ + _ + GA^ 4 + _ . - + _ 5 + _ + • + + + 6 + _ + _ _ + GA? 4 + _ _ _ + _ 5 + _ + + + + 6 + _ + _ _ + GA^ 2 _ + + + + _ + _ + 3 + + + . _ + _ + _ _ GA 1 3 15 _ _ _ + _ _ + _ + 16 _ _ _ _ + _ _ _ _ Table 4.6. Presence (+) or absence (-) of GLC peaks with similar retention times as those of standard GAs, from f r a c t i o n s i n which the standards were expected. - 128 -None of the spectra from scans at peaks with similar retention times corresponded exactly to any of the standard GAs, nor could they be ident i f i e d as spectra of other GAs with the use of published data. The lack of fraction purity and the low concentrations of GAs contained in the extracts probably contributed to this f a i l u r e to identify individual GAs. Many of the spectra showed features common to those of some gibberellins published by Binks et al.,(1969). As an example, Figures 4 . 4 , 4 . 5 . and 4 . 6 . show the mass spectra of s i l y l a t e d GA^ and the mass spectra from scans at two closely situated GLC peaks with similar retention times as GA^ from s i l i c a gel fraction No. 15 of female bud extracts. Background was subtracted from the l a t t e r two. A l l three show peaks at the mass-to-charge ratio (m/e) 73 and m/e 75 which are associated with the trimethy1sily1 (TMSi) ether group. Significant peaks at m/e 207/208 are cha r a c t e r i s t i c of 7_TMSi ethers of GAs. Also, v i c i n a l alcohol functions in GAs produce strong ions at m/e 147. Other features associated with the TMSi ether group are fragment ions at M-15 and M-89/90. If in the spectrum in Figure 4 . 5 - , the mass of the compound is taken as 526, the M-15 and M_90 are represented by m/e 511 and m/e 436 respectively. S i m i l a r l y , in the spectrum i l l u s t r a t e d in Figure 4 . 6 . , M-90 would be represented by m/e 438 i f the mass is taken as 528. However, none of these masses correspond to the theoretical values of the TMSi ethers of GAs whose reference spectra have been published. The intensity of these s i g n i f i c a n t peaks suggests that GAs were in low concentrations. Important features of their spectra could also have been masked by those of contaminants with similar retention times (Gaskin and MacMillan, 1978) . -129 -73 GAj STANDARD 562 147 \ 205 203 V I 4-90 M-15 50 100 150 250 300 m/e 350 400 450 500 5 50 600 Figure h.k Mass spectrum of the TMSi ether of GA - 130 -Figure k.$. Mass spectrum of scan at peak whose retention time was similar to that of the TMSi ether of GA (Fraction No. 15-Replicat ion 3 of female bud extract). 3 - 131 -FEMALE Figure k.6. Mass spectrum of scan at peak whose retention time was similar to that of the TMSi ether of GA (Fraction No. 15~Replication 3 of female bud extract). - 132 -Di scuss ion The small quantities of standard GAs which were available made i t impossible to subject them to s i l i c a gel p a r t i t i o n chromatography. The positions of standard GAs shown are thus based on data from experiments (Durley et al., 1972; Pharis, unpublished data) in which conditions differed and may have caused s l i g h t s h i f t i n g of elution positions. The experiments reported here were done under their own identical conditions and so some comparison between vegetative, male and female bud GAs is legitimate. These comparisons are made for materials which emerge in positions where the standards were expected. The inconsistency of results between replications, and the f a i l u r e to identify endogenous gibberellins meant that few def i n i t e conclusions were possible concerning the different GAs present in the vegetative, male and female buds. Different GAs have been associated with the sex of s t r o b i l i and with various aspects of vegetative development in conifers (Pharis, 1976b; Pharis and Kuo, 1977). It is possible that a similar situation exists in red alder. Thus, i t would not be surprising i f the apparent differences in the GAs present in the extracts from the different bud types (Table k.6.) were found tp be real. In this case, GA^ w o u ' d appear to play a minor role in vegetative development whereas GA^ and GA^ would appear to be of l i t t l e significance in the development of female buds. - 133 -There were indications in the GLC phase of this study, that many fractions of the different extracts contained several components. The dwarf rice seedling bioassay is s p e c i f i c for GAs though a l l GAs do not respond equally. The r e l a t i v e l y low biological a c t i v i t y obtained in these bioassays could have resulted from the presence of inhibitory impurities and/or very low concentrations of GAs in the fractions. The results of the combined GLC-MS show, as expected, the continuing need for improved p u r i f i c a t i o n techniques and publication of more spectra. So many GAs occur in such small quantities that development of high y i e l d p u r i f i c a t i o n procedures remains a top p r i o r i t y . The most recently developed high resolution technique is high performance li q u i d chromatography (Reeve and Crozier, 1978) . This was not available for this study. In GLC, many naturally occurring substances chromatograph with similar retention times as GAs and as such, GLC alone does not conclusively identify the lat t e r (Gaskin and MacMillan, 1978) . Contaminants, not usually detected in bioassays, can complicate GLC-MS analyses and i d e a l l y , p u r i f i c a t i o n before this procedure should separate GAs from a l l other components (Hedden, 1979) . It is evident that this was not rigorously achieved in this study although the pa r t i t i o n and bioassay procedures do show that much contamination was removed. The lim i t s of bioassays are well appreciated. Higher concen-trations of GAs might have been obtained using larger sample sizes in the extraction process. However, this would have correspondingly increased the amount of impurities and/or inhibitors which as noted before, are not usually detected in and may in h i b i t bioassays. Hence, the problems that were en-countered in the GLC and GLC-MS phases would have s t i l l prevailed. - 134 -P a r t i t i o n chromatography on gradient e luted Woelm s i l i c a gel columns appears to be one of the most s u i t a b l e methods of p u r i f i c a t i o n (Durley et al., 1972). For extracts from plant materials such as red a lder buds which appear to contain a wide v a r i e t y of substances other than GAs, the use of one or a combination of other chromatographic methods (Hedden, 1979) p r i o r to the s i l i c a gel p a r t i t i o n method, might, achieve the degree of p u r i f i c a t i o n necessary. The disadvantage of these a d d i t i o n a l steps is that they generate several more f r a c t i o n s , each r e q u i r i n g separate a n a l y s i s and hence c o n t r i b u t i n g to the loss of f i n a l substance f o r i d e n t i f i c a t i o n . M e t h y l a t i o n , p r i o r to s i l y l a t i o n , increases the v o l a t i l i t y of GAs for GLC and GLC-MS. In a d d i t i o n , mass spectra and other GLC-MS data f o r the TMSi ethers of the methyl esters of several GAs have already been published (eg. by Binks et al., 1969 and Metzger and Zeevaart , 1980) and t h i s ready reference should f a c i l i t a t e i d e n t i f i c a t i o n i f standard GAs are not a v a i l a b l e . Conclusions A low but s i g n i f i c a n t level of b i o l o g i c a l a c t i v i t y was detected in the dwarf r i c e bioassay for g i b b e r e l l i n s performed on f r a c t i o n s of extracts from v e g e t a t i v e , male and female buds of red a l d e r . GLC peaks from f r a c t i o n s with s i m i l a r retent ion times as those of standard GAs, a l s o suggested the presence of GAs in these e x t r a c t s . Combined GLC-MS confirmed the presence of several elements which were c h a r a c t e r i s t i c of pure g i b b e r e l l i n s . Low concentrations and/or inadequate p u r i f i c a t i o n of extracts combined to stop c r i t i c a l i d e n t i f i c a t i o n of any unknown GA from the material examined. - 135 -CHAPTER V GENERAL DISCUSSION It is clear from the evidence presented in Chapter I that more and more interest is being directed towards red alder in the P a c i f i c Northwest. The l i t e r a t u r e surveyed showed that there are very large gaps in our knowledge of the biology of red alder. Many of these gaps must be f i l l e d before a strategy for forest management of this species can be formulated. Knowledge of i t s seed biology, seedling growth and regulation of reproductive processes are three related subjects which were selected for research in these studies. With attention now being focused on the p o s s i b i l i t y of red alder's intensive management and the s i 1 v i c u l t u r a l systems needed to achieve t h i s , the need for as much relevant information as possible cannot be over emphasized. The germination studies have shown that the percentage of viable seeds in individual collections of red alder can vary widely, from as low as 7.3% to as high as 73-0%; that physiological dormancy can exist in a s i g n i f i c a n t number of seeds although i t is not common; that low temperature s t r a t i f i -cation and presoaking in tap water can remove dormancy when present, and can also increase the rate of germination over that of untreated seeds. This type of information w i l l be of great assistance to seed c o l l e c t o r s , nurserymen and s i 1 v i c u l t u r a l i s t s who employ direct seeding. They can use the data on the y i e l d of viable seeds to plan quantities of seed to be collected in - 136 -order to obtain a desirable number of sound seeds. Allowances can be made for the different types of abnormalities described when deciding on the quantities of seed to be sown so as to obtain a required level of germination. Operations could also be hastened by pretreating seeds to overcome any dormancy which may occur. Poor root development in seedlings raised in nurseries is often the cause of poor survival after outplanting. The results of the seedling studies suggest that root development which is adequate to ensure survival after outplanting can be achieved by varying the texture and/or f e r t i l i t y of the growth medium. The rate of height growth in the seedling stage is often a desirable c h a r a c t e r i s t i c in the selection of tree stocks. The lack of provenance differences observed in the analysis for height growth indicates that no one provenance (of those tested) is superior to others in this regard. The preliminary conclusion is that selection for height growth at the provenance level is not worthwhile unless provenances from the extremes of the species' range are included (De Bell and Wilson, 1978). The results of the f e r t i l i z a t i o n treatments have shown the type of growth responses that can be expected from seedlings with the use of f e r t i l i z e r s in nurseries or in the f i e l d . These are increased height growth and dry matter accumulation, and reduced root-shoot dry weight ratios. Growers of seedlings can thus achieve desirable seedling sizes with suitable growth dis t r i b u t i o n s . The apparent absence of GA, in female buds and that of GA in vegetative - 1 3 7 -buds suggest that there are differences in the GA syndromes for male, female and vegetative buds of red alder. This indicates that the study should be pursued. Although no d e f i n i t e conclusions were possible from this study, the published procedures were tested and extraction, bioassay, p a r t i t i o n chromatography, gas-liquid chromatography and combined gas-liquid chromatography-mass spectrometry were established as effective methods for the analysis of GAs. The weak point in the procedure was the use of p a r t i t i o n chromatography for the f i r s t p u r i f i c a t i o n stages. Many workers have t r i e d to resolve this procedure. Materials need d i s t i l l a t i o n and often, batches of s i l i c a gel are unreliable. Clearly, there is room for improvement in the steps involved in this procedure. The recent development and application of high performance l i q u i d chromatography (HPLC) seems to present a real but expensive means of obtaining p u r i f i c a t i o n of GAs and providing the chance to detect currently unknown types. The results of the studies described add to our basic knowledge of red alder. There are many possible future projects. The area of nitrogen f i x a t i o n i s being studied extensively in alders and other hardwood species (Becking, 1975 ; Gordon and Dawson, 1979). The information thus accumulating w i l l give a much clearer picture of this phenomenon in red alder. The preliminary s o i l and f e r t i l i z e r data presented in Chapter III requires much expansion using isotopes to determine the s p e c i f i c ionic requirements and uptake kinetics for red alder's survival and establishment of nitrogen f i x a t i o n . - 138 -It is also important to establish clear guidelines for the selection of suitable trees for tree breeding programs as early as possible and f i e l d t r i a l s which were o r i g i n a l l y a part of these studies, should also be conducted. Studies on endogenous gibberellins w i l l require more attention and with the advances in technology, i t should be possible to understand their action and metabolism, and thus influence red alder's reproductive biology. - 139 -LITERATURE CITED A l l e n , E.K. and O.N. A l l e n , 1958. B i o l o g i c a l a s p e c t s o f s y m b i o t i c n i t r o g e n f i x a t i o n . In W. Ruhland ed. E n c y c l o p e d i a o f p l a n t p h y s i o l o g y , V o l . 8 , S p r i n g e r - V e r l a g : 48-118. A t k i n s o n , W.A. and W.I. H a m i l t o n , 1978. The v a l u e o f red a l d e r as a s o u r c e o f n i t r o g e n i n D o u g l a s - f i r / a l d e r mixed s t a n d s . In D.G. B r i g g s et al. ed. U t i l i z a t i o n and management of a l d e r . P r o c . 1977 Symp., Ocean S h o r e s , Washington. Pac. N.W. For. and Range Exp. S t . P u b l . : 337-351. A t k i n s o n , W.A., B.T. Bormann and D.S. D e B e l l , 1979. Crop r o t a t i o n o f D o u g l a s - f i r and red a l d e r : a p r e l i m i n a r y b i o l o g i c a l and economic assessment. Bot.Gaz. 140 ( S u p p l . ) : S102-S107. A t t e r b u r y , T., 1978. A l d e r c h a r a c t e r i s t i c s as they a f f e c t u t i l i z a t i o n . In D.G. B r i g g s e t al. ed. U t i l i z a t i o n and management o f a l d e r . P r o c . 1977 Symp., Ocean S h o r e s , Washington. Pac. N.W. For. and Range Exp. S t . Publ . : 71-81 . B e a d l e , N.C.W., 1954. S o i l phosphate and the d e l i m i t a t i o n o f p l a n t communities i n e a s t e r n A u s t r a l i a . E c o l o g y 35 : 370 - 3 7 5 . B e c k i n g , J.H., 1970. P1ant-endophyte s y m b i o s i s i n non-leguminous p l a n t s . P I . and S o i 1 3 2 : 611 - 6 5 4 . , 1972. The i mportance o f Alnus rubra f o r the c u l t i v a t i o n o f - Pseudotsuga menziesii-. N e d e r l a n d s Bosbouw T i j d s c h r i f t 4 4:132 - 1 3 7 -, 1975. Root n o d u l e s i n non-legumes. In J.G. T o r r e y and D.J. C l a r k s o n ed. The development and f u n c t i o n o f r o o t s . T h i r d Cabot Symp., Acad. P r e s s : 507~566. Behm, R.D., 1978. D e v e l o p i n g new a l d e r m arkets. In D.G. B r i g g s et al. ed. U t i l i z a t i o n and management o f a l d e r . P r o c . 1977 Symp., Ocean S h o r e s , Washington. Pac. N.W. For. and Range Exp. St. P u b l . : 157-161. Bene, J . , 1950. D o u g l a s - f i r s u b s t i t u t e s f o r B r i t i s h Columbia plywood i n d u s t r y . B.C. Lumberman 34: 49~50 and 145. B e r n s t e n , CM., 1961 a. Growth and development o f red a l d e r compared w i t h c o n i f e r s i n 30 y e a r o l d s t a n d s . Res. Pap., Pac. N.W. For. and Range Exp. S t . #38: 20 pp. , 196lb. P r u n i n g and e p i c o r m i c b r a n c h i n g i n red a l d e r . J . F o r . 5 9 : 6 7 5 - 6 7 6 . Bhatnagar, S.P. and B.M. J o h r i , 1972. Development of arigiosperm seeds. In T.T. K o z l o w s k i ed. Seed b i o l o g y , V o l . 1, Acad. P r e s s : 77-149. B i n k s , R. , J . M a c m i l l a n and R.J. P r i c e , I969. P l a n t Hormones V I I I : Combined gas chromatography-mass s p e c t r o m e t r y o f the methyl e s t e r s o f g i b b e r e l l i n s A 1 t o A ^ and t h e i r t r i m e t h y l e t h e r s . P h y t o c h e m i s t r y 8: 271-284. - 140 -Bollen, W.B. and E. Wright, 1961. Microbes and nitrate in s o i l s from vi r g i n and young growth forests east and west of the Cascade mountains and along the P a c i f i c coast. Can. J. Microbiol. 1: 785-792. Bollen, W.B., C.S. Chen, K.C. Lu and R.F. Tarrant, 1967- Influence of red alder on f e r t i l i t y of a forest s o i l . Microbial and chemical effects. Res. B u l l . , Ore. For. Res. Lab. #12: 61 pp. Bollen, W.B. and K.C. Lu, 1968. Nitrogen transformations in s o i l s beneath red alder and conifers. In J.M. Trappe et al. ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967: 141-148. Bond, G., 1958. Symbiotic nitrogen f i x a t i o n by non-legumes. In E.G. Hallsworth ed. Nutrition of legumes. Buttersworths Sc. Publ.: 216-231. : , 1959- Fixation of nitrogen by non-legume root-nodule plants. Symp. Soc. Exp! B i o l . 13, Camb. Uni. Press: 59~72. , 1963. The root nodules of non-leguminous angiosperms. Symp. Soc. Gen. Microbiol. 13: 72-91. , 1967. Fixation of nitrogen by higher plants other than legumes. Ann. Rev. PI. Phys. 18: 107-126. , 1974. Root-nodule symbiosis with Actinomycete-1ike organisms. In A Quispel ed. The biology of nitrogen f i x a t i o n , North Hall Publ. Co.: 342-378. Bonner, F.T., 1974. Seed testing. In Seeds of woody plants in the United States. U.S.D.A. Agr. Handbook 450: 136-152. Briggs, D.G., J.S. Bethel and G.F. Schreuder, 1978. An approach for comparing the relative value of alder with other species from forest to end product. In D.G. Briggs et al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 35 _46. Buchanan, T.S., 1948. Poria weivii: i t s occurrence and behaviour on species other than the cedars. N.W. Sc. 22: 7 -12. C a l i f o r n i a Department of Water Resources Report, 1977- The continuing Californian drought. State of Cal. Res. Agency Publ.: 138 pp. Chi Ids, T.W. and K.R. Shea, 1967- Annual losses from diseases in P a c i f i c Northwest forests. U.S.D.A. For. Serv. , Pac. N.W. For. and Range Exp. St., Res. B u l l . PNW-20: 19 pp. Chow, S., 1972. Thermal reactions and industrial uses of bark. Wood and fibre 4: 130-138. Chow, S. and K.J. Pickles, 1971. Thermal softening and degradation of wood and bark. Wood and fibre 3: 166-178. - 141 -C l a r k , D.H., 1957- Developments i n the use o f red a l d e r . For. P r o d . J . 7: 17-20A. C o l e , D.W., S.P. G e s s e l and J . T u r n e r , 1978. Comparative m i n e r a l c y c l i n g i n red a l d e r and D o u g l a s - f i r . In D.G. B r i g g s et al. ed. U t i l i z a t i o n and management o f a l d e r . P r o c . 1977 Symp., Ocean F a l l s , W ashington. Pac. N.W. For. and Range Exp. S t . P u b l . : 327~336. C r o z i e r , A., R. Kuo, R.C. D u r l e y and R.P. P h a r i s , 1970. The b i o l o g i c a l a c t i v i t i e s o f 26 g i b b e r e l l i n s i n n i n e p l a n t b i o a s s a y s . Can. J . Bot. 4 8 : 8 6 7 - 8 7 7 . C z a b a t o r , F . J . , 1962. G e r m i n a t i o n v a l u e : an i n d e x f o r c o m b i n i n g speed and c o m p l e t e n e s s o f p i n e seed g e r m i n a t i o n . For Sc. 8 : 3 8 6 - 3 9 6 . De B e l l , D.S. and M.A. Radwan, 1979- Growth and n i t r o g e n r e l a t i o n s o f c o p p i c e d b l a c k cottonwood and red a l d e r i n pure and mixed p l a n t a t i o n s . Bot. Gaz. 140 ( S u p p l . ) : S97-S101. De B e l l , D.S. and B.C. W i l s o n , 1978. N a t u r a l v a r i a t i o n i n red a l d e r . In D.G. B r i g g s et al. ed. U t i l i z a t i o n and management o f a l d e r . P r o c . 1977 Symp., Ocean S h o r e s , Washington. Pac. N.W. For. and Range Exp. S t . P u b l . : 193 -208 . De B e l l , D.S., R.F. S t r a n d and D.L. Reukema, 1978. S h o r t - r o t a t i o n p r o d u c t i o n o f red a l d e r : some o p t i o n s f o r f u t u r e f o r e s t management. In D.G. B r i g g s et al. ed. U t i l i z a t i o n and management o f a l d e r . P r o c . 1977 Symp., Ocean S h o r e s , Washington. Pac. N.W. For. and Range Exp. S t . P u b l . : 2 3 1 - 2 4 4 . De Rop, H.C., 1969. T i l l a g e and r o o t growth. In W.J. W h i t t i n g t o n ed. Root growth. P r o c . E a s t e r Sch. Agr. Sc., 1968. B u t t e r w o r t h and Co.: 339 - 358 . D u r l e y , R . C , A. C r o z i e r , R.P. P h a r i s and G.E. M c L a u g h l i n , 1972. Chromato-graphy o f 33 g i b b e r e l l i n s on a g r a d i e n t e l u t e d s i l i c a g e l p a r t i t i o n column. P h y t o c h e m i s t r y 11: 3 0 2 9 " 3 0 3 3 . E a v i s B.W. and D. Payne, 1969- S o i l p h y s i c a l c o n d i t i o n s and r o o t growth. In W.J. W h i t t i n g t o n ed. Root growth. P r o c . E a s t e r Sch. Agr. S c . , 1968. B u t t e r w o r t h and Co.: 315~338. Espenas, L.D., 1951- The s e a s o n i n g o f Oregon hardwoods. I n f . C i r c , Ore. F o r . Prod. Lab. #6: 35 pp. _, 1971. S h r i n k a g e o f D o u g l a s - f i r , w e s t e r n hemlock and red a l d e r as a f f e c t e d by d r y i n g c o n d i t i o n s . For. Prod. J . 2 1 : 4 4 - 4 6 . Evans, R.S., 1974. Energy p l a n t a t i o n s : s h o u l d we grow t r e e s f o r power p l a n t f u e l . West. Fo r . Prod. Lab. I n f . Rep. VP-X - 129 - Can. For. S e r v . : 15 pp. -•142 -Evans, H.J. and L.E. Barber, 1977. Biological nitrogen f i x a t i o n for food and fiber production. Science 197: 332-339-Feddern, E.T., 1978. Harvesting of red alder. In D.G. Briggs et al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 61-69 . Finnis, J.M., 1964. Chemical f r i l l treatment of alder. Res. Prog. Rep., West Weed Contr. Conf. 1964: 47. F o r r i s t a l , F.F. and S.P. Gessel, 1955- Soil properties related to forest cover type and productivity on the Lee Forest, Snohomish County, Washington. Proc. Soil Sc. Soc. Am. 19: 35^-359-Franklin, J.F., C.T. Dryness, D.G. Moore and R.F. Tarrant, 1968. Chemical s o i l properties under coastal Oregon stands of alder and conifers. In J.M. Trappe et al. ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967: 157-172. Freese, F., 1974. Elementary s t a t i s t i c a l methods for foresters. U.S.D.A. Agr. Handbook 317: 87 pp. Gaskin, P. and J. MacMillan, 1978. GC and GC-MS techniques for gibbere11ins. In J.R. Hillman ed. Isolation of plant growth substances. Camb. Uni. Press, Camb.: 79"95. Gavrielit-Gelmond, H., 1971. Growth and development of the peanut plant (Arachis hypogaea) in relation to seedling evaluation in germination. Proc. Int. Seed Test. Ass. 36: 121 -130 . Gessel, S.P. and J. Turner, 1974. L i t t e r production by red alder in western Washington. For. Sc. 20 : 325-330 . Gordon, J.C. and J.O. Dawson, 1979- Potential uses of nitrogen-fixing trees and shrubs in commercial forestry. Bot. Gaz. 140 (Suppl.): S 8 8 -S90 . Haddock, P.G., 19^9- A problem ch i l d reforms: new perspectives in the management of red alder. For. CI. Quart. 22: 9 - 1 5 -H a l l , J.W., 1952. The comparative anatomy and phylogeny of the Betulaceae. Bot. Gaz. 113: 235-270 . Hansen, E., 1975- Phellinus (Poria) weirii root rot in Douglas-fir-alder stands 10-17 years old. U.S.D.A. For. Serv. Res. Note, Pac. N.W. For. and Range Exp. St. , PNW-250: 5 pp. Hedden, P., 1979- Aspects of gibbe r e l l i n chemistry. In N.B. Mandava ed. Plant growth substances. Symp. 13 th . Mid. At 1. Reg. meeting, Am. Chem. Soc. , N. J., 1979: 19~56. Hedden, P., J. MacMillan and B.O. Phinney, 1978. The metabolism of the gibberel 1'ins. Ann. Rev. PI. Phys. 29 : 149-192. -•143 -Hether i ngton, J.C, 1964. Brush control in coastal B r i t i s h Columbia. Res. Note, B.C. For. Serv., #38: 56 pp. Hildenbrand, H., 1972. Economics of marketing alder, in A.B. Berg ed. Managing young forests in the Douglas-fir region. Ore. State Uni., Sch. For. Symp. 1970: 53~6l. Hillstrom, W.A., 1974. Chip barking of several western species. U.S.D.A. For. Serv. Res. Note, N. Cent. For. Esp. St., NC 164: 4 pp. Hoyer, G., W. Fergerson, M. Newton and D.R.M. Scott, 1978. A comparison of red alder, Douglas-fir and western hemlock productivities as related to s i t e - a panel discussion. In D.G. B;riggs et al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington, Pac. N.W. For. and Range Exp. St. Publ.: 175-182. Hrutfiord, B.F., 1978. Red alder as plywood species. In D.G. Briggs et al. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 135-138. Hughes, D.R., S.P. Gessel and R.P. Walker, 1968. Red alder deficiency symptoms and f e r t i l i z e r t r i a l s . In J.M. Trappe et a l . ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967: 225-237. Johnson, F.D., 1968a. Disjunct populations of red alder in Idaho. In J.M. Trappe et al. ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967: 1-8. , 1968b. Taxonomy and d i s t r i b u t i o n of northwestern alders. In J.M. Trappe et al. ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967: 9-22. Johnson. H.M., 1917- Alnus oregona: i t s value as a forest type on the Siuslaw National Forest. J. For. 15: 981-987. Johnson, H.M., E.J. Hanzlik and W.H. Gibbons, 1926. Red alder of the P a c i f i Northwest: i t s u t i l i z a t i o n with notes on growth and management. U.S.D.A., Dept. B u l l . #1437: 44 pp. Justice, O.L., 1972. Essentials of seed testing. In T.T. Kozlowski ed. Seed biology, Vol. 3, Acad. Press: 301-370. Kenady, R.M., 1978. Regeneration of red alder. In D.G. Briggs et al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 183-191-Kennedy, R.W. and G.K. E l l i o t t , 1957- Spiral grain in red alder. For. Chr. 33: 238-251. Kolosov, I.I., 1974. Absorptive a c t i v i t y of root systems in plants. U.S.D.A Publ. (translated from Russian), Ind. Nat. Sc. Doc. Cent., New Delhi: 525 PP-Kozlik, C.J., 1962. Seasoning red alder lumber. For. Prod. Res. Lab., Ore. #06: 20 pp. - 1 4 4 -, I967. Establishing colour in red alder lumber. Ore. For. Res. Lab. Rep. #D8: 11 pp. , 1978. S t a b i l i z i n g color and drying red alder. In D.G. Briggs et al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 9 3 - 1 0 1 . Kozlowski, T.T., 1971- Growth and development of.trees, Vol. 2 . Acad. Press: 51^ PP-Kramer, P.J., 19^9- Plant and s o i l water relationships. McGraw H i l l Book Co. : 3^7 pp. Kramer, P.J. and T.T. Kozlowski, i960. Physiology of trees. McGraw H i l l Book Co.: 642 pp. , 1979- Physiology of woody plants. Academic Press: 811 pp. Krisnamoorthy, H.N., 1975- Gibberel1ins and plant growth. John Wiley and Sons: 356 pp. Krueger, K.W. and R.H. Ruth, 1968. Photosynthesis of red alder, Douglas-f i r , Sitka spruce and western hemlock seedlings. In J.M. Trappe et al. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967:239. , 1969. Comparative photosynthesis of red alder, Douglas-fir, Sitka spruce and western hemlock seedlings. Can. J. Bot. 47: 519"527-Kurth, E.F. and E.L. Becker, 1953. The chemical nature of extractives from red alder. Tappi 36: 461-466. Lang, A., 1970. Gibberel1ins: structure and metabolism. Ann. Rev. PI. Phys. 2 1 : 5 3 7 - 5 7 0 . Ledig, F.T. , F.H. Bormann and K.F. Wenger, 1970. The distribution of dry matter between shoot and roots in l o b l o l l y pine. Bot. Gaz. 131 : 3 4 9 - 3 5 9 . Leney, L., A. Jackson and D.H. Erickson, 1978. Properties of red alder {Alnus rubra Bong.) and i t s comparison to other hardwoods. In D.G. Briggs et al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ . : 25-33. Leonard, O.A., 1961. Undesirable woody plants. Res. Prog. Rep., West Weed Contr. Conf. I96I: 1 6 - 2 6 . L i , C.Y., K .C. Lu, W.B. Bollen and J.M. Trappe, 1969- Effect of phenolic and other compounds on growth of Poria weirii in vitro. Microbios 3: 305-311. - 145 -L i , C.Y., K.C. Lu, J.M. Trappe and W.B. Bollen, 1967- Selective nitrogen assimilation in Poria weirii. Nature, Lond. 213: 814. , 1968. Enzyme systems of red alder and Douglas-fir in relation to infection by Poria weirii. In J.M. Trappe et al. ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967= 241-250. , 1970a. Separation of phenolic compounds in a l k a l a i hydrolysates of a forest s o i l by thin layer chromatography. Can. J. Soil Sc. 50: 4 5 8 - 4 6 0 . , 1970b. Inhibition of Poria weirii and Pomes annosus by l i n o l e i c acid. For. Sc. 16: 329~330. , 1972a. Nitrate-reducing capacity of roots and nodules of Alnus rubra and roots of Pseudotsuga menziesii. PI. and Soil 37: 409_414. > 1972b. Poria weirii - i n h i b i t i n g and other phenolic compounds in roots of red alder and Douglas-fir. Microbios 5: 6 5 - 6 8 . L i t t l e , G.R., 1978. Supply of western alder stumpage, i t s quantity and quality, 1976-1996 and trends in alder stumpage and product markets. In D.G. Briggs et al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 19-23 . Lu, K.C, C.S. Chen and W.B. Bollen, 1968. Comparison of microbial populations between red alder and conifer s o i l s . In J.M. Trappe et al. ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967: 173-178. Maloney, T.M., 1978. Alder: one of tomorrow's important structural raw materials. In D.G. Briggs &t al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 125-132. Manning, G.H., 1975- B r i t i s h Columbia's neglected hardwood resource. Env. Can., Pac. For. Res. Cent. Rep. BC-X-118: 18 pp. McGuane, H., 1978. Alder as a wood for veneer and plywood. In D.G. Briggs et al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 133. McVean, D.N., 1955. Ecology of Alnus glutinosa (L.) Gaertn. II. Seed di s t r i b u t i o n and germination. J. Ecol. 4 3 : 6 1 - 7 1 . Metzger, J.D. and J.A.D. Zeevaart ( I98O) . Identification of six endogenous gibberellins in spinach shoots. PI. Phys. 65: 623 -626 . M i l l e r , R.E. and M.D. Murray, 1978. The effects of red alder on growth of Douglas-fir. In D.G. Briggs et al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 283~306. - 146 -Minore, D.j 1968. Effects of a r t i f i c i a l flooding on seedling survival of six northwestern tree species. Res. Note, Pac. N.W. For. and Range Exp. St., PNW-92: 12 pp. , 1970. Seedling growth of eight northwestern tree species over three water tables. Res. Note, Pac. N.W. For. and Range Exp. St. PNW-115: 8 pp. Minore, D. and C.E. Smith, 1971. Occurrence and growth of four northwestern tree species over shallow water tables. U.S.D.A. For. Serv. Note, Pac. N.W. For. and Range Exp. St. #160: 9 pp. Minore, D., C.E. Smith and R.F. Woollard, I969. Effects of high s o i l density on seedling growth of seven northwestern tree species. Res. Note. Pac. N.W. For. and Range Exp. St., PNW-12: 6 pp. Murikami, Y., 1970. New rice seedling test for gibberel1ins-microdrop method. Jap. Agr. Res. Quart. 5: 9 PP-Neal J r . , J.L., K.C. Lu, W.B. Bollen and J.M. Trappe, 1968a. A comparison of rhizosphere microfloras associated with mycorrhizae of red alder and Douglas-fir. In J.M. Trappe et al. ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967: 57~71 -Neal J r . , J.L., J.M. Trappe, K.C. Lu and W.B. Bollen, 1968b. Some ecto-trophic mycorrhizae of Alnus rubra. In J.M. Trappe et al. ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967: 179-184. Nelson, E.E., I967. Factors affecting s u r v i v a l o f Poria weirii in buried cubes of Douglas-fir heartwood. For. Sc. 13: 78-84. , 1968. Survival of Poria weirii in conifer, alder and mixed conifer-alder stands. U.S.D.A. For. Serv. Note, Pac. N.W. For. and Range Exp. St., PNW-83: 5 PP-, 1970. Effects of nitrogen f e r t i l i z e r on survival of Poria weirii and populations of s o i l fungi and aerobic Actinomycetes. N.W. Sc. 44: 102-107. , 1975a. Survival of Poria weirii in wood buried in urea-ammended forest s o i l . Phytopathology. 65: 501-502. , 1975b. Survival of Poria weirii on paired plots in alder and conifer stands. Microbios 12: 155-158. Nelson, E.E., E.M. Hansen, C.Y. Li and J.M. Trappe, 1978. The role of red alder in reducing losses from laminated root rot. In D.G. Briggs et al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 273 - 2 8 2 . - 147 -Newton, M., P. El Hassen and J. Zavitkowski, 1968. Role of red alder in western Oregon forest succession. In J.M. Trappe et al. ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1 9 6 7 : 7 3 - 8 4 . Nielson, R.W., 1977- Red alder u t i l i z a t i o n in B r i t i s h Columbia. West For. Prod. Lab. Inf. Rep. #VP-X-164, Dept. Fish, and Env., Van.: 2 9 pp. Panshin, A.J. and C. de Zeeuw, 1 9 6 4 . Textbook of wood technology, V o l . 1 . McGraw H i l l Book Co.: 7 0 5 pp. Perry, D.A., 1 9 7 6 . Seed vigour and seedling establishment. In J.R. Thomson ed. Advances in research and technology of seeds, Part 2 . Cent. Agr. Publ. and Doc, Wageningen: 62-85-Pharis, R.P., 1 9 7 6 a . Probable roles of plant hormones in regulating shoot elongation, diameter growth and crown form of coniferous trees. In M.G.R. Cannell and F.T. Last ed. Tree physiology and y i e l d improvement. Acad. Press: 2 9 1 - 3 0 6 . , 1 9 7 6 b . Manipulation of flowering in conifers through the use of plant hormones. In J.P. Mikscheed. Modern methods in forest genetics. Springer-Verlag: 265~282. Pharis, R.P. and CG. Kuo, 1977- Physiology of gibberellins in conifers. Can. J. For. Res. 7: 299"325-Plank, M.E. 1 9 7 1 . Red alder. U.S.D.A., American Woods F - S 2 1 5 : 7 PP-Pollock, B.M. and E.E. Roos, 1 9 7 2 . Seed and seedling vigour. In T.T. Kozlowski ed. Seed biology, Vol. 1 Academic Press: 3 1 3 - 3 8 7 -Quick, R.H., 1 9 5 7 - Neutral s u l f i t e semi-chemical pulping of red alder (Alnus rubra) For. Prod. J. 7= 2 4 6 - 2 4 7 -Reeve, D.R. and A. Crozier, 1 9 7 8 . The analysis of gibberellins by high performance l i q u i d chromatography. In J.R. Hillman ed. Isolation of plant growth substances. Camb. Uni. Press, Camb.: 41-77. Rediske, J.H. and G.R. Staebler, 1962. Herbicidal s e l e c t i v i t y of chloro-phenobutyrics on Douglas-fir. For. Sc. 8: 3 5 3 - 3 5 9 . Remington, R.L., 1 9 7 8 . Alder.... should we chip i t or saw i t ? In D.G. Briggs et al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocan Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 1 0 3 - 1 0 9 . Rogers, W.S. and G.C. Head, I969. Factors affecting the d i s t r i b u t ion and growth of roots of perennial woody species. In W.J. Whittington ed. Root growth. Proc. Easter Sch. Agr. Sc., I968, Butterworth and Co.: 2 8 0 - 2 9 5 -Ruth, R.H., I968. First-season growth of red alder seedlings under gradients in solar radiation. In J.M. Trappe et al. ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967: 9 9 - 1 0 5 . - 148 -Ruth, R.H. and CM. Bernsten, 1956. Chemical basal treatment to control red alder. Res. Note, Pac. N.W. For. and Range Exp. St., PNW-128: 6 pp. Rymer, K.W., 1951. Red alder in B r i t i s h Columbia. B u l l . For. Branch, Can. Dept. Res. and Dev. #98: 19 pp. Saunders, P., 1978. Phytohormones and bud dormancy. In D.S. Letham et al. ed. Phytohormones and related compounds - a comprehensive tre a t i s e , Vol. 2. Elsevier/North-Hol1 and Biom. Press: 423 -445 . Schalin, I., 1968. Germination analysis o f grey alder (Alnus inoana) and black alder (Alnus glutinosa) seeds. In J.M. Trappe et al.ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967: 107-113-Schopmeyer, C.S., 1974. Alnus B. Ehrh. In Seeds of woody plants in the United States. U.S.D.A., For. Serv., Agr. Handbook 450: 2 0 6 - 2 1 1 . Schroeder, H.A. and E.D. Hansen, I968. Two-stage high y i e l d pulping of red alder. Tappi 51 : 1-7-Smith, J.H.G., 1972. Tree size and yields in juvenile red alder stands. Paper prepared from N.W. Sc. Ass. meeting, 1972: 35 pp. , 1974. Biomass of some young red alder stands. IUFR0 Biomass Studies, 1974. C o l l . L i f e Sc. and Agr., Uni. Maine: 399 -410 . , 1978 . Growth and yi e l d of red alder: effects o f spacing and thinning. In D.G. Briggs et al. ed. U t i l i z a t i o n and management o f alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 2 4 5 - 2 6 3 . Smith, J.H.G. and D.S. DeBell, 1974. Some effects o f stand density on biomass o f red alder. Can. J. For. Res. 4 : 335~340. Smith, N.J., 1978. Red alder as a potential source o f energy. In D.G. Briggs et al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 139-155. S t e t t l e r , R.F., 1978. Biological aspects of red alder pertinent to potential breeding programs. In D.G. Briggs et al. ed. U t i l i z a t i o n and management o f alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 209 -222 . Stewart, R.E., 1972a. Field screening o f stem applied herbicides on Coast Range brush species. Res. Prog. Rep., West Soc. Weed Sc.: 18 -19 . , 1972b. Field screening o f granular herbicides on pole size red alder. Res. Prog. Rep., West Soc. Weed Sc.: 19 -20 . - 149 -, 1974. Bud break sprays for s i t e preparation and release from six coastal brush species. Res. Pap., Pac. N.W. For. and Range Exp. St., PNW-176: 20 pp. Stewart, W.D.P., 1966. Nitrogen f i x a t i o n in plants. Athlone Press, Uni. Lond.: 168 pp. Tarrant, R.F., 1961. Stand development and so i l f e r t i l i t y in a Douglas-fir/ red alder plantation. For. Sc. 7: 2 3 8 - 2 4 6 . ______ 1972. The role of red alder in improving s o i l f e r t i l i t y and growth of associated trees. In A.B. Berg ed. Managing young forests in the Douglas-fir region. Ore. State Uni., Sch. For Symp., 1970: 17-34. , 1978. Attitudes toward red alder in the Douglas-fir region. In D.G. Briggs et al. ed. U t i l i z a t i o n and management of alder. Proc. 1977 Symp., Ocean Shores, Washington. Pac. N.W. For. and Range Exp. St. Publ.: 1-7. Tarrant, R.F., L.A. Isaac and R.F. Chandler J r . , 1951- Observations on l i t t e r f a l l and foliage nutrient content of some P a c i f i c Northwest tree species. J. For. 4 9 : 914-915-Tarrant, R.F. and R.E. M i l l e r , 1963 - Accumulation of organic matter and so i l nitrogen beneath a plantation of red alder and Douglas-fir. Proc. Soil Sc. Soc. Am. 2 7 : 231-234. Tarrant, R.F., K.C. Lu, W.B. Bollen and J.P. Franklin, 1969- Nitrogen enrichment of two forest ecosystems by red alder. Res. Pap., Pac. N.W. For. and Range Exp. St., PNW-76: 8 pp. Tarrant, R.F. and J.M. Trappe, 1971- The role of Alnus in improving the forest environment. PI. and S o i l , Special Vol. 1971: 3 3 5 _ 3 4 8 . Trappe, J.M., 1972. Regulation of s o i l organisms by red alder: a potential biological system for control of Poria weirii. In A.B. Berg ed. Managing young forests in the Douglas-fir region. Ore. State Uni., Sch. For. Symp., 1970: 35-51 . Trappe, J.M., C.Y. L i , K.C. Lu and W.B. Bollen, 1973- D i f f e r e n t i a l response of Poria weirii to phenolic acids from Douglas-fir and red alder. For. Sc. 19: 191-196. Tschirley, F.H., 1956. Undesirable woody plants. Res. Prog. Rep., West Weed Contr. Conf. 1956 : 2 6 - 4 2 . Vaadi, Y. and C. I t a i , 1969- Interrelationships of growth with reference to the di s t r i b u t i o n of growth substances. In W.J. Whittington ed. Root growth. Proc. Easter Sch. Agr. Sc., 1968 : 65"79-- 150 -Wallis, G., 1968. Resistance of Alnus rubra to infection by root rot fungus Poria weirii. In J.M. Trappe et al. ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967: 195-, 1976. Phellinus (Poria) weirii root rot. Detection and management proposals in Douglas-fir stands. Env. Can. For. Tech. Rep. 12: 16 pp. Wallis, G. and G. Reynolds, 1965- The i n i t i a t i o n and spread of Poria weirii root rot of Douglas-fir. Can. J. Bot. 43: 1 _9-, 1974. Urea and nitrate f e r t i l i z e r s f a i l to inh i b i t root rot. Env. Can., For. Serv. Bi-mon. Res. Note 30: 25~26. Wareing, P.F. and I.D.J. P h i l l i p s , 1978. The control of growth and dif f e r e n t i a t i o n in plants, 2nd. ed. Pergamon Press: 347 PP-Warrack, G., 19^9- Treatment of red alder in the coastal region of B r i t i s h Columbia. Res. Note, Br. Col. For. Serv. #14: 7 pp. Wicklow, M.C., W.B. Bollen and W.C. Dennison, 197**- Comparison of s o i l microfungi in 40 year old stands of pure alder, pure conifer and alder-conifer mixtures. Soil B i o l , and Biochem. 6: 73~78. Wilson, B.F., 1975- Distribution of secondary thickening in tree root systems. In J.G. Torrey and D.T. Clarkson ed. The development and function of roots. Academic Press: 197-219. Wollum 11, A.G. and C.T. Youngberg, 1-964. The influence of nitrogen fi x a t i o n by non-1eguminous woody plants on the growth of pine seedlings. J. For. 62: 316-321. Worthington, N.P., 1965- Red alder. In H.A. Fowells ed. Si Ivies of forest trees of the United States. U.S.D.A. Agr. Handbook #271: 8 3 - 8 8 . Worthington, N.P., R.H. Ruth and E.E. Matson, 1962. Red alder: i t s management and u t i l i z a t i o n . Misc. Publ., U.S.D.A., #881: 44 pp. Wright, J.W., 1976. Introduction to forest genetics. Academic Press: 463 pp. Yoho, J.G., D.E. Chappelle and D.L. Schweitzer, 1969a. The economics of converting red alder to Douglas-fir. Res. Pap., Pac. N.W. For. and Range Exp. St., PNW-81: 31 pp. , 1969b. The marketing of red alder pulpwood and sawlogs. Res. Note, Pac. N.W. For. and Range Exp. St., PNW-96: 7 pp. Zavitkowski, J. and M. Newton, 1968. Effect of organic matter and combined nitrogen on nodulation and nitrogen f i x a t i o n in red alder. In J.M. Trappe et al. ed. Biology of alder. Proc. N.W. Sc. Ass. meeting, 1967: 209 - 2 2 3 . , 1971. L i t t e r f a l l and l i t t e r accumulation in red alder stands in western Oregon. PI. and Soil 35: 257-268. Zavitkowski, J. and R.D. Stevens, 1972. Primary productivity of red alder ecosystems. Ecol. 53: 235~242. 

Cite

Citation Scheme:

        

Citations by CSL (citeproc-js)

Usage Statistics

Share

Embed

Customize your widget with the following options, then copy and paste the code below into the HTML of your page to embed this item in your website.
                        
                            <div id="ubcOpenCollectionsWidgetDisplay">
                            <script id="ubcOpenCollectionsWidget"
                            src="{[{embed.src}]}"
                            data-item="{[{embed.item}]}"
                            data-collection="{[{embed.collection}]}"
                            data-metadata="{[{embed.showMetadata}]}"
                            data-width="{[{embed.width}]}"
                            data-media="{[{embed.selectedMedia}]}"
                            async >
                            </script>
                            </div>
                        
                    
IIIF logo Our image viewer uses the IIIF 2.0 standard. To load this item in other compatible viewers, use this url:
https://iiif.library.ubc.ca/presentation/dsp.831.1-0095116/manifest

Comment

Related Items