UBC Theses and Dissertations
Puccinia graminis f.sp. tritici, race c17 : physiology of uredospore germination and germtube differentiation Hopkinson, Sarah J.
Germinating uredospores of race C17 of Puccinia graminis f.sp. tritici form characteristic infection structures (appressorium, infection peg, vesicle, infection hypha) in response to a 1.5 h heat shock at 29° C administered 2 h after germination at 19° C. The proportion of sporelings forming infection structures was augmented by nutrients, n-nonyl alcohol and, an appropriately timed heat shock. The heat shock temperature required to induce maximum differentiation had a very precise optimum which varied slightly for each spore lot. Variations one degree above or below this optimum reduced the percent differentiation by greater than 40%. The presence of an inhibitor of protein synthesis, puromycin, in the germination medium: (1) prevented uredosporeling differentiation but had no effect on germination, (2) significantly reduced the proportion of germtubes forming appressoria, and (3) in most cases prevented the division of germtube nuclei. It was concluded that essential differentiation-specific proteins are synthesized from the onset of germination, throughout the formation of appressoria and to the completion of differentiation. These results were consistent with the observed effects of heat shock on the rate of protein hydrolysis. During germination there was a net hydrolysis of protein leading to an increase in size of the endogenous pool of free amino acids and to an increased leakage of amino acids to the germination medium. Heat shock effectively reduced the amount of endogenous free amino acids and the extent to which amino acids were lost to the medium. It was concluded that in heat shocked sporelings protein synthesis was increased relative to protein hydrolysis by comparison with the relative rates of these two processes in germinating (non-shocked) uredosporelings. Moreover, there was no net protein synthesis during the formation of infection structures induced by heat shock. The loss of amino acids to the germination medium was selective, particularly in heat shocked sporelings.