ZHANG Hai-ping,CHEN Yan,YAN Kai.Selection of SSR Markers for SCN4 Resistance and Identification of Excellent Resistant Germplasm[J].Soybean Science,2020,39(01):1-11.[doi:10.11861/j.issn.1000-9841.2020.01.0001]
大豆抗胞囊线虫4号生理小种SSR标记筛选及优异种质鉴定
- Title:
- Selection of SSR Markers for SCN4 Resistance and Identification of Excellent Resistant Germplasm
- 文献标志码:
- A
- 摘要:
- 大豆抗胞囊线虫的表型鉴定工作量较大，鉴定结果易受环境影响，是抗源筛选和抗病品种选育的限制因素之一。不受时间、环境限制的分子标记鉴定为抗病鉴定提供了一种高效快捷准确简单的鉴定方法。为筛选可用于抗大豆胞囊线虫4号生理小种分子标记引物，本研究采用基因型鉴定和人工接种鉴定相结合的方法鉴定193份大豆资源的大豆胞囊线豆抗性。以部分高抗大豆资源和高感资源为材料，对1 000对SSR引物进行初筛，用初筛获得的引物扩增193份抗、感大豆资源，筛选抗胞囊线虫4号生理小种（SCN 4）SSR标记引物，用于大豆抗胞囊线虫4号生理小种分子标记辅助鉴定。结果表明：筛选出4个与大豆胞囊线虫4号生理小种相关的SSR标记，分别为Satt400、Satt680、Satt533和Satt504。Satt400、Satt680、Satt533和Satt504对感病资源的检出率均为100%，对抗病资源的检出率分别为70.58%、63.15%、92.3%和57.14%。结合人工接种鉴定结果显示，4个标记组合鉴定可提高对胞囊线虫4号生理小种抗性的选择效率，达100%。经人工接种鉴定，从193份资源中筛选出12份抗病资源，其中10份资源为高抗，2份资源为中抗。这12份抗源的4个SSR标记带型均为抗病带型，其余感病和高感资源的4个SSR标记带型则至少有一个为感病带型。因此，利用分子标记评价抗感特性的标准为被1个或1个以上的感病带型检出的资源为感病资源，当4个标记均为抗病带型，该资源为抗病资源。利用该引物组合对山西省种质库提供的100份资源进行鉴定。筛选出4份抗源，人工接种鉴定结果表明，黑豆、小颗黑和晋1265（茶）3份资源抗性级别为高抗，大黑豆抗性级别为中抗。因此，该引物组合可有效辅助鉴定大豆对胞囊线虫4号生理小种的抗性。
- Abstract:
- The workload of phenotype identification of soybean resistance to cyst nematode is large, and the identification results are easily affected by the environment, which is one of the limiting factors for resistance source selection and resistance variety selection. Molecular marker identification without time and environment constraints provides an efficient, rapid, accurate and simple method of disease resistance identification. In order to screen molecular markers for distinguishing resistant resources from susceptible resources, identified the resistance to soybean cyst nematode of 193 soybean resource, part of the resistant resources and the high susceptible resources were genotyped by 1 000 SSR markers and 7 SSR markers that can distinguish SCN4 resistance sources from susceptible sources were found. Then, 193 soybean resources were genotyped by the 7 SSR markers. The results showed that 4 markers, Satt400, Satt680, Satt533 and Satt504, were selected. The selection efficiency of susceptible resources with Satt400, Satt680, Satt533 and Satt504 was all 100%. The selection efficiency of resistant resources with Satt400, Satt680, Satt533 and Satt504 was 70.58%, 63.15%, 92.3% and 57.14% respectively. The combination of 4 markers could add the selection efficiency to resistant resources, and the selection efficiency was up to 100%. Combined with artificial inoculation identification, 12 of the 193 soybean resources were identified as resistant resources. These 12 resistant resources were all detected with resistant alleles at four loci and other resources were detected with at least one susceptible allele of the four loci. Therefore, the resources were considered as resistant resources which were all detected with resistant alleles of the four loci and the resources were considered as susceptible resources which were detected with at least one susceptible allele of the four loci. 100 resources from Shanxi crop gene bank were genotyped by the 4 markers. Four resources, Heidou, Xiaokehei, Jin1265 and Daheidou, were identified as resistant resources. Then, the resistance to SCN4 of the 4 resources was identified by artificial inoculation. The results showed that the resistance of Heidou, Xiaokehei and Jin1265 were high, and the resistance of Daheidou was middle. It is considered that the combinated markers could be used in marker-assisted selection of resistant germplasm.
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备注/Memo
收稿日期：2019-08-05