UBC Theses and Dissertations

UBC Theses Logo

UBC Theses and Dissertations

Some aspects of boron, copper and iron nutrition of lodgepole pine and Douglas-fir Majid, Nik Muhamad 1984

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

Item Metadata

Download

Media
831-UBC_1984_A1 M34_4.pdf [ 13.02MB ]
Metadata
JSON: 831-1.0096652.json
JSON-LD: 831-1.0096652-ld.json
RDF/XML (Pretty): 831-1.0096652-rdf.xml
RDF/JSON: 831-1.0096652-rdf.json
Turtle: 831-1.0096652-turtle.txt
N-Triples: 831-1.0096652-rdf-ntriples.txt
Original Record: 831-1.0096652-source.json
Full Text
831-1.0096652-fulltext.txt
Citation
831-1.0096652.ris

Full Text

SOME ASPECTS OF BORON, COPPER AND IRON NUTRITION OF LODGEPOLE PINE AND DOUGLAS-FIR  by NIK MUHAMAD MAJID (Dip. A g r i c , Malaya, 1971) (B.S., Louisiana State University, 1973) (M.F., Louisiana State University, 1975)  A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE FACULTY OF GRADUATE STUDIES (Department of S o i l Science)  We accept this thesis as conforming to the required standard  THE UNIVERSITY OF BRITISH COLUMBIA June 1984 © Nik Muhamad Majid, 1984  In  presenting  this  requirements  for  an  of  British  it  freely available  agree for  that  I  by  understood  that  his  or  of  reference  and  study.  I  extensive be  her or  July  3,  1984  copying  granted  by  the  of  publication  not  be  allowed  Columbia  of  make  further this  head  representatives.  Science  The U n i v e r s i t y o f B r i t i s h 1956 Main M a l l Vancouver, Canada V6T 1Y3  Date  University shall  shall  Soil  the  Library  permission.  Department  at  the  the  may  copying  f i n a n c i a l gain  degree  f u l f i l m e n t of  that  for  purposes  or  partial  agree  for  permission  scholarly  in  advanced  Columbia,  department  for  thesis  It  this  without  thesis  of  my  is  thesis my  written  ii  ABSTRACT  T h i s t h e s i s r e p o r t s the f i n d i n g s of two complementary  investiga-  t i o n s on boron, copper and i r o n n u t r i t i o n of l o d g e p o l e p i n e and Douglasfir.  The primary aim of the f i r s t study was  l e v e l s of boron, copper, t o t a l i r o n and  t o determine the  " a c t i v e " i r o n f o r lodgepole pine  grown under c o n t r o l l e d c o n d i t i o n s i n the greenhouse. conducted a t f i v e d i f f e r e n t f i e l d  critical  The second study,  locations i n interior British  Columbia,  i n v o l v e d f o l i a r a p p l i c a t i o n of copper s u l p h a t e , f e r r o u s s u l p h a t e and urea to  l o d g e p o l e p i n e ; and f e r r o u s s u l p h a t e and urea to D o u g l a s - f i r .  main o b j e c t i v e o f the f i e l d  study was  t o a s s e s s t r e e growth and  n u t r i e n t responses to f o l i a r - a p p l i e d n u t r i e n t s . the  The foliar  Both s t u d i e s i n v o l v e d  a p p l i c a t i o n of n i t r o g e n t o i n v e s t i g a t e i t s s i g n i f i c a n c e f o r the  micronutrients. The f i n d i n g s from the greenhouse experiments i n d i c a t e d l o d g e p o l e p i n e , the c r i t i c a l 16 ppm is are  range, expressed as c o n c e n t r a t i o n i s 7 to  f o r boron and 2 t o 3 ppm  the c r i t i c a l v a l u e . 32 and 44 ppm,  n u t r i e n t element.  f o r copper.  The lower v a l u e i n each case  For " a c t i v e " and t o t a l i r o n , the c r i t i c a l v a l u e s  respectively.  v a l u e , i n each case, may  C o n c e n t r a t i o n s a t or below the  critical  be a s s o c i a t e d w i t h acute d e f i c i e n c y of the  C o n c e n t r a t i o n s above the c r i t i c a l v a l u e imply that the  n u t r i e n t i s a t a l e v e l s u f f i c i e n t f o r optimum or near-optimum R e s u l t s from the f i e l d fairly  that f o r  experiments w i t h l o d g e p o l e p i n e i n d i c a t e d  comparable v a l u e s to those from the greenhouse.  v a l u e f o r copper was  growth.  found to be 4 ppm  The  f o r copper and 29 ppm  critical for active  iii  iron.  I t was  a l s o suggested  t h a t copper t o x i c i t y i n l o d g e p o l e pine might  occur whenever f o l i a r copper c o n c e n t r a t i o n exceeds 17 F o l i a r n u t r i e n t a p p l i c a t i o n proved  ppm.  to be a q u i c k and  effective  means of r a i s i n g copper and i r o n c o n c e n t r a t i o n s to adequate l e v e l s i n l o d g e p o l e p i n e f o l i a g e ; and i r o n i n D o u g l a s - f i r f o l i a g e where these n u t r i e n t s were d e f i c i e n t .  F o l i a r a p p l i c a t i o n of urea was  effective i n  r a i s i n g f o l i a r n i t r o g e n c o n c e n t r a t i o n i n l o d g e p o l e p i n e , but not i n Douglas-fir.  Combined n u t r i e n t a p p l i c a t i o n s were more e f f e c t i v e  individual nutrient application.  However, these e f f e c t s on  n u t r i e n t s were o n l y temporary and  d i d not l a s t beyond the year of  c a t i o n , except  than  foliar appli-  i n D o u g l a s - f i r which seemed a b l e to r e t a i n more a p p l i e d  i r o n i n the f o l i a g e than d i d l o d g e p o l e p i n e , d u r i n g the second year  of  growth• Shoot growth and blomass p r o d u c t i o n i n both s p e c i e s responded g r e a t l y to treatments fertilization.  o n l y d u r i n g the second growing season f o l l o w i n g  Lodgepole p i n e needle  l e n g t h a t one  showed a s i g n i f i c a n t p o s i t i v e response of  growth.  The h i g h e s t response  minimal f o l i a r was  scorching.  was  to treatments  i n the second  o b t a i n e d from treatments  No s i g n i f i c a n t p o s i t i v e t r e e growth  d e t e c t e d d u r i n g the year of f e r t i l i z e r The  of the s i t e s a l s o  f e r r o u s s u l p h a t e to l o d g e p o l e pine was  percent, r e s p e c t i v e l y .  No f o l i a r  i n j u r y was  f e r r o u s s u l p h a t e a p p l i e d to D o u g l a s - f i r .  observed  burn i n e i t h e r s p e c i e s .  1 percent  extremely  f e r r o u s s u l p h a t e caused moderate f o l i a r  response  0.1  e v i d e n t ) of and  2  with 4 percent  N i t r o g e n a p p l i e d a t 2 percent  urea d i d not cause any f o l i a r copper s u l p h a t e was  caused  application.  s a f e a p p l i c a t i o n dosage (where no f o l i a r i n j u r y was  copper s u l p h a t e and  that  year  The a p p l i c a t i o n of  t o x i c to l o d g e p o l e p i n e ; 4 percent burn.  iv  N i t r o g e n absorbed appeared  by the r o o t system  (greenhouse  experiment)  to have an a n t a g o n i s t i c e f f e c t on f o l i a r boron, copper,  and a c t i v e  i r o n i n lodgepole pine.  these m i c r o n u t r i e n t s i n the f o l i a g e  The c o n c e n t r a t i o n and content of decreased as f o l i a r n i t r o g e n  i n c r e a s e d as a r e s u l t of i n c r e a s i n g the n i t r o g e n s u p p l y . of urea  total  F o l i a r feeding  ( f i e l d e x p e r i m e n t ) , on the o t h e r hand, d i d not seem to have any  physiological synergistic  interaction  with f o l i a r  e f f e c t of u r e a on f o l i a r  copper. iron.  In f a c t , t h e r e was a  The l e v e l of t o t a l and  forms of i r o n i n the f o l i a g e was i n c r e a s e d as a r e s u l t of u r e a application.  active  v  TABLE  OF  CONTENTS Page  ABSTRACT  i i  TABLE OF CONTENTS  v  LIST OF TABLES  ix  LIST OF FIGURES  xi  LIST OF APPENDICES  xiv  ACKNOWLEDGEMENTS  xvi  CHAPTER 1.  GENERAL INTRODUCTION  1  CHAPTER 2.  LITERATURE REVIEW  3  A.  Nutrient Requirement Evaluations  3  1.  Fertilizer Field Trials  3  2.  F e r t i l i z e r Pot T r i a l s  4  3.  Visual Symptoms  4  4.  S o i l Analysis  5  5.  F o l i a r Analysis  6  CHAPTER 3.  B.  Expression of F o l i a r Nutrient Composition  C.  Nutrient Requirements of Conifers  9 12  GREENHOUSE EXPERIMENTS  15  A.  Introduction  15  B.  Methods and Materials  15  1.  Experimental Design  15  2.  Pot Arrangement  16  3.  Greenhouse Environment  17  4.  Preparation and Maintenance of Growth Medium  17  vi  Page  5.  Germination and Thinning  19  6.  Composition of Nutrient Solutions  20  7.  Nutrient A p p l i c a t i o n  21  8.  Harvesting, Sample Measurement and  9. 10.  Preparation  24  Chemical Analysis  24  S t a t i s t i c a l Analysis  26  C. Results and Discussion 1.  Boron Experiment (a) Visual Symptoms (b) C r i t i c a l Level  26 26 33  2.  Copper Experiment  33  (a)  V i s u a l Symptoms  33  (b)  C r i t i c a l Level  41  3.  CHAPTER 4.  26  Iron Experiment  41  (a)  Visual Symptoms  41  (b)  C r i t i c a l Level  43  D. Summary of Greenhouse Experiments  51  FIELD EXPERIMENT  52  A. Introduction  52  B. Methods and Materials  54  1.  S i t e Descriptions  54  (a)  Site Locations  54  (b)  Stand Characteristics  56  (c)  S o i l Characteristics  57  2.  Experimental Design  57  3.  F e r t i l i z e r Application  60  vii  Page  4.  F i e l d Sampling and Measurements  64  5.  Sample P r e p a r a t i o n and Measurements  65  6.  Chemical A n a l y s i s of F o l i a r Samples  65  7.  S o i l Sample P r e p a r a t i o n  8.  Assessment of F e r t i l i z e r Response (a)  and A n a l y s i s  •  66 66  E v a l u a t i o n o f Shoot Length and F o l i a r Mass Response  9. C.  67  (b)  E v a l u a t i o n of F o l i a r N u t r i e n t Response  (c)  F o l i a r Nutrient Status  S t a t i s t i c a l Analysis  .  68  I n t e r p r e t a t i o n ..  69 69  R e s u l t s and D i s c u s s i o n  70  1.  71  2.  Foliar (a)  Copper Treatments  71  (b)  I r o n Treatments  72  (c)  Nitrogen  73  (d)  Causes of Needle Burn  Treatments  73  Tree Growth Responses  75  (a)  Shoot Growth  75  (i)  Lodgepole Pine  75  (ii)  Douglas-fir  81  (b)  3.  Scorching  F o l i a r Mass  83  (i)  Lodgepole Pine  83  (ii)  Douglas-fir  85  F o l i a r N u t r i e n t Responses  87  (a)  Copper  87  (b)  A c t i v e Iron  101  (i)  Lodgepole Pine  101  (ii)  Douglas-fir  107  viii  Page (c)  D. CHAPTER 5.  109  (i)  Lodgepole Pine  109  (ii)  Douglas-fir  115  Summary of F i e l d Experiment  CONCLUSIONS  REFERENCES CITED  Nitrogen  116 118 122  XX  LIST  OF  TABLES  Table  1  2  3  4  5  6  7  8  9  10  11  12  13  14  Page  F o l i a r n u t r i e n t values u s e f u l i n i n d i c a t i n g nutrient status of conifers  13  Chemical a n a l y s i s of the l e a c h a t e from the sand used as the growth medium  18  T h i n n i n g f r e q u e n c y , dates and number o f s e e d l i n g s remaining a f t e r each t h i n n i n g  20  C o n c e n t r a t i o n of manipulated solutions  21  elements i n n u t r i e n t  C o n c e n t r a t i o n and source of elements i n standard n u t r i e n t s o l u t i o n f o r l e v e l 1 n i t r o g e n supply (10 ppm)  22  C o n c e n t r a t i o n and source o f elements i n s t a n d a r d nutrient s o l u t i o n f o r l e v e l 2 n i t r o g e n supply (100 ppm)  23  F o l i a r boron c o m p o s i t i o n , mass o f 100 needles and seedling height  31  A n a l y s i s of v a r i a n c e o f f o l i a r n u t r i e n t composit i o n w i t h v a r y i n g supply l e v e l s of boron and nitrogen  32  F o l i a r copper c o m p o s i t i o n , mass o f 100 needles and seedling height  39  A n a l y s i s of v a r i a n c e of f o l i a r n u t r i e n t composit i o n w i t h v a r y i n g supply l e v e l s of copper and nitrogen  40  T o t a l and a c t i v e i r o n c o m p o s i t i o n , mass o f 100 needles and s e e d l i n g h e i g h t  47  A n a l y s i s o f v a r i a n c e o f f o l i a r n u t r i e n t composit i o n w i t h v a r y i n g supply l e v e l s o f i r o n and nitrogen  48  Some c h e m i c a l c h a r a c t e r i s t i c s o f s o i l p r o f i l e s o f the study s i t e s  58  Some p h y s i c a l s o i l c h a r a c t e r i s t i c s o f t h e study sites  59  X  Table  Page  15  Fertilizer  treatments f o r the main (1981) t r i a l  16  Fertilizer  treatments f o r the repeat  17  Chemical a n a l y s i s of water used f o r f e r t i l i z e r solution  18  19  20  21  22  23  24  25  26  ....  (1982) t r i a l  ..  61 62  preparing 63  A n a l y s i s of v a r i a n c e of t r e e growth response v a r i a b l e s f o r l o d g e p o l e p i n e and D o u g l a s - f i r  76  A n a l s y s i s of v a r i a n c e f o r f o l i a r n u t r i e n t response of l o d g e p o l e pine (main t r i a l )  88  Percentage v a r i a n c e components i n r e l a t i o n to f o l i a r n u t r i e n t responses f o r l o d g e p o l e p i n e (main t r i a l )  89  A n a l y s i s of v a r i a n c e f o r f o l i a r n u t r i e n t responses of l o d g e p o l e pine ( r e p e a t t r i a l )  90  Percentage v a r i a n c e components i n r e l a t i o n f o l i a r n u t r i e n t responses f o r l o d g e p o l e pine (repeat trial)  90  A n a l y s i s of v a r i a n c e f o r f o l i a r n u t r i e n t responses of D o u g l a s - f i r (main t r i a l )  91  Percentage v a r i a n c e components i n r e l a t i o n f o l i a r n u t r i e n t responses f o r D o u g l a s - f i r (main t r i a l ) ....  91  A n a l y s i s of v a r i a n c e f o r f o l i a r n u t r i e n t responses of D o u g l a s - f i r (repeat t r i a l )  92  Treatment and concentration  year e f f e c t s on f o l i a r (%) of D o u g l a s - f i r  nitrogen 115  xi  LIST  OF  FIGURES  Figure  1  Page  G e n e r a l i z e d r e l a t i o n s h i p between growth and nutrient concentration  tissue 7  2  Lodgepole  3  D i f f e r e n t i a l growth response of l o d g e p o l e pine s e e d l i n g s to i n c r e a s i n g boron supply a t low n i t r o g e n level  29  D i f f e r e n t i a l growth response of l o d g e p o l e pine s e e d l i n g s to i n c r e a s i n g boron supply a t h i g h n i t r o g e n level  30  R e l a t i o n s h i p between f o l i a r boron c o m p o s i t i o n s e e d l i n g growth ( l o d g e p o l e p i n e )  34  4  5  p i n e s e e d l i n g s without boron supply  Lodgepole  7  D i f f e r e n t i a l growth response of l o d g e p o l e p i n e s e e d l i n g s t o i n c r e a s i n g copper supply a t low n i t r o g e n level  37  D i f f e r e n t i a l growth response of l o d g e p o l e pine s e e d l i n g s to i n c r e a s i n g copper supply a t h i g h n i t r o g e n level  38  R e l a t i o n s h i p between f o l i a r copper s e e d l i n g growth ( l o d g e p o l e p i n e )  42  9  supply  composition  36  and  10  Lodgepole  11  D i f f e r e n t i a l growth response of l o d g e p o l e pine s e e d l i n g s to i n c r e a s i n g i r o n supply a t low n i t r o g e n level  45  D i f f e r e n t i a l growth response of l o d g e p o l e pine s e e d l i n g s t o i n c r e a s i n g i r o n supply a t h i g h n i t r o g e n level  46  R e l a t i o n s h i p between f o l i a r t o t a l i r o n c o m p o s i t i o n s e e d l i n g growth ( l o d g e p o l e p i n e )  49  12  13  14  15  p i n e s e e d l i n g s without  copper  and  6  8  p i n e s e e d l i n g s without  28  i r o n supply  R e l a t i o n s h i p between f o l i a r a c t i v e i r o n c o m p o s i t i o n s e e d l i n g growth ( l o d g e p o l e p i n e ) Map  showing l o c a t i o n of the study s i t e s  44  and  and 50 55  xii  Figure  16  17  18  19  20  21  22  23  24  25  26  27  28  29  30  31  Page  Second-year shoot growth r a t i o i n r e l a t i o n t o t r e a t ments a t S i t e 1 ( l o d g e p o l e p i n e )  77  Second-year shoot growth r a t i o i n r e l a t i o n t o t r e a t ments a t S i t e 2 ( l o d g e p o l e p i n e )  78  Second-year shoot growth r a t i o i n r e l a t i o n to t r e a t ments a t S i t e 3 ( l o d g e p o l e p i n e )  79  Second-year shoot growth r a t i o i n r e l a t i o n t o t r e a t ments a t S i t e 4 ( l o d g e p o l e p i n e )  80  Second-year shoot growth r a t i o i n r e l a t i o n t o t r e a t ments a t S i t e 5 ( D o u g l a s - f i r )  82  F o l i a r mass o f l o d g e p o l e p i n e i n the second season i n r e l a t i o n t o treatments  84  F o l i a r mass of D o u g l a s - f i r i n the second season i n r e l a t i o n t o treatments  growing  growing 86  F o l i a r copper c o n c e n t r a t i o n i n r e l a t i o n t o treatments at S i t e 2 ( l o d g e p o l e p i n e )  93  F o l i a r copper c o n c e n t r a t i o n i n r e l a t i o n t o treatments at S i t e 3 (lodgepole pine)  94  F o l i a r copper c o n c e n t r a t i o n i n r e l a t i o n to treatments at S i t e 4 ( l o d g e p o l e p i n e )  95  F o l i a r copper c o n c e n t r a t i o n i n r e l a t i o n t o treatments at S i t e 1 ( l o d g e p o l e p i n e )  97  F o l i a r copper c o n c e n t r a t i o n i n r e l a t i o n t o treatments at S i t e 5 ( D o u g l a s - f i r )  98  F o l i a r a c t i v e i r o n concentration i n r e l a t i o n to t r e a t ments a t S i t e 1 ( l o d g e p o l e p i n e )  102  F o l i a r active iron concentration i n r e l a t i o n to treatments a t S i t e 2 ( l o d g e p o l e p i n e )  103  F o l i a r a c t i v e i r o n concentration i n r e l a t i o n to t r e a t ments a t S i t e 3 ( l o d g e p o l e p i n e )  104  F o l i a r a c t i v e i r o n concentration i n r e l a t i o n to t r e a t ments a t S i t e 4 ( l o d g e p o l e p i n e )  105  xiii  Figure  32  33  34  35  36  Page  F o l i a r active i r o n concentration i n r e l a t i o n to t r e a t ments a t S i t e 5 ( D o u g l a s - f i r )  108  F o l i a r nitrogen concentration i n r e l a t i o n to t r e a t ments a t S i t e 1 ( l o d g e p o l e p i n e )  110  F o l i a r n i t r o g e n c o n c e n t r a t i o n i n r e l a t i o n t o treatments at S i t e 2 ( l o d g e p o l e p i n e )  I l l  F o l i a r n i t r o g e n c o n c e n t r a t i o n i n r e l a t i o n to treatments at S i t e 3 (lodgepole pine)  112  F o l i a r n i t r o g e n c o n c e n t r a t i o n i n r e l a t i o n to treatments at S i t e 4 ( l o d g e p o l e p i n e )  113  xiv  LIST OF APPENDICES Appendix  A.l  A.2  A. 3  B. l  B.2  B. 3  Page  Modified Parkinson tissue analysis  and A l l e n D i g e s t i o n f o r p l a n t 132  N i t r i c a c i d d i g e s t i o n f o r a n a l y s i s of copper i r o n i n plant tissue Procedure f o r a c t i v e i r o n d e t e r m i n a t i o n tissue  and 133  i n plant 134  F o l i a r elemental experiment  c o n c e n t r a t i o n s i n the boron  F o l i a r elemental experiment  c o n c e n t r a t i o n s i n the copper  135  136  F o l i a r e l e m e n t a l c o n c e n t r a t i o n s i n the  iron  experiment  137  C. l  S o i l p r o f i l e d e s c r i p t i o n of s i t e  1  138  C.2  S o i l p r o f i l e d e s c r i p t i o n of s i t e 2  140  C.3  S o i l p r o f i l e d e s c r i p t i o n of s i t e 3  142  C.4  S o i l p r o f i l e d e s c r i p t i o n of s i t e 4  144  C. 5  S o i l p r o f i l e d e s c r i p t i o n of s i t e 5  D. l  Copper treatment: pine)  D.2  D.3  146  c u r r e n t - y e a r growth  (lodgepole 148  Copper and n i t r o g e n treatment: (lodgepole pine)  c u r r e n t - y e a r growth 149  Copper, i r o n and n i t r o g e n treatment:  current-year  growth ( l o d g e p o l e p i n e )  150  D.4  Copper treatment:  151  D.5  Copper and n i t r o g e n treatment: (lodgepole pine)  D.6  D.7  second-year growth ( l o d g e p o l e p i n e ) second-year growth  Copper, i r o n and n i t r o g e n treatment growth ( l o d g e p o l e p i n e ) C o n t r o l tree (lodgepole pine)  152 second-year 153 154  XV  Appendix  Page  D.8  I r o n treatment:  c u r r e n t - y e a r growth ( l o d g e p o l e p i n e ) .  D.9  I r o n and n i t r o g e n treatment:  current-year  155  growth  (lodgepole pine)  155  D.10  I r o n treatment:  second-year growth ( l o d g e p o l e p i n e )  D.ll  I r o n and n i t r o g e n treatment:  .  156  second-year growth  (lodgepole pine)  157  D.12  Iron treatment:  D.13  I r o n and n i t r o g e n treatment: c u r r e n t - y e a r growth (Douglas-fir) N i t r o g e n treatment: c u r r e n t - y e a r growth ( l o d g e p o l e  158  pine)  159  D.14  c u r r e n t - y e a r growth ( D o u g l a s - f i r ) ...  D. 15  Nitrogen  E. l  Mean v a l u e s f o r f o l i a r copper c o n c e n t r a t i o n (ppm) o f l o d g e p o l e p i n e and D o u g l a s - f i r i n r e l a t i o n t o treatments (main t r i a l ) Mean v a l u e s f o r f o l i a r a c t i v e i r o n c o n c e n t r a t i o n (ppm) o f l o d g e p o l e p i n e and D o u g l a s - f i r i n r e l a t i o n to treatments (main t r i a l )  E.2  E.3  E.4  treatment:  c u r r e n t - y e a r growth ( D o u g l a s - f i r )  158  160  161  164  Mean v a l u e s f o r f o l i a r t o t a l i r o n c o n c e n t r a t i o n (ppm) o f l o d g e p o l e pine and D o u g l a s - f i r i n r e l a t i o n to treatments (main t r i a l )  167  Mean v a l u e s f o r f o l i a r n i t r o g e n c o n c e n t r a t i o n (%) of l o d g e p o l e p i n e and D o u g l a s - f i r i n r e l a t i o n t o treatments (main t r i a l )  170  xvi  ACKNOWLEDGEMENTS  I am deeply indebted to my supervisor, Dr. T.M. Ballard for h i s guidance and wise counsel which have contributed greatly to this research and also my study at U.B.C. my Supervisory Committee:  I am also grateful to the other members of  Dr. A.A. Bomke, Dr. L.M. Lavkulich, Dr. L.E.  Lowe and Dr. J . Otchere-Boateng for their help and constructive c r i t i c i s m of this t h e s i s . During the course of conducting this research, I was helped i n a variety of ways by a number of other people and friends.  In p a r t i c u l a r ,  I wish to acknowledge the help given by Ms. P. Carbis, Mr. J . Emanuel, Mr. D.G. Giles, Mr. F.M. K e l l i h e r , Ms. J . Lansiquot, Mr. B. Von Spindler, Ms. E. Wolterson and Mr. B. Wong. I would l i k e to acknowledge Dr. A. Kozak and Dr. H.E. Schreier for their advice i n s t a t i s t i c a l analysis; Dr. R.E. M i l l e r of the U.S. Forest Service f o r his advice on f o l i a r spray application; the B.C. M i n i s t r y of Forests f o r help i n locating the research s i t e s and providing seed; and the Natural Sciences and Engineering Research Council of Canada for f i n a n c i a l support of the project. I would l i k e to thank U n i v e r s i t i Pertanian Malaysia f o r granting me educational leave, and the Government of Malaysia f o r f i n a n c i a l a i d . L a s t l y , many thanks to my wife Fatimah f o r her love, support and encouragement.  To my son, Azlan, who missed many happy moments with his  father, I dedicate this thesis.  1  CHAPTER 1.  GENERAL INTRODUCTION  Plants are composed of a large number of chemical elements, several of which are categorized as "micronutrients" because they are required only i n small amounts. molybdenum and zinc.  These include boron, copper, iron, manganese,  The e s s e n t i a l i t y of these micronutrients f o r  healthy growth has been so well established f o r so many plant species that i t can be considered applicable for a l l . In forestry, micronutrient n u t r i t i o n a l research i s s t i l l i n i t s infancy.  Our present knowledge on this subject i s limited and fragmen-  tary compared with that available either for cultivated plants or f o r macronutrients  i n forest trees.  The problem one encounters i n reviewing  the l i t e r a t u r e on micronutrients i n forest trees i s , therefore, mainly that of synthesizing the fragmentary reports from various f i e l d s of study including a g r i c u l t u r e , h o r t i c u l t u r e , botany, ecology, biogeochemistry and forestry. One area of basic research that has gained some attention i n forestry i s the determination of mineral requirements of various tree species.  Most of these investigations, however, pertained to macro-  nutrient requirements,  with r e l a t i v e l y very l i t t l e e f f o r t directed  towards micronutrients.  This i s despite the fact that micronutrient  deficiencies have been i d e n t i f i e d i n forest situations i n many parts of the world  (Stone 1968).  experimental  Perhaps this i s partly because of the s p e c i a l  d i f f i c u l t i e s involved i n the study of micronutrients  (Fortesque and Marten 1969).  2  Another area o f f o r e s t r y r e s e a r c h t h a t has a t t r a c t e d much a t t e n t i o n i n recent years i s f o r e s t f e r t i l i z a t i o n . fertilizer  There a r e numerous r e p o r t s on  t r i a l s and o p e r a t i o n a l f e r t i l i z a t i o n  A u s t r a l i a , New Zealand  and the t r o p i c s .  i n North America, Europe,  The m a j o r i t y o f t h e s e , however,  i n v o l v e d s o i l a p p l i c a t i o n o f f e r t i l i z e r s and t o some extent a e r i a l c a t i o n of g r a n u l a r m a c r o n u t r i e n t D e B e l l 1981; M o r r i s o n mation on f i e l d f o l i a r  fertilizers  1981; P u l s f o r d 1981).  appli-  ( M i l l e r and F i g h t 1979; There i s v e r y l i m i t e d  a p p l i c a t i o n of l i q u i d f e r t i l i z e r s  infor-  on f o r e s t t r e e s .  T h i s method o f f e r t i l i z e r a p p l i c a t i o n has been used i n a g r i c u l t u r e f o r a l o n g time and has been an accepted m i c r o n u t r i e n t s f o r many crops Some f o l i a r  p r a c t i c e i n supplying various  (Murphy and Walsh 1972; Traynor  1980).  a n a l y s i s work i n i n t e r i o r B r i t i s h Columbia l e d to  i n f e r e n c e o f p o s s i b l e m i c r o n u t r i e n t d e f i c i e n c i e s i n some f o r e s t ( B a l l a r d 1981).  stands  I n o r d e r t o improve d i a g n o s i s of such problems and  e x p l o r e p o s s i b l e r e m e d i a l measures, t h i s t h e s i s r e s e a r c h was c a r r i e d out i n the form of two complementary s t u d i e s .  The f i r s t  study, r e p o r t e d i n  Chapter 3, i n v o l v e s t h r e e greenhouse experiments on m i c r o n u t r i e n t requirements  o f l o d g e p o l e p i n e (Pinus c o n t o r t a D o u g l . ) .  r e p o r t s f i n d i n g s from f i e l d e x p e r i m e n t a t i o n fertilizer menziesii  Chapter 4  on f o l i a r a p p l i c a t i o n o f  s o l u t i o n s t o l o d g e p o l e p i n e and D o u g l a s - f i r (Pseudotsuga [Mirb.] F r a n c o ) .  S p e c i f i c o b j e c t i v e s o f these two i n v e s t i g a -  t i o n s a r e g i v e n i n the r e s p e c t i v e c h a p t e r s . l i t e r a t u r e i s g i v e n i n Chapter 2.  A g e n e r a l review of  3 CHAPTER 2.  A•  Nutrient Requirement  LITERATURE REVIEW  Evaluations  Evaluation of nutrient requirements of forest trees and  identifica-  tion of mineral d e f i c i e n c i e s can be conducted by a number of d i f f e r e n t methods.  Each has i t s own  applications, merits, and drawbacks.  (1974) reviewed f i v e commonly employed methods for conifers.  Morrison  These  include f e r t i l i z e r f i e l d t r i a l s , f e r t i l i z e r pot t r i a l s , v i s u a l symptoms, s o i l analysis  and f o l i a r analysis.  This section gives a general review  of these methods, four of which ( f e r t i l i z e r f i e l d t r i a l , f e r t i l i z e r t r i a l , v i s u a l symptoms and f o l i a r analysis) were used i n the investigations reported 1.  pot  two  i n this thesis.  Fertilizer Field Trials  Tamm (1964) affirmed  that "no single method other than direct f i e l d  experiments can give complete and r e l i a b l e information status of a forest stand."  on the nutrient  He also pointed out i t might not always be  economically f e a s i b l e to conduct large scale f i e l d experiments.  The  use  of single-tree plot technique suggested by Viro (1967) has been used to overcome this problem.  It permits a large number of treatments to be  tested, reduces costs, provides faster and reasonably r e l i a b l e indications of treatment responses (Gessel et a l . 1960;  Viro 1967,  1970).  If  the danger of f e r t i l i z e r f i x a t i o n i n the s o i l exists, as i n the case for micronutrients,  the use of f o l i a r spraying i s the preferred method  (Murphy and Walsh 1972).  4  2.  F e r t i l i z e r Pot T r i a l s  Pot t r i a l s or greenhouse techniques have been widely used i n forest n u t r i t i o n research because they allow the use of more complicated under less variable s o i l and c l i m a t i c conditions.  designs  In addition, results  could be obtained i n a much shorter time and are generally less d i f f i c u l t to  evaluate. Naturally there are several l i m i t a t i o n s to greenhouse studies.  main disadvantage i s the problem of extrapolating results to f i e l d tions.  The  condi-  For instance, the nutrient requirements of seedlings are  d i f f e r e n t from those of adult trees (Tamm 1964;  Mead and P r i t c h e t t 1971).  Despite the a r t i f i c i a l environment, much valuable information could be obtained 1979).  for diagnostic purposes (Walker et a l . 1955;  W i l l 1961;  Ingestad  For micronutrients, Bar rows (1959) indicated that because of the  complexity of nutrient interactions i n plants, the establishment nutrient requirements and c r i t i c a l ranges of the micronutrients  of could be  done only when a l l other e s s e n t i a l elements are present i n the plant i n the amounts and r a t i o s appropriate for maximum growth.  This condition  can be best attained under controlled conditions with sand or solution culture, from which the complexities 3.  of s o i l are excluded.  V i s u a l Symptoms The use of v i s u a l symptoms i s perhaps the most d i r e c t method  employed i n plant n u t r i t i o n research, and often gives the f i r s t i n d i c a tions of any n u t r i t i o n a l disorder.  The advantages of this method reside  i n i t s s i m p l i c i t y and i n the fact that laboratory f a c i l i t i e s are not required.  Although v i s u a l symptoms can be u s e f u l , they have some  5  limitations. symptoms.  For instance, not a l l d e f i c i e n c i e s show d i s t i n c t i v e  Those of N, Ca, S, Fe or Mn tend to produce nonspecific  general chloroses  (Morrison 1974).  i n a plant, the symptoms of one may symptoms may  If there are two or more d e f i c i e n c i e s mask those of the others, or the  not appear to be t y p i c a l of any one element.  Also, by the  time v i s u a l symptoms have appeared, the supply of the element i n the plant has reached the deficiency l e v e l and growth has already been reduced (Barrows 1959).  Another l i m i t a t i o n i n using v i s u a l symptoms i s  what Z o t t l (1973) termed "hidden hunger."  It i s a s i t u a t i o n where tree  growth can be severely reduced without any clear v i s u a l symptom.  It i s  therefore desirable to have a diagnostic t o o l to detect an adequate l e v e l of any e s s e n t i a l element before i t drops to the point where v i s u a l symptoms begin to appear. 4.  S o i l Analysis  The usefulness of s o i l chemical data for tree nutrient status i n t e r p r e t a t i o n and for prediction of nutrient supply to forest trees i s limited (Armson 1973;  van den Burg 1976;  this f a i l u r e to technical and conceptual  Khanna 1981).  Khanna attributed  difficulties.  The technical  d i f f i c u l t i e s involve obtaining a representative s o i l sample. i n the d i s t r i b u t i o n of tree roots and heterogeneity  of forest s o i l s make  i t extremely d i f f i c u l t to c o l l e c t samples that could represent nutrient-supplying status of the entire s o i l volume. l y c r i t i c a l with micronutrients  Variability  the  This i s p a r t i c u l a r -  because, some of them, such as boron, are  very soluble and move r e a d i l y through the s o i l whereas others, such as copper and zinc, are immobilized (Barrows 1959).  and tend to accumulate near the surface  6  The conceptual problem involves the d i f f i c u l t y i n determining the available or mobilisable forms of nutrients i n nature.  Total analysis  for s o i l nutrients does not o f f e r meaningful values for a v a i l a b i l i t y of any p a r t i c u l a r nutrient to the plant.  Many extractants have been used to  measure the 'available' portion of the s o i l nutrient, but the values obtained are seldom comparable (van den Burg 1976).  For micronutrients,  Cox and Kamprath (1972) stated that s o i l tests have often proved unreliable because of i n s u f f i c i e n t information about the chemistry of micronutrients  i n s o i l and absorption mechanisms by plant roots.  These  d i f f i c u l t i e s have led, i n recent years, to a prefe rence for the use of f o l i a r analysis over s o i l analysis. 5.  F o l i a r Analysis  F o l i a r analysis i s a technique that measures not the supply of nutrients available i n the s o i l but an index of the amount taken up by the tree (Morrison  1974).  It has been used to diagnose nutrient  deficiencies (Ballard 1981; Carter ejt al^., 1984), i n interpreting responses i n f e r t i l i z e r t r i a l s (Richards and Bevege 1969; Lea et a l . , 1980), and to e s t a b l i s h basic relationships between nutrient concentration and supply (van den Driessche 1969; Swan 1972; Ingestad 1979). B a s i c a l l y , the use of f o l i a r analysis f o r the f i r s t two objectives mentioned above (diagnosis of nutrient deficiencies and interpretation of f e r t i l i z e r response) i s based on the assumption that a quantitative relationship exists between plant growth and f o l i a r nutrient l e v e l s . This basic relationship i s referred to by Armson (1973) as a growth response curve.  Figure 1 i l l u s t r a t e s this r e l a t i o n s h i p .  When a nutrient  i s l i m i t i n g growth, i n i t i a l input of that nutrient w i l l increase growth  TISSUE NUTRIENT CONC. OF AN ELEMENT F i g u r e 1.  G e n e r a l i z e d r e l a t i o n s h i p between growth and t i s s u e nutrient concentration.  8 but a further increase i n the supply of that nutrient tends to cause the growth to l e v e l o f f .  Additional input beyond this point may  cause a  decline i n growth. A central concept used for i n t e r p r e t a t i o n of this growth response curve i s that there i s a " c r i t i c a l range" (BC i n Figure 1) or  "critical  percentage" (B i n Figure 1) of each nutrient for each kind of plant (Macy 1936;  Barrows 1959).  Below this range (AB i n Figure 1) trees w i l l  likely  show deficiency symptoms and w i l l respond substantially i n increased growth to applications of the element i n question.  Above this range,  (CD i n Figure 1) there i s luxury consumption, where no b e n e f i c i a l response to applications of the element would be expected. DE represents the t o x i c i t y range.  In Figure 1,  The " c r i t i c a l percentage" i s defined  as associated with 90 percent of maximum y i e l d (Richards and Bevege  1970;  Swan 1972). The use of f o l i a r analysis, hence the c r i t i c a l l e v e l s , to assess n u t r i t i o n a l d e f i c i e n c i e s i s not free from l i m i t a t i o n s .  In many instances,  the c r i t i c a l ranges or levels were determined under controlled greenhouse conditions (Morrison 1974).  It i s therefore questionable whether the  values obtained could be applied to actual f i e l d conditions where environmental and physiological factors may (Leaf 1968,  1973).  modify tree nutrient status  Armson (1973) also warned that the greatest u n r e l i a -  b i l i t y of comparative values may  be expected when these values are extra-  polated from one growing l o c a t i o n to another or to stands at d i f f e r e n t stages of development. The d i f f i c u l t y  i s further aggravated i n the case of micronutrients.  One main l i m i t a t i o n i n using c r i t i c a l values for micronutrients i s  9  associated with elemental  i n t e r a c t i o n s (Watanabe e t a l . , 1965).  d e f i c i e n c y and optimum ranges have been found response to one (DeKock 1955, Lambert  The  to o v e r l a p because the  n u t r i e n t o f t e n depends upon the l e v e l of o t h e r n u t r i e n t s  1981).  A second l i m i t a t i o n , as i n d i c a t e d by H i l l  and  (1981) i s t h a t sometimes a C-shaped response curve e x i s t s betwen  growth and m i c r o n u t r i e n t s c o n c e n t r a t i o n s i n the f o l i a g e , compared to the c u r v i l i n e a r curve i n F i g u r e 1.  In t h i s case, the n u t r i e n t present i n  v e r y l i m i t i n g amount i n the p l a n t may  first  show a decrease  and  i n c r e a s e i n percentage c o n c e n t r a t i o n w i t h i n c r e a s e i n growth. commonly a s s o c i a t e d w i t h c e r t a i n m i c r o n u t r i e n t s ( S t e e n b j e r g However, U l r i c h and v a l u e s of f o l i a r  Hills  (1973) s t a t e d t h a t one  a n a l y s i s i s being a b l e to prevent  than c o r r e c t them a f t e r they o c c u r .  These authors  the c r i t i c a l c o n c e n t r a t i o n i s e s t i m a t e d c u l t u r e and  then  an  This i s  1954).  of the g r e a t e s t  d e f i c i e n c i e s rather a l s o mentioned t h a t  best through the use of  to a l e s s e r e x t e n t by s o i l c u l t u r e or f i e l d  solution  experiment  techniques.  B.  E x p r e s s i o n of F o l i a r N u t r i e n t N u t r i e n t composition  Composition  i n p l a n t t i s s u e i s commonly expressed  as  c o n c e n t r a t i o n on a dry mass b a s i s , f o r example, percentage of dry weight of  the t i s s u e  (for macronutrients),  present i n s m a l l amounts). a b s o l u t e content (1973) noted  or i n p a r t s per m i l l i o n  I t has a l s o been expressed  i n the t i s s u e  (e.g. gram per shoot  t h a t the terms c o n c e n t r a t i o n and  ( f o r elements  i n terms of  or per n e e d l e ) .  content have o f t e n been  used synonymously, even though the d i s t i n c t i o n between the two q u i t e obvious  (Farhoomand and  Peterson  1968).  Leaf  should  be  10  Concentration has been widely used because of the assumption that concentration alone can s u f f i c i e n t l y indicate whether a particular element i s at a d e f i c i e n t , adequate or toxic l e v e l i n the plant (Farhoomand and Peterson 1968).  Van den Driessche (1974) mentioned that  nutrient measurements should be independent of growth parameters such as weight, i f the objective i s to i n f e r whether a nutrient i s growthlimiting. Generally, i f a p a r t i c u l a r element that i s l i m i t i n g growth i s added i n a f e r t i l i z e r treatment, an increase i n the concentration of the element i n the tissue occurs. and unequivocal (Figure 1). relationship may (1972).  This relationship i s regarded as positive However i t i s possible that a negative  occur (Steenbjerg 1954), as demonstrated by Ebbel  This has been described as " d i l u t i o n e f f e c t " (Munson and  Nelson  1973) or "Steenbjerg e f f e c t " , where a decrease i n concentration occurs due to a r e l a t i v e l y large increase i n size or dry matter production which d i l u t e s the nutrient i n the plant tissue. interpretations. continuous uptake.  This may confuse diagnostic  Content, on the other hand, may  increase because of the  Therefore, expressing f o l i a r nutrient data on a  content (uptake) basis would help overcome diagnostic imprecision due to the d i l u t i o n effect (Farhoomand and Peterson 1968). Another method of expressing nutrient composition, as suggested  by  Stachurski and Zimka (1975), i s i n terms of content per unit of leaf area.  The supporting argument i s that expressing nutrient composition on  a dry weight basis may  confound interpretations because of major changes  in tissue dry weight resulting from fluctuations i n stored carbohydrates (Bradbury and Malcolm 1978).  Gholz (1978) and Smith e t ^ a l . (1981) found  11  this method to be superior to expression on a dry weight basis.  It i s ,  however, uncertain whether d i l u t i o n e f f e c t would be reduced i n using leaf area (van den Driessche  1974).  The fourth method of examining f o l i a r nutrient data i s to use c r i t i c a l and optimum nutrient r a t i o s (Ingestad  1970).  The basis f o r  using r a t i o s i s that f o l i a r composition i s associated with an i d e a l equilibrium among elements i n the plant tissue (Bonneau 1973). notion has been supported by van den Driessche  This  (1974) who noted that  maximum y i e l d can only be achieved when a l l nutrients are at optimum concentration and balance. Interactions between nutrients can also confound diagnostic i n t e r pretations.  According  to Olsen (1972), the effects of nutrient i n t e r -  action could either be mutual (synergism) or r e c i p r o c a l (antagonism). In a s y n e r g i s t i c i n t e r a c t i o n , the uptake or translocation of one element by a plant i s stimulated by another element. antagonistic i n t e r a c t i o n .  The opposite takes place i n  Interations, therefore, tend to m i l i t a t e  against d i r e c t i n t e r p r e t a t i o n of nutrient concentrations content.  or absolute  This has l e d to the use of nutrient ratios to r a t i o n a l i z e such  interactions.  Some of the commonly used r a t i o s i n forest n u t r i t i o n  research are N/S, N/P, N/K, K/Ca, Ca/Mg, and P/Al (Leyton 1958; Tamm 1964;  Waring 1972; van den Driessche  1974).  The methods of expressing nutrient composition mentioned above used techniques that measure t o t a l concentration or content i n the plant tissue.  Leece (1976) c r i t i c i z e d this approach because i t does not  distinguish the p h y s i o l o g i c a l l y active from the inactive f r a c t i o n of a p a r t i c u l a r nutrient element under study.  This concept was already  being  12  a p p l i e d to " a c t i v e " i r o n i n f r u i t (1933).  t r e e s a h a l f - c e n t u r y ago  by Oserkowsky  He d e f i n e d a c t i v e i r o n as t h a t form of i r o n t h a t i s a c t i v e i n  c h l o r o p h y l l formation.  Zech (1970) a p p l i e d t h i s concept  to d i f f e r e n t i a t e  between c h l o r o t i c and normal t r e e s of Scots pine (Pinus s i l v e s t r i s L . ) . He found  t h a t the a c t i v e i r o n l e v e l i n c h l o r o t i c needles was  t h a t of green ones though the t o t a l i r o n content was c h l o r o t i c needles.  less  than  sometimes h i g h e r i n  Hence, t o t a l i r o n i n the f o l i a g e i s sometimes not  i n d i c a t i v e of i r o n d e f i c i e n c y . The  p r e s e n t study employs f o u r of the f i v e methods mentioned to  express n u t r i e n t c o m p o s i t i o n . element per 100  C.  These a r e c o n c e n t r a t i o n , content  n e e d l e s ) , some n u t r i e n t r a t i o s , and a c t i v e  (mg  iron.  N u t r i e n t Requirements of C o n i f e r s Some n u t r i e n t c o n c e n t r a t i o n v a l u e s f o r a s e l e c t e d number of c o n i -  f e r o u s s p e c i e s are summarized i n T a b l e 1.  More d e t a i l e d reviews were  g i v e n by Stone (1968) and M o r r i s o n (1974).  A more r e c e n t review f o r  c o n i f e r o u s s p e c i e s i n B r i t i s h Columbia was  done by B a l l a r d and C a r t e r  (1983), but as c a u t i o n e d by the a u t h o r s , many of the v a l u e s g i v e n are not species-specific.  Some of the v a l u e s o b t a i n e d i n the l i t e r a t u r e were  used i n the t e x t of t h i s t h e s i s f o r comparative  purposes.  The  potential  l i m i t a t i o n s d i s c u s s e d i n p r e v i o u s s e c t i o n s were r e c o g n i z e d when i n t e r p r e t i n g and e x t r a p o l a t i n g these v a l u e s .  13 TABLE 1.  F o l i a r n u t r i e n t values of c o n i f e r s .  Type o f culture  Deficient (critical value/range)  useful i n i n d i c a t i n g nutrient status  Adequate  High  Reference  NITROGEN, I d r y weight Pinus c o n t o r t s  1.20 - 1.70  Sand  Pinus c o n t o r t a  Field  1.0  Pseudotsuga • e n z l e s l l  Field  1.0  1.7 - 3.0  >3.0  Swan 1972  1.50  Everard 1973  1.40  Everard 1973  PHOSPHORUS, X d r y weight >0.40  Swan 1972  Pinus c o n t o r t a  Sand  0.10 - 0.17  0.17- 0.40  Plnua c o n t o r t a  Field  0.12 - 0.15  0.15  Everard 1973  Pseudotsuga n e n z l e s l l  Field  0.14  0.20  Everard  1973  POTASSIUM, I dry weight Pinus c o n t o r t a  Sand  0.30 - 0.50  Pinus c o n t o r t a  Field  0.40 - 0.50  Pseudotsuga m e n z l e s l l  Field  0.50  0.50 - 1.10  >1.10  Swan 1972  0.50  E v e r a r d 1973  0.70  Everard  1973  CALCIUM, I d r y weight Pinus  contorta  Pseudotsuga  Sand  0.06 - 0.06  menzlesll  0.08 - 0.30  >0.30  0.25  Swan 1972 B a l l a r d and C a r t e r 1983  MAGNESIUM, I dry weight Pinus c o n t o r t a Pseudotsuga  Sand  0.07 - 0.09  menzlesll  0.09 - 0.16  >0.16  0.10  Swan 1972 B a l l a r d and C a r t e r 1983  SULPHUR, Z dry weight Pinus s l l v e s t r l s  Solution  0.04 - 0.15  0.15 - 0.20  Ingested  1960  BORON, ppm  7-18  14 - 135  Pinus ponderosa  Field  Pinus p a t u l a  Field  Pinus p i n a s t e r  Field  8  16  Pinus r a d i a t a  Field  8  Pinus r a d i a t a  Field  5-7  -  Pinus realnoaa  Field  Pinus r a d i a t a  Pot  -  Pinus taeda  Nursery  Pinus s l l v e s t r l s  (soil)  Field  6  1.8 - 4.2 4  -  -  54-75  -  Parker 1956 V a i l e t a l . 1961 Stone and W i l l 1965 W i l l 1971 Windsor and K e l l y 1972 Neary e t a l . 1975 Snowdon 1982 Stone e t a l ^ 1982 Aronsson 1983  14 IABLE 1.  Type o f culture  Deficient (critical value/range)  (cont'd)  Adequate  High  Reference  COPPER, ppm Picea s i t c h e n s i s  Nursery  Pinus ponderosa  Field  Pseudotsuga  Nursery  Pinus r a d i a t a  Field  1.0  3.0  Pinus r a d i a t a  Field  3.0  -  menziesii  2.3 - 2.8  7.8 -10.8  -  2.5 - 7.9  2.6  4.5  -  Pinus e l l i o t t i  S o i l (pot)  Pinus r a d i a t a  Nursery  Pseudotsuga  menziesii  Field  4  -  Pseudotsuga  menziesii  Field  1.5  -  Pseudotsuga  menziesii  Vermiculite  1.7 - 2.6  4  2.0  14-30  -  fienzian  and Warren 1956  Parker 1956 Oldenkamp and Smllde 1966 R u l t e r 1969 W i l l 1971 van Lear and Smith 1972 Knight 1975 S t r u l l u and Bonneau 1978 Binns e_t a_l. 1980 Lambert and Weidensaul 1982  IRON, ppm Pinus s i l v e s t r i s  Solution  50  Pinus s i l v e s t r i s  Field  50  Pinus r a d i a t a  Field  34  50 - 70  -  -  Ingestad 1960 Zech 1970 R u i t e r 1983  MANGANESE, ppm Pinus ponderosa  Field  -  Picea abies  Field  4-20  Pinus r a d i a t a  Field  20 - 30  38 -102 20  -  -  Parker 1956 Ingestad 1958 Woods 1983  ZINC,, ppm Pinus r a d i a t a  Field  10  -  Pinus r a d i a t a  Nursery  15  15 - 50  Pinus r a d i a t a  Field  10 - 15  Pinus r a d i a t a  Field  10 - 12  -  -  R u i t e r 1972 Knight 1975 McGrath 1978 Woods 1983  15  CHAPTER 3.  A.  GREENHOUSE EXPERIMENTS  Introduction There i s very limited information from the l i t e r a t u r e on various  aspects of micronutrient n u t r i t i o n of forest trees.  The extensive review  by Stone (1968) on this subject emphasized this lack of information, but l i t t l e progress has been achieved since Stone published his review.  As  evident from the l i t e r a t u r e review section, there has been no research reported i n Canada on the determination of the c r i t i c a l l e v e l s of micronutrients for any of the coniferous species.  In fact, there i s no  published information on this subject f o r lodgepole pine, which i s the most wide-ranging of the American pines ( C r i t c h f i e l d 1980). This chapter reports findings from a greenhouse study conducted with lodgepole pine as the test species.  The main objective of the  experiments was to bracket the c r i t i c a l levels of boron, copper, t o t a l i r o n (Fe) and active iron (AFe) for lodgepole pine.  B.  Methods and Materials 1.  Experimental  Design  E s s e n t i a l l y the study consisted of three separate experiments.  In  each of these experiments, the supply of one of the three micronutrients (boron, copper and iron) was given at three levels and associated with two levels of nitrogen.  The other eleven essential nutrients were  supplied at levels known to be s u f f i c i e n t for optimum growth.  16  The reason f o r s u p p l y i n g n i t r o g e n a t two l e v e l s i s because  o f the assumption  (low and adequate)  that there i s a p h y s i o l o g i c a l  interation  between n i t r o g e n and some m i c r o n u t r i e n t s . A p p l i c a t i o n s o f n i t r o g e n f e r t i l i z e r s have been shown t o induce and/or boron  ( M o l l e r 1983) and copper  a c c e n t u a t e d e f i c i e n c i e s of  ( R u i t e r 1969) i n Pinus spp. growing on  s o i l s m a r g i n a l i n terms o f a v a i l a b i l i t y o f these n u t r i e n t s .  Both  n i t r o g e n and i r o n p l a y an e s s e n t i a l r o l e i n c h l o r o p h y l l f o r m a t i o n and K i r k b y  (Mengel  1982).  The c r i t i c a l l e v e l cannot be determined d e s i g n because response c u r v e .  p r e c i s e l y from t h i s  study  t h e r e a r e too few d a t a p o i n t s f o r p r e c i s e e s t i m a t i o n of a When maximum observed growth i s a s s o c i a t e d w i t h the  i n t e r m e d i a t e n u t r i e n t supply l e v e l , d a t a a s s o c i a t e d w i t h t h i s l e v e l can be used f o r p r e l i m i n a r y a p p r o x i m a t i o n o f " c r i t i c a l " v a l u e s . r e s e a r c h w i l l be needed f o r c o n f i r m a t i o n and/or  Subsequent  f u r t h e r refinement o f  such e s t i m a t e s . Each experiment  b a s i c a l l y consisted of a 3 x 2 f a c t o r i a l  combination r e p l i c a t e d f o u r t i m e s .  treatment  Each r e p l i c a t e was r e p r e s e n t e d by one  pot, g i v i n g a t o t a l o f 72 pots f o r the three experiments. were arranged i n a c o m p l e t e l y randomized  design.  The treatments  Experimental d u r a t i o n  was s i x months. 2.  Pot Arrangement  The p o s i t i o n o f each pot on the bench was r o t a t e d each n u t r i e n t a p p l i c a t i o n was made. mental d i f f e r e n c e s  time  T h i s was t o reduce e f f e c t s of e n v i r o n -  ( f o r example, s h o r t wave r a d i a t i o n and temperature  d i f f e r e n c e s ) a l o n g the greenhouse bench.  17  3.  Greenhouse  Environment  The greenhouse was cooled during the summer months with automatic v e n t i l a t o r s , and steam-heated during the f a l l . from 14°C to as high as 35°C during the summer.  The temperature range was Relative humidity was  about 75 percent. At the top of the seedlings under natural l i g h t i n g i n mid-March, quantum flux density i n the 0.4 to 0.7 \im wave band was only 150-200 umol m~^s-'-, as measured with a Quantum Sensor. -  I t was  necessary to increase the l i g h t i n t e n s i t y to about 300 pmol m~^s~^ for rapid and continuous growth of seedlings. tary l i g h t i n g from fluorescent lamps.  This was done with supplemen-  A 16-hour photoperiod was main-  tained f o r the entire duration of the study. 4.  Preparation and Maintenance of Growth Medium  Pure white sand (trade name "20-30 V S i l i c a Ottawa Sand") was used as the growth medium.  The sand was acid washed, using the method by  Hewitt (1966), modified s l i g h t l y because of high impurities.  The proce-  dure involved submerging the sand i n a mixture of 2N HCI and 1 percent oxalic acid f o r one week.  This was repeated f o r three consecutive weeks,  with fresh acid for each weekly washing.  The sand was then washed with  demineralized water to remove any of the surplus acid from the growth medium.  A t o t a l of 35 washings for each container of 30 kg sand was  necessary to raise the pH from 0.5 to 5.5, which was a suitable pH for seed germination. Analysis of the f i n a l leachate showed no trace of the micronutrient metal elements to be tested, and n e g l i g i b l e amounts of some macronutrients (Table 2).  This laborious but thorough washing procedure  18  TABLE 2.  Chemical analysis of the leachate from the sand used as the growth medium.  Element  Concentration  K  0.02%  Ca  0.01%  Mg  0.0%  Cu  0.0 ppm  Fe  0.0 ppm  Mn  0.0 ppm  Zn  0.02 ppm  was necessary to prepare a satisfactory batch of pure sand c r u c i a l f o r the micronutrient studies. About 4 kg of the sand was placed i n each of the 8 - l i t e r p l a s t i c pots.  Drainage holes at the bottom of each pot were covered with a piece  of polyethylene c l o t h , to prevent any loss of sand.  Each pot was placed  i n a 10-inch diameter p l a s t i c tray, used to c o l l e c t leachate for weekly pH measurements.  Because the sand and nutrient solution were e s s e n t i a l l y  unbuffered, large and rapid pH changes could occur.  It was found that,  a f t e r every three nutrient applications, the pH dropped from about 5.5 to 2.0.  This pH d r i f t due to formation of acids i n the sand occurred  throughout the study period of s i x months.  I t was necessary to raise the  pH back to 5.5, and this was done by flushing each pot with at least s i x l i t r e s of demineralized water once every week. to remove algae that grew on the sand culture.  This washing also tended  19  The moisture content of the sand was maintained at 10 percent by weight.  At this l e v e l , the water content i s approximately two-thirds of  f i e l d capacity (Allen 1968).  The pots were occasionally weighed as the  seedlings grew bigger, to check that the water content was at this l e v e l . It was necessary to increase the frequency of watering during the summer months. 5.  Germination and Thinning  The seeds were provided by the S i l v i c u l t u r e Branch, B r i t i s h Columbia Ministry of Forests.  These seeds were collected from two  locations i n the Prince George region:  Willow River area (Seedlot No.  2093) and Vanderhoof area (Seedlot No. 2313). Prior to sowing, the seeds were placed i n demineralized water f o r imbibition and then were placed i n a ring of shallow indentations made on the surface of the wet sand.  Pots were seeded on March 18, 1982.  Seeds  from both seedlots were sown i n each pot but separated by a 2-cm p l a s t i c divider placed across the center of the pot. One hundred seeds were sown i n each pot.  A p l a s t i c cover was placed over the pots during the  germination period i n order to maintain high humidity.  Emergence of  about 85 percent was achieved a f t e r eight days. Five thinnings were done during the duration of the experiment. Details are outlined i n Table 3.  At the end of s i x months of growth,  s a t i s f a c t o r y inducement of deficiences was very noticeable and s u f f i c i e n t biomass was produced from each r e p l i c a t e (pot) f o r chemical analysis.  20  TABLE 3.  Thinning frequency, dates and number of seedlings remaining after each thinning.  Number of thinning  1  A p r i l 18  40  2  May  18  24  3  June  18  18  4  July  18  16  5  August 18  F i n a l harvest  6.  Number of remaining seedlings per pot  Date  14  September 18  Composition of Nutrient Solutions  The three experiments were designated as boron, copper and iron experiments, each having s i x treatments (three l e v e l s of the test micronutrients and two levels of nitrogen). The composition of the nutrient solution was based on that used by Swan (1972). tions of the "manipulated" elements 4.  The solution concentra-  (B, Cu, Fe and N) are given i n Table  In each of the three separate experiments, the composition of the  standard nutrient solutions was the same f o r the non-manipulated elements.  Tables 5 and 6 give solution composition information about  these, f o r the lower (10 ppm) N and higher (100 ppm) N treatments, respectively.  21  TABLE 4.  Concentration of manipulated elements i n nutrient solutions.  Test element  Nutrient  l e v e l (ppm)  1  2  3  Boron  0  0.04  0.40  Copper  0  0.002  0.02  Iron  0  0.05  5.0  10  100  Nitrogen  7.  Nutrient Application  The nutrient solutions were prepared i n demineralized Before application, the pH of the solution was adjusted 0.025N NaOH.  water.  to pH 5.5 with  Treatment was done two weeks after sowing and subsequently  once every two days f o r the entire duration of the experiment. nutrient solution was applied d i r e c t l y to the sand surface, contact with the upper portions of the seedlings.  The  avoiding  The volume applied was  one l i t r e per pot. Supplementary watering was done only i n July when I t was extremely warm and necessitated the addition of 500 mL of deminera l i z e d water to each pot the day following treatment a p p l i c a t i o n . Preparation and application of nutrient solutions were a l l done i n p l a s t i c measuring cylinders and containers.  A l l work was manually done  and every possible e f f o r t was taken to avoid contamination.  22  TABLE 5. Concentration and source of elements i n standard nutrient solution f o r l e v e l 1 nitrogen supply (10 ppm).  Element  Element ppm  Source mg/L  Associated element  Element ppm  N  4.969  NH C1 KNO3  18.98 23.56 16.16  Cl Ca K  12.57 4.0 6.24  2.795  Ca(N0 ) .4H 0  2.238 10.0  KH2PO4  43.90  K  12.61  6.24 12.61 56.15  KNO3 Cl  51.07  Mg  50.0  MgSO4.7H 0  Ca  4.0 36.0  Ca(N0 ) .4H 0  65.98 2.87  MgS0 .7H 0  K  Cl  12.57 51.07 63.89  Source  4  3  2  2  KH P04 KC1 2  107.25 506.72  2  3  CaCl  2  2  99.68  2  4  Cl  63.89  2  FeS04.7H 0 2  NH4CI KC1 CaCl  2  Fe  5.0  FeS04.7H 0  24.90  Mn  0.20  MnCl .4H 0  0.72  Cu  0.02  CuS0 .5H 0  0.079  Zn  0.05  ZnS04-7H 0  0.22  0.40  H3BO3  2.29  0.03  Na Mo04.2H 0  Mo  65.98  2  2  4  2  2  2  2  2  0.071  2.87  23  TABLE 6.  Concentration and source of elements i n standard nutrient solution f o r l e v e l 2 nitrogen supply (100 ppm).  Source mg/L  Associated element  189.8  CI  125.7  Ca(N0 )2.4H 0  235.6  Ca  40.0  22.38  KNO3  161.6  K  62.39  10.0  KH2PO4  43.9  K  12.61  62.39  KNO-3  12.61  KH P0  Mg  50.0  MgS0 .7H 0  Ca  40.0  Ca(N0 ) .4H 0  65.98  MgS04.7H 0  2.87  FeS04.7H 0  Element  Element ppm  N  49.69  NH C1  27.95  K  Source  4  3  2  2  Element ppm  4  4  506.72  2  3  2  65.98  2  2  2  CI  125.7  Fe  5.0  FeS04.7H 0  Mn  0.20  MnCl .4H 0  0.72  Cu  0.02  CuS04.5H 0  0.079  Zn  0.05  ZnS04.7H 0  0.22  B •  0.40  H3BO3  0.29  Mo  0.03  Na Mo04.2H 0  NH4CI  24.9  2  2  2  2  2  2  2  0.071  2.87  24  8.  Harvesting, Sample Measurement and Preparation  V i s u a l symptoms f o r a l l 18 treatments were recorded before harvesting.  A l l seedlings were harvested by cutting off the stems at  sand l e v e l .  The seedlings i n each pot from the two d i f f e r e n t seedlots  (provenances) were processed separately, except f o r the roots, which were impossible to separate. A l l seedlings were thoroughly washed with demineralized water after harvest.  The needles were then stripped from the stems and  heights were measured on f i v e seedlings per provenance per r e p l i c a t e . Needles, stems and roots were dried at 70°C i n a forced-draft oven. Drying of needles took at least 10 hours, u n t i l they could be snapped cleanly into two when bent.  (This was the c r i t e r i o n used to estimate  whether samples were dry enough for e f f i c i e n t grinding).  It took about  24 hours to dry the stems and roots to get to a constant weight. following weights were taken:  The  stem, root, t o t a l foliage and one hundred  (randomly selected) needles. Needle, stem and root samples were ground separately i n a Waring blender stainless s t e e l cup, and kept i n a i r t i g h t screw-cap bottles.  plastic  Before chemical analyses, s u f f i c i e n t portions of each sample  were re-dried i n the oven at 65°C for three hours, cooled i n a desiccator, and the required amount was then weighed for analysis. 9.  Chemical Analysis  The wet digestion method of Parkinson and A l l e n (1975), s l i g h t l y modified by Ballard (1981), was employed to prepare foliage, stem and  25  root samples for the determination of t o t a l N, P, K, Ca, Mg, A l , Cu, Fe, Mn and Zn.  Total N and P were simultaneously analysed on the o r i g i n a l  digest by the Technicon Autoanalyzer I I .  Potassium, Ca, Mg, A l , Cu, Fe,  Mn and Zn concentrations i n the digest were determined by atomic absorption spectophotometry. Appendix A . l .  Details of the procedure are outlined i n  The values f o r Cu and Fe were noticeably low, especially  for Cu where the concentration In the digest approached the instrument's sensitivity limit.  Therefore foliage samples were re-analysed for these  two elements using a more accurate and sensitive method involving HNO3 digestion.  Details of this procedure proposed by Ballard (personal  communication^-) are described i n Appendix A.2.  The values obtained by  this method were used i n this thesis. The concept that a f r a c t i o n of the t o t a l i r o n i s metabolically active (Oserkowsky 1933) warranted that this form of iron to be determined.  Appendix A.3 outlines Oserkowsky's procedure for active iron  determination as modified s l i g h t l y by Ballard (1981). Boron was determined by the azomethine-H colorimetric procedure of Wolf (1971, 1974), as modified by Gaines and M i t c h e l l (1979).  This  procedure involves dry ashing; this element can be v o l a t i l i z e d during wet ashing (Jones and Steyn 1973). Determination of t o t a l sulphur was done using a Fisher Model 475 Sulphur Analyzer.  The procedures used were those i n the manufacturer's  manual, with some modifications as described by Lowe and Guthrie (1981).  T.M. Ballard. Professor, Faculty of Forestry/Department of S o i l Science, University of B r i t i s h Columbia.  10.  S t a t i s t i c a l Analysis  Comparison between provenances was research.  not an o b j e c t i v e of t h i s  T h e r e f o r e , the d a t a o b t a i n e d f o r the two  thesis  provenances were  pooled to g i v e a s i n g l e v a l u e f o r each v a r i a b l e measured f o r each replicate.  R e s u l t s of stem and r o o t a n a l y s i s are not d i s c u s s e d i n t h i s  thesis. The assessment of c r i t i c a l on the assumption  l e v e l s f o r B, Cu, AFe and Fe was  t h a t the c r i t i c a l  l e v e l on the growth response  based curve  ( F i g u r e 1) i s a t a p o i n t where 90 p e r c e n t of maximum growth o c c u r s (Richards and Bevege 1970; All  Swan 1972).  d a t a were s u b j e c t e d to a n a l y s i s of v a r i a n c e and means were  s e p a r a t e d by the Duncan's New  C.  M u l t i p l e Range t e s t .  R e s u l t s and D i s c u s s i o n 1.  Boron  Experiment  (a)  V i s u a l Symptoms  Complete withdrawal  of boron (B^) f o r both supply l e v e l s of  n i t r o g e n (N^ and N )  produced  Growth r e d u c t i o n was  more severe a t the lower n i t r o g e n s u p p l y  2  severe r e d u c t i o n i n s e e d l i n g growth. (N^)  i n d i c a t i n g n i t r o g e n d e f i c i e n c y as w e l l .  The stems were t h i n and  Needles  or orange c o l o r a t i o n .  were s h o r t and  s t i f f w i t h bronze  needles formed a c l u s t e r due with r e s i n .  crooked Terminal  to adhesion of needle t i p s t o each other  These symptoms are q u i t e s i m i l a r to those d e s c r i b e d by  Snowdon (1982) f o r r a d i a t a pine s e e d l i n g s and Stone et a l . (1982) f o r s l a s h p i n e i n the n u r s e r y , both s u f f e r i n g from boron d e f i c i e n c y .  27  There was higher  no a p p r e c i a b l e d i f f e r e n c e i n needle c o l o u r between the  l e v e l s of boron ( B 2 and B 3 ) , except t h a t the needles  s l i g h t l y greener a t the N  than the  2  levels.  Seedling height  biomass were d i s t i n c t i v e l y h i g h e s t f o r treatment B 3 N 2 . d e s c r i p t i o n s are i l l u s t r a t e d i n F i g u r e s 2, 3 and A l t h o u g h s e e d l i n g growth was  The  4.  than at the difference supply  level  (Table 7).  (P <^ 0.01)  l e v e l s were about 1 and  There was  above f i n d i n g was  0.05)  a highly significant between the  F o l i a r n i t r o g e n a t the  2 percent,  foliar  s i g n i f i c a n t l y lower (P <^  i n f o l i a r n i t r o g e n composition  l e v e l s of N (Table 8 ) .  The  l e v e l was  B2^i,  d i s c o l o r a t i o n seemed  more severe w i t h B 2 N 2 - 'At t h i s same l e v e l of boron supply, 2  and  above  g r e a t e r w i t h B 2 N 2 than w i t h  such d e f i c i e n c y symptoms as needle t w i s t i n g and  boron c o n c e n t r a t i o n a t the N  were  and N 2  two supply  r e s p e c t i v e l y (Appendix B . l ) .  consistent with  the h y p o t h e s i s  that better  s o i l n i t r o g e n s t a t u s , a l l o w i n g more biomass p r o d u c t i o n ,  results in  i m p o s i t i o n of a g r e a t e r demand f o r boron (Stone 1968).  Such a r e s u l t  important  p r a c t i c a l i m p l i c a t i o n s i n B r i t i s h Columbia where n i t r o g e n  d e f i c i e n c y i s f r e q u e n t l y s e r i o u s and contemplated.  On  nitrogen f e r t i l i z a t i o n i s often  s i t e s where no boron d e f i c i e n c y problems have p r e v i o u s l y  been e v i d e n t , n i t r o g e n f e r t i l i z e r might induce s o i l s boron supply has the occurrence with  has  nitrogen.  been m a r g i n a l .  of induced  boron d e f i c i e n c y , i f the  In Sweden, M o l l e r (1983) r e p o r t e d  boron d e f i c i e n c y i n Scots p i n e when f e r t i l i z e d  Figure 2.  Lodgepole pine s e e d l i n g s without  boron  supply.  29  Figure 3.  Differential growth response of lodgepole pine seedlings to increasing boron supply at low nitrogen level.  Figure 4.  D i f f e r e n t i a l growth response of lodgepole pine seedlings to increasing boron supply at high nitrogen l e v e l .  TABLE 7.  F o l i a r boron composition, mass of 100 needles and seedling height.  Treatment B  N (ppm)  0  0.01a (+0.0001)  10  43.4b (+5.42)  0.05b  2  10  (B N!) 3  0  100 (BiN ) 2  0.04  100  (B N ) 2  2  0.40  100  (B N ) 3  2  Content (mg/100 needles)  11.7a (+1.68)  (B Ni) 0.40  Concentration (ppm)  10 (B]_N]_)  0.04  Foliar B  107.3d  (+0.001)  Mass of 100 needles (g) 1.06a (+0.27) 1.16a (+0.29)  0.13c  1.22a  (+0.03)  (+0.25)  7.3a  0.01a  2.01b  (+3.04)  (+0.005)  (+7.72)  15.5a  0.04b  (+0.26) 2.14b  Average seedling height (mm) 91a (+1.13) 100a (+1.03) 98a (+0.95) 141b (+1.33) 158c  (+2.05)  (+0.005)  (+0.17)  (+0.79)  85.5c (+9.83)  0.18d (+0.03)  2.03b (+0.14)  153c (+1.22)  In each column, means with a different s u f f i x are s i g n i f i c a n t l y different at the 5-percent level. Numbers i n parenthesis are standard deviations.  32  TABLE 8.  Source of variation  Analysis of variance of f o l i a r nutrient composition with varying supply levels of boron and nitrogen.  Degrees of freedom  N  P  K  Ca  Mg  S  Al  B  Cu  AFe  Fe  Mn  Zn  Boron  2  *  ns  **  ns  **  **  *  **  ns  ns  ns  **  **  Nitrogen  1  **  ns  **  **  **  ns  **  **  *  **  **  **  **  Boron x Nitrogen  2  ns  ns  ns  ns  *  ns  *  **  ns  ns  ns  ns  ns  Error  42  Total  48  >  Significant  at the 0.05 and 0.01 l e v e l s , respectively.  ns = Not s i g n i f i c a n t .  33  (b)  C r i t i c a l Level  The estimation of the c r i t i c a l l e v e l for boron was done at the higher nitrogen supply (100 ppm N) where boron was the only l i m i t i n g element.  Highest observed one hundred needle mass and height were  associated with a f o l i a r boron concentration and content of 15.5 ppm 0.04 mg/100 needles, respectively (Table 7).  and  If these growth data repre-  sent the growth response plateau of Figure 1, 90 percent of maximum growth would correspond to f o l i a r boron values of 7.3 ppm and 0.01 mg/100 needles (Table 7).  Rounding o f f these values, the c r i t i c a l boron concen-  t r a t i o n could be assumed to be 7 ppm and the c r i t i c a l range from 7 to 16 ppm B.  Figure 5 i l l u s t r a t e s the relationships between f o l i a r boron and  growth, where the l a t t e r i s expressed as either mass of 100 needles or seedling height. The above concentration values are within the range reported by V a i l et a l . (1961), Stone and W i l l (1965) and W i l l (1971) on other pine species under f i e l d conditions. The value obtained i n this study might be used as an approximate and preliminary guideline to assess the boron status of lodgepole pine growing under f i e l d conditions. Actual f i e l d experimentation i s necessary, however, to confirm or refine this guideline • 2.  Copper Experiment (a)  Visual Symptoms  There was s l i g h t difference i n needle colour among the three copper treatments.  Some degree of needle t i p burn and chlorosis  was  34  2001  N  N  2  2  I50H  SEEDLING HEIGHT (mm) l(XH  50'  N,  —i 120  — i  40  CONC.  80  (ppm)  FOLIAR Figure 5.  N,  0  0.06  0.12  CONTENT  BORON  Relationship between f o l i a r boron composition and seedling growth (lodgepole pine).  (mg)  0.18  35  observed on seedlings that received low nitrogen but no copper (C^N^). This i s probably due to both copper and nitrogen d e f i c i e n c i e s . The most pronounced symptoms where copper was element ( C ] ^ ) were dark blue-green tender needles, and bushiness.  the only l i m i t i n g  coloration, drooping and long  There was also a marked depression i n  height growth and biomass production.  These symptoms are i d e n t i c a l to  those described by Binns et^ a l . (1980) for copper deficiency i n Sitka spruce and Scots pine. One  of the functions of copper i n plants i s i n c e l l wall metabo-  lism, especially for l i g n i n synthesis (Bussler 1981).  Inhibition of  l i g n i f i c a t i o n due to copper deficiency w i l l lead to bending and loss of plant vigour.  This probably explains drooping and soft needles observed  i n this experiment.  Figures 6 to 8 i l l u s t r a t e the d i f f e r e n t i a l v i s u a l  symptoms explained above. F o l i a r nutrient composition Appendix B.2.  There was  for this experiment i s given i n  a highly s i g n i f i c a n t difference (P £ 0.01)  in  f o l i a r nitrogen between the two levels of nitrogen supply (Table 10). Average f o l i a r nitrogen at the 2.0  and N  2  levels were about 1.1  and  percent, respectively. F o l i a r copper concentration at the C3N2  l e v e l (at which copper and nitrogen were supplied i n adequate amounts) was  s i g n i f i c a n t l y lower than at the CgN^  case of boron, this result has important  l e v e l (Table 9).  As i n the  p r a c t i c a l implications i n that  nitrogen f e r t i l i z a t i o n might induce copper deficiency i n areas where no copper deficiency problems have previously been evident.  Ruiter (1969)  found that low copper concentrations i n radiata pine foliage appeared to have been nitrogen-induced.  36  F i g u r e 6.  Lodgepole p i n e s e e d l i n g s w i t h o u t copper s u p p l y .  Figure 7.  D i f f e r e n t i a l growth response of lodgepole pine seedlings to increasing copper supply at low nitrogen l e v e l .  38  Figure 8-  D i f f e r e n t i a l growth response of lodgepole pine seedlings to increasing copper supply at high nitrogen l e v e l .  TABLE 9.  F o l i a r copper composition, mass of 100 needles and seedling height.  Foliar  Treatment Cu  10  1.1a  0.001a  (+0.48)  (+0.0001)  (+0.15)  (+1.04)  2.5bc (+1.08)  0.003ab (+0.001)  1.08a (+0.27)  (+1.33)  5.6d (+1.73)  0.007c (+0.002)  1.24a (+0.15)  121b (+1.02)  0.002a  1.78b  (+0.001)  (+0.27)  10  (C Ni) 2  0.02  10  (C3N1) 0  100 (CiN ) 2  0.002  100  (C N ) 2  2  0.02  100  (C N ) 3  2  Average seedling height (mm)  Concentration (ppm)  (ClNi) 0.002  Mass of 100 needles (g)  N (ppm)  0  Cu  0.9a (+0.60)  Content (mg/100 needles)  1.24a  99a  102a  128b (+1.47)  1.8ab  0.004b  (+1.57)  (+0.003)  (+0.25)  (+1.01)  3.0c (+0.63)  0.007c (+0.001)  2.33c (+0.14)  171c  In each column, means with a different level.  2.15c  s u f f i x are s i g n i f i c a n t l y d i f f e r e n t  Numbers i n parenthesis are standard deviations.  161c  (+0.86)  at the 5-percent  40  TABLE 10.  Source of variation  Analysis of variance of f o l i a r nutrient composition with varying supply levels of copper and nitrogen.  Degrees of freedom  N  P  K  Ca  Mg  S  Al  B  Cu  AFe  Fe  Mn  Zn  Copper  2  **  **  **  **  **  ns  ns  ns  **  **  **  *  ns  Nitrogen  1  **  **  **  **  **  **  **  **  **  **  **  **  **  Copper x Nitrogen  2  **  **  ns  *  **  **  ns  ns  *  ns  *  **  *  and  0.01  Error  42  Total  47  *, ** = Significant at the 0.05 ns = Not  significant.  levels,  respectively,  41  (b)  C r i t i c a l Level  Table 9 shows seedling growth response to varying levels of copper and nitrogen supply.  At the N  2  (100 ppm N) l e v e l , biomass production  and seedling height were highest when f o l i a r copper concentration 3.0 ppm  (or 0.007 mg Cu/100 needles).  i n Figure 9.  was  These relationships are depicted  Ninety percent of the maximum observed biomass and height  growth was attained by seedlings having an average of 1.8 mg Cu/100 needles) i n their f o l i a g e .  ppm Cu (0.004  Rounding off these values, the  c r i t i c a l copper concentration could be assumed to be 2 ppm and the c r i t i c a l range from 2 to 3 ppm  Cu i f the maximum observed l e v e l approximates  the actual maximum growth. The above concentration values are consistent with the findings of other investigators for conifers (Oldenkamp and Smilde 1966; 1969;  S t r u l l u and Bonneau 1978; 3.  Iron Experiment  (a)  V i s u a l Symptoms  Ruiter  and Lambert and Weidensaul 1982).  The most obvious symptoms when iron was completely withdrawn were general chlorosis, long yellow needles, stunted bud development and severe retardation i n height growth.  When the seedlings were three  months old, the needles turned to a whitish colour which i s an indication of extreme iron deficiency ( H i l l and Lambert 1981).  This symptom did  not p e r s i s t and the needles turned yellow when seedlings were f i v e months old.  With the F N^ 2  treatment (0.05 ppm Fe + 10 ppm N) the  needles  42  3.0-1  50 H 0  1  1  3  6  CONC.  (ppm)  FOLIAR Figure 9 .  H 0  1  1  0.005  0.01  CONTENT  COPPER  R e l a t i o n s h i p between f o l i a r copper composition s e e d l i n g growth ( l o d g e p o l e p i n e ) .  and  (mg)  43  showed bright yellow d i s c o l o r a t i o n .  The above descriptions are  i l l u s t r a t e d i n Figures 10 to 12. F o l i a r nitrogen at the N  2  supply l e v e l was s i g n i f i c a n t l y higher  (P < 0.01) than the N]_ l e v e l (Table 12, Appendix B.3).  At the higher  l e v e l of iron supply ( F 3 ) , both t o t a l and active i r o n were s i g n i f i cantly lower at the N  2  than  l e v e l (Table 11).  I t appears that an  increase i n f o l i a r nitrogen due to increased nitrogen supply caused a substantial decrease i n f o l i a r i r o n . values at the F N 3  2  Even though the actual f o l i a r iron  l e v e l were r e l a t i v e l y high (about 146 ppm), this  finding implies an important p r a c t i c a l consideration.  I f this decreasing  trend i n f o l i a r i r o n continues, repeated or high rate of nitrogen f e r t i l i z a t i o n might eventually induce iron deficiency. (b)  C r i t i c a l Level  The highest biomass and seedling height for the high nitrogen treatment  (100 ppm N) were associated with f o l i a r t o t a l and active iron  concentrations of 146.3 and 147 ppm, respectively (Table 11, Figures 13 and 14).  Ninety percent of this maximum growth was attained by seedlings  having 44.0 ppm t o t a l i r o n and 31.6 ppm active iron i n their f o l i a g e . Rounding off these values, the c r i t i c a l concentration of t o t a l and active i r o n could be assumed to be 44 and 32 ppm, respectively. The concentrations associated with maximum growth are very high, and may well represent "luxury" l e v e l s . c r i t i c a l range.  Thus further study may be needed to estimate the  The c r i t i c a l concentration estimates are highly compara-  ble to those reported by Ingestad for active iron i n Scots pine.  (1960) for t o t a l iron, and Zech (1970)  Zech found that green needles had about  44  Figure 10.  Lodgepole pine seedlings without  iron supply.  45  Figure 11.  D i f f e r e n t i a l growth response of lodgepole pine seedlings to increasing iron supply at low nitrogen l e v e l .  46  F>N1  Figure  F l r  12.  j2.  FiSZ  D i f f e r e n t i a l growth response seedlings to i n c r e a s i n g i r o n nitrogen level.  of lodgepole pine supply at high  TABLE 11.  Total and active iron composition, mass of 100 needles and seedling height.  Total f o l i a r Fe  Treatment Fe  N  Mass of 100 needles (g)  Average seedling height (mm)  Content (mg/100 needles)  Concentration (ppm)  48.3a (+6.54)  0.05a (+0.01)  37.7a. (+13.17)  (+0.01)  1.09a (+0.16)  108a (+1.16)  0.05 10 (F N )  81.4a (+30.31)  0.08a (+0.04)  54.8a (+14.33)  0.05a (+0.01)  1.01a (+0.29)  106a (+1.09)  5.0  359.4c  0.47c (+0.17)  351.1c  (ppm) 0  10 (FiNi)  2  L  10 (F Ni) 3  0  100 (F]N )  Concentration (ppm)  Active Fe  (+126.89) 44.0a  0.08a  (+120.16) 31.6a  Content (mg) 0.04a  0.46c (+0.16)  1.34ab (+0.19)  0.06a  1.79c  116a (+0.86) 163c  (+10.85)  (+0.02)  (+5.70)  (+0.01)  (+0.29)  (+1.90)  0.05 100 (F N )  52.7a (+6.25)  0.12a (+0.01)  45.3a (+3.46)  0.10a (+0.02)  2.27d (+0.31)  161c (+0.99)  5.0  146.3b  0.36b (+0.09)  147.0b  0.36b  2.45d  (+0.08)  (+0.29)  2  2  2  100 (F N ) 3  2  (+28.96)  (+26.19)  In each column, means with a different s u f f i x are s i g n i f i c a n t l y different at the 5-percent level. Numbers i n parenthesis are standard deviations.  177d (+3.70)  48  TABLE 12.  Source of variation  Analysis of variance of f o l i a r nutrient composition with varying supply levels of iron and nitrogen.  Degrees of freedom  N  P  K  Ca  Mg  S  Al  B  Cu  AFe  Fe  Mn  Zn  A  A  AA  AA  AA  AA  AA  n  AA  AA  AA  AA  AA  AA  ug  AA  Iron  2  ns  *  ns  **  **  AA  Nitrogen  1  AA  AA  AA  AA  AA  AA  Iron x Nitrogen  2  **  ns  ns  **  ns  ns  Error  42  Total  47  A  >  AA  n  s  **  = Significant at the 0.05 and 0.01 l e v e l s ,  ns = Not s i g n i f i c a n t .  AA  ns  s  ns  respectively.  N  2  2.0 H  MASS OF 100 NEEDLES (g)  i.oi  200 n  I50-| SEEDLING HEIGHT (mm)  100 H  50  200 CONC.  (ppm)  FOLIAR Figure  13.  0.25 CONTENT ( m  TOTAL IRON  R e l a t i o n s h i p between f o l i a r t o t a l i r o n c o m p o s i t i o n and s e e d l i n g growth ( l o d g e p o l e p i n e ) .  50  2.0  MASS OF 100 NEEDLES (g)  H  1.0  0 200  150  SEEDLING HEIGHT (mm)  100  r  50 200  CONC.  400  (ppm)  FOLIAR Figure 14.  0  0.25  CONTENT  0.5  (mg)  ACTIVE IRON  Relationship between f o l i a r active iron composition and seedling growth (lodgepole pine).  51  30 or more ppm active iron, while those which were c h l o r o t i c because of i r o n deficiency, had less than 30  D.  ppm.  Summary of Greenhouse Experiments The primary aim of the greenhouse experiments was to bracket the  c r i t i c a l deficiency value or range of boron, copper and iron f o r lodgepole pine through the use of f o l i a r analysis data and seedling growth response.  While direct extrapolation of the findings to f i e l d  situa-  tions has to be approached with caution, the results obtained s i g n i f y some important p r a c t i c a l applications. The findings suggest that the c r i t i c a l ranges expressed as concent r a t i o n (ppm) are 7 to 16 for boron, 2 to 3 f o r copper, with the lower values representing c r i t i c a l levels for these elements.  The  critical  l e v e l s f o r iron are estimated at 44 and 32 ppm f o r t o t a l and active iron, respectively.  Concentrations below the c r i t i c a l l e v e l may be associated  with acute deficiency of that p a r t i c u l a r element.  Values above the  c r i t i c a l range imply optimum or near optimum growth.  The levels at which  t o x i c i t y symptoms would appear have not been reached i n this study. There was a r e c i p r o c a l relationship between nitrogen supply and foliar  composition of boron, copper, active iron and t o t a l i r o n .  The  composition of these three micronutrients i n the foliage decreased as f o l i a r nitrogen concentration was increased by increasing nitrogen supply.  This has a s i g n i f i c a n t p r a c t i c a l implication i n that nitrogen  fertilization  might induce deficiencies of these micronutrients.  52  CHAPTER 4.  A.  FIELD EXPERIMENT  Introduction It i s well known that foliage of plants can rapidly absorb plant  nutrients.  There are two main paths of entry for f o l i a r nutrients:  c u t i c l e and the stomata. f o l i a r absorption  the  A detailed description of the mechanism of  has been described  by a number of authors including  Boynton (1954), van Overbeek (1956), Wittwer and Teubner (1959), Jyung and Wittwer (1965) and Franke (1967). F o l i a r application of f l u i d f e r t i l i z e r s has been used i n a g r i c u l ture for a long time and has been an accepted practice i n various micronutrients 1980).  supplying  for many crops (Murphy and Walsh 1972;  This method of applying  advantages for f o r e s t r y .  Traynor  f e r t i l i z e r s offers many t h e o r e t i c a l  These are mainly related to avoiding  soil  reactions which reduce the a v a i l a b i l i t y of applied nutrients to the trees (Knight 1978;  Mengel and Kirkby 1982).  F o l i a r sprays can also  correct nutrient d e f i c i e n c i e s faster or maintain optimum n u t r i t i o n of a particular nutrient better than could be accomplished by s o i l application.  M i l l e r and Young (1976) also indicated that f o l i a r nutrient  application has more f l e x i b i l i t y i n application timing, reduces the potential for nutrient leaching and water p o l l u t i o n , improves l o g i s t i c s of delivery and application, and allows more options i n f e r t i l i z e r formulations.  53  The main l i m i t a t i o n i n the use of f o l i a r application i s that i t can cause severe f o l i a r scorching.  This method, therefore, has not been  generally successful i n applying macronutrients because of the d i f f i c u l t y i n getting s i g n i f i c a n t quantities of these nutrients into the plants without causing serious injury to the foliage (Engelstad and Russel 1975). The use of weak solutions of macronutrient f e r t i l i z e r s i n forestry has produced discouraging r e s u l t s .  For example, Paavelainen (1972) i n  Finland reported no remarkable growth response and no s i g n i f i c a n t nitrogen uptake by Scots pine sprayed with 0.7 percent urea solution.  In  the southeastern United States, the application of diammonium phosphate solution, containing 1.4 percent nitrogen, to slash pine seedlings did not increase nutrient uptake or growth of seedlings (Schultz 1968).  In  another greenhouse experiment with slash pine, Eberhardt and Pritchett (1971) observed needle burning at 0.4  percent nitrogen concentration, and  the severity increased as nitrogen supply was increased.  The only  encouraging r e s u l t was reported by M i l l e r and Young (1976) who  found i t  feasible to apply urea-ammonium solutions containing 32 percent nitrogen to Douglas-fir/western hemlock stands at rates up to 224 kg of the f e r t i l i z e r per hectare.  They reported an increase of 106 percent i n  volume growth r e l a t i v e to untreated trees over a four-year period. There i s very l i t t l e published information on f i e l d  experimentation  involving the use of micronutrient f e r t i l i z e r s as f o l i a r sprays i n forestry.  One isolated example i s i n New Zealand where micronutrient  sprays have been used to correct boron, copper, i r o n and zinc d e f i c i e n cies of radiata pine i n the nursery (Knight 1978; Mead and Gadgil 1978).  54  N u t r i t i o n a l surveys by f o l i a r analysis of several forest stands i n i n t e r i o r B r i t i s h Columbia by Ballard (1981) indicated presumptive evidence of nitrogen, copper and iron d e f i c i e n c i e s .  This chapter reports  findings from preliminary f i e l d investigation on the use of urea, ferrous sulphate and copper sulphate solutions as f o l i a r sprays on lodgepole pine and Douglas-fir.  (A Douglas-fir stand on calcareous s o i l was selected  because no l o g i s t i c a l l y suitable lodgepole pine stand on such a s o i l had been found by the time this part of the study commenced).  The main aims  of the study were: 1.  To assess tree growth response to f o l i a r application of fertilizers.  2.  To determine f o l i a r nutrient responses to f o l i a r applied nutrients.  3.  To evaluate the potential for f o l i a r f e r t i l i z a t i o n to r e l i e v e copper and iron d e f i c i e n c i e s .  B.  Methods and Materials 1.  Site Descriptions  (a)  Site Locations  The experiment was conducted at f i v e different s i t e s i n the Prince George and Kamloops Forest Regions i n i n t e r i o r B r i t i s h Columbia (Figure 15).  Two of the s i t e s , Tsus Creek (53°45' N, 121°49' W, elevation 854 m)  and Opatcho Lake (53°44' N, 122°18' W, elevation 900 m) are i n Prince George Forest Region.  The remaining three s i t e s are at Marshall Lake  (50°56' N, 122°34' W, elevation 1540 m), Shulaps Creek (50°57* N, 122°18' W, elevation 1235 m) and P a v i l i o n Lake (50°52' N, 121°43*W, elevation 905  135*  130*  Figure 15.  123'  120*  Map showing location of the study s i t e s .  56  m) i n Kamloops Forest Region.  For brevity, Tsus Creek, Opatcho Lake,  Marshall Lake, Shulaps Creek and P a v i l i o n Lake s i t e s w i l l be referred to as Sites 1, 2, 3, 4 and 5, respectively, f o r the remainder of the text. The two forest regions are generally characterized by dry cold climates.  Normal p r e c i p i t a t i o n f o r the year and f o r the growing season  recorded at the nearest weather stations f o r Prince George area are 628 mm and 300 mm respectively, and that for the Kamloops area are 440 mm and 198 mm.  Frost-free period (30 years average, 1951-1980) f o r Sites 1, 2,  3, 4 and 5 are 85, 85, 131, 198 and 56 days, respectively.  In the Prince  George area the coldest month i s January with a normal d a i l y minimum temperature of -12.1°C.  The warmest month i s July with a normal d a i l y  maximum temperature of 22.0°C.  The minimum and maximum d a i l y temperature  for the Kamloops s i t e s are -13.8°C (January) and 21.6°C (July), respect i v e l y (Environment Canada 1982). (b)  Stand Characteristics  Sites 1, 2, 3 and 4 consisted of mainly young even-aged lodgepole pine of natural o r i g i n .  In 1982, the estimated stand age (at stump) f o r  Sites 1, 2, 3 and 4 were 23, 23, 33 and 13 years, respectively. f i r i s the only tree species at Site 5.  Douglas-  The estimated age of t h i s stand  i s about 100 years. The lodgepole pine s i t e s , with the exception of Site 3, are severely overstocked.  For example, stand densities average 50,000 and  122,000 stems/ha at Sites 1 and 2, respectively (Dickey 1981). and 5, on the hand, are c r i t i c a l l y understocked.  Sites 3  Tree heights at the  lodgepole pine s i t e s ranged from 1.5 to 5 m and diameter  (dbh) varied  57  from 16 to 64 mm.  The Douglas-fir trees have heights and diameters  ranging from 2.7 to 6.5 m, and 38 to 124 mm, (c)  respectively.  S o i l Characteristics  S o i l p r o f i l e descriptions according to the Canadian system (Canada S o i l Survey Committee 1978) are given i n Appendix C.  S o i l s at Sites 1,  2, 3, 4 and 5 are c l a s s i f i e d as Orthic Humo-Ferric Podzol, eluviated Dystric Brunisol, Orthic Gray L u v i s o l , Brunisolic Gray L u v i s o l , and Orthic Regosol, respectively. Table 13 shows some chemical s o i l c h a r a c t e r i s t i c s of the s o i l p r o f i l e excavated at each of the s i t e s .  Available water storage capaci-  t i e s f o r the f i v e s o i l s were calculated based on Clapp and Hornberger (1978), using f i e l d estimates of rooting depth, s o i l texture, and coarse fragments content (Table 14).  The chemical and physical s o i l character-  i s t i c s were used only f o r s i t e characterization and are not discussed In d e t a i l i n the text. 2.  Experimental Design  The experiment was carried out based on single-tree plot technique (Viro 1967) using completely randomized design with f i v e r e p l i c a t e trees per  treatment at each s i t e .  tions.  There are a t o t a l of 14 treatment combina-  The nature and number of treatments varied from one s i t e to  another (Table 15). Within each s i t e , careful selection of test trees was done to ensure homogeneity i n stand c h a r a c t e r i s t i c s and freedom from any deformity.  The distance between any two test trees was as f a r apart as  TABLE 13.  Site  1  2  3  4  5  Some chemical characteristics of s o i l p r o f i l e s at the study s i t e s .  Horizon  pH H0  Total C %  Total N %  Available P ppm  LFH Ae Bf BC  4.0 4.2 5.0 5.1  19.85 1.32 0.29 0.13  0.82 0.02 0.02 0.01  62.8 38.0 36.0 36.0  24.2 22.0 14.5 13.0  LFH Ae Bf BC  4.6 4.7 5.5 5.6  19.83 1.23 0.64 0.11  0.75 0.05 0.04 0.01  52.2 17.0 56.0 6.0  LFH Ae Bt BC  4.9 5.7 5.7 5.7  18.90 0.79 0.48 0.24  0.75 0.03 0.02 0.01  LF(H) AE Bm Bt  5.7 5.9 5.6 5.3  19.81 0.67 0.62 0.47  LFH C  6.5 7.5  19.68 3.05  2  .  DTPA-extractable cations Cu Fe Mn  Zn  1.44 0.40 0.26 0.16  327.0 76.6 4.6 6.2  711.0 8.0 7.0 2.1  32.0 1.0 0.4 0.1  26.4 24.6 16.0 11.0  1.12 0.88 0.28 0.22  399.2 106.0 33.4 18.6  993.4 45.0 4.9 11.8  20.2 1.9 0.3 o.o:  53.0 90.0 9.0 2.0  25.2 26.3 24.0 24.0  0.48 0.22 1.56 1.38  167.6 37.0 41.4 23.4  281.8 22.6 25.4 14.8  15.4 0.5 0.3 0.3  0.84 0.02 0.02 0.02  37.4 46.0 300.0 15.0  23.6 33.5 31.0 23.5  0.66 0.62 0.42 1.14  243.2 51.6 59.6 60.6  501.8 26.9 8.1 8.4  41.4 0.6 0.5 0.3  0.73 0.18  11.4 2.0  27.0 16.9  0.82 1.00  22.0 9.4  70.0 7.9  74.8 1.4  C/N  59  TABLE 14.  Site  Some physical s o i l c h a r a c t e r i s t i c s of the study s i t e s .  Rooting depth  S o i l texture  (mm)  Coarse fragment content 3 —3 (mm mm '  Available water capacity (mm)  1  330  sandy loam  0.05  37  2  550  sandy loam  0.05  52  3  400  sandy loam  0.05  42  4  800  loam  0.15  88  5  500  sandy loam  0.50  28  60  possible to avoid any treatment effect of one tree upon another.  Trees  were i d e n t i f i e d with p l a s t i c tags to eliminate any contamination from tagging. 3.  F e r t i l i z e r Application  The f e r t i l i z e r s used were copper sulphate, ferrous sulphate and urea solutions, applied singly and also i n combination with each other at d i f f e r e n t levels of concentrations and frequency of applications.  There  was a t o t a l of 14 treatment combinations involving 250 trees. The main t r i a l was i n i t i a t e d i n spring, 1981 with the f i r s t a p p l i cation made just as the current year needles emerged.  Sites 2, 3 and 4  received copper, iron and nitrogen treatments, whereas only iron and nitrogen treatments were applied at Sites 1 and 5. cation i n 1982 for the main t r i a l . f e r t i l i z e r application.  There was no r e a p p l i -  Table 15 outlines d e t a i l s of the  Treatment numbers as designated i n Table 15 w i l l  be used throughout the text.  Iron and nitrogen treatments were repeated  on d i f f e r e n t trees i n 1982 at Sites 1, 2, 4 and 5 (Table 16). Preparation of the f e r t i l i z e r solutions was done at the s i t e . involved dissolving and d i l u t i n g to the required dosage i n water.  This Tap  water from Vancouver was used to prepare the solutions f o r treatments at the Prince George s i t e s , whereas for the Kamloops s i t e s , water was taken from Shulaps Creek.  Chemical analysis of the water from both sources i s  given i n Table 17. A commercial household detergent (trade name 'Joy'), at a concentration of 0.5 percent by volume was added to the f e r t i l i z e r solution as a surfactant to improve solution contact with needle surfaces and thereby enhance absorption.  61 TABLE 15. F e r t i l i z e r treatments for the main (1981) t r i a l .  Site  1 (Tsus Creek)  Treatment No.  2  3 ( M a r s h a l l Lake)  Control  5  4% F e S 0  7  2% urea  June 18, J u l y 8  4% FeS04 + 2% urea  June 18, J u l y 8  1  Control  2  1% CuS0  3  5  0  0 2  J u l y 28  1  4  0.1% CuS0  J u l y 28  1  5  4% F e S 0  June 18, J u l y 8  2  A  4  4  6  2% FeSO^  J u l y 28  1  7  2% urea  June 18, J u l y 8  2  8  1% CuS0  June 18, J u l y 8  2  9  0.1% CuSO^ + 2% urea  J u l y 28  1  10  0.2% CUSO4 + 2% urea  J u l y 28  1  11  4% FeSO^ + 2% urea  June 18, J u l y 8  2  12  1% CuS0  June 18, J u l y 8  2  13  0.1% CUSO4 + 2% FeS04 + 2% urea  J u l y 28  1  14  0.2% CUSO4  J u l y 28  1  4  4  + 2% urea  + 4% FeS04 + 2% urea + FeS04 + 2% urea  1  Control  2  1% CUSO4  June 12, J u l y 30  5  4% FeS04  June 12, J u l y 30  2  7  2% urea  June 12, J u l y 30  2  2% urea  June 12, J u l y 30  2  4% FeS04 + 2% urea  June 12, J u l y 30  2  0  1% CUSO4 +  0  0 2  1  Control  2 3  1% CUSO4 0.2% CUSO4  June 11, J u l y 29 J u l y 29  4  0.1% CuS0  J u l y 29  1  5  4% FeS04  2  6  2% F e S 0  June 19, J u l y 29 J u l y 29  7  2% urea  June 11, J u l y 29 June 11  2  8  ( P a v i l i o n Lake)  June 18, J u l y 8  4  0.2% CuSO^  8  (Shulpas Creek  Frequency of application  June 18, J u l y 8  11  4  Date o f application  1  11  (Opatcho Lake)  Treatment  4  4  1% CUSO4 +  2% urea  0 2 1  1 1  11  4% FeS04 + 2% urea  June 19  1  12  1% CUSO4 +  June 19  1  0  1  Control  5  4% FeS0  7 11  4% FeS04 + 2% urea  0  2% urea  June 11. J u l y 10 June 11, J u l y 10  2  4% FeS04 + 2% urea  June 11, J u l y 10  2  4  2  62  TABLE 16.  Site  F e r t i l i z e r treatments f o r the repeat (1982) t r i a l .  Treatment No.  Date of application  Frequency of application  1  Control  0  0  5  4% FeSO-4  July 4, July 29  2  7  2% urea  July 4, July 29  2  4% FeSC-4 + 2% urea  July 4, July 29  2  11  1  Control  0  0  5  4% FeSO-4  July 4, July 29  2  7  2% urea  July 4, July 29  2  11  4% FeS0  July 4, July 29  2  1  Control  5  4% FeS0  June 29, July 30  2  7  2% urea  June 29, July 30  2  4% FeS04 +2% urea  June 29 July 30  1  0  0 2 2 2  11  1  Control  5  4%  7 11  4  + 2% urea  0 4  FeS04  June 28, July 30  2% urea  June 28, July 30  4% F e S 0 4 + 2% urea  June 28, July 30  TABLE 17.  Element  Chromium Cobalt Calcium Copper  Chemical analysis of water used f o r preparing f e r t i l i z e r solution.  Vancouver tap water* (Capilano system) (ppm)  Shulaps Creek (ppm)  <0.001  0.017  -  0.001  1.4  19.0  <0.001  0.002  Iron  0.13  0.038  Magnesium  0.19  8.8  Sodium  0.45  0.2  Potassium  0.12  0.2  Chloride  0.54  0.3  * Greater Vancouver Regional D i s t r i c t , Water D i s t r i c t Data.  64  Each f e r t i l i z e r solution was applied with a backpack p l a s t i c sprayer.  The entire crown of a l l lodgepole pine trees was  sprayed;  Douglas-fir crowns were sprayed up to a height of about 6.5 m (using a ladder).  Spraying continued u n t i l dripping from the canopy began.  For  combined f e r t i l i z e r treatments, the following sequence of application was followed: ferrous sulphate, copper sulphate and urea.  The amount of  f e r t i l i z e r solution needed f o r each treatment of f i v e trees was between 1.5 and 2 l i t e r s . 4.  F i e l d Sampling and Measurements  Foliage samples from each tree of the main t r i a l were collected i n October, 1981.  Both current and previous-year foliage were sampled.  Samples were taken by cutting some shoots with a pole pruner from the sprayed zone, approximately around the upper one-third of the tree crown. Each sample was kept i n a p l a s t i c bag, transported to the laboratory and stored i n a deep cold-storage room at -10°C before further processing. Collection of second-year  foliage samples of the main t r i a l and  year samples of the repeat t r i a l was done i n October, 1982.  first-  Tree height  and diameter measurements were also taken at this time for description of stand c h a r a c t e r i s t i c s . A s o i l p i t was excavated at each s i t e for p r o f i l e description. S o i l samples were collected f o r each horizon f o r chemical analyses. Results of s o i l chemical analysis were used only for s i t e characterization; they were not s t a t i s t i c a l l y analysed.  65  5.  Sample Preparation and Measurements  F o l i a r samples collected i n 1981 were separated into the current and previous-year's (1980) growth portions.  F o l i a r sample preparation i s  similar to that f o r the greenhouse samples, and has already been described i n Chapter  3.  The same procedure was followed for samples collected i n 1982 except f i v e shoots from each sample tree were saved f o r shoot length measurement.  The shoot length f o r each year's growth f o r the three  consecutive years was measured to the nearest millimeter.  In the main  t r i a l , these three values represent p r e - f e r t i l i z a t i o n , and f i r s t - y e a r and second-year post f e r t i l i z a t i o n shoot growth.  For the repeat t r i a l , only  the previous and current-year shoot length were taken.  The shoot length  measurement gave a data base of 3750 values for the entire experiment. Dry mass of 100 randomly selected needles was also measured f o r each sample collected i n 1982.  This provided a measure of f i r s t - y e a r  p o s t - f e r t i l i z a t i o n biomass response for the repeat t r i a l . trial,  For the main  (assuming a possible increase but no decrease i n number of needle  primordia, as a result of treatments) i t provided an estimate of minimum second-year biomass response. for  A t o t a l of 604 f o l i a r samples was  prepared  chemical analyses. 6.  Chemical Analysis of F o l i a r Samples  The f o l i a r samples were analysed for t o t a l N, P, K, Ca, Mg, A l , B, Cu, Fe, AFe, Mn and Zn.  Total sulphur was done only on some samples to  assess whether the trees were sulphur-deficient. analyses have been described i n Chapter  3.  The procedures for the  66  7.  S o i l Sample Preparation and Analysis  Mineral s o i l and forest f l o o r samples were a i r - d r i e d at room temperature mm)  (22°C).  The s o i l samples were then sieved through a #10  s t a i n l e s s s t e e l sieve.  blender  (2.0  Forest f l o o r samples were ground i n a Waring  to pass a 20-mesh sieve.  The samples were analysed for pH, t o t a l N, available P, organic C, available Cu, Fe, Mn and Zn. electrode pH meter.  S o i l pH was measured i n water using a glass  A 1:2 and 1:8  soil:water suspension was  used for  samples from the mineral and organic s o i l horizons, respectively.  Total  N determination was  and  by Kjeldahl digestion procedure (Bremner 1965)  the digests were analysed for N c o l o r i m e t r i c a l l y on the  Autoanalyzer.  Available P was measured i n Bray's No. 1 solution and organic C by the Walkley-Black method ( A l l i s o n 1965).  Copper, Fe, Mn and Zn were  extracted with DTPA extracting solution and the f i l t r a t e s analysed  by  atomic absorption spectrophotometer (Lindsay and Norvell 1978). 8.  Assessment of F e r t i l i z e r Response  Several variables can be used to evaluate response to f e r t i l i z e r application.  Some common examples include height, diameter, basal area,  t o t a l volume, merchantable volume, biomass production and f o l i a r nutrient status.  Needle weight and f o l i a r nutrient composition  have been used by  Timmer and Stone (1978), Morrow and Timmer (1981), and Weetman and Fournier (1982). Whichever variable i s used, Armson (1974) stated that the basic reason for applying f e r t i l i z e r s i s to bring about a s i g n i f i c a n t increase i n net photosynthate production which i s translated into a desired form  67  of  growth.  Armson a l s o suggested t h a t v a r i a b l e s s e l e c t e d s h o u l d be most  s e n s i t i v e and a l s o most l i k e l y  to g i v e the i n v e s t i g a t o r the i n s i g h t  into  what i s g o i n g on i n b i o l o g i c a l terms when f e r t i l i z e r s are a p p l i e d . For  these r e a s o n s , f e r t i l i z e r response i n t h i s s t u d y was e v a l u a t e d  i n two ways: as t r e e growth response and as f o l i a r n u t r i e n t Tree growth response was dry  mass o f 100 n e e d l e s .  e v a l u a t e d i n terms of shoot ( t w i g ) l e n g t h and I t i s assumed t h a t these two parameters  be used as p r e d i c t o r s of subsequent Fournier  response.  volume response.  could  Weetman and  (1982) used needle weight and f o l i a r n u t r i e n t s as response  variables. For  f o l i a r n u t r i e n t response, the e l e m e n t a l c o n c e n t r a t i o n s were  e x p r e s s e d as p e r c e n t f o r m a c r o n u t r i e n t s , and ppm n u t r i e n t s , both on an oven-dry b a s i s . emphasis i n the t e x t i s on elements  (= mg/kg), f o r m i c r o -  Twelve elements were a n a l y s e d but  t h a t were a p p l i e d  (copper, i r o n and  nitrogen). (a)  E v a l u a t i o n o f Shoot Length and F o l i a r Mass Response  Shoot growth response to f e r t i l i z e r treatments was  expressed as an  average of r a t i o s between p o s t - f e r t i l i z a t i o n to p r e - f e r t i l i z a t i o n lengths.  To compare d i f f e r e n c e s i n response among treatments the shoot  l e n g t h growth was  c a l c u l a t e d w i t h the f o l l o w i n g formula, and expressed as  percentage.  [av(Af/Bf) -  where:  shoot  av(Ac/Be)]100  av = the average f o r a l l r e p l i c a t e s A = increment a f t e r B = increment b e f o r e  fertilization fertilization  68  f = fertilized c = control ( u n f e r t i l i z e d ) The r a t i o Af/Bf i s an index of f e r t i l i z e r response as well as climatic (environmental) e f f e c t s , whereas Ac/Be i s an index of only c l i m a t i c (environmental) influence.  Hence the above formula provides an  index of f e r t i l i z e r response alone. As indicated e a r l i e r , f o l i a r mass measurement was taken only i n 1982.  This gives an estimate of second-year f o l i a r mass response f o r the  main t r i a l and current-year response f o r the repeat t r i a l .  Comparison  among treatments was calculated and expressed as a percentage with the formula:  mf - mc mc where:  m = f o l i a g e mass f = after  fertilization  c = control ( u n f e r t i l i z e d ) . The above formula eliminates climatic (environmental) influence and permits expression of f e r t i l i z e r response only. (b)  Evaluation of F o l i a r Nutrient Response  The following formula was used to compare f o l i a r nutrient response among treatments within the same year:  ( v  where:  n f  " ) 100 nc  n = nutrient element  n c  69  f = after f e r t i l i z a t i o n c = control ( u n f e r t i l i z e d ) . The  c a l c u l a t i o n of year-to-year v a r i a t i o n of f o l i a r nutrient  response for the same treatment was based on the formula:  nf„  nc„  {-fnf^ where:  -) 100 nc^  n = nutrient element f  = second year after f e r t i l i z a t i o n  2  fl c  = f i r s t year after f e r t i l i z a t i o n  l> 2 c  (c)  =  f i r s t and second year control, respectively,  F o l i a r Nutrient  Status  Interpretation  For interpretation, concentrations and some ratios of f o l i a r nutrients evaluated i n this study were compared to those reported i n the literature.  In some cases, especially with copper, active iron and  boron, results obtained from the greenhouse experiment were also used f o r the assessment. 9.  Statistical  The  data for a l l parameters were subjected to a multi-way analysis  of variance.  Analysis  The UBC Genlin program (Greig and Bjerring 1980) was used  to perform this t e s t .  This program was found to be suitable for perform-  ing tests i n an unbalanced analysis of variance.  The program i s based on  analysis of variance for unequal subclass numbers using the least-squares method (Steel and Torrie 1960).  70  Where effects of main factors (treatment, year and s i t e ) were s i g n i f i c a n t , the Duncan New Multiple Range test was performed for mean separation and to evaluate the order of magnitude of the difference.  In  situations where f i r s t - and second-order interactions were s i g n i f i c a n t , variance components were partitioned and trend analysis was applied ( L i t t l e 1981).  This involved descriptive interpretation of the results  based on graphical data representation.  Calculations were done using  equations described i n section 8 of this chapter.  C.  Results and Discussion In the following sections, effects of the main factors (treatment,  year and s i t e ) as evident from f i e l d observations, and results from laboratory analysis and measurements w i l l be presented.  Shoot growth  response for the main t r i a l includes current (1981) and second-year (1982) assessment, and for f o l i a r mass i t i s only a second-year response evaluation.  F o l i a r analysis results for the main t r i a l represent  previous-year responses.  (1980), current-year (1981), and second-year (1982)  In the repeat t r i a l , only current-year (1982) measurement i s  presented for shoot growth and f o l i a r mass production. results represent previous-year  F o l i a r analysis  (1981) and current-year (1982) response.  It must be remembered that the samples of previous-year foliage of both main and repeat t r i a l s also received f e r t i l i z e r treatments; no sample c o l l e c t i o n was  done prior to f e r t i l i z a t i o n .  71  1.  F o l i a r Scorching  (a)  Copper Treatments  Among the 14 d i f f e r e n t combinations  of treatments  (Table 15), those  involving the use of 1 percent copper sulphate solution applied alone or i n combination with ferrous sulphate and/or urea caused very severe scorching and burning of the needles (Appendices  D.l, D.2, D.3).  This  was observed at a l l the three s i t e s (Sites 2, 3 and 4) receiving the above treatments.  The observation was made s i x weeks a f t e r the f i r s t  application, but presumably the injury happened quite soon a f t e r t r e a t ments were applied since f o l i a r absorption of nutrient elements i s a rapid process (Boynton 1954; Wittwer and Tuebner 1959). The scorching i s probably due to copper sulphate.  Apparently even  a single application of 0.2 percent copper sulphate solution (5000 ppm Cu) i s toxic for lodgepole pine.  Oldenkamp and Smilde (1966) made no  mention of any toxic effect when two applications of 1 percent copper sulphate solution were made to eight-year old Douglas-fir.  In the  nursery, concentrations of 0.05 to 0.125 percent copper sulphate spray are common rates of application (Stone 1968).  Lyle (1972) recommended a  higher rate of 0.8 percent copper sulphate spray for l o b l o l l y seedlings i n the nursery.  pine  It i s obvious that d i f f e r e n t tree species  d i f f e r i n their tolerance to copper. However, at the end of the f i r s t growing season following f e r t i l i zation i n 1981, new and healthier foliage was produced on these treated trees (Appendices D.l, D.2, D.3). i n 1982,  There was increased vegetative growth  the second year a f t e r f e r t i l i z a t i o n (Appendices D.4, D.5, D.6).  72  For both years, the f o l i a r colour was much greener than the foliage of the control tree (Appendix D.7). (b)  Iron Treatments  There was minimal scorching of needles following the f i r s t applicat i o n of 4 percent ferrous sulphate alone or i n combination with 2 percent urea on lodgepole pine (Appendices D.8, D.9). Needle burn was s l i g h t l y more serious for ferrous sulphate plus urea treatment  than ferrous  sulphate alone, but less severe than treatments that included copper sulphate (discussed i n previous section).  Recovery from the injury  during the current-year growing season was better than f o r copper-treated trees but increased vegetative growth took place during the 1982 growing season (Appendices D.10, D . l l ) . There was no apparent f o l i a r injury with 2 percent ferrous sulphate treatment.  This would seem to be a suitable dosage f o r lodgepole pine.  For Douglas-fir, 4 percent ferrous sulphate applied alone or with urea did not cause any v i s i b l e f o l i a r damage (Appendices D.12, D.13). In this study, Douglas-fir apparently could tolerate a higher l e v e l of iron i n the f o l i a g e than lodgepole pine.  In contrast, Korstian et a l .  (1921) found that two sprayings of 2 percent ferrous sulphate on western yellow pine and Douglas-fir seedlings caused very severe injury to the needles.  Lyle (1972) recommended a spray solution of 2.8 percent ferrous  sulphate to correct i r o n deficiency i n the nursery.  (c)  Nitrogen Treatment  There was no evident f o l i a r  injury from repeated applications of 2  percent urea for both lodgepole pine and Douglas-fir (Appendices D.15).  D.14,  This dosage of two applications per growing season therefore  seems to be a safe dosage f o r these two species.  M i l l e r and Young (1976)  applied 32 percent nitrogen as urea-ammonium solution to Douglas-fir/ western hemlock stand and experienced serious scorching of the needles. However, they indicated that burning of up to about 30 percent of the needle surface i s acceptable. (d)  Causes of Needle Burn  It seems appropriate at this stage to explain the differences i n severity of scorching observed among treatments, especially i n the case of lodgepole pine.  As mentioned e a r l i e r , the most severe needle burn  resulted from combined application of copper sulphate, ferrous sulphate and urea; moderate burn from ferrous sulphate with urea; minimal  burn  from ferrous sulphate alone, and no burning from urea alone. Needle burning caused by f o l i a r  treatments of nutrient solutions  has been referred to as "osmotic burning" by M i l l e r and Young (1976).  It  i s a c e l l rupture due to large difference i n osmotic pressure across the c e l l wall due to the concentrated f e r t i l i z e r  solution outside the c e l l .  To equalize the pressure, f l u i d tends to move out of the c e l l and the f e r t i l i z e r solute moves i n .  If this transport process cannot occur fast  enough, due to large amounts of solute moving i n , the c e l l ruptures ( M i l l e r and Young, 1976).  It i s probable that copper sulphate and  ferrous sulphate solutions were too concentrated and this caused a build-up of pressure outside the c e l l .  74  Furthermore, the sulphate forms of f e r t i l i z e r s (which were used i n this study) are considered as low-analysis f e r t i l i z e r s which have high s a l t index (Tisdale and Nelson 1975).  High s a l t index f e r t i l i z e r s have a  higher tendency to cause osmotic burning.  Another related possible  reason why copper sulphate and/or ferrous sulphate caused higher Injury when used with urea i s that urea improves the permeability of the c u t i c l e and thus favours d i f f u s i o n of solutes (Franke 1967).  This increased the  pressure d i f f e r e n t i a l across the c e l l wall, and with high concentration of copper, iron and sulphate ions, l e d to c e l l rupture. "Osmotic burning" i s presumably not the only reason f o r the severe needle damage caused by copper sulphate. The concentration (mole fraction) of solute i n 1 percent copper sulphate i s much lower than that i n the much less injurious 4 percent ferrous sulphate solution.  Thus i t  appears that copper t o x i c i t y occurred where solute concentration was quite tolerable.  In f a c t , copper sulphate has been used as an e f f e c t i v e  herbicide (Reuther and Labanauskas 1966).  The mechanism of micronutrient  t o x i c i t y i s not well understood, but according to Kramer and Kozlowski (1979), t o x i c i t y of these elements, including copper, i s mainly related to their injurious effects on enzyme systems. The addition of detergent as a surfactant was to enable the spray solution to form a f i l m over the leaf surface, Instead of forming droplets.  The p r i n c i p a l advantage of this i s enhanced nutrient uptake by the  leaf.  A second advantage, according to Neumann and Prinz (1975) i s that  burn damage may be reduced.  However, i t was noticed i n this thesis  research that the spray solution formed droplets hanging from the needle  75  surfaces.  This led to l o c a l i z a t i o n of the f e r t i l i z e r solution and might  have aggravated f o l i a r i n j u r y . 2.  Tree Growth Responses  (a)  Shoot Growth  (i)  Lodgepole pine  There was no s i g n i f i c a n t response to treatments i n lodgepole pine shoot growth during the current-growing season for both the main and repeat t r i a l s (Table 18). Treatment-site i n t e r a c t i o n was also not s i g n i ficant.  There was, however, a highly s i g n i f i c a n t difference (P _< .01)  among treatments and s i t e s f o r the second-year shoot growth of the main trial.  Treatment-site i n t e r a c t i o n was also highly s i g n i f i c a n t .  Trend  analysis w i l l therefore be employed to interpret treatment responses. Figures 16 to 19 i l l u s t r a t e and treatments.  the relationship between shoot growth  In general, there was a d e f i n i t e relationship between  shoot growth and f o l i a r burning discussed i n previous section.  The  highest response i n shoot increment during the second-year of growth at a l l four s i t e s was from treatments that caused l i t t l e or no f o l i a r burn, mainly treatments 4, 7 and 11 (Figures 17 to 19). There was an average of 49 percent increase f o r these treatments.  Treatment 5, which also  caused minimal f o l i a r burning, gave inconsistent response from one s i t e to another.  At Sites 1, 2 and 3 i t increased shoot length by 44, 4 and  42 percent, respectively, but gave no response at Site 4.  Shoot incre-  ment f o r treatments that caused severe needle burn i n fact was lower than that of untreated trees.  This e f f e c t was more pronounced  from treatments  76  TABLE 18. Analysis of variance of tree growth response variables f o r lodgepole pine and Douglas-fir.  Species  Source of v a r i a t i o n  Treatment  Site  Treatment x site  First-year shoot growth Second-year shoot growth  ns **  **  ns **  Second-year foliage mass  A*  ns  ns  First-year shoot growth  ns  ns  ns  *  ns  First-year foliage mass  First-year shoot growth Second-year shoot growth Second-year f o l i a g e mass  ns **  First-year shoot growth First-year f o l i a g e mass  ns ns  Lodgepole pine: Main t r i a l  Repeat t r i a l  Douglas-fir: Main t r i a l  Repeat t r i a l  **  *,** = S i g n i f i c a n t at the 0.05 and 0.01 l e v e l s , respectively, ns = Not s i g n i f i c a n t . - = Not available (only one Douglas-fir  site).  ns  FeSO  TREATMENT -- No. - -  Urea  4  %  1 .5  4  11  4  7  2 2  1.6 i  SHOOT  GROWTH RATIO  1.2 -  0.8  /  1  1  r  I  5  7  TREATMENT Figure 16.  NUMBER  Second-year shoot growth r a t i o i n r e l a t i o n treatments at Site 1 (lodgepole pine).  78  TREATMENT -- No.  CuSO  FeSO„  u  %  1  .  2 3  1 0.1  4 5  0.2  Urea ------.  .  4  6  2  7  2  8  1  0  9  -  .  1  2  -  2  10  0.2  -  2  11  .  12  1  4  2  13  0.1  2  2  14  0.2  2  2  4  2  1.5 1 / \  SHOOT GROWTH RATIO 1.0  / /  \  \ \  \  /  \  \  /  \ \  i  w  H  i  \  /  \  /  \ / \ / \ / \/  \ l \ I  M  v 0.5  T 2  1 3  1  4  1 5  1 6  TREATMENT Figure 17.  1 7  1 8  ' 9  « 10  1 II  NUMBER  Second-year shoot growth r a t i o i n r e l a t i o n to treatments at Site 2 (lodgepole pine).  r  12  1  1  13  14  79  TREATMENT — No.  CuS0\  FeS0\  1 2 5 7 8 11  Urea  4 4  2 2 2  3.5n  SHOOT  GROWTH RATIO  2.5-  N  1.5  /  T  I  2  5  TREATMENT Figure 18.  7  8  NUMBER  Second-year shoot growth r a t i o i n r e l a t i o n to treatments at Site 3 (lodgepole pine).  II  80  TREATMENT -- No. - -  CuS0 -  1  FeSO,, %  4  -  2 3 4 5 6 7 8  1 0.2 0.1 4 2 1  11 12  1  4  \  GROWTH  2 2  4  i  i \  / / / •  1.6 -  2 ?  -  i  SHOOT  Urea  \  \  \V \ \  \  \ \  RATIO  1.2 -  \ /  \  0.8  — i  1  1  2  1  3  1  4  1  5  TREATMENT Figure 19.  1  6  1  7  1  8  NUMBER  Second-year shoot growth r a t i o i n r e l a t i o n to treatments at Site 4 (lodgepole pine).  1  »  II  12  81  2, 8 and 12 at Site 2 (Figure 17) and also treatment 2 at Site 3 (Figure 18).  There was an average decrease of about 26 percent i n shoot length  from these treatments. Phytotoxicity effects could be the reason for the negative response. This could also be the reason for the non-significant or delayed response during the first-growing season following f e r t i l i z a t i o n .  There  was,  however, a highly s i g n i f i c a n t (P _< 0.01) f i r s t - y e a r nutrient uptake (section 3 of this chapter), and one would expect a corresponding s i g n i f i c a n t growth response, which did not take place.  It might have been  possible that certain physiological mechanisms involving the enzyme systems i n the tree did not increase the tree's photosynthetic rate u n t i l the second year.  This was also reflected by needle length increment at  Site 1, which did not respond i n the f i r s t year but responded greatly i n the second year (Ballard and Majid 1984). As for s i t e differences, the highest response occurred at Site 3. As mentioned e a r l i e r , Site 3 i s sparsely populated compared to the severely overstocked stands at Sites 1, 2 and 4.  Therefore, there was  less competition for l i g h t , moisture and other growth requirements for trees at Site 3. (ii)  Douglas-fir  As i n the case f o r lodgepole pine, there was no s i g n i f i c a n t effect of treatments on Douglas-fir shoot growth during the f i r s t - y e a r growing season, but there was a highly s i g n i f i c a n t (P <^ .01) response i n the second-growing  season (Table 18).  Figure 20 compares i n d i v i d u a l treat-  ment means r e l a t i v e to shoot growth.  Treatment  11 (a combination of  TREATMENT  No.  FeSC\  --  Urea  %  1.5 i  SHOOT GROWTH RATIO 1.0  H  0.5 I  5  TREATMENT Figure 20.  7  II  NUMBER  Second-year shoot growth r a t i o i n r e l a t i o n to treatments at Site 5 (Douglas-fir). (The value outside the range delineated by the v e r t i c a l bar i s s i g n i f i c a n t l y d i f f e r e n t , P < 0.05).  83  i r o n and nitrogen) produced the highest response, an increase of 44 percent i n shoot length compared to the 1980 p r e - f e r t i l i z a t i o n growth. In summary, the highest response i n shoot growth took place during the second-year of growth, and from treatments that caused minimal f o l i a r burn.  Among treatments tested, the combined iron and nitrogen applica-  tion, and copper applied as 0.1 percent copper sulphate were most promising. (b)  F o l i a r Mass  (i)  Lodgepole pine  Analysis of v a r i ance revealed highly s i g n i f i c a n t (P K. .01)  differ-  ence among treatments i n f o l i a r mass production during the second-year of growth following f e r t i l i z a t i o n i n the main t r i a l (Table 18). no s i g n i f i c a n t treatment-site i n t e r a c t i o n .  There was  No s i g n i f i c a n t f i r s t - y e a r  treatment response was detected i n the repeat t r i a l . Figure 21 compares i n d i v i d u a l treatment means i n r e l a t i o n to f o l i a r mass of lodgepole pine.  Treatment 4 was s i g n i f i c a n t l y different  from a l l other treatments (P <^ .05).  It produced the highest positive  response, an increase of 77 percent i n needle mass r e l a t i v e to the untreated trees.  A l l the other treatments also resulted i n s i g n i f i c a n t  positive response, but 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 one another (P <^ .05).  The average increase i n f o l i a r mass for these other treat-  ments ranged from 34 to 61 percent. For  the nine copper treatment combinations, the general trend seems  to indicate that a lower copper dosage applied once per growing  season  (treatments 3, 4 and 13) resulted i n higher f o l i a r mass production than  84 CuSC\  TREATMENT -- No. --  l  FeSO  Urea  u  1 0.1 0.2  2 3 4 5 6 7 8 9 10 11 12 13 14  4 2 1 0.1 0.2  2 2 2 2 2 2 2 2  4 4 2 2  1 0.1 0.2  2.0T FOLIAGE MASS (g)  \ \  I .5  1.0  -i  2  1 3  r  4  5  6  ~T~  7  TREATMENT Figure 21.  n  r  8  9  ~T  r n  10 II  1 1— 12 13 14  NUMBER  F o l i a r mass of lodgepole pine i n the second growing season i n r e l a t i o n to treatments. (Values outside the range delineated by the v e r t i c a l bar are s i g n i f i c a n t l y d i f f e r e n t , P < 0.05).  85  higher copper dosage applied twice during the growing season, as i n treatments 2 and 8. earlier.  This might be due to phytotoxicity as discussed  A d i f f e r e n t trend was observed i n the case of iron treatments.  Instead, a higher dosage (treatment 5) gave better response than lower iron application as i n treatment 6.  Presumably  because f o l i a r damage was  minimal f o r the higher iron dosage.  Iron and nitrogen treatments each  applied alone gave better response than when applied together as i n treatment 11, but these were less e f f e c t i v e than treatment 13 which combined a low copper dose. (ii)  Douglas-fir  There was also a highly s i g n i f i c a n t difference (P <^ .01) among treatments f o r second-year Douglas-fir f o l i a r mass production i n the main trial  (Table 18).  and treatments.  Figure 22 shows the relationship between f o l i a r mass  Treatment  5 produced the highest response, an increase  of 67 percent over that of untreated trees, followed by a 58 percent increase from treatment 7.  F o l i a r mass from treatment 11 (which produced  the highest response i n shoot increment) was, however, not s i g n i f i c a n t l y d i f f e r e n t from that of untreated trees.  There was no s i g n i f i c a n t treat-  ment e f f e c t on f o l i a r mass production i n the repeat t r i a l (Table 18).  TREATMENT -- No. - -  FeSC\  Urea %  0.5 1  FOLIAGE MASS (g)  0.4-  0.3 -  0  J  — i I  1  1  r~  5  7  II  TREATMENT Figure 22.  NUMBER  F o l i a r mass of Douglas-fir i n the second growing season i n r e l a t i o n to treatments. (Values outside the range delineated by the v e r t i c a l bar are s i g n i f i c a n t l y d i f f e r e n t , P < 0.05).  87  3.  F o l i a r Nutrient Responses  Treatment, s i t e , year and t h e i r interactions a l l had a highly s i g n i f i c a n t e f f e c t (P <^ 0.01)  on f o l i a r nutrients and aluminium i n the  lodgepole pine main t r i a l (Table 19).  The only exception i s zinc.  20 shows the contribution of each factor i n terms of percentage component.  variance  In the repeat t r i a l , treatments had a s i g n i f i c a n t e f f e c t on  most f o l i a r nutrients except potassium, magnesium, boron and zinc 21).  Table  Treatment-site i n t e r a c t i o n was  t o t a l iron and active i r o n .  (Table  s i g n i f i c a n t for aluminium, copper,  Table 22 gives the percentage  variance  component for each factor. For the Douglas-fir main t r i a l (1981  treatments), Table 23 shows  the s t a t i s t i c a l differences among treatments, year and i n t e r a c t i o n i n f o l i a r nutrients.  treatment-year  Treatment-year interaction was  s i g n i f i c a n t only for t o t a l and active i r o n .  highly  Table 24 gives the r e l a t i v e  contribution of each of the factors to the t o t a l source of v a r i a t i o n for these two variables.  In the repeat t r i a l , nitrogen, aluminium, copper,  t o t a l iron and active i r o n were s i g n i f i c a n t l y affected by treatments (Table  25). In the following sections, emphasis i s on elements that were  applied:  copper, i r o n and nitrogen.  on the main t r i a l .  The  The main focus of the discussion i s  trends of current year f o l i a r nutrient response  to treatments for the above nutrients were similar i n the repeat t r i a l . (a)  Copper  Treatments that included copper sulphate resulted i n substantial increase  i n f o l i a r copper i n both the previous and current-year foliage  88  TABLE 19.  Analysis of variance for f o l i a r nutrient responses of lodgepole pine (main t r i a l ) .  Source of variation  N  P  K  Ca  Mg  Al  B  Cu  Fe  AFe  Mn  Zn  Treatment  **  **  **  **  ns  **  **  **  **  **  ns  ns  Year  **  **  **  **  ns  **  **  **  **  **  **  *  Site  *  **  **  **  **  **  **  **  **  **  **  **  **  **  *  ns  ns  ns  ns  **  **  **  ns  *  *  **  **  **  **  **  **  **  **  **  **  jjg  Year x Site  ns  **  *  **  **  **  **  **  **  **  **  ns  Treatment x Year x Site  ns  ns  ns  ns  ns  *  ns  **  **  **  ns  ns  Treatment x Year Treatment x Site  *, ** = Significant at the 5 and 1 percent l e v e l s , ns = Not s i g n i f i c a n t .  respectively,  TABLE 20.  Percentage variance components i n r e l a t i o n to f o l i a r nutrient responses f o r lodgepole pine (main t r i a l ) .  Percentage v a r i a t i o n * Source of variation  N  P  K  Ca  Mg  Al  B  Cu  Fe  AFe  Mn  Zn  Treatment  10.82  21.43  7.69  7.84  1.20  1.90  3.17  14.99  36.90  40.28  1.40  4.50  Year  28.25  21.43  22.44  32.03  0.86  3.86  1.19  6.07  22.84  16.75  8.25  1.51  Site  0.95  7.14  2.56  6.54  49.33  63.39  45.21  4.18  2.29  1.56  45.47  2.99  14.30  7.14  5.45  2.61  0.93  1.37  2.91  12.98  22.03  25.18  1.50  10.91  3.20  7.14  6.09  3.59  4.00  1.84  3.76  13.04  2.34  2.07  3.02  2.10  0.54  1.79  1.92  1.96  2.67  3.92  5.69  5.96  1.75  1.08  3.45  1.52  3.54  2.86  3.21  2.29  2.67  2.36  1.68  14.38  2.27  2.01  2.28  3.79  35.94  28.57  43.27  36.27  28.00  17.67  25.60  30.18  9.90  11.79  25.43  70.38  Treatment x Year Treatment x Site Year x Site Treatment x Year x Site Residual  * Percent variance components were calculated  for variables where interactions were s i g n i f i c a n t .  TABLE 21. Analysis of variance f o r f o l i a r nutrient of lodgepole pine (repeat t r i a l ) .  responses  Source of variation  N  P  K  Ca  Mg  Al  B  Cu  Fe  AFe  Treatment  AA  AA  ns  **  ns  **  ns  AA  AA  AA  ns  ns  AA  AA  AA  AA  A  AA  AA  A  ns  ns  ns  AA  ns  AA  A  AA  ns  ns  Site Treatment x Site  A  AA =  ns  ns  S i g n i f i c a n t at the 5 and 1 percent l e v e l s ,  Mn  Zn  ns  respectively.  ns = Not s i g n i f i c a n t ,  TABLE 22.  Percentage variance components i n r e l a t i o n to f o l i a r nutrient response f o r lodgepole pine (repeat t r i a l ) .  Percent Source of variation Treatment Site Treatment x Site Residual  variation*  Al  Cu  Fe  AFe  8.37  25.54  77.65  70.46  68.92  17.42  2.65  7.14  8.30  32.09  5.43  8.62  14.41  24.93  14.27  13.78  * = Calculated only where main factors interaction were s i g n i f i c a n t .  91  TABLE 23.  Analysis of variance f o r f o l i a r nutrient of Douglas-fir (main t r i a l ) .  responses  Source of variation  N  P  K  Ca  Mg  Al  B  Cu  Fe  AFe  Mn  Zn  Treatment  **  **  ns  ns  **  ns  *  ns  **  **  ns  **  Site  **  **  **  **  ns  **  **  **  **  **  ns  **  Treatment x year  ns  ns  ns  ns  ns  ns  ns  ns  **  **  ns  ns  *, ** = S i g n i f i c a n t at the 5 and 1 percent l e v e l s ,  respectively,  ns = Not s i g n i f i c a n t .  TABLE 24.  Percentage variance components i n r e l a t i o n to f o l i a r nutrient responses f o r Douglas-fir (main t r i a l ) .  „ . Source of v a r i a t i o n c  Percentage v a r i a t i o n * ° Fe  AFe  69.83  64.95  Year  9.27  12.02  Treatment x year  9.66  10.94  11.42  12.47  Treatment  Residual  * Calculated only where interactions were s i g n i f i c a n t .  92  TABLE 25.  Analysis of variance f o r f o l i a r nutrient responses of Douglas-fir (repeat t r i a l ) .  Source of variation  N  P  K  Ca  Mg  Al  B  Cu  Fe  AFe  Mn  Zn  Treatment  **  ns  ns  ns  ns  *  ns  **  **  **  ns  ns  *, ** = Significant at the 5 and 1 percent, respectively, ns = Not s i g n i f i c a n t .  (Figures 23 to 25).  This undoubtedly proves the e f f i c i e n c y of nutrient  absorption of copper by leaves as indicated by Rukovac and Wittwer (1957). The amount of copper absorbed was mainly related to the l e v e l of copper applied, though treatment differences existed from one s i t e to another.  At Site 2, the highest f o l i a r copper l e v e l was attained from  treatment 2, followed i n decreasing order by treatments 8, 12, 10, 3, 14, 9, 4 and 13 (Figure 23).  Copper absorption was higher from treatment  8 than treatment 2 at Site 3 (Figure 24).  At Site 4, the trend i n  decreasing order was treatments 12, 8, 2, 3 and 4.  The common feature,  however, was that the application of 1 percent copper sulphate resulted i n higher f o l i a r copper than the lower rates of 0.1 and 0.2 percent. At Site 2, f o r instance, the actual f o l i a r copper concentration of the current-year foliage from treatment 2 i s 112 ppm compared to about 10 ppm  93  TREATMENT --  No.  25 H  2 2 2 2 2 2 2 2  4 4 2 2  1 0.1 0.2  t  I I I•  5i A !\ /  3  4  \  /  V  T  1  1  1  1  1  1  1  6  7  8  9  10  II  12  13  TREATMENT Figure 23.  Urea  -  4 2  1 0.1 0.2  I \  2  u  95  112  FOLIAR Cu CONC. (ppm)  1 0.1 0.2  7 8 9 10 11 12 13 14  1982  l  %  5 6  1981  lf I I I  FeSO  H  --  1 2 3 4  1980  50-1  CuS0  NUMBER  Foliar copper concentration in relation to treatments at Site 2 (lodgepole pine).  1  14  1982  7  8  1  2 2 2  -  4  11  20-|  ft '  \  '  FOLIAR  \  '  Cu  \  /  \  /  CONC.  \  '  \  '  (ppm) 7  1  IOH  \ \  i  / .•••.\\  Ni—-  / 4  *T"  2  5  TREATMENT F i g u r e 24.  A  *  *  / \ \x  I / \\ \ i/ \\ 1  1/  t  A. W T  7  8  NUMBER  F o l i a r copper c o n c e n t r a t i o n i n r e l a t i o n to treatments a t S i t e 3 ( l o d g e p o l e p i n e ) .  -i  II  95  TREATMENT — No.  1980  CuS0  1  1981  2 3 4 5 6 7 R 11 12  1982  FeSO,, %  u  .  Urea  .  .  1 0.2 0.1 4 2 1  2 2 2 2  4  1  4  51.5  A  20 n  FOLIAR Cu CONC. (ppm) 10  A  If  T  1 2  I At--  1  1  3  4  r-  8  5  TREATMENT Figure 25.  / /  / \  \ \  1  / /  -i  II  NUMBER  F o l i a r copper concentration i n r e l a t i o n to treatments at Site 4 (lodgepole pine).  1  12  96  for treatment 4.  Appendix E . l gives the mean f o l i a r copper values for  a l l treatments i n the main t r i a l . Iron and nitrogen applied i n d i v i d u a l l y or i n combination with each other had no marked effect on f o l i a r copper l e v e l . - This was consistent at a l l the f i v e s i t e s , that i s , for both lodgepole pine and Douglas-fir (Figures 23 to 27).  The f o l i a r copper concentration ranged from 1.5 to  4.2 ppm (Appendix E . l ) . level.  The control trees had a similar range of copper  This suggests that there i s no obvious external (treatment) or  physiological i n t e r a c t i o n of either iron or nitrogen on copper when nutrients are fed to the f o l i a g e .  Such interaction, however, was shown  i n the greenhouse experiments where nutrients were absorbed through the root system. As for yearly v a r i a t i o n , there was a sharp decline i n copper l e v e l i n the second-year f o l i a g e of copper-treated trees (Figures 23 to 25). The general trend shows that treatments which produced a higher increase i n both the, previous and current-year f o l i a g e tend to cause a bigger decrease i n the second-year f o l i a g e . and 12.  These include treatments 2, 3, 8  The lowest decrease was from treatments 4, 9 and 13.  For  instance, f o l i a r copper concentration for treatment 2 at Site 2 decreased from 112 ppm to 7 ppm  (151 percent decrease) as compared to a decrease of  6 ppm (119 percent reduction) for treatment 4 (Figure 23).  The actual  f o l i a r copper concentration of copper-treated trees i n the second- year foliage at the three s i t e s ranged from about 4 to 8  ppm.  There are two implications that could be made here on the behaviour of foliar-absorbed copper.  F i r s t , even though i t i s considered not  readily mobile i n the plant (Mengel and Kirkby 1982), results from this  TREATMENT - - No. --  1980  1 5 7 11  1981 1982  3.0-  •*  FOLIAR Cu CONC. (ppm)  /  /  /  4 4  ' / / / / / / / /  Urea  2 2  \ \ \ \ \ \ \ \ •  "I—  5 TREATMENT  Figure 26.  u  -A  2.0-  1.0  FeSO  7  NUMBER  F o l i a r copper concentration i n r e l a t i o n to treatments at Site 1 (lodgepole pine).  -i  II  1980  TREATMENT -- No. - -  FeSO,  1 5 7 11  1981 1982  Urea  4 2 2  4  4.0-.  FOLIAR Cu CONC. (ppm)  \ \  3.0-  2.0 J  V  , I  ,  ,  5  7  TREATMENT Figure 27.  /  NUMBER  F o l i a r copper concentration i n r e l a t i o n to treatments at Site 5 (Douglas-fir).  , II  99  experiment indicate that i t can be translocated to the newly produced, current-year foliage following f e r t i l i z a t i o n .  This i s evident from the  high copper concentration i n the newly-formed foliage sampled from trees that suffered severe f o l i a r burning (Figures 23 to 25). effect of f o l i a r application of copper i s short-lived.  Secondly, the Loneragan (1981)  hypothesized that copper entering leaves i s bound by nitrogen compounds such as proteins which are retained against transport even during development of copper deficiency.  As shown i n Appendix E . l , the f o l i a r  copper values i n the second-year f o l i a g e are mostly within the deficiency range reported i n the l i t e r a t u r e and also with that determined i n the greenhouse experiment. It seems appropriate at this stage to estimate the c r i t i c a l and toxic l e v e l s of copper f o r lodgepole pine under actual f i e l d conditions. Before making this assessment, i t i s f i r s t necessary to establish that tree growth response was not due to sulphur i n copper sulphate. An organic N/organic S mass r a t i o of 14.6 was determined f o r radiata pine (Kelly and Lambert 1972) and f o r Douglas-fir (Turner et a l . 1977). value presumably also applies to lodgepole pine.  This  Tree growth i s not  sulphur-limited i f the N/S r a t i o i s below this value.  If i t i s near or  above 14.6 there might be sulphur deficiency, and as such, growth response might be due to sulphur application.  In the three copper-  treated stands the N/S r a t i o ranged from 9.6 to 12.6.  Therefore, any  growth response should be attributable only to copper e f f e c t s . As mentioned e a r l i e r , a l l copper treatments produced s i g n i f i c a n t positive response i n biomass production. Among treatments involving the use of copper sulphate alone, the highest 100-needle mass was 2.04 g,  100  obtained from treatment 4 (Figure 21).  Ninety percent of this would then  be a 100-needle mass of 1.84 g, which i s approximately the mass obtained from treatment 3 (Figure 21).  The corresponding 1982 f o l i a r copper  concentration of trees receiving this treatment averaged 4.1 ppm (Appendix E . l ) .  Rounding o f f this value, i t i s suggested that lodgepole  pine growing under similar f i e l d situations with f o l i a r copper below 4 ppm be considered as copper d e f i c i e n t .  This value i s s l i g h t l y greater  than that determined from the greenhouse experiment (2 to 3 ppm). Even though Stone (1968) says that copper t o x i c i t y i s uncommon i n forest trees, high f o l i a r copper levels have been found i n trees at several locations i n B r i t i s h Columbia (Ballard, personal communication), presumably because of copper ore associated with the s o i l parent material.  Also, copper t o x i c i t y can be a serious problem i n forest areas  neighbouring mineral ore processing plants (Lozano and Morrison 1981). In this experiment, the l e v e l of copper t o x i c i t y could only be inferred from the reduction of current-year shoot growth.  The corresponding  f o l i a r copper concentrations at Sites 2, 3 and 4 (Figures 23 to 25), where treatments started to depress shoot growth, ranged from 10.2 to 28.9 ppm, with an average value of 16.6 ppm.  Therefore, i t i s suggested  that copper t o x i c i t y i n lodgepole pine may occur whenever f o l i a r copper concentration exceeds about 17 ppm.  This i s f a i r l y consistent with those  values reported by Reuther and Labanauskas (1972) and, Lozano and Morrison (1982).  (1966), van Lear and Smith  101  (b)  Active  (i)  Lodgepole p i n e  The and  Iron  r e l a t i o n s h i p between a c t i v e i r o n c o n c e n t r a t i o n i n the f o l i a g e  treatments a t the f o u r l o d g e p o l e  28 t o 31. sulphate  pine s i t e s i s i l l u s t r a t e d i n Figures  As i n the case f o r copper, f o l i a r a p p l i c a t i o n of f e r r o u s  proved t o be an e f f i c i e n t means o f r a p i d l y i n c r e a s i n g the a c t i v e  i r o n l e v e l i n the f o l i a g e .  Both p r e v i o u s  and c u r r e n t - y e a r i r o n - t r e a t e d  f o l i a g e had h i g h amounts o f a c t i v e i r o n . I r o n a b s o r p t i o n , however, d i f f e r e d among d i f f e r e n t i r o n combinations.  A pronounced and i n t e r e s t i n g d i f f e r e n c e i s t h a t  a p p l i e d w i t h copper and n i t r o g e n (treatment a c t i v e i r o n l e v e l i n the c u r r e n t - y e a r n i t r o g e n (treatment  treatment iron  12) produced the h i g h e s t  f o l i a g e compared to i r o n  plus  11) or i r o n a p p l i e d alone as i n treatment 5 ( F i g u r e s  29 and 3 1 ) . These t h r e e treatments used the same l e v e l o f f e r r o u s phate (4 p e r c e n t ) .  sul-  Where copper was not a p p l i e d , treatment 11 r e s u l t e d  i n h i g h e r a b s o r p t i o n o f i r o n than treatments 5 or 6 ( F i g u r e s 28 t o 3 1 ) . T h i s t r e n d i s c o n s i s t e n t a t a l l the f o u r s i t e s .  For i r o n applied  alone,  as i n treatments 5 and 6, h i g h e r a b s o r p t i o n o c c u r r e d w i t h treatment 5 ( F i g u r e s 29 and 3 1 ) . T h i s might be due t o h i g h e r dosage from treatment 5.  However, h i g h i r o n supply d i d not n e c e s s a r i l y r e s u l t i n h i g h e r  iron.  foliar  F o r i n s t a n c e , a c t i v e i r o n c o n c e n t r a t i o n from treatments 13 and 14  which used 2 percent  f e r r o u s s u l p h a t e w i t h copper and n i t r o g e n was h i g h e r  than from treatment 5 ( F i g u r e 2 9 ) . According cuticle.  t o Franke (1967) urea improves the p e r m e a b i l i t y of the  The extent  o f urea p e n e t r a t i o n through the c u t i c l e exceeds t h a t  102  TREATMENT — NO.  1980  FeSO^  1 5 7 11  1981 1982  Urea  4 4  3001 FOLIAR ACTIVE Fe CONC. (ppm) 2  0  //  0  il  / 100-  £ I  -V5  7  TREATMENT NUMBER Figure 28.  F o l i a r active iron concentration i n r e l a t i o n to treatments at Site 1 (lodgepole pine).  II  FeS0\ -- % --  TREATMENT - - No. --  1 2 3 4 5 6 7 8 9 10 11 12 13 14  1980 1981 1982  1 0 .1 0.2  4  2  1 0.1 0.2  2 2 2 2 2 2 2 2  4 4 2 2  1 0.1 0.2  300 n  FOLIAR ACTIVE Fe CONC. (ppm) 150 -  0 - -i 1  I  1—i 2  3  1—i 4  5  1—i  1  1—i  6  8  9  7  TREATMENT Figure 29.  10  1 II  1—i—i 12  NUMBER  F o l i a r active iron concentration i n r e l a t i o n to treatments at Site 2 (lodgepole pine).  13  14  104  TREATMENT -- No. --  1980  CuSO^  1 2 5 7 8 11  1981 1982  FeSO^  Urea  4 4  200  FOLIAR A C T I V E Fe -I CONC. (ppm) 100-  \  'I ^•• • • •  I  2  5  TREATMENT Figure 30.  7  8  NUMBER  F o l i a r active iron concentration i n r e l a t i o n to treatments at Site 3 (lodgepole pine).  II  105  TREATMENT No.  CuS0  --  -  1 2 3 4  1980 1981  5  1982  7 R 11 12  FeS0  4  -  Urea  4  %  1 0.2 0.1  4 2  6 1  4 4  1  '  300-|  ;/  FOLIAR ACTIVE Fe . CONC.  !  A  (ppm) 2 0 0  / \ / \ / \ / \ / \  100  1  / /  •••  *  ii  x  "/ • • T  3  4  5  •  ^5t=H=.^ 6  7  -r 8  TREATMENT NUMBER F i g u r e 31.  2 2 2 2  F o l i a r active iron concentration i n r e l a t i o n treatments a t S i t e 4 ( l o d g e p o l e p i n e ) .  T  II  1  12  106  of ions by 10 to 20 f o l d and this increased permeability for urea also favours f o l i a r absorption of ions applied with i t .  This might explain  the higher l e v e l of active iron when ferrous sulphate was applied with urea. Active iron l e v e l i n the current-year foliage showed a s i g n i f i c a n t increase resulting from copper a p p l i c a t i o n as 1 percent copper sulphate (treatment  2) but decreased  for both the lower copper dosage as i n treat-  ments 3 and 4 (Figures 29 and 31).  There was a higher increase i n f o l i a r  active iron from nitrogen applied either alone or together with copper. Average active i r o n concentrations from these two treatments were 44 and 34 ppm, respectively, compared to 24 ppm for the control trees (Appendix E.2).  This indicates that there i s a p o s i t i v e interaction of nitrogen  and nitrogen plus copper on active i r o n .  This i s contrary to the  findings from the greenhouse experiments where increasing nitrogen supply decreased  the active iron concentration i n the f o l i a g e .  As for yearly v a r i a t i o n , there was a s i g n i f i c a n t drop i n active i r o n concentration i n the second-year foliage (Figures 28 to 31).  The  actual active iron values of iron-treated trees at the four lodgepole pine s i t e s ranged from 25 to 46 ppm. probably due to i t s mobility.  The sharp decline i n iron l e v e l i s  Iron has been known to be fixed i n the  leaves with l i t t l e or no translocation to the growing region (Boynton 1954,  Bukovac and Wittwer 1957, Hsu et a l . , 1982). Trends of t o t a l iron concentration w i l l not be discussed i n d e t a i l ,  as they are the same as the trend for active iron, described above. (Appendix E.3 gives actual t o t a l iron values for a l l treatments and s i t e s for the three years of foliage age).  107  The estimation of c r i t i c a l threshold value f o r active iron under f i e l d situations i s also done by f i r s t ascertaining that responses were due to iron alone and not to sulphur i n ferrous sulphate.  The use of N/S  r a t i o discussed i n previous section on copper also applies for i r o n .  A  similar range of N/S values was found for the four iron-treated lodgepole pine stands.  Therefore, as f a r as treatments are concerned, tree growth  response that occurred can be attributed solely to iron, and not to sulphur. Among the two treatments using only ferrous sulphate (treatments 5 and 6), treatment 6 at Site 2 produced the highest shoot growth response (Figure 17).  Ninety percent of this maximum shoot growth would then be  1.22 which i s approximately the response obtained i n 1982 by both t r e a t ments 5 and 6 at Sites 2 and 4 (Figures 17 and 19).  The corresponding  f o l i a r active iron concentration ranged from 25.0 to 32.8 ppm (Appendix E.2).  Averaging these values, one could estimate that the c r i t i c a l  f o l i a r active iron concentration i s about 29 ppm, which agrees with the value reported by Zech (1970) for Scots pine.  This i s s l i g h t l y below the  c r i t i c a l range f o r active iron (32 to 45 ppm) determined i n the greenhouse experiment (ii)  (Chapter 3).  Douglas-fir  Figure 32 i l l u s t r a t e s the relationship between f o l i a r active iron and treatments for Douglas-fir.  There was also a substantial increase i n  active iron concentration i n both the previous and current-year f o l i a g e as a result of iron treatments. As f o r yearly v a r i a t i o n i n active iron l e v e l , there was also a subs t a n t i a l decline from the f i r s t to second-year growing seasons (Figure  108  TREATMENT  1980  No.  1981  1  1982  7  FeSO,,  --  Urea  %  5  4  11  4  2  2  FOLIAR ACTIVE CONC. (ppm)  i I  1  1  5  7  TREATMENT Figure 32.  NUMBER  F o l i a r active iron concentration i n r e l a t i o n to treatments at Site 5 (Douglas-fir).  1  II  109  32).  However, the mean active iron concentration i n the Douglas-fir  second-year foliage f o r treatments 5 and 11 were 79.3 and 89.5 ppm, respectively, and that of the control trees was 31.3 ppm (Appendix E.2). As mentioned e a r l i e r , the corresponding pine ranged from 25.0 to 46.3 ppm.  values f o r iron-treated lodgepole  It appears t h a f Douglas-fir needles  were able to r e t a i n the applied iron longer than lodgepole  pine.  No s a t i s f a c t o r y explanation could be given f o r the above d i f f e r ences.  It i s obvious that d i f f e r e n t species reacted d i f f e r e n t l y to  foliar-applied iron.  The p o s s i b i l i t y that iron was rendered more a v a i l -  able to tree roots during the second-growing season has to be excluded since s o i l iron status i s very low (Table 13).  It could have been  possible that some of the i r o n i n the 1981 foliage was translocated to the 1982 f o l i a g e .  Even though iron i s considered not readily mobile  within the plant (Mengel and Kirkby 1982), i t can be translocated i n the form of iron c i t r a t e to the growing regions of the plant ( T i f f i n 1972, Brown 1978). (c)  Nitrogen  (i)  Lodgepole pine  There was a highly s i g n i f i c a n t difference (P _< .01) among treatments involving the use of 2 percent urea i n terms of f o l i a r nitrogen concentration (Table 19).  In general, a l l treatments caused an increase  i n f o l i a r nitrogen concentration of both the previous and current-year f o l i a g e of the four lodgepole pine stands, but with higher increase i n current-year foliage (Figures 33 to 36). The only exception was at Site 4 where treatments 5 and 8 caused a decrease i n nitrogen concentration of  TREATMENT - - No. - 1980  1 5 7 11  1981 1982  1.6  FeSO,  Urea  4 4  n  FOLIAR N CONC. (%) 1.2 -  0.8  J  1  I  1  1  5  7  TREATMENT NUMBER Figure 3 3 .  F o l i a r nitrogen concentration i n r e l a t i o n to treatments a t S i t e 1 ( l o d g e p o l e p i n e ) .  Ill  TREATMENT -- No.  1 2 3 4 5 6 7 8 9 10 11 12 13 14  1980 1981 1982  0.5-"-i  I  CuSO,,  FeSO % -  1 0.1 0.2  - , 4 2  0.2 1 0.1 0.2  1 .  0  4  1  1  1  1  1  1  "  1  1  '  2  3  4  5  6  7  8  9  10  II  TREATMENT Figure 34.  Urea  H  4  1  -  -  2 2  1  12  NUMBER  F o l i a r nitrogen concentration i n r e l a t i o n to treatments at Site 2 (lodgepole pine).  1  13  2 2 2 2 2 2 2 2  '  14  112  1980 1981 1982  TREATMENT — No.  CuS0  4  FeSO^  Urea  %  1 2 5 7 8  2 2  11  TREATMENT Figure 35.  NUMBER  F o l i a r nitrogen concentration i n r e l a t i o n to treatments at Site 3 (lodgepole pine).  113  TREATMENT  1980  —  CuSO,,  N  FeSO\  -  o  i  1 9 8 1  1982  .  .  0.2 0.1  3  4 5 6 7 8 11 12  Urea  %  .  4 2 1  1  4  4  - . 2 2 2 2  -  1.5-1  /  FOLIAR  /  N CONC.  /  / ^ \  1.2 1  f  /  '  \  /  •  \.  / • •• •  •  •  •• •  0.9 H -i 2  1  1  1  1  1  3  4  5  6  7  TREATMENT Figure .36.  r 8  NUMBER  F o l i a r nitrogen concentration i n r e l a t i o n to treatments at Site 4 (lodgepole pine).  12  114  the previous-year f o l i a g e (Figure 36).  In r e l a t i o n to the untreated  trees, the increase i n the current-year foliage ranged from 9 to 60 percent of the control values. The trend indicates that the application of urea alone  (treatment  7) was less e f f e c t i v e i n increasing f o l i a r nitrogen concentration compared to when i t was applied with copper and/or i r o n , as i n treatments 8, 11 and 12.  For instance, f o l i a r nitrogen concentration i n current-year  foliage f o r treatment 7 ranged from 1.2 to 1.4 percent, and that from treatment 11 ranged from 1.4 to 1.5 percent  (Appendix E.4).  In f a c t ,  treatment 7 resulted i n the lowest f o l i a r nitrogen i n the current-year foliage as compared to the other treatments (Figures 35 and 36).  It  appears that both copper and iron have a s y n e r g i s t i c effect on f o l i a r uptake of nitrogen i n lodgepole pine. There was a decline i n f o l i a r nitrogen concentration for a l l the treated trees i n the second-year f o l i a g e (Figures 33 to 36).  As i n the  case for copper and active iron, the highest decrease was from treatments that o r i g i n a l l y caused higher nitrogen absorption, that i s , treatments 8, 11 and 12. The second-year f o l i a r nitrogen concentration of a l l treated trees at the four s i t e s averaged 1.1 percent. current-year f o l i a g e was 1.3.  The average value for the  Using the c r i t i c a l value of 1.2 percent  suggested by Swan (1972) and Binns e_t a l . (1980) for lodgepole pine, i t can be concluded  that urea a p p l i c a t i o n was e f f e c t i v e i n increasing f o l i a r  nitrogen to above the deficiency l e v e l during the year of a p p l i c a t i o n , but not i n the following year.  This indicates that the effect of f o l i a r  application of urea i s also s h o r t - l i v e d .  115  (ii)  Douglas-fir  There were highly s i g n i f i c a n t (P j< .01) treatment and year effects on f o l i a r nitrogen concentration i n Douglas-fir foliage (Table 23). Treatment-year i n t e r a c t i o n was not s i g n i f i c a n t (P <^ .05). Table 26 compares i n d i v i d u a l treatments means as well as yearly means f o r each treatment. As f o r treatment differences, treatments 5, 7 and 11 s i g n i f i c a n t l y increased f o l i a r nitrogen concentration i n the current year f o l i a g e . There was, however, no s i g n i f i c a n t difference among these three treatments.  As f o r yearly v a r i a t i o n , there was an apparent increase i n f o l i a r  nitrogen from the f i r s t to second-year f o l i a g e f o r treatments 5 and 11, and a decrease f o r treatment 7.  However, accounting f o r v a r i a t i o n i n the  control trees by using the formula i n Section B.8 of this chapter there was an actual decrease i n f o l i a r nitrogen f o r treatments 5, 7 and 11. TABLE 26. Treatment and year effects on f o l i a r nitrogen concentration (%) of Douglas-fir.  Year/  Treatment l  1980  a  1981  a  0.76  0.76  0.88  5b  0.86  0.91  0.98  7  0.80  0.93  0.90  0.86  0.90  0.95  a  b  lib  Each value i s a mean of f i v e trees. In each column and row, values designated 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 at the 5-percent l e v e l .  116  Therefore, the e f f e c t of applying urea to the foliage also did not l a s t beyond the current-growing  season.  Based on the f o l i a r c r i t i c a l value of 1.2 percent for Douglas-fir (Binns et a l . , 1980) and on the f o l i a r nitrogen range from 0.8 to percent attained by urea application, i t could be concluded  0.98  that f o l i a r  application of 2 percent urea f a i l e d to increase f o l i a r nitrogen l e v e l of Douglas-fir to above the c r i t i c a l value for nitrogen deficiency.  D.  Summary of F i e l d Experiment F e r t i l i z e r solutions of copper sulphate, ferrous sulphate and urea  were applied at d i f f e r e n t rates and various combinations lodgepole pine and Douglas-fir at various s i t e s . was  to the crowns of  The primary objective  to determine the effects of f o l i a r application of f e r t i l i z e r s on the  growth and f o l i a r nutrient status. The following results were obtained: 1.  The application of 1 percent copper sulphate alone or i n combination with ferrous sulphate and urea caused very severe needle burn i n lodgepole pine.  The optimum copper dosage for this species was  at 0.1 percent copper sulphate.  Moderate needle burn was  observed  on lodgepole pine treated with 4 percent ferrous sulphate, but not on Douglas-fir. Nitrogen applied as 2 percent urea did not cause any f o l i a r injury for either species. 2.  There was a highly s i g n i f i c a n t positive response i n shoot growth and biomass production i n both species during the second-growing season.  At one of the lodgepole s i t e s , needle length responded  greatly during the second-year of growth.  No positive tree growth  117  response was detected during the year of f e r t i l i z e r application. Treatments that caused minimal f o l i a r scorching gave the best response.  These were 0.1 percent copper sulphate, 4 percent  ferrous sulphate plus 2 percent urea, and urea applied alone at 2 percent. 3.  F o l i a r application of copper and/or iron was highly e f f e c t i v e i n a l l e v i a t i n g deficiencies of these elements i n lodgepole pine, and i r o n deficiency i n Douglas-fir. F o l i a r application was  similarly  highly e f f e c t i v e i n a l l e v i a t i n g nitrogen deficiency i n lodgepole pine, but not i n Douglas-fir. Combined applications of f e r t i l i z e r s were more e f f e c t i v e than i n d i v i d u a l application.  However, the  effects were only temporary and did not l a s t beyond the current growing season.  F o l i a r composition of these three elements was  either within or approaching growing season.  the deficiency range i n the second  Douglas-fir, however, showed a tendency to be able  to r e t a i n more iron than lodgepole pine i n the second year after fertilization. 4.  It i s suggested that the c r i t i c a l l e v e l for copper i n lodgepole pine growing under f i e l d conditions be tentatively set at 4  ppm,  and the c r i t i c a l l e v e l for active iron at about 29 ppm.  estima-  No  tion of the threshold values of these nutrients was made f o r Douglas-fir. 5.  Nitrogen and/or iron treatments did not seem to have any physiol o g i c a l i n t e r a c t i o n with copper.  The application of urea alone and  urea with copper sulphate, however, increased the active iron concentration i n the f o l i a g e .  118  CHAPTER 5:  CONCLUSIONS  Two complementary studies, one i n the greenhouse with lodgepole pine seedlings and the other i n the f i e l d with lodgepole pine and Douglas-fir, were conducted to investigate some aspects of boron, and iron n u t r i t i o n of these two species.  copper,  The greenhouse experiments were  primarily designed to determine the threshold values of boron, copper and iron i n lodgepole pine.  The f i e l d investigation involved f o l i a r  applica-  tion of copper, iron and nitrogen to lodgepole pine and Douglas-fir: ( i ) to determine the c r i t i c a l values of copper and i r o n for lodgepole pine under f i e l d conditions, ( i i ) to evaluate the potential of f o l i a r  fertili-  zation i n r e l i e v i n g deficiencies of copper and i r o n , and ( i i i ) to assess tree growth and f o l i a r nutrient responses to f o l i a r  fertilization.  For lodgepole pine grown under greenhouse conditions, the f o l i a r concentration c r i t i c a l ranges are 7 to 16 ppm f o r boron, 2 to 3 ppm f o r copper; f o r active and t o t a l i r o n , c r i t i c a l levels are 32 and 44 ppm, respectively.  Findings from the f i e l d experiments with lodgepole pine  indicated a c r i t i c a l value of 4 ppm for copper and 29 ppm for active iron.  The threshold values for copper and i r o n estimated f o r seedlings  grown i n the greenhouse are f a i r l y comparable to those determined f o r trees growing under actual f i e l d conditions. These findings support the hypothesis made by Swan (1972) that f o l i a r nutrient concentrations associated with best growth are largely independent  of tree age. I t  would not be grossly inaccurate to state that the p r a c t i c a l significance  119  of the results obtained would be useful to c o r r e c t l y diagnose d e f i c i e n cies of copper and i r o n i n lodgepole pine growing under f i e l d s i t u a t i o n s . For boron, however, f i e l d experimentation i s needed to v e r i f y the results of the greenhouse study. Another important greenhouse finding that might have s i g n i f i c a n t p r a c t i c a l implications relates to the fact that an increase i n f o l i a r nitrogen, as a r e s u l t of increasing nitrogen supply, decreases f o l i a r composition of boron, copper and i r o n .  In many forest stands i n B r i t i s h  Columbia, nitrogen deficiency i s frequently a serious problem, and nitrogen f e r t i l i z a t i o n i s one of the management tools often contemplated. Such a practice might induce deficiencies of these three  micronutrients,  which otherwise would have been adequate for tree growth. In the f i e l d experiment, f o l i a r application of copper and iron proved to be an e f f i c i e n t means of a l l e v i a t i n g d e f i c i e n c i e s of these nutrient elements i n lodgepole pine, and iron i n Douglas-fir.  two  Applica-  tion of 2 percent urea also proved to be e f f e c t i v e i n r e l i e v i n g nitrogen deficiency i n lodgepole pine, but not i n Douglas-fir.  Combined nutrient  applications were more e f f e c t i v e than i n d i v i d u a l nutrient application. This i s highly desirable economically since i t would reduce the costs of applying several elements. The above effects were, however, only temporary and did not  last  beyond the year of application, except for Douglas-fir, which showed a tendency to be able to r e t a i n more of the applied iron than lodgepole pine.  In the case of lodgepole pine, f o l i a r copper, i r o n and  concentrations  nitrogen  were within or approaching the deficiency range during  the  120  second growing season.  This warrants repeated applications of at least  once every growing season, which might not be economically a t t r a c t i v e . As f a r as dosage of application i s concerned,  the use of 0.1  per-  cent copper sulphate for lodgepole pine seemed to be the safest l e v e l , with no f o l i a r injury observed.  Iron applied as 4 percent ferrous  sulphate caused moderate f o l i a r burning for lodgepole pine but not i n Douglas-fir.  A l e v e l of 2 percent ferrous sulphate was  suitable and equally e f f e c t i v e f o r lodgepole pine.  found to be  Nitrogen applied as 2  percent urea did not cause any f o l i a r injury for both species.  It seems  appropriate to conduct further tests with higher levels of urea to determine whether f o l i a r nitrogen response would l a s t for a longer period of time. Tree growth response  to f o l i a r f e r t i l i z a t i o n measured i n terms of  shoot increment and biomass production were s i g n i f i c a n t l y positive for both species only during the second-year of growth.  The non-significant  response during the year of f e r t i l i z e r application could be due to either f o l i a r injury or possibly because of physiological processes that delay growth response.  For lodgepole pine, the highest response was from  treatments that caused minimal or no f o l i a r burning.  These are  0.1  percent copper sulphate, 2 percent urea and 4 percent ferrous sulphate plus 2 percent urea.  Shoot growth response f o r Douglas-fir was highest  from 4 percent ferrous sulphate plus 2 percent urea but foliage mass was affected most by 4 percent ferrous sulphate followed by 2 percent urea. A longer term experiment i s necessary to indicate i f tree growth responses associated with f o l i a r f e r t i l i z i n g are b e n e f i c i a l over long as well as short periods of time.  Despite the short-lived effect on f o l i a r  121 nutrient status, f o l i a r f e r t i l i z a t i o n may deserve further consideration for f e r t i l i z e r application to forested lands. Another important area of research i n f o l i a r f e r t i l i z a t i o n  that  warrants further investigation i s the effect of increasing the dosage of urea on the f o l i a r composition of boron, copper and i r o n .  As indicated  e a r l i e r f o r the greenhouse experiments, where nutrients were absorbed v i a the root system, an increase i n f o l i a r nitrogen was associated with a decrease i n f o l i a r boron, copper and t o t a l and active forms of i r o n . F o l i a r feeding of 2 percent urea i n the f i e l d experiment, on the contrary, did not appear to induce copper deficiency, and i n f a c t , increased the iron l e v e l i n the f o l i a g e .  If such a trend holds true with  increasing supply of urea, f o l i a r f e r t i l i z a t i o n may, i n the future, become an important and promising forest management t o o l i n the P a c i f i c northwest where nitrogen deficiency i s often encountered.  In situations  where both nitrogen and iron d e f i c i e n c i e s occur, the application of urea alone, by f o l i a r feeding, could possibly a l l e v i a t e deficiencies of both elements. It may also be worthwhile to conduct f i e l d experimentation on the effects of s o i l application of nitrogen f e r t i l i z e r s on f o l i a r micronutrient status to test the hypothesis that nitrogen f e r t i l i z a t i o n to induce micronutrient d e f i c i e n c i e s .  tends  If this hypothesis i s confirmed,  f e r t i l i z i n g trees through the foliage might be an alternative method.  122 REFERENCES CITED A l l e n , S.E. 1968. Techniques and problems i n greenhouse evaluation of f e r t i l i z e r sources. In: Bengston, G.W. (ed.) Forest f e r t i l i z a tion: theory and practice. Tenn. V a l . Auth., Muscle Shoals, Alabama, pp. 255-263. A l l i s o n , L.E. Part 2.  1965. Organic carbon. In: Methods of s o i l analysis. Agronomy 9. Am. Soc. Agron., Madison, Wis. pp. 1367-1396.  Armson, K.A. 1973. S o i l and plant analysis techniques as diagnostic c r i t e r i a for evaluating f e r t i l i z e r needs and treatment response. In: Forest f e r t i l i z a t i o n symp. proc. USDA For. Serv. Gen. Tech. Rep. NE-3. pp. 155-166. Armson, K.A. 1974. Predictive techniques i n r e l a t i o n to growth and development of conifers. In: Forest f e r t i l i z a t i o n i n Canada Workshop Proc. Can. For. Serv., Great Lakes For. Res. Cen. For. Tech. Rep. 5:17-21. Aronsson, A. 1983. Growth disturbances caused by boron deficiency i n some f e r t i l i z e d pine and spruce stands on mineral s o i l s . Commun. Inst. For. Fenn. 116:116-122. Ballard, T.M. 1981. F o l i a r analysis research. B.C. Min. For., Res. Branch. 132 pp. Ballard, T.M., status.  and R.E. Carter.- 1983. B.C. Min. For. 76 pp.  Contract Res. Rep. to  Evaluating forest stand nutrient  Ballard, T.M., and N. Majid. 1984. Iron deficiency of lodgepole pine stands on non-calcareous s o i l s i n B r i t i s h Columbia. For. Chron. ( i n review). Barrows, H.L. 1959. Evaluating the micronutrient requirements of trees. In: Mineral n u t r i t i o n of trees symp. Duke Univ. Sch. For. B u l l . 15:18-31. Benzian, B., and R.G. Warren. 1956. Copper deficiency i n Sitka spruce seedlings. Nature 178:864-865. Binns, W.O., G.J. Mayhead, and J.M. MacKenzie. 1980. Nutrient d e f i c i e n cies of conifers i n B r i t i s h Forests: an i l l u s t r a t e d guide. For. Comm. L e a f l e t 76. 23 pp. Bonneau, M. 1973. The state of research i n forest n u t r i t i o n . In: FA0/IUFR0 Int. Symp. on Forest F e r t i l i z a t i o n Proc. Paris, pp. 1-22.  Boynton, D. 1954. N u t r i t i o n by f o l i a r a p p l i c a t i o n . Ann. Rev. Physiol. 5:31-54.  Plant  Bradbury, I.K., and D.C. Malcolm. 1978. Dry matter accumulation by Picea sitchensis seedlings during winter. Can. J . For. Res. 8:207-213. Bremner, J.M. Part 2. 1178.  1965. Total nitrogen. In: Methods of S o i l Analysis. Agronomy 9. Am. Soc. Agron., Madison, Wis. pp. 1149-  Brown, J.C. 1978. Mechanism of iron uptake by plants. Environment 1:249-257.  Plant, C e l l and  Bukovac, M.J., and S.H. Wittwer. 1957. Absorption and mobility of f o l i a r applied nutrients. Plant Physiol. 32:428-435. Bussler, W. 1981. Physiological functions and u t i l i z a t i o n of copper. In: Loneragan, J.F., A.D. Robson, and R.D. Graham (eds.) Copper i S o i l s and Plants. Academic Press, A u s t r a l i a , pp. 213-234. Canada S o i l Survey Committee. 1978. The Canadian system of s o i l c l a s s i f i c a t i o n . Can. Dept. Agric. Publ. 1646, Supply and Services Canada, Ottawa, Ont. 164 pp. Carter, R.E., J . Otchere-Boateng, and K. Klinka. 1984. Dieback of a 30-year-old Douglas-fir plantation i n the B r i t t a i n River Valley, B r i t i s h Columbia: symptoms and diagnosis. For. E c o l . Manage. 7:249-263. Clapp, R.B., and G.M. Hornberger. 1978. Empirical equations for some s o i l hydraulic properties. Water Resour. Res. 14:601-604. Cox, F.R., and E.J. Kamprath. 1972. Micronutrient s o i l s t e s t s . In: Mortvedt, J . J . , P.M. Giordano, and W.L. Lindsay (eds.) Micronutrients i n A g r i c u l t u r e . S o i l S c i . Soc. Am., Madison, Wis. pp. 289-317. C r i t c h f i e l d , W.B. 1980. Genetics of lodgepole pine. Res. Pap. WO-37. 57 pp.  USDA For. Serv.  DeBell, D.S. 1981. Increasing forest productivity through nutrient management. In: Increasing Forest Productivity Proc. National Convention of the Soc. Am. Foresters, pp. 51-55. DeKock, P.C. 1955. 79:167-175.  Iron n u t r i t i o n of plants at high pH.  Soil Sci.  DeKock, P.C. 1981. Iron n u t r i t i o n under conditions of s t r e s s . J . Plant Nutrition 3:513-521.  124  Dickey, B. 1981. The effect of stand density on f o l i a r nutrient concentrations of lodgepole pine stands i n the B.C. I n t e r i o r . B.S.F. Thesis. Univ. of B r i t i s h Columbia, Vancouver, B.C. 41 pp. (Unpubl.). Ebbel, L.F. 1972. Cone-production and stem-growth response of Douglasf i r to rate and frequency of nitrogen f e r t i l i z a t i o n . Can. J . For. Res. 2:327-338. Eberhardt, P.J., and W.L. P r i t c h e t t . 1971. F o l i a r applications of nitrogen to slash pine seedlings. Plant S o i l 34:731-739. Engelstad, O.P., and D.A. Russel. 1975. F e r t i l i z e r s for use under t r o p i c a l conditions. Adv. Agron. 27:175-208. Environment Canada. 1982. Canadian climate normals. climate program, v. 6. 268 pp.  A publ. of Can.  Everard, J . 1973. F o l i a r analysis. Sampling methods, interpretation and application of r e s u l t s . Quart. J . For. 67:51-66. Farhoomand, N.B., and L.A. Peterson. Agron. J . 60:708-709.  1968. Concentration and content.  Fortesque, J.A.C., and G.G. Marten. 1969. Micronutrients: forest ecology and systems a n a l y s i s . Petawawa For. Exp. Stn., Chalk River, Ont. Infor. Rep. PS-X-8. 36 pp. Franke, W. 1967. Mechanisms of f o l i a r penetration of solutions. Ann. Rev. Plant Physiol. 18:281-300. Gaines, T.P., and G.A. M i t c h e l l . 1979. Boron determination i n plant tissue by the azomethine-H method. Commun. S o i l S c i . Plant Anal. 10:1099-1108. Gessel, S.P., K.J. Turnbull, and F.T. Tremblay. 1960. How to f e r t i l i z e trees and measure response. National Plant Food Inst. Washington, D.C. 67 pp. Gholz, H.L. 1978. Assessing stress i n Rhododendron macrophyllum through an analysis of leaf physical and chemical c h a r a c t e r i s t i c s . Can. J . Bot. 56:546-556. Greig, M., and J . B j e r r i n g . 1980. UBC Genlin. A general least squares analysis of variance program. Computing Centre, Univ. B r i t i s h Columbia, Vancouver, B.C. 55 pp. Hewitt, E.J. 1966. Sand and water culture methods used i n the study of plant n u t r i t i o n . Common. Agric. Bur. Tech. Commun. 22. pp. 404416.  125  H i l l , J . , and M.J. Lambert. 1981. Physiology and management of micronutrients i n forest trees i n Australasia. In: Australian Forest N u t r i t i o n Workshop P r o c , Canberra, pp. 93-103. Hsu, H.H., H.D. Ashmead, and D.J. Graff. 1982. Absorption and d i s t r i b u tion of f o l i a r applied i r o n by plants. J . Plant N u t r i t i o n 5:969-974. Ingestad, T. 1958. Studies on manganese deficiency i n forest stands. Medd. Stat. Skogsforsk. 48. 20 pp. Ingestad, T. 1960. Studies on the n u t r i t i o n of forest tree seedlings. I I I . Mineral n u t r i t i o n of pine. Physiol. Plant. 13:513-533. Ingestad, T. 1970. A d e f i n i t i o n of optimum nutrient requirements i n Birch seedlings. I . Physiol. Plant. 23:1127-1138. Ingestad, T. 1979. Mineral nutrient requirements of Pinus s i l v e s t r i s and Picea abies seedlings. Physiol. Plant. 45:373-380. Jones, J.B., and W.J.A. Steyn. 1973. Sampling, handling, and analyzing plant tissue samples. In: Walsh, L.M., and J.D. Beaton (eds.) S o i l Testing and Plant Analysis. S o i l S c i . Soc. Am., Madison, Wis. pp. 249-270. Jyung, W.H., and S.H. Wittwer. 1965. Pathways and mechanisms f o r f o l i a r absorption of mineral nutrients. Agric. S c i . Rev. 3:26-36. K e l l y , J . , and M.J. Lambert. 1972. The relationship between sulphur and nitrogen i n the f o l i a g e of Pinus radiata. Plant S o i l 37:395-407. Khanna, P.K. 1981. S o i l analysis f o r evaluation of forest nutrient supply. In: Australian Forest Nutrition Workshop Proc. pp. 231-238. Knight, P.J. 1975. Copper deficiency i n Pinus radiata i n a peat s o i l nursery. N.Z. J . For. S c i . 5:209-218. Knight, P.J. 1978. F e r t i l i z e r practice i n New Zealand forest nurseries. N.Z. J . For. S c i . 8:27-53. Korstian, C.F., C. Hartley, L.F. Watts, and G.G. Hahn. 1921. A chlorosis of conifers corrected by spraying with ferrous sulphate. J . Agric. Res. 21:153-171. Kramer, P.J., and T.T. Kozlowski. 1979. Academic Press, New York. 811 pp.  Physiology  of Woody Plants.  Lambert, M.J., and T.C. Weidensaul. 1982. Copper requirements of container-grown conifer seedlings. Can. J . For. Res. 12:848-852.  126  Lea, R., W.C. Tierson, D.H. Bickelhaupt, and A.L. Leaf. 1980. Different i a l f o l i a r response of northern hardwoods to f e r t i l i z a t i o n . Plant S o i l 54:419-439. Leaf, A.L. 1968. K, Mg and S deficiencies i n forest trees. In: Bengston, G.W. (ed.) Forest F e r t i l i z a t i o n : Theory and Practice. Ten. V a l . Auth., Muscle Shoals, Alabama, pp. 88-122. Leaf, A.L. 1973. Plant analysis as an aid i n f e r t i l i z i n g f o r e s t s . Walsh, L.M., and J.D. Beaton (eds.) S o i l Testing and Plant Analysis. S o i l S c i . Soc. Am., Madison, Wis. pp. 427-454.  In:  Leece, D.R. 1976. Diagnosis of n u t r i t i o n a l disorders of f r u i t trees by leaf and s o i l analyses and biochemical indices. J . Aust. Inst. Agric. S c i . 42:3-19. Leyton, L. 1958. The relationship between the growth and mineral composition of c o n i f e r s . In: Thimann, K.V. (ed.) The Physiology of Forest Trees. Ronald Press, New York. pp. 323-345. Lindsay, W.L., and W.A. Norvell. 1978. Development of a DTPA s o i l test for zinc, i r o n , manganese and copper. S o i l S c i . Soc. Am. J . 42:421-428. L i t t l e , T.M. 1981. Interpretation and presentation of r e s u l t s . Sci. 16:637-640.  Hort.  Loneragan, J.F. 1981. D i s t r i b u t i o n and movement of copper i n plants. In: Loneragan, J.F., A.D. Robson, and R.D. Graham (eds.) Copper i n S o i l s and Plants. Academic Press, A u s t r a l i a , pp. 165-188. Lowe, L.E., and T. Guthrie. 1981. The development of an a n a l y t i c a l method to determine sulphur content of f o l i a g e . Contract Res. Rep. to B.C. Min. For., Res. Branch. Lozano, F.C., and I.K. Morrison. 1981. Disruption of hardwood n u t r i t i o n by sulphur dioxide, n i c k e l , and copper a i r p o l l u t i o n near Sudbury, Canada. J . Environ. Qual. 10:198-204. Lozano, F.C., and I.K. Morrison. 1982. Growth and n u t r i t i o n of white pine and white spruce seedlings i n solutions of various n i c k e l and copper concentrations. J . Environ. Qual. 11:437-441. Lyle, E.S. 1972. Diagnosing mineral defiency by f o l i a r f e r t i l i z a t i o n . Tree Plant. Notes 23:23-24. Macy, P. 1936. The quantitative mineral requirements Physiol. 11:749-764.  of plants.  Plant  127  McGrath, J.F. 1978. Phosphate and zinc n u t r i t i o n of young Pinus radiata D. Don i n the Donnybrook Sunkland. Western Aust. For. Dept. Res. Pap. 34. 8 pp. Mead, D.J., and R.L. Gadgil. 1978. F e r t i l i z e r use i n established radiata pine stands i n New Zealand. N.Z. J . For. S c i . 8:105-134. Mead, D.J., and W.L. P r i t c h e t t . 1971. A comparison of tree responses to f e r t i l i z e r s i n f i e l d and pot experiments. S o i l S c i . Soc. Am. Proc. 35:346-349. Mengel, K., and E.A. Kirkby. 1982. P r i n c i p l e s of plant n u t r i t i o n . International Potash Inst., Switzerland. 655 pp. M i l l e r , R.E., and R.D. Fight. 1979. F e r t i l i z i n g Douglas-fir f o r e s t s . USDA For. Serv. Gen. Tech. Rep. PNW-83. 25 pp. M i l l e r , R.E., and D.C. Young. 1976. Forest f e r t i l i z a t i o n : f o l i a r a p p l i cation of nitrogen solutions proves e f f i c i e n t . Fert. Solutions 20:36-60. Moller, G. 1983. Variation of boron concentration i n pine needles from trees growing on mineral s o i l i n Sweden and response to nitrogen f e r t i l i z a t i o n . Commun. Inst. For. Fenn. 116:111-115. Morrison, I.K. 1974. Mineral n u t r i t i o n of conifers with s p e c i a l reference to nutrient status interpretation: a review of literature. Can. For. Serv. Pub. No. 1343. 74 pp. Morrison, I.K. 1981. Assessment of the current state of forest f e r t i l i zation research i n Canada. In: Thompson, K.M. (ed.) An Industrial Assessment of Forestry Research i n Canada. Pulp Pap. Res. Inst. Can. 2:115-141. Morrow, L.D., and V.R. Timmer. 1981. Intraseasonal growth and nutrient composition of jack pine needles following f e r t i l i z a t i o n . Can. J . For. Res. 11:696-702. Munson, R.D., and W.L. Nelson. 1973. P r i n c i p l e s and practices of plant analysis. In: Walsh, L.M., and J.D. Beaton (eds.) S o i l Testing and Plant Analysis. S o i l S c i . Soc. Am., Madison, Wis. pp. 223-248. Murphy, L.S., and L.M. Walsh. 1972. Correction of micronutrient deficiencies with f e r t i l i z e r s . In: Mortvedt, J . J . , P.M. Giordano, and W.L. Lindsay (eds.) Micronutrients i n Agriculture. S o i l S c i . Soc. Am., Madison, Wis. pp. 347-387. Neary, D.G., G. Schneider, and D.P. White. 1975. Boron t o x i c i t y i n red pine following municipal waste water i r r i g a t i o n . S o i l S c i . Soc. Am. Proc. 39:981-982.  128  Neumann, P.M., and R. Prinz. 1975. F o l i a r iron spray potentiates growth of seedlings on iron-free media. Plant Physiol. 55:988-990. Oldenkamp, L., and K.W. Smilde. 1966. Copper deficiency i n Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco). Plant S o i l 25:150-152. Olsen, S-R. 1972. Micronutrient i n t e r a c t i o n s . In: Mortvedt, J . J . , P.M. Giordano, and W.L. Lindsay (eds.) Micronutrients i n A g r i c u l ture. S o i l S c i . Soc. Am., Madison, Wis. pp. 243-264. Oserkowsky, 1933. Quantitative r e l a t i o n between chlorophyll and i r o n i n green and c h l o r o t i c pear leaves. Plant Physiol. 8:449-468. Paavelainen, E. 1972. Reaction of Scots pine on various nitrogen f e r t i l i z e r s on drained peatlands. Commun. Inst. For. Fenn. 77. 44 pp. Parker, J . 1956. Variations i n copper, boron and manganese i n leaves of Pinus ponderosa. For. S c i . 2:190-198. Parkinson, J.A., and S.E. A l l e n . 1975. A wet oxidation procedure suitable f o r the determination of nitrogen and mineral nutrients i n b i o l o g i c a l material. Commun. S o i l S c i . Plant Anal., 6:1-11. Pulsford, J . S . 1981. F e r t i l i z e r technology. Nutrition Workshop Proc. pp. 239-252.  In: Australian Forest  Reuther, W., and C.K. Labanauskas. 1966. Copper. In: Chapman, H.D. (ed.) Diagnostic C r i t e r i a f o r Plants and S o i l s . Univ. C a l i f . Div. Agric. S c i . pp. 157-179. Richards, B.N., and D.I. Bevege. 1969. C r i t i c a l f o l i a g e concentrations of nitrogen and phosphorus as a guide to the nutrient status of Araucaria underplanted to Pinus. Plant S o i l 31:328-336. Richards, B.N., and D.I. Bevege. 1972. P r i n c i p l e s and practice of f o l i a r analysis as a basis f o r crop-logging i n pine plantations. I. Basic considerations. Plant S o i l 36:109-119. Ruiter, J . H . 1969. Suspected copper deficiency i n radiata pine. S o i l 31:197-200.  Plant  Ruiter, J.H. 1983. Establishment of Pinus radiata on calcareous Commun. Inst. For. Fenn. 116:91-104.  soils.  Schultz, R.P. 1968. S o i l and f o l i a r f e r t i l i z a t i o n of well drained and flooded slash pine seedlings. USDA For. Serv. Res. Pap. SE-32. 8 pp. Smith, R.B., R.H. Waring, and D.A. Perry. 1981. Interpreting f o l i a r analyses from Douglas-fir as weight per unit of leaf area. Can. J . For. Res. 11:593-598.  129  Snowdon, P. 1982. Diagnosis of boron deficiency i n s o i l s by pot experiments with Pinus r a d i a t a . Aust. For. Res. 12:217-229. Stachurski, A., and J.R. Zimka. 1975. Methods of studying forest ecosystems: leaf area, leaf production, and withdrawal of nutrients from leaves of trees. Ekol. P o l . 23:637-648. Steel, R.G.D., and J.H. T o r r i e . 1960. P r i n c i p l e s and procedures of s t a t i s t i c s . McGraw-Hill Book Co., New York. 481 pp. Steenbjerg, F. 1954. Manuring, plant production and the chemical composition of the plant. Plant S o i l 5:226-242. Stone, E.L. 1968. Microelement n u t r i t i o n of forest trees: a review. In: Bengston, G.M. (ed.) Forest F e r t i l i z a t i o n : Theory and Practice. Tenn. V a l . Auth., Muscle Shoals, Alabama, pp. 132-175. Stone, E.L., C A . H o l l i s , and E.L. Barnard. 1982. Boron deficiency i n a southern pine nursery. South. J . Applied For. 6:108-112. Stone, E.L., and J.M. W i l l . 1965. Boron deficiency i n Pinus radiata and P_. pinaster. For. S c i . 11:425-433. S t r u l l u , D.G., and M. Bonneau. 1978. Contribution al'etude des carences en cuivre chez l e s Abietacees. Can. J . Bot. 56:2648-2659. Swan, H.S.D. 1972. F o l i a r nutrient concentrations i n lodgepole pine as indicators of tree nutrient status and f e r t i l i z e r requirement. Pulp Pap. Res. Inst. Can. Wdlds. Rep. 42. 19 pp. Tamm, C O . 1964. Determination of nutrient requirements of forest stands. In: Romberger, J.A., and P. Mikola (eds.) Int. Rev. Forestry Res. 1:115-170. Academic Press, New York. T i f f i n , L.O. 1972. Translocation of micronutrients i n plants. In: Mortvedt, J . J . , P.M. Giordano, and W.L. Lindsay (eds.) Micronutrients i n Agriculture. S o i l S c i . Soc. Am., Madison, Wis. pp. 199-229. Timmer, V.R., and E.L. Stone. 1978. Comparative f o l i a r analysis of young balsam f i r f e r t i l i z e d with nitrogen, phosphorus, potassium and lime. S o i l S c i . Soc. Am. J . 42:125-130. Tisdale, S.L., and W.L. Nelson. 1975. S o i l F e r t i l i t y and F e r t i l i z e r s . Macmillan Publ. Co., New York. 694 pp. Traynor, J . 1980. Ideas i n S o i l and Plant Nutrition. 120 pp.  Kovak Bks. C a l i f .  130  Turner, J . , M.J. Lambert, and S.P. Gessel. 1977. Use of foliage sulphate concentrations to predict response to urea application by Douglas-fir. Can. J . For. Res. 7:476-480. U l r i c h , A., and F.J. H i l l s . 1973. Plant analysis as an aid i n f e r t i l i z i n g sugar crops: Part 1. Sugar beets. In: Walsh, L.M., and J.D. Beaton. S o i l Testing and Plant Analysis. S o i l S c i . Soc. Am., Madison, Wis. pp. 271-288. V a i l , J.W., M.S. Parry, and W.E. Carlton. 1961. back i n pines. Plant S o i l 14:393-398. van den Burg, J . fertility. 163.  Boron deficiency d i e -  1976. Problems related to analysis of forest s o i l In: Proc. XVI IUFRO World Congress, Norway, pp. 148-  van den Driessche, R. 1969. Tissue nutrient concentrations i n Douglasf i r and Sitka spruce i n sand cultures, and value of nutrient concentration l e v e l s for interpreting f o l i a r analyses. B.C. For. Serv. Res. Note 47. 42 pp. van den Driessche, R. 1974. Prediction of mineral nutrient status of trees by f o l i a r analysis. Bot. Rev. 40:347-394. van Lear, D.H., and W.H. Smith. 1972. Relationships between macro- and micronutrient n u t r i t i o n of slash pine on three coastal p l a i n s o i l s . Plant S o i l 36:331-347. van Overbeek, J.' 1956. regulators. Ann.  Absorption and translocation of plant growth Rev. Plant Physiol. 7:355-372.  Viro, P.J. 1967. One-tree plots i n manuring forest stands. XIV IUFRO Congress 4:597-607. V i r o , P.J. 1970. Time and effect of forest f e r t i l i z a t i o n . Inst. For. Fenn. 70. 17 pp.  In:  Proc.  Commun.  Walker, R.B., S.P. Gessel, and P.G. Haddock. 1955. Greenhouse studies i n mineral requirements of conifers: western red cedar. For. S c i . 1:51-60. Waring, H.D. 1972. Pinus radiata and the nitrogen-phosphorus i n t e r action. In: Boardman, R. (ed.) Australian Forest Tree Nutrition Conf., Canberra, pp. 144-161. Watanabe, F.S., W.L. Lindsay, and S.R. Olsen. 1965. Nutrient balance involving phosphorus, iron and z i n c Proc. S o i l S c i . Soc. Am. 29:562-565. Weetman, G.F., and R. Fournier. 1982. pine response to f e r t i l i z a t i o n . 1289.  Graphical diagnoses of lodgepole S o i l S c i . S o c Am. J . 46:1280-  131  W i l l , G.M. 1961. Mineral requirements J . A g r i c . Res. 4:309-327.  of radiata pine seedlings.  N.Z.  W i l l , G.M. 1971. Copper deficiency i n radiata pine planted on sands at Mangawhai forest. N.Z. J . For. S c i . 2:217-221. W i l l , G.M. 1971. The occurrence and treatment of boron deficiency i n New Zealand pine forests. N.Z. For. Serv. For. Res. Inst. Res. L e a f l e t 32:1-4. Windsor, G.J., and J . K e l l y . 1972. The effects of f e r t i l i z a t i o n on shoot dieback and f o l i a r boron and sulphur concentrations i n several clones of Pinus radiata. In: Boardman, R. (ed.) A u s t r a l i a n Forest Tree Nutrition Conf., Canberra, pp. 241-256. Wittwer, S.H., and F.C. Teubner. 1959. F o l i a r absorption of mineral nutrients. Ann. Rev. Plant Physiol. 10:13-32. Wolf, B. 1971. The determination of boron i n s o i l extracts, plant materials, composts, manures, water and nutrient solutions. Commun. S o i l S c i . Plant Anal. 2:363-374. Wolf, B. 1974. Improvement s i n the azomethine—H method for the determination of boron. Commun. S o i l S c i . Plant Anal. 5:39-44. Woods, R.W. 1983. Trace element problems induced by heavy nitrogen f e r t i l i z a t i o n of Pinus radiata i n South A u s t r a l i a . Commun. Inst. For. Fenn. 116:178-182. Zech, W. 1970. Nadelanalytische Untersuchungen uber die Kalkchlorose der Waldkiefer (Pinus s i l v e s t r i s ) . z.f. Pflanzenernahr. u. Bodenk. 125:1-6. Z o t t l , H.W. 1973. Diagnosis of n u t r i t i o n a l disturbance i n forest stands. In: FAO/IUFRO Int. Symp. on Forest F e r t i l i z a t i o n P r o c , Paris, pp. 75-96.  132  APPENDIX A . l MODIFIED PARKINSON AND ALLEN DIGESTION FOR PLANT TISSUE ANALYSIS  1.  Weigh dried 30-35 blank  1 g (to the nearest mg) subsample of ground plant tissue (oven at 70°C for 3 hours) and place i n 100 mL digestion tube. tubes per set could be prepared, with a reference sample and a i n each set.  2.  Add 5 mL of cone. H2SO4 (reagent grade) to each sample, and mix on a mechanical vibrator immediately.  3.  Dispense 1 mL of Li2S0^ - H2O2 mixture (prepared by mixing 7.0 g Li2S04, 0.21 g selenium powder i n 175 mL 30% H2O2) into each tube. Wait u n t i l reaction (foaming and spattering) ceases before continuing.  4.  Repeat step 3.  5.  Heat the rack of tubes on the digestion block at 360°C. Use discontinuous heating to overcome i n i t i a l foaming; that i s , 20-40 seconds on block, cool for about 2 minutes, 40-50 seconds of heating and cool, 1-2 minutes on block and cool f o r 5-10 minutes.  6.  Add another 1 mL LIS04 - H 0 reaction ceases.  7.  Repeat step 6.  8.  Digest on block f o r 1 1/2 hours at 360°C.  9.  After 1 1/2 hours, remove rack from block. Add 0.5 mL H2O2 to each tube, return rack to block and digest for another 30 minutes.  10.  Repeat step 9.  11.  Remove rack from block and allow digests to cool (approximately 1 hour). Samples should be pale yellow to milky white i n colour.  12.  Add about 80 mL of demineralized water. Allow to cool to room temperature before making to f i n a l volume (100 mL) with demineralized water.  13.  Cover tubes with parafilm or inert stopper, invert 3-4 times to mix, and pour contents into a labelled 125 mL p l a s t i c b o t t l e .  14.  The o r i g i n a l digest solutions are analyzed by atomic absorption spectrophotometry f o r Fe, Mn, Zn, Cu and A l . Total N and P are analysed on the auto-analyzer. A 25x d i l u t i o n of the o r i g i n a l digest solution i s made up for analysis of Ca, Mg and K on the atomic absorption spectrophotometer.  2  2  mixture to each tube.  Wait  till  Total digestion time i s 2 1/2 hours.  133  APPENDIX A.2 NITRIC ACID DIGESTION FOR ANALYSIS OF COPPER AND  IRON IN PLANT TISSUE  1.  Weigh 0.7 g (to the nearest mg) subsample of ground sample (oven dried at 70°C for 3 hours) and place sample into digestion tube. A set of 30-35 tubes could be prepared for one run, each set having a reference sample and a blank.  2.  Add 5 mL cone. HNO3 to sample, mix by swirling and add an addit i o n a l 5 mL of HNO3.  3.  Cover tubes with glass marbles to prevent s p i l l i n g and heat on digestion block at 40°C for 1 hour.  4.  Increase heat up to 140°C and continue heating for 2 hours, counting from time the block reaches 140°C  5.  Remove tubes from block and allow to cool.  6.  Add about 7 mL demineralized water to each tube and mix by swirling. Allow to cool. Pour sample into a 25 mL measuring cylinder. Rinse digestion tube with demineralized water and add rinsings to the cylinder. Make volume up to 25 mL with demineralized water. Cover cylinder with parafilm and mix content by inverting at least three times. Pour contents into a 60 mL p l a s t i c b o t t l e .  7.  Analyze the solution for copper and i r o n by atomic absorption spectrophotometer.  134  APPENDIX A.3 PROCEDURE FOR ACTIVE IRON DETERMINATION IN PLANT TISSUE  1.  Weigh 0.20 g of ground sample (oven-dried at 70°C for 3 hours) into a 60 mL screw-capped p l a s t i c b o t t l e .  2.  Add 10 mL IN HCI (reagent grade) i n demineralized water to each sample. Tightly cap the bottle to prevent leakage. (70 samples could be prepared i n one run).  3.  Shake the bottle horizontally f o r 24 hours on a reciprocating shaker at room temperature. Have a blank and reference samples for each set.  4.  F i l t e r the extract through Whatman No. 41 f i l t e r paper and c o l l e c t the f i l t r a t e i n a p l a s t i c bottle (previously calibrated and marked at 25 mL) or i n a 25 mL volumetric f l a s k .  5.  Add 10 mL IN HCI to the 60 mL sample bottle, shake b r i e f l y by hand, and wash through the same f i l t e r paper into the p l a s t i c bottle or volumetric f l a s k .  6.  Make to 25 mL volume with IN HCI.  7.  Analyze for active i r o n on atomic absorption spectrophotometer. Analysis should be done within 48 hours.  APPENDIX B . l FOLIAR ELEMENTAL CONCENTRATIONS  IN THE BORON EXPERIMENT  Elements Treatment N  P  K  Ca %  Mg  S  Al  B  Cu  BlNl  1.10 (+0.08)  0.21 (+0.04)  0.67 (+0.03)  0.36 (+0.05)  0.25 (+0.04)  0.25 (+0.03)  0.006 (+0.001)  11.7 (+1.7)  4.4 (+0.5)  B2Nl  1.12 (+0.06)  0.21 (+0.03)  0.76 (+0.06)  0.38 (+0.03)  0.26 (+0.02)  0.32 (+0.03)  0.006 (+0.001)  43.4 (+5.4)  B3N1  1.03 (+0.1)  0.21 (+0.02)  0.82 (+0.08)  0.37 (+0.03)  0.26 (+0.02)  0.32 (+0.04)  0.007 (+0.003)  BlN  2  2.00 (+0.1)  0.22 (+0.02)  0.86 (+0.05)  0.25 (+0.02)  0.27 0.16 (+0.008) (+0.02)  B N 2  2  2.01 (+0.1)  0.20 (+0.01)  0.97 (+0.06)  0.29 (+0.01)  0.20 (+0.01)  B N  2  1.93 (+0.07)  0.19 (+0.02)  1.00 (+0.1)  0.26 (+0.03)  0.20 (+0.01)  3  Each value i s the mean of 8 samples. The number i n parenthesis i s the standard  deviation.  Fe  AFe ppm  Mn  Zn  332 (+90)  294 (+59)  47 (+14.9)  4.2 (+0.7)  352 332 (+119) (+106)  300 (+55)  50 (+15)  107.3 (+7.7)  4.2 (+0.8)  424 442 (+178) (+162)  245 (+33)  39 (+5.2)  0.003 (+0.0005)  7.3 (+3.0)  5.1 (+0.3)  218 (+62)  207 (+80)  107 (+11)  27 (+29)  0.33 (+0.01)  0.004 (+0.0005)  15.5 (+2.1)  4.6 (+0.6)  165 (+32)  136 (+38)  75 (+6)  0.34 (+0.04)  0.003 (+0.001)  85.5 (+9.8)  4.3 (+0.8)  205 (+33)  171 (+25)  339 (+91)  16 (+2.8)  14 68 (+6.4) (+2.0)  APPENDIX B.2 FOLIAR ELEMENTAL CONCENTRATIONS  IN THE COPPER EXPERIMENT  Elements Treatment N  K  P  Ca V  _  Mg  S  Al  B  Cu  Fe  AFe  Mn  Zn  ppm  ClNl  1.11 (+0.08)  0.21 (+0.02)  0.85 (+0.08)  0.31 (+0.04)  0.22 (+0.02)  0.28 (+0.02)  0.006 (+0.002)  103.9 (+10.8)  1.1 (+0.5)  349 (+130)  323 (+126)  198 (+29)  43 (+11)  C N!  1.10 (+0.07)  0.20 (+0.02)  0.84 (+0.08)  0.39 (+0.06)  0.27 (+0.03)  0.34 (+0.04)  0.007 (+0.003)  111.7 (+7.3)  2.5 (+1.1)  349 (+66)  297 (+45)  309 (+52)  50 (+12)  C3N1  1.05 (+0.1)  0.19 (+0.01)  0.75 (+0.07)  0.40 (+0.06)  0.26 (+0.02)  0.33 (+0.06)  0.006 (+0.003)  109.2 (+11.7)  5.6 (+1.7)  190 (+129)  174 (+111)  280 (+55)  41 (+6.5)  l«2  2.23 (+0.1)  0.29 (+0.04)  1.1 (+0.01)  0.27 (+0.01)  0.20 (+0.02)  0.36 (+0.04)  0.005 (+0.002)  77.0 (+5.3)  0.9 (+0.6)  304 (+76)  289 (+81)  102 (+21)  24 (+4.6)  1.97 (+0.1)  0.21 (+0.02)  1.1 (+0.01)  0.27 (+0.01)  0.19 (+0.02)  0.34 (+0.03)  0.003 (+0.001)  77.1 (+5.9)  1.8 (+1.6)  160 (+37)  139 (+29)  68 (+7.8)  16 (+3.1)  1.83 (+0.07)  0.18 (+0.01)  0.9 (+0.05)  0.30 (+0.02)  0.22 (+0.01)  0.37 (+0.02)  0.003 (+0.001)  83.6 (+8.7)  3.0 (+0.6)  162 (+52)  132 (+31)  69 (+7.8)  14 (+1.5)  2  C  C N 2  2  C N  2  3  Each value i s the mean of 8 samples. The number i n parenthesis i s the standard d e v i a t i o n .  APPENDIX B.3 FOLIAR ELEMENTAL CONCENTRATIONS  IN THE IRON EXPERIMENT  Elements Treatment N  P  F1N1  1.14 (+0.1)  0.20 (+0.02)  0.83 (+0.07)  0.39 (+0.04)  F Ni  0.98 (+0.08)  0.20 (+0.02)  0.79 (+0.06)  F3Nl  1.09 (+0.09)  0.18 (+0.01)  1.74 (+0.1)  2  F  1 2 N  F N 2  2  F3N2  K  Ca  S  Al  Cu  Fe  AFe  0.30 (+0.02)  0.37 (+0.04)  0.007 (+0.001)  100.0 (+7.3)  4.7 (+1.2)  48.2 (+6.5)  37.7 (+13.2)  374 (+53)  86 (+11.9)  0.36 (+0.04)  0.28 (+0.03)  0.29 (+0.03)  0.006 (+0.001)  97.2 (+12.8)  3.2 (+0.4)  81.4 (+30.3)  54.8 (+14.3)  338 (+87)  71 (+17.5)  0.75 (+0.08)  0.43 (+0.05)  0.27  0.35  (+0.03)  0.008 (+0.001)  103.5 (+8.2)  3.7 (+0.5)  359.4 (+127)  351.1 (+120)  287  41  (+0.05)  (+39)  (+6.3)  0.19 (+0.02)  0.90 (+0.06)  0.36 (+0.04)  0.26 (+0.02)  0.40 (+0.04)  0.007 (+0.002)  99.1 (+8.5)  3.9 (+0.9)  44.0 (+10.9)  31.6 (+5.7)  136 (+26)  24 (+6.7)  1.81 (+0.08)  0.18 (+0.02)  0.94 (+0.09)  0.31 (+0.02)  0.22 (+0.01)  0.36 (+0.04)  0.006 (+0.001)  84.6 (+6.4)  3.4 (+0.3)  52.7 (+6.2)  45.3 (+3.5)  83 (+6)  15 (+2.8)  1.84 (+0.06)  0.18 (+0.01)  0.90 (+0.09)  0.30 (+0.01)  0.22 (+0.01)  0.36 (+0.03)  0.006 (+0.001)  89.6 (+5.5)  3.5 (+0.6)  146.3 (+28.9)  147.0 (+26.3)  66 (+7.6)  13 (+1.6)  Each value i s the mean of 8 samples. The number i n parenthesis i s the standard d e v i a t i o n .  Mg  B  Mn  Zn  138  APPENDIX C l SOIL PROFILE DESCRIPTION OF SITE 1  Horizon  LFH  Depth (cm)  5-0  Description  Very dark gray (7.5 YR 3/0, moist); fresh and p a r t i a l l y decomposed organic matter, fibrous, few, f i n e and medium roots; abrupt, smooth boundary; 4-6 cm thick; pH 4.0 (1/8 s o i l / H 0 ) . 2  Ae  0-2  Pinkish gray (7.5 YR 6/2, moist); sandy loam; single grain; loose, f r i a b l e ; s l i g h t l y s t i c k y , non-plastic; few, f i n e and medium horizontal roots; very few, f i n e , vesicular pores; abrupt, wavy boundary; 0-4 cm thick; about 5% coarse fragments; pH 4.2 (1/2 soil/H 0). 2  Bf  2-12  Reddish yellow (5 YR 4/6, moist); loam; weak, f i n e to medium subangular blocky; f r i a b l e , s l i g h t l y s t i c k y , non-plastic; few, f i n e , and p l e n t i f u l medium horizont a l roots; very few, medium, oblique, matrix, vesicul a r pores; clear, smooth boundary; 8-11 cm thick; about 5% coarse fragments; pH 5.0 (1/2 s o i l / H 0 ) . 2  BC  12-33  Brownish yellow (10 YR 4/6, moist); loamy sand, single grain; loose, f r i a b l e ; non-sticky, p l a s t i c ; few, f i n e and p l e n t i f u l , medium horizontal roots; very few, f i n e oblique, matrix, vesicular pores; gradual smooth boundary; 8-24 cm thick; about 7% coarse fragments; pH 5.1 (1/2 s o i l / H 0 ) . 2  C  33+  Dark yellowish brown (10 YR 3/6, moist); sand, single grain; loose, f r i a b l e ; non-sticky, non-plastic; few, f i n e , oblique, matrix, vesicular pores; about 15% coarse fragments, pH 5.6 (1/2 s o i l / H 0 ) . 2  CLASSIFICATION:  ORTHIC HUMO-FERRIC P0DZ0L  139  Appendix C.l (Cont'd).  A s o i l p r o f i l e of S i t e 1 .  140  APPENDIX C.2 SOIL PROFILE DESCRIPTION OF SITE 2  Horizon  Depth (cm)  Description  LFH  5-0  Very dark gray (7.5 YR 3/0, moist); semidecomposed organic matter; fibrous, abundant f i n e roots; abrupt, smooth boundary; 3-6 cm thick; pH 4.6 (1/8 S 0 H / H 2 O ) .  Ae  0-6  Brown (7.5 YR 5/2, moist); loam, weak, medium, subangular blocky; f r i a b l e , s l i g h t l y sticky, nonp l a s t i c ; abundant f i n e horizontal roots; few, f i n e , oblique vesicular pores; abrupt, wavy boundary; 4-7 cm thick; about 5% coarse fragments; pH 4.7 (1/2 soil/H 0). 2  Bf  6-12  Brown to dark brown (5 YR 4/4, moist); sandy loam; weak, f i n e to medium subangular blocky; f r i a b l e , s l i g h t l y s t i c k y , non-plastic; p l e n t i f u l f i n e , horizontal roots; p l e n t i f u l , medium, oblique, vesicular pores; clear wavy boundary; 5-8 cm thick; about 5% coarse fragments; pH 5.5 (1/2 s o i l / ^ O ) .  BC  12-30  Strong brown (10 YR 5/6, moist); gravelly sandy loam; weak, f i n e to medium subangular blocky; f r i a b l e , s l i g h t l y s t i c k y , non-plastic; few f i n e , horizontal roots; p l e n t i f u l , medium, oblique, vesicular pores; clear, wavy boundary, 16-20 cm thick; about 25% coarse fragments; pH 5.6 (1/2 S 0 H / H 2 O ) .  C  30-55+  Dark yellowish brown (10 YR 4/6, moist); gravelly sand, single grain; loose, f r i a b l e ; non-sticky, non-plastic; few, f i n e horizontal roots; few f i n e , oblique, vesicular pores; about 25% coarse fragments, pH 5.7 (1/2 s o i l / H 0 ) . 2  CLASSIFICATION:  ELUVIATED DYSTRIC BRUNISOL  Appendix C.2 ( C o n t ' d ) .  A s o i l p r o f i l e of S i t e 2.  142  APPENDIX C.3 S o i l P r o f i l e Description of Site 3  Horizon  LFH  Depth (cm)  3-0  Description  Very dark gray (7.5 YR 3/2, moist); fresh and p a r t i a l l y decomposed organic matter; fibrous, fine roots; abrupt boundary; 4-5 cm thick; pH (1/8 s o i l / H 0 ) .  few 4.9  2  Ae (volcanic ash)  0-9  Light gray (10 YR 7/2, moist); very gravelly sand; single grain; loose, f r i a b l e , non-sticky, nonp l a s t i c ; few fine horizontal roots; gradual smooth boundary; 3-5 cm thick; about 40% coarse fragments; pH 5.7 (1/2 s o i l / H 0 ) . 2  Bt  9-14  Greyish brown (10 YR 5/2, moist); clay loam; moderate medium, subangular blocky; s l i g h t l y hard, firm, sticky, s l i g h t l y p l a s t i c ; few fine and medium h o r i zontal roots; very few, very f i n e , oblique vesicular pores; gradual wavy boundary; 8-10 cm thick; about 15% coarse fragments; pH 5.7 (1/2 s o i l / H 0 ) . 2  BC  14+  Brown (10 YR 5/3, moist); gravelly clay loam; weak to moderate, medium, subangular blocky; s l i g h t l y hard, firm, s l i g h t l y sticky, s l i g h t l y p l a s t i c ; few, f i n e and medium horizontal roots; few, f i n e , oblique, vesicular pores; about 25% coarse fragments; pH 5.7 (1/2 s o i l / H 0 ) . 2  CLASSIFICATION:  ORTHIC GRAY LUVISOL  143  Appendix C.3 ( C o n t ' d ) .  A s o i l p r o f i l e o f S i t e 3.  144  APPENDIX C.4 S o i l P r o f i l e Description of Site 4  Horizon  LF(H)  Depth (cm)  3-0  Description  Black (10 YR 2/1, moist); fresh and p a r t i a l l y decomposed organic matter; clear, wavy boundary, 3-6 cm thick; pH 5.7 (1/8 s o i l / H 0 ) . 2  Ae  0-7  Light brownish gray (10 YR 5/2, moist); loamy sand; single grain; loose, f r i a b l e , non-sticky, nonp l a s t i c ; very f i n e and few, f i n e , horizontal roots; p l e n t i f u l very f i n e , oblique, vesicular pores; clear wavy boundary; 6-8 cm thick; about 5% coarse f r a g ments; pH 5.9 (1/2 s o i l / H 0 ) . 2  Bm  7-25  Brown (10 YR 5/2, moist); loamy sand, single grain; loose, f r i a b l e , non-sticky, p l e n t i f u l , very f i n e and few, f i n e horizontal roots; p l e n t i f u l , very f i n e , oblique vesicular pores; clear, wavy boundary; 6-8 cm thick; about 5% coarse fragments; pH 5.6 (1/2 s o i l / H 0 ) , 2  Bt  25-80  Brown to dark brown (10 YR 4/3, moist); clay loam; moderate, medium, prismatic; s l i g h t l y hard, firm, sticky, s l i g h t l y p l a s t i c ; few f i n e horizontal roots; very few, very f i n e , oblique, vesicular pores; c l e a r , wavy boundary; 17-25 cm thick; about 15% coarse fragments; pH 5.3 (1/2 s o i l / H 0 ) . 2  CLASSIFICATION:  BRUNISOLIC GRAY LUVISOL  145  Appendix C.4 ( C o n t ' d ) .  A s o i l p r o f i l e of S i t e 4.  146  APPENDIX C.5 S o i l P r o f i l e Description of Site 5  Horizon  Depth (cm)  LFH  5-0  Description  Dusky red (2.5 YR 3/2, moist); fresh and p a r t i a l l y decomposed organic matter; abrupt, smooth boundary; 0-8 cm thick; pH 6.5 (1/8 s o i l / H 0 ) . 2  Ck  0-50+  Dark grayish brown (10 YR 4/2, moist); gravelly sandy loam; single grain; loose, f r i a b l e , s l i g h t l y sticky, non-plastic; few, very f i n e to f i n e horizontal roots; very few and f i n e , oblique, vesicular pores; about 50 % coarse fragments; pH 7.5 (1/2 s o i l / H 0 ) . 2  CLASSIFICATION:  ORTHIC REGOSOL  Appendix C.5 (Cont'd).  A s o i l p r o f i l e of Site 5.  148  Appendix D.2.  Copper and nitrogen treatment: current-year growth (lodgepole pine).  Appendix D.3.  Copper, i r o n and n i t r o g e n treatment: c u r r e n t - y e a r growth ( l o d g e p o l e p i n e ) .  151  Appendix D.4.  Copper treatment: (lodgepole p i n e ) .  second-year growth  152  Appendix D.5.  Copper and n i t r o g e n t r e a t m e n t : second-year growth ( l o d g e p o l e p i n e ) .  153  Appendix D . 6 .  Copper, iron and nitrogen treatment: second-year growth (lodgepole pine).  155  Appendix D.8.  Appendix D.9.  Iron treatment: current-year growth (lodgepole pine).  Iron and nitrogen treatment: currentyear growth (lodgepole pine).  156  Appendix D.10.  Iron treatment: second-year growth (lodgepole pine).  157  Appendix D . l l .  Iron and n i t r o g e n treatment: second-year growth ( l o d g e p o l e  pine).  158  Appendix D.13.  Iron and nitrogen treatment: growth (Douglas-fir).  current-year  Appendix D.14.  Nitrogen treatment: current year growth (lodgepole pine)  Appendix D.15.  Nitrogen treatment: (Douglas-fir).  current-year growth  APPENDIX E . l .  MEAN VALUES FOR FOLIAR COPPER CONCENTRATION (PPM) OF LODGEPOLE PINE AND DOUGLAS-FIR IN RELATION TO TREATMENTS (MAIN TRIAL).  Treatment  1980  Site  1. C o n t r o l  2. 1Z C11SO4  3. 0.2Z C11SO4  4. 0.1Z C11SO4  5. 4Z FeS04  1  2  3  1981 4  5  1  2  1982  3  4  5  1  2  3  4  X  1.50  1.64  0.71  2.86  2.64  2.42  2.00  2.21  3.28  3.50  3.07  3.14  2.00  1.21  3.43  0  0.30  0.48  0.00  1.69  0.41  0.53  0.60  0.58  0.64  0.69  0.32  0.30  0.78  0.41  0.20  n  5  5  5  5  5  5  5  5  5  5  5  5  5  5  5  X  NA  _  19.02  9.05  NA  NA  112.14  10.21  10.57  NA  NA  6.50  5.14  3.65  NA  a  NA  6.08  NA  NA  73.95  3.97  4.79  NA  NA  1.17  2.43  1.05  NA  n  0  -  6.50 4  3  0  0  5  5  0  0  5  5  5  0  X  NA  19.11  NA  13.40  NA  NA  28.93  NA  9.21  NA  NA  3.86  NA  4.33  NA  a  NA  1.58  NA  2.55  NA  NA  16.14  NA  2.37  NA  NA  0.53  NA  0.47  NA  n  0  4  0  4  0  0  0  5  0  0  5  0  5  0  0  X  NA  7.98  NA  6.25  NA  NA  0  NA  2.58  NA  3.56  NA  NA  n  0  3  0  4  0  0  3  5  -  10.36  NA  5.86  NA  NA  3.93  NA  3.86  NA  4.69  NA  2.31  NA  NA  0.72  NA  0.59  NA  5  0  5  0  0  5  0  5  0  X  1.50  2.22  3.29  1.07  2.79  3.14  3.71  2.21  3.21  3.93  3.43  2.71  2.86  4.00  4.00  a  0.16  0.64  0.59  0.0  0.69  0.30  0.99  0.30  0.98  0.84  0.54  0.41  1.04  0.59  0.78  n  5  5  5  3  5  5  5  5  5  5  5  5  5  5  5  APPENDIX E . l .  Treatment  1980  Site  6.  1  2% FeS04  7.  2% urea  8.  1% CuS0  +  9.  LO.  4  2% urea  0.1%  +  CUSO4  2% urea  0.2% CuS0  +  (cont'd)  2% urea  4  2  3  1981 4  5  1  2  1982  3  4  5  1  2  3  4  5  X  NA  1.97  NA  1.86  NA  NA  2.71  NA  3.43  NA  NA  3.36  NA  3.21  a  NA  0.20  NA  0.96  NA  NA  0.41  NA  0.54  NA  NA  1.15  NA  0.72  NA  n  0  4  0  5  0  0  5  0  5  0  0  5  0  5  0  X  3.29  2.95  3.07  2.07  2.57  3.29  2.71  2.64  3.57  2.86  3.28  3.64  2.28  3.71  3.71  0  2.04  0.18  0.19  0.92  0.39  0.46  0.96  0.32  0.36  0.36  0.30  1.39  0.54  1.03  0.82  n  5  4  5  5  5  5  5  5  5  5  5  5  5  5  5  X  NA  _  20.83  4.29  NA  NA  95.09  a  NA  -  10.00  0.0  NA .  NA  87.0  n  0  1  0  0  0  3  4  NA  12.93  11.64  NA  NA  7.86  6.07  4.00  NA  4.83  3.14  NA  NA  1.54  2.30  0.64  NA  5  5  0  0  5  5  5  0  X  NA  13.21  NA  NA  NA  NA  14.71  NA  NA  NA  NA  4.50  NA  NA  NA  a  NA  0.0  NA  NA  NA  NA  8.00  NA  NA  NA  NA  1.67  NA  NA  NA  n  0  2  0  0  0  0  0  0  0  0  0  5  0  0  0  X  NA  13.21  NA  NA  NA  NA  33.22  NA  NA  NA  NA  5.00  NA  NA  NA .  a  NA  0.0  NA  NA  NA  NA  12.84  NA  NA  NA  NA  1.43  NA  NA  NA  n  0  1  0  0  0  0  0  0  0  0  4  0  0  0  5  APPENDIX E . l .  Treatment  1980 1  Site  4% FeS0  11.  1% CuS0  + 4% FeS0 + 2% urea 13.  14.  5  1  2  1982 4  3  5  1  2  3  4  5  1.79  2.50  2.59  4.20  3.14  3.36  2.57  2.68  3.29  3.64  3.71  2.43  3.57  3.36  0.0  0.62  0.18  1.00  0.30  0.78  0.93  1.14  0.46  0.64  0.60  0.16  0.80  0.82  0.59  n  1  3  4  3  5  5  5  5  5  5  5  5  5  5  5  4  X  NA  _  NA  51.50  NA  NA  48.84  NA  15.79  NA  NA  4.91  NA  5.71  NA  4  a  NA  -  NA  0.0  NA  NA  11.03  NA  8.94  NA  NA  0.73  NA  1.52  NA  n  0  0  1  0  0  4  0  5  0  0  4  0  5  0  4  4  0.2% CuS0  + 2% FeS0 + 2% urea  1981 4  3  X  0.1% CuS0  + 2% FeS0 + 2% urea  2  0  4  + 2% urea  12.  (cont'd)  4  4  0  3.29  X  NA  4.82  NA  NA  NA  NA  8.86  NA  NA  NA  NA  4.07  NA  NA  NA  0  NA  0.25  NA  NA  NA  NA  2.01  NA  NA  NA  NA  1.30  NA  NA  NA  n  0  2  0  0  0  0  5  0  0  0  0  5  0  0  0  X  NA  4.29  NA  NA  NA  NA  16.64  NA  NA  NA  NA  3.64  NA  NA  NA  a  NA  0.0  NA  NA  NA  NA  11.25  NA  NA  NA  NA  0.64  NA  NA  NA  n  0  1  0  0  0  0  0  0  0  0  0  0  0  NA =  Treatment not a p p l i e d .  -  =  M i s s i n g value. Average concentration value.  x  =  a  =  Standard d e v i a t i o n ,  n  =  Number of samples.  5  5  APPENDIX E.2.  MEAN VALUES FOR FOLIAR ACTIVE IRON CONCENTRATION (PPM) OF LODGEPOLE PINE AND DOUGLAS-FIR IN RELATION TO TREATMENTS (MAIN TRIAL).  Treatment  1980  Site  1. C o n t r o l  1  3. 0.2% C11SO4  4. 0.1Z C11SO4  4  5  1  2  3  1982 4  5  1  2  3  4  51.25  51.00  33.75  31.25  34.05  24.25  25.75  30.20  15.50  37.00  33.30  18.50  23.75  31.50  o  6.56  4.45  5.38  3.19  7.09  11.94  6.82  3.84  5.27  10.48  7.65  3.58  2.34  12.10  5  5  5  5  5  5  5  5  5  5  5  5  5  5  5  31.25 5.99 5  x  NA  -  35.31  70.42  NA  NA  49.25  34.50  20.25  NA  NA  20.00  22.25  38.50  NA  a  HA  -  4.83  46.07  NA  NA  26.75  2.88  4.87  NA  NA  5.30  6.02  8.90  NA  n  0  0  3  0  4  3  0  5  5  0  0  5  5  5  0  x  NA  35.00  NA  21.88  NA  NA  23.25  NA  11.25  NA  NA  20.30  NA  32.05  NA  a  NA  0.0  NA  6.65  NA  NA  5.97  NA  2.65  NA  NA  1.85  NA  5.62  NA  n  0  4  0  4  0  0  5  0  5  0  0  5  0  5  0  x  NA  40.42  NA  34.38  NA  NA  25.50  NA  14.50  NA  NA  26.00  NA  38.00  NA  0  NA  5.05  NA  16.85  NA  NA  11.06  NA  5.77  NA  NA  3.58  NA  6.94  NA  3  0  0  0  5  0  5  n  5. 4Z FeS04  1981  3  x n  2. 1% CuS04  2  O  4  X  207.75  213.75  127.75  214.17  199.75  a  34.62  58.18  26.08  63.84  61.83  n  5  5  5  3  5  0  5  0  223.25 161.25 111.75 101.25 178.25 105.14 5  23.78 5  16.24 5  5  31.46 5  36.79  0  5  0  41.50  25.00  35.75  42.00  79.25  3.79  6.25  9.30  9.42  13.39  5  5  5  5  5  APPENDIX E.2.  Treatment  1980  Site  6.  2%  7.  1  FESO4  2% urea  8.  1%  +  CUSO4  2% urea  0.1%  +  CUSO4  2% urea  0.2% CuS0  10.  +  2% urea  4  2  3  1981 4  5  1  2  3  1982 4  5  1  2  3  4  X  NA  153.75  NA  78.75  NA  NA  165.50  NA  76.50  NA  NA  32.75  NA  30.75  NA  a  NA  23.32  NA  33.92  NA  NA  60.93  NA  40.46  NA  NA  8.50  NA  11.10  NA  n  0  0  0  0  0  5  0  4  0  5  5  0  5  5  0  X  38.00  24.06  27.81  30.00  40.25  26.00  35.05  38.50  29.25  23.76  36.00  26.00  23.00  29.75  a n  27.82  7.86  3.89  11.69  9.32  8.72  3.41  6.75  13.71  2.65  2.71  3.89  6.22  2.85  4.:  4  5  5  5  5  5  5  5  5  5  5  5  5  5  5  30.!  X  NA  _  28.33  34.25  NA  NA  54.50  37.00  33.40  NA  NA  25.75  30.75  27.75  NA  a  NA  -  9.71  0.0  NA  NA  15.58  2.09  2.23  NA  NA  3.14  3.14  9.24  NA  3  1  0  0  5  5  0  0  5  5  5  0  0  9.  (cont'd)  0  4  X  NA  45.00  NA  NA  NA  NA  45.94  NA  NA  NA  NA  28.50  NA  NA  NA  a  NA  0.0  NA  NA  NA  NA  2.71  NA  NA  NA  NA  7.68  NA  NA  NA  n  0  2  0  0  0  0  5  0  0  0  0  5  0  0  0  X  NA  20.00  NA  NA  NA  NA  47.25  NA  NA  NA  NA  28.44  NA  NA  NA  a  NA  0.0  NA  NA  NA  NA  13.27  NA  NA  NA  NA  3.59  NA  NA  NA  n  0  1  0  0  0  0  0  0  0  0  4  0  0  0  5  APPENDIX E.2. (cont'd)  Treatment  1980  Site  11.  4% F e S 0  1  4  + 2% urea  2  3  1981 4  5  1  2  3  1982 4  5  1  2  3  x  336.25  287.50  192.50  285.42  210.50  240.00  233.50  123.25  222.75  178.00  46.25  36.50  38.50  33.00  89.50  a  0.0  12.31  53.60  56.02  51.28  79.26  93.17  27.31  69.49  44.89  3.42  4.18  14.93  5.42  24.54  n  1  5  5  3  4  3  5  5  5  5  5  5  5  5  5  1% CUSO4  x  NA  -  NA  315.50  NA  NA  279.06  NA  287.00  NA  NA  29.06  NA  26.25  NA  + 4% FeSO^  a  NA  -  NA  0.0  NA  NA  104.76  NA  77.15  NA  NA  2.37  NA  7.23  NA  + 2% urea  n  0  0  0  1  4  0  5  0  12.  13.  0.1% CUSO4 + 2% F e S 0  4  + 2% urea  14.  0.2% CUSO4 + 2% F e S 0  4  0  0  4  0  5  0  0  x  NA  138.75  NA  NA  NA  NA  196.25  NA  NA  NA  NA  32.00  NA  NA  NA  a  NA  10.61  NA  NA  NA  NA  76.72  NA  NA  NA  NA  5.63  NA  NA  NA  n  0  2  0  0  0  0  0  0  0  0  5  0  0  0  5  x  NA  183.75  NA  NA  NA  NA  176.75  NA  NA  NA  NA  33.75  NA  NA  NA  o  NA  0.0  NA  NA  NA  NA  78.88  NA  NA  NA  NA  3.64  NA  NA  NA  n  0  0  0  0  0  0  0  0  0  5  0  0  0  + 2% urea  NA  =  Treatment  -  =  Missing value.  x  =  Average c o n c e n t r a t i o n v a l u e ,  o  =  Standard d e v i a t i o n ,  n  =  Number of samples.  1  5  not a p p l i e d .  ON ON  APPENDIX E.3.  MEAN VALUES FOR FOLIAR TOTAL IRON CONCENTRATION (PPM) OF LODGEPOLE PINE AND DOUGLAS-FIR IN RELATION TO TREATMENTS (MAIN TRIAL).  Treatment  1980  Site  1. C o n t r o l  2. 1% CuS0  4  3. 0.2% C11SO4  4. 0.1% C11SO4  1  4  1981  3  4  5  1  2  3  1982 4  5  1  2  3  4  5  x  72.21  46.71  56.57  51.79  61.07  55.93  43.21  30.71  31.85  38.79  35.36  24.07  31.43  35.36  49.50  o  7.06  13.17  7.38  7.75  6.96  9.73  13.67  6.47  1.46  4.10  9.65  1.92  5.16  14.23  10.34  n  5  5  5  5  5  5  5  5  5  5  5  5  5  5  5  x  NA  -  56.75  111.07  NA  NA  56.47  36.57  38.13  NA  NA  25.14  35.86  31.36  NA  o  NA  -  5.28  36.29  NA  NA  24.14  7.55  7.22  NA  NA  3.69  6.47  7.36  NA  n  0  0  3  0  4  3  0  5  5  0  0  5  5  5  0  x  NA  47.41  NA  49.11  NA  NA  44.57  NA  32.57  NA  NA  26.25  NA  27.15  NA  a  NA  2.72  NA  10.36  NA  NA  14.11  NA  5.40  NA  NA  1.39  NA  2.01  NA  n  0  0  5  x o  NA  48.33  NA  56.34  NA  NA  39.71  NA  40.28  NA  NA  28.14  NA  29.86  NA  NA  15.09  NA  17.41  NA  NA  20.83  NA  12.02  NA  NA  3.80  NA  4.54  NA  0  5  n  5. 4% F e S 0  2  O  4  0  3  4  0  0  4  0  x  259.21  241.07  161.93  236.66  235.93  a  42.55  72.66  35.71  67.30  62.79  n  5  5  5  3  5  0  0  5  5  0  0  0  0  270.43 155.50 117.43 132.64 219.43 107.58 5  24.92 5  15.92 5  5  41.98 5  51.04  5  5  0  0  5  5  0  0  40.28  22.50  46.43  36.50  116.93  8.38  5.67  9.42  5.95  15.93  5  5  5  5  5  APPENDIX E.3.  Treatment  1980  Site  2% F e S 0  1  4  2% urea  4  + 2% urea  0.1% CuS0  4  0.2% CuS0  + 2% urea  4  3  1981 4  5  1  2  3  1982  4  5  1  2  3  4  5  X  NA  152.77  NA  116.00  NA  NA  149.57  NA  102.14  NA  NA  29.71  NA  33.29  NA  23.20  NA  36.18  NA  NA  52.56  NA  48.04  NA  NA  11.23  NA  5.71  NA  n  0  0  0  0  0  0  5  0  4  0  5  5  0  5  5  NA  X  66.43  49.28  42.12  50.64  52.00  53.79  38.70  30.29  41.29  37.14  32.64  27.57  27.11  28.78  46.:  a  19.50  9.51  4.20  6.56  6.95  11.06  7.20  5.25  13.18  9.25  2.54  4.53  1.69  2.58  ii.;  4  5  5  5  5  5  5  5  5  5  5  5  5  5  5  X  NA  _  40.95  39.86  NA  NA  94.37  30.22  38.93  NA  NA  23.29  32.64  32.07  NA  a  NA  -  4.32  0.0  NA  NA  30.01  4.41  6.64  NA  NA  5.25  4.25  8.16  NA  3  1  0  0  5  5  0  0  5  5  5  0  0  + 2% urea  2  0  n  1% CuS0  (cont'd)  0  4  X  NA  66.18  NA  NA  NA  NA  36.16  NA  NA  NA  NA  25.00  NA  NA  NA  a  NA  0.86  NA  NA  NA  NA  5.32  NA  NA  NA  NA  5.25  NA  NA  NA  n  0  2  0  0  0  5  0  0  0  0  5  0  0  0  0  X  NA  25.48  NA  NA  NA  NA  42.50  NA  NA  NA  NA  20.90  NA  NA  NA  a  NA  0.0  NA  NA  NA  NA  22.51  NA  NA  NA  NA  2.36  NA  NA  NA  n  0  1  0  0  0  0  0  0  0  0  4  0  0  0  5  APPENDIX E.3. (cont'd)  Treatment  1980  Site  4% FeS0  11.  1  12.  1% CuS0  + 4% F e S 0 + 2% urea  4  4  0.1% CuS0  13.  + 2% FeS0 + 2% urea  4  0.2% CuS0  14.  + 2% FeS0 + 2% urea  4  4  4  3  1981 4  5  1982  1  2  3  4  5  .  1  2  3  396.43  261.54  243.12  347.27  226.79  279.71  200.28  123.57  234.36  231.85  38.64  29.57  50.50  46.00  117.1  0  0.0  11.90  48.74  49.99  45.69  79.27  74.90  42.50  71.35  47.38  6.29  5.97  2.31  5.74  33.  n  1  5  5  5  X  NA  a n  NA  X  4  + 2% urea  2  4  3  5  5  5  5  5  5  5  -  NA  325.71  NA  NA  200.18  NA  302.78  NA  NA  23.57  NA  44.22  -  NA  0.0  NA  NA  38.51  NA  85.34  NA  NA  1.78  NA  11.03  5  0  0  4  0  3  0  0  0  1  0  0  4  0  5  NA NA  5  0  -  NA  156.43  NA  NA  NA  NA  161.21  NA  NA  NA  NA  27.36  NA  NA  NA  a  NA  12.63  NA  NA  NA  NA  64.69  NA  NA  NA  NA  6.66  NA  NA  NA  n  0  0  0  0  0  0  0  0  0  5  0  0  0  2  5  X  NA  185.00  NA  NA  NA  NA  184.43  NA  NA  NA  NA  22.22  NA  NA  NA  a  NA  0.0  NA  NA  NA  NA  37.33  NA  NA  NA  NA  2.15  NA  NA  NA  n  0  1  0  0  0  0  0  0  0  0  5  0  0  0  NA  =  Treatment not a p p l i e d .  -  =  M i s s i n g value. Average concentration value,  x  =  a  =  Standard d e v i a t i o n ,  n  »  Number of samples.  5  ON  APPENDIX E.4.  Site  2. 1Z C11SO4  3. 0.2Z &1SO4  4. 0.1Z CUSO4  5. 4Z FeS04  DOUGLAS-FIR  1981  1980  Treatment  1. C o n t r o l  MEAN VALUES FOR FOLIAR NITROGEN CONCENTRATION (Z) OF LODGEPOLE PINE AND IN RELATION TO TREATMENTS (MAIN TRIAL).  1  2  3  4  5  1  2  3  1982 4  5  1  2  3  4  5  X  0.91  0.86  0.95  1.01  0.76  0.94  0.94  0.95  1.06  0.76  1.07  1.07  1.06  1.09  0  0.03  0.06  0.08  0.07  0.12  0.05  0.07  0.11  0.10  0.10  0.04  0.04  0.12  0.06  0.07  n  5  5  5  5  5  5  5  5  5  5  5  5  5  5  5  X  NA  0  NA  a  0  0  -  0.88  1.01  1.16  NA  NA  1.15  1.25  1.40  NA  NA  1.03  1.01  1.06  NA  0.09  0.03  NA  NA  0.26  0.06  0.11  NA  NA  0.20  0.06  0.02  NA  4  3  0  0  3  5  5  0  0  5  5  5  0  NA  I  NA  1.01  NA  1.05  NA  NA  1.18  NA  1.18  NA  NA  1.06  NA  1.14  a  NA  0.07  NA  0.07  NA  NA  0.04  NA  0.13  NA  NA  0.16  NA  0.09  NA  n  0  4  0  4  0  0  5  0  5  0  0  5  5  0  0  X  NA  1.08  NA  1.01  NA  NA  1.25  NA  1.16  NA  NA  1.12  NA  1.20  NA  0  NA  0.04  NA  0.07  NA  NA  0.07  NA  0.10  NA  NA  0.12  NA  0.12  NA  n  0  3  0  4  0  0  5  0  5  0  0  5  5  0  0  X  1.06  0.98  1.11  0.96  0.86  1.26  1.33  1.39  1.26  0.91  1.15  0.96  1.06  1.13  0.98  a  0.08  0.13  0.03  0.03  0.08  0.09  0.14  0.13  0.10  0.05  0.13  0.11  0.14  0.07  0.06  n  5  5  5  3  5  5  5  5  5  5  5  5  5  5  5  •  APPENDIX E.4.  Treatment  1980  Site  6.  7.  8.  2% FeS0  1  4  2% urea  1% CuS0  4  + 2% urea  9.  0.1% CuS0  4  + 2% urea  10.  0.2% CuS0  + 2% urea  (cont'd)  4  2  3  1981 4  5  1  2  3  1982  4  5  1  2  3  X  NA  0.98  NA  1.10  NA  NA  1.18.  NA  1.24  ' NA  NA  1.11  NA  1.09  NA  a  NA  0.06  NA  0.09  NA  NA  0.06  NA  0.10  NA  NA  0.06  NA  0.10  NA  n  0  4  0  5  0  0  5  0  5  0  0  5  0  5  0  X  0.96  0.98  1.03  1.02  0.80  1.37  1.26  1.19  1.14  0.93  1.07  1.09  0.96  1.12  0.'  a  0.10  0.06  0.06  0.08  0.06  0.19  0.13  0.07  0.10  0.06  0.17  0.07  0.06  0.11  0.1  n  5  5  5  5  5  5  5  5  5  5  5  5  5  5  5  X  NA  -  0.94  0.96  NA  NA  1.29  1.50  1.31  NA  NA  0.96  1.11  0.93  NA  a  NA  -  0.06  0.0  NA  NA  0.26  0.20  0.06  NA  NA  0.11  0.16  0.07  NA  3  1  0  0  4  5  5  0  0  5  5  5  0  0  0  X  NA  1.02  NA  NA  NA  NA  1.15  NA  NA  NA  NA  1.01  NA  NA  NA  a n  NA  0.06  NA  NA  NA  NA  0.13  NA  NA  NA  NA  0.07  NA  NA  NA  0  2  0  0  0  0  5  0  0  0  0  5  0  0  • 0  X  NA  1.14  NA  NA  NA  NA  1.26  NA  NA  NA  NA  1.21  NA  NA  NA  0  NA  0.0  NA  NA  NA  NA  0.16  NA  NA  NA  NA  0.21  NA  NA  NA  n  0  1  0  0  0  0  5  0  0  0  0  4  0  0  0  APPENDIX E.A. (cont'd)  Treatment  1980  Site  1  4% FeSO^  2  3  1981  4  5  1  2  1982  3  4  5  1  2  3  X  0.94  1.03  1.04  1.15  0.86  1.50  1.38  1.40  1.39  0.90  1.14  1.06  1.06  1.09  0.!  a n  0.0  0.08  0.07  0.16  0.09  0.19  0.05  0.13  0.17  0.07  0.25  0.26  0.08  0.21  0.1  1  3  4  3  5  5  5  5  5  5  5  5  5  5  5  1% CuS04  X  NA  1.08  NA  NA  1.45  NA  1.51  NA  NA  0.99  NA  1.09  NA  0  NA  NA  0.0  NA  NA  0.29  NA  0.08  NA  NA  0.09  NA  0.08  NA  n  0  0  NA  + 4% FeS04 + 2% urea  0  1  0  0  4  0  5  0  0  4  0  5  0  11.  + 2% urea  12.  13.  0.1%  CUSO4  + 2% FeS0 + 2% urea 14.  0.2%  4  CUSO4  + 2% FeS04 + 2% urea  X  NA  0.99  NA  NA  NA  NA  1.22  NA  NA  NA  NA  1.09  NA  NA  NA  a n  NA  0.09  NA  NA  NA  NA  0.11  NA  NA  NA  NA  0.14  NA  NA  NA  0  2  0  0  0  0  5  0  0  0  0  5  0  0  0  X  NA  0.91  NA  NA  NA  NA  1.14  NA  NA  NA  NA  1.12  NA  NA  NA  0  NA  0.0  NA  NA  NA  NA  0.14  NA  NA  NA  NA  0.22  NA  NA  NA  n  0  1  0  0  0  0  5  0  0  0  0  5  0  0  0  NA  =  Treatment  -  =  M i s s i n g value.  not a p p l i e d .  Average c o n c e n t r a t i o n value,  x  •  o  =  Standard d e v i a t i o n ,  n  =  Number of samples.  

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}]}"
                            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:
http://iiif.library.ubc.ca/presentation/dsp.831.1-0096652/manifest

Comment

Related Items