College of Horticulture, Northeast Agricultural University, Mucai Street 59, Xiangfang District Harbin, 150030, China.
BGI, Shenzhen, China.
BMC Plant Biol. 2017 Dec 29;17(1):266. doi: 10.1186/s12870-017-1215-z.
Tomato gray leaf spot disease caused by Stemphylium lycopersici (S. lycopersici) is considered one of the major diseases of cultivated tomatoes. The only S. lycopersici resistance gene, Sm, was derived from the wild tomato species S. pimpinellifolium. Sm has been identified as an effective source of gray leaf spot resistance in tomatoes and has been mapped to tomato chromosome 11. In this study, the first bulked segregant analysis (BSA) combined with genome resequencing for the mapping and screening of the Sm candidate gene was performed.
Based on the resequencing results, we identified 50,968 Diff-markers, most of which were distributed on chromosome 11. A total of 37 genes were located in the interval of 0.26-Mb. The gene loci of resistant and susceptible lines were sequenced successfully using PCR products. The relative expression levels of candidate genes in resistant and susceptible lines were confirmed via qRT-PCR, Solyc11g011870.1.1 and Solyc11g011880.1.1 were identified through qRT-PCR. A marker, D5, which was cosegregated with the resistant locus, was identified according to the mutation of the Solyc11g011880.1.1 trait in the resistant line.
The Sm gene was mapped to the short arm of chromosome 11. The candidate genes Solyc11g011870.1.1 and Solyc11g011880.1.1 displayed expression patterns related to the resistance response. This study will be valuable for Sm cloning and Sm gene breeding in tomato.
由筋孢菌(Stemphylium lycopersici)引起的番茄灰叶斑病被认为是栽培番茄的主要病害之一。唯一的筋孢菌抗性基因 Sm 来自野生番茄物种 S. pimpinellifolium。Sm 被鉴定为番茄灰叶斑病抗性的有效来源,并已被定位到番茄第 11 号染色体上。本研究首次采用 bulked segregant analysis (BSA) 与基因组重测序相结合的方法,对 Sm 候选基因进行定位和筛选。
基于重测序结果,我们鉴定了 50968 个 Diff-markers,大多数分布在第 11 号染色体上。共鉴定到 37 个基因位于 0.26-Mb 的区间内。利用 PCR 产物成功对抗性和感病系的基因座进行测序。通过 qRT-PCR 验证了候选基因在抗性和感病系中的相对表达水平,确定了 Solyc11g011870.1.1 和 Solyc11g011880.1.1 这两个候选基因。根据抗性系中 Solyc11g011880.1.1 性状的突变,鉴定出与抗性基因座共分离的标记 D5。
Sm 基因被定位到第 11 号染色体的短臂上。候选基因 Solyc11g011870.1.1 和 Solyc11g011880.1.1 的表达模式与抗性反应有关。本研究将为番茄 Sm 基因的克隆和 Sm 基因的育种提供有价值的信息。