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结合单核苷酸多态性基因分型阵列与集群分离分析法定位小麦品系03031-1-5 H62中一个控制成株期抗条锈病的基因

Combining Single Nucleotide Polymorphism Genotyping Array with Bulked Segregant Analysis to Map a Gene Controlling Adult Plant Resistance to Stripe Rust in Wheat Line 03031-1-5 H62.

作者信息

Wu Jianhui, Wang Qilin, Xu Liangsheng, Chen Xianming, Li Bei, Mu Jingmei, Zeng Qingdong, Huang Lili, Han Dejun, Kang Zhensheng

机构信息

First, second, third, seventh, eighth, and tenth authors: State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, P. R. China; fifth, sixth, and ninth authors: State Key Laboratory of Crop Stress Biology for Arid Areas, College of Agronomy, Northwest A&F University, Yangling, Shaanxi 712100, P. R. China; and fourth author: U.S. Department of Agriculture, Agricultural Research Service, Wheat Health, Genetics, and Quality Research Unit and the Department of Plant Pathology, Washington State University, Pullman.

出版信息

Phytopathology. 2018 Jan;108(1):103-113. doi: 10.1094/PHYTO-04-17-0153-R. Epub 2017 Nov 9.

Abstract

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most devastating diseases of wheat worldwide. Growing resistant cultivars is considered the best approach to manage this disease. In order to identify the resistance gene(s) in wheat line 03031-1-5 H62, which displayed high resistance to stripe rust at adult plant stage, a cross was made between 03031-1-5 H62 and susceptible cultivar Avocet S. The mapping population was tested with Chinese P. striiformis f. sp. tritici race CYR32 through artificial inoculation in a field in Yangling, Shaanxi Province and under natural infection in Tianshui, Gansu Province. The segregation ratios indicated that the resistance was conferred by a single dominant gene, temporarily designated as YrH62. A combination of bulked segregant analysis (BSA) with wheat 90K single nucleotide polymorphism (SNP) array was used to identify molecular markers linked to YrH62. A total of 376 polymorphic SNP loci identified from the BSA analysis were located on chromosome 1B, from which 35 kompetitive allele-specific PCR (KASP) markers selected together with 84 simple sequence repeat (SSR) markers on 1B were used to screen polymorphism and a chromosome region associated with rust resistance was identified. To saturate the chromosomal region covering the YrH62 locus, a 660K SNP array was used to identify more SNP markers. To develop tightly linked markers for marker-assisted selection of YrH62 in wheat breeding, 18 SNPs were converted into KASP markers. A final linkage map consisting of 15 KASP and 3 SSR markers was constructed with KASP markers AX-109352427 and AX-109862469 flanking the YrH62 locus in a 1.0 cM interval. YrH62 explained 63.8 and 69.3% of the phenotypic variation for disease severity and infection type, respectively. YrH62 was located near the centromeric region of chromosome 1BS based on the positions of the SSR markers in 1B deletion bins. Based on the origin, responses to P. striiformis f. sp. tritici races, and marker distances, YrH62 is likely different from the other reported stripe rust resistance genes/quantitative trait loci on 1B. The gene and tightly linked KASP markers will be useful for breeding wheat cultivars with resistance to stripe rust.

摘要

条锈病由条形柄锈菌小麦专化型(Puccinia striiformis f. sp. tritici)引起,是全球小麦最具毁灭性的病害之一。种植抗病品种被认为是防治这种病害的最佳方法。为了鉴定小麦品系03031 - 1 - 5 H62中的抗病基因,该品系在成株期对条锈病表现出高抗性,将03031 - 1 - 5 H62与感病品种Avocet S进行杂交。通过在陕西省杨凌的田间人工接种以及甘肃省天水的自然发病条件下,用中国条形柄锈菌小麦专化型小种CYR32对作图群体进行测试。分离比例表明该抗性由一个单显性基因控制,暂时命名为YrH62。采用混合分组分析法(BSA)与小麦90K单核苷酸多态性(SNP)芯片相结合来鉴定与YrH62连锁的分子标记。从BSA分析中鉴定出的总共376个多态性SNP位点位于1B染色体上,从中选择了35个竞争性等位基因特异性PCR(KASP)标记以及1B染色体上的84个简单序列重复(SSR)标记用于筛选多态性,并鉴定出一个与抗锈性相关的染色体区域。为了使覆盖YrH62位点的染色体区域饱和,使用660K SNP芯片来鉴定更多的SNP标记。为了开发紧密连锁的标记用于小麦育种中YrH62的分子标记辅助选择,将18个SNP转化为KASP标记。构建了一个由15个KASP和3个SSR标记组成的最终连锁图谱,KASP标记AX - 109352427和AX - 109862469在YrH62位点两侧,间隔为1.0 cM。YrH62分别解释了病情严重程度和感染类型表型变异的63.8%和69.3%。基于1B染色体缺失区间内SSR标记的位置,YrH62位于1BS染色体的着丝粒区域附近。基于其来源、对条形柄锈菌小麦专化型小种的反应以及标记距离,YrH62可能与1B染色体上其他已报道的条锈病抗性基因/数量性状位点不同。该基因和紧密连锁的KASP标记将有助于培育抗条锈病的小麦品种。

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