Karthikeyan Adhimoolam, Li Kai, Jiang Hua, Ren Rui, Li Cui, Zhi Haijian, Chen Shouyi, Gai Junyi
Soybean Research Institute, Nanjing Agricultural University, Nanjing, 210095, China.
MOA National Center for Soybean Improvement, Nanjing Agricultural University, Nanjing, 210095, China.
Mol Genet Genomics. 2017 Aug;292(4):811-822. doi: 10.1007/s00438-017-1310-8. Epub 2017 Mar 28.
Soybean mosaic virus (SMV) is one of the most devastating pathogens for soybeans in China. Among the country-wide 22 strains, SC5 dominates in Huang-Huai and Changjiang valleys. For controlling its damage, the resistance gene was searched through Mendelian inheritance study, gene fine-mapping, and candidate gene analysis combined with qRT-PCR (quantitative real-time polymerase chain reaction) analysis. The parents F, F, and RILs (recombinant inbred lines) of the cross Kefeng-1 (Resistance, R) × NN1138-2 (Susceptible, S) were used to examine the inheritance of SC5-resistance. The F was resistant and the F and RILs segregated in a 3R:1S and 1R:1S ratio, respectively, indicating a single dominant gene conferring the Kefeng-1 resistance. Subsequently, the genomic region conferring the resistance was found in "Bin 352-Bin353 with 500 kb" on Chromosome 2 using the phenotyping data of the 427 RILs and a high-density genetic map with 4703 bin markers. In the 500 kb genomic region, 38 putative genes are contained. The association analysis between the SNPs in a putative gene and the resistance phenotype for the 427 RILs prioritized 11 candidate genes using Chi-square criterion. The expression levels of these genes were tested by qRT-PCR. On infection with SC5, 7 out of the 11 genes had differential expression in Kefeng-1 and NN1138-2. Furthermore, integrating SNP-phenotype association analysis with qRT-PCR expression profiling analysis, Glyma02g13495 was found the most possible candidate gene for SC5-resistance. This finding can facilitate the breeding for SC5-resistance through marker-assisted selection and provide a platform to gain a better understanding of SMV-resistance gene system in soybean.
大豆花叶病毒(SMV)是中国大豆最具毁灭性的病原体之一。在全国范围内的22个菌株中,SC5在黄淮和长江流域占主导地位。为了控制其危害,通过孟德尔遗传研究、基因精细定位、候选基因分析并结合qRT-PCR(定量实时聚合酶链反应)分析来寻找抗性基因。以杂交组合科丰1号(抗性,R)×NN1138-2(感病,S)的亲本F、F和重组自交系(RILs)来研究对SC5抗性的遗传。F表现为抗性,F和RILs分别以3R:1S和1R:1S的比例分离,表明科丰1号的抗性由单个显性基因控制。随后,利用427个RILs的表型数据和具有4703个bin标记的高密度遗传图谱,在2号染色体上“Bin 352-Bin353,500 kb”区域发现了赋予抗性的基因组区域。在500 kb的基因组区域中包含38个推定基因。利用卡方检验标准,对427个RILs中一个推定基因的单核苷酸多态性(SNPs)与抗性表型进行关联分析,确定了11个候选基因。通过qRT-PCR检测这些基因的表达水平。在感染SC5后,11个基因中有7个在科丰1号和NN1138-2中表现出差异表达。此外,将SNP-表型关联分析与qRT-PCR表达谱分析相结合,发现Glyma02g13495是最有可能的SC5抗性候选基因。这一发现有助于通过标记辅助选择培育抗SC5的品种,并为更好地理解大豆中的SMV抗性基因系统提供一个平台。