Identification and structure of the mating-type locus and development of PCR-based markers for mating type in powdery mildew fungi
Identification and structure of the mating-type locus and development of PCR-based markers for mating type in powdery mildew fungi
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DOI：
10.1016/j.fgb.2011.04.004
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发表时间：
2011-07-01
影响因子：
3
通讯作者：
Milgroom, Michael G.
中科院分区：
文献类型：
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作者：
Brewer, Marin Talbot;Cadle-Davidson, Lance;Milgroom, Michael G.
在子囊菌中，交配相容性由交配型基因座 MAT1 调节。本研究的目的是对葡萄白粉病真菌 Erysiphe necator 中 MAT1 基因座的基因进行鉴定和测序，开发基于 PCR 的标记来确定 E. necator 的交配类型，并开发简并引物，用于通过 PCR 扩增其他白粉病真菌交配型个体型的保守区域。我们根据 Blumeria graminis f 基因组中的同源序列鉴定了 E. necator 中 MAT1-2 异型的 MAT1-2-1。 sp。我们从 E. necator 的转录组序列中发现了 MAT1-1 形态的 MAT1-1-1 和 MAT1-1-3。我们开发并应用了可靠的基于 PCR 的多重标记来确认基因型与交配表型相关，交配表型是通过与交配型测试分离株配对来确定的。此外，我们使用该标记对来自不同葡萄属的 E. necator 群体进行基因分型。来自美国各地。我们发现这两种交配类型都存在于所有种群中，并且交配类型比例没有偏离 1:1。 E. necator 的交配型基因与其他Leotiomycetes 相似；然而，E. necator 中的 MAT1 基因座的结构与 B. graminis 的 MAT1-2 形态一样，与其他子囊菌明显不同，因为它的结构大大扩展，并且可能包含大量重复 DNA。结果，我们无法对任一个体形态进行完整的扩增和测序。我们设计了简并引物，可扩增代表白粉病主要分支的 E. necator、Podosphaera xanthii、Microsphaera syringae 和 B. graminis 中 MAT1-1 和 MAT1-2 的保守区域。本研究中从这些物种获得的这些简并引物或序列也可用于识别和测序其他白粉病真菌中的 MAT1 基因或设计交配型标记。 (C) 2011 Elsevier Inc. 保留所有权利。
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.