Identification and structure of the mating-type locus and development of PCR-based markers for mating type in powdery mildew fungi

被引:47
作者
Brewer, Marin Talbot [1 ]
Cadle-Davidson, Lance [2 ]
Cortesi, Paolo [3 ]
Spanu, Pietro D. [4 ]
Milgroom, Michael G. [1 ]
机构
[1] Cornell Univ, Dept Plant Pathol & Plant Microbe Biol, Ithaca, NY 14853 USA
[2] USDA ARS, Grape Genet Res Unit, Geneva, NY 14456 USA
[3] Univ Milan, Dipartimento Protez Sistemi Agroalimentare & Urba, I-20133 Milan, Italy
[4] Univ London Imperial Coll Sci Technol & Med, Dept Biol, London SW7 2AZ, England
基金
英国生物技术与生命科学研究理事会;
关键词
Erysiphe necator; Erysiphales; Mating type; MAT locus; Powdery mildew; Transcriptome; UNCINULA-NECATOR; ERYSIPHE-NECATOR; FLAG SHOOT; CLEISTOTHECIA; NEUROSPORA; INOCULUM; BIOLOGY; CLONES; DNA;
D O I
10.1016/j.fgb.2011.04.004
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
In ascomycetes, mating compatibility is regulated by the mating-type locus, MAT1. The objectives of this study were to identify and sequence genes at the MAT1 locus in the grape powdery mildew fungus, Erysiphe necator, to develop a PCR-based marker for determining mating type in E. necator, and to develop degenerate primers for amplification by PCR of conserved regions of mating-type idiomorphs in other powdery mildew fungi. We identified MAT1-2-1 of the MAT1-2 idiomorph in E. necator based on the homologous sequence in the genome of Blumeria graminis f. sp. hordei and we found MAT1-1-1 and MAT1-1-3 of the MAT1-1 idiomorph from transcriptome sequences of E. necator. We developed and applied a reliable PCR-based multiplex marker to confirm that genotype correlated with mating phenotype, which was determined by pairing with mating-type tester isolates. Additionally, we used the marker to genotype populations of E. necator from different Vitis spp. from throughout the USA. We found both mating types were present in all populations and mating-type ratios did not deviate from 1:1. The mating-type genes in E. necator are similar to those of other Leotiomycetes; however, the structure of the MAT1 locus in E. necator, like the MAT1-2 idiomorph of B. graminis, is markedly different from other ascomycetes in that it is greatly expanded and may contain a large amount of repetitive DNA. As a result, we were unable to amplify and sequence either idiomorph in its entirety. We designed degenerate primers that amplify conserved regions of MAT1-1 and MAT1-2 in E. necator, Podosphaera xanthii, Microsphaera syringae, and B. graminis, representing the major clades of the Erysiphales. These degenerate primers or sequences obtained in this study from these species can be used to identify and sequence MAT1 genes or design mating-type markers in other powdery mildew fungi as well. (C) 2011 Elsevier Inc. All rights reserved.
引用
收藏
页码:704 / 713
页数:10
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