Microsatellite markers for the grapevine pathogen, Eutypa lata

Abstract We isolated and characterized nine polymorphic microsatellite markers for Eutypa lata, a fungal pathogen responsible for Eutypa dieback of grapevine, in populations from two California vineyards (24 isolates per vineyard). Allele frequency ranged from two to 11 alleles per locus and haploid gene diversity ranged from 0.33 to 0.83. All samples comprised unique haplotypes. Our results suggest that there is sufficient allelic polymorphism to estimate fine-scale spatial structure, and to identify possible sources of inoculum from habitats outside of vineyards. Keywords: Ascomycota, Diatrypaceae, Eutypa dieback, Eutypa lata, plant pathogen, Vitis vinifera Received 19 May 2008; revision accepted 10 June 2008 The means of spread of Eutypa dieback from vine-to-vine within vineyards is likely due to dispersal of Eutypa lata sexual spores (ascospores), and not asexual spores (conidia), based on evidence of distributions of vegetative compatibility groups, reproductive structures (perithecia), and symptomatic grapevines Genomic DNA was extracted from an isolate of E. lata from Switzerland [isolate 208.87; Centraalbureau voor Schimmelcultures, Utrecht, the Netherlands], purified (GENECLEAN III Kit, MP Biomedicals), digested with Taq αI (New England BioLabs), and enriched for both a trinucleotide, CAC 10 , and a tetranucleotide mixture (AAAC 6 , AAAG 6 , AAAT 8 , AGAT 8 ; Integrated DNA Technologies). Digested DNA was ligated to linker oligonucleotides 20B (5′-GCGGTTCCCGGTCGAGTTGG-3′) and 22B (5′-pCGCCAACTCGACCGGGAACCGC-3′