DNA markers identify variation in Australian populations of Uncinula necator
Evans, KJ and Whisson, DL and Stummer, BE and Scott, ES, DNA markers identify variation in Australian populations of Uncinula necator, Mycological Research, 101 pp. 923-932. ISSN 0953-7562 (1997) [Refereed Article]
Restriction fragment length polymorphisms (RFLPs) were identified among total DNA from clonal lines of Uncinula necator when cloned sequences of total U. necator DNA were used as probes. Four probes, pUnP14, pUnP27, pUnE21 and pUnE4, hybridized to multiple-copy sequences and, with the exception of pUnE4, detected genetic variation among clonal lines of U. necator. Clones pUnP14, pUnP27 and pUnE4 produced banding patterns that were stable for DNA extracted from different asexual generations of U. necator clonal lines over at least 15 months. In addition, clones were evaluated for species specificity. Clones pUnP27 and pUnE4 detected only U. necator sequences in total DNA from infected grapevine leaves. Clones pUnPI4 and pUnE4 did not hybridize to total DNA from a range of fungi. Genetic diversity in a sample of the Australian U. necator population was investigated; 15 genotypes were identified among 29 U. necator clonal lines examined. Genetic variation was detected in samples collected within micro-geographical areas, for example, different genotypes representing both mating types of U. necator were detected on a single plant of Vitis amurensis. RFLP analysis of the banding patterns produced using pUnP14, pUnP27 and pUnE21, identified two broad genetic groups, designated A and B. Analysis of DNA fragment patterns obtained using the polymerase chain reaction (PCR) and the plant intron splice junction (ISJ) primer R1 also supported the allocation of U. necator clonal lines into groups A and B.