Regulation of hyphal growth rate of Hypoxylon mammatum by amino acids: Stimulation by proline

Abstract

Variation among isolates of Hypoxylon mammatum caused by amino acids added as sole nitrogen sources demonstrated genetic heterogeneity among the population in the mechanisms regulating hyphal growth rates. Isolates were obtained by a random sample consisting of 22 single ascospore isolates from a local population of hypoxylon cankers on P. tremuloides. Hyphal growth rates were determined from colony diameter measurements on defined media containing either alanine, asparagine, glycine, leucine, lysine, or proline. All isolates but two grew faster on proline than the other amino acids tested. The mean growth rate of the population sample was 3.9 mm day −1 on proline compared to 1.96 mm day −1 on asparagine, the second fastest mean growth rate. Growth rates of the 22 isolates on these six amino acids were largely uncorrelated, indicating independent mechanisms of regulation. Only asparagine gave significant correlations with more than two other amino acids. A more detailed examination of selected isolates representing a range of growth rates on proline showed that the rapid growth rates were also dependent on other materials present in Difco Bacto-Agar. Growth of the proline stimulated isolates was considerably slower on media gelled with Noble agar. Other nitrogen sources, especially glutamate, added with proline stimulated the growth of these isolates. Isolate dependent variation in the stimulatory effect of the N source added with proline was demonstrated. Stimulation of growth rate by proline may be related to drought stress-enhanced canker elongation in vivo, since drought stress caused proline accumulation in the host, Populus tremuloides.