Molecular Phylogenetic Analysis of Ribosomal DNA Internal Transcribed Spacer Regions and Comparison of Fertility in Phomopsis Isolates from Fruit Trees

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Abstract

Sequences of the ribosomal DNA(rDNA)internal transcribed spacer(ITS)regions were examined to infer a molecular phylogeny of small-spored Phomopsis isolates, designated W-type(mainly white colony, weakly virulent, bearing both alpha and beta conidia at 25℃ on PDA)and G-type(mainly gray colony, highly virulent, bearing only alpha conidia at 25℃ on PDA), and P.amygdali from fruit trees. Phomopsis G-type and P.amygdali were a monophyletic group distinct from the W-type. The W-type isolates were divided into two monophyletic groups. Diaporthe citri, D.tanakae, P.asparagi, P.viticola, P.vitimegaspora and D.nomurai, which are morphologically distinguishable from W- and G-types, differed from the W- and G-types in molecular phylogenetic analyses. PCR-RFLP analysis of rDNA ITS regions was useful to distinguish each of the Phomopsis species and groups using three restriction enzymes. In mating tests, W-type isolates from fruit trees were heterothallic and inter-fertile even between isolates belonging to the different monophyletic groups. Isolates of the G-type and P.amygdali collected in Japan were cross-fertile. Some isolates from Lunaria annua, Ulmus glabra and Juglans regia belonged to one of the two monophyletic groups of the W-type and were cross-fertile with W-type isolates from Rosaceous fruit trees.

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