Characterization of Sphaeropsis sapinea Isolates from South Africa, Mexico, and Indonesia

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Abstract
Post-hail-associated dieback of Pinus patula and P. radiata trees, induced by Sphaeropsis sapinea (=Diplodia pinea), is a common and important disease in commercial pine plantations. Two morphotypes, A and B, have been described for this fungus based on differences in cultural characteristics, conidial morphology, and virulence among isolates from the north central United States. The existence of the two described morphotypes was later verified through the use of random amplified polymorphic DNA (RAPD) analyses, and the morphotypes were designated as the A and B RAPD marker groups. The objective of this study was to characterize a set of S. sapinea isolates from South Africa, Indonesia, and Mexico using RAPD analysis and DNA sequencing of the internal transcribed spacer (ITS) region of the ribosomal DNA operon. Sizes of conidia and culture morphology were, furthermore, used to compare the three groups of S. sapinea isolates that emerged from the RAPD analysis. Two of the RAPD groups included isolates from the United States, representing the A and B morphotypes, while the third RAPD group accommodated Indonesian and one Mexican isolate. ITS sequences of all the S. sapinea isolates were highly homologous and resolved only the A and B RAPD groups. The ITS sequences of the isolates in the third RAPD group were the same as those of the A RAPD group. Conidia of isolates representing the A and B morphotypes were approximately the same size, but those of the third RAPD group were significantly longer. RAPD analysis enabled us to identify a third group of S. sapinea that is different from the well-recognized A and B groups. Isolates in this third RAPD group also have a distinct morphological characteristic and thus represent a third discrete morphological group, which we refer to as the C morphotype of S. sapinea.