利用BSA-Seq方法定位一个水稻早衰相关基因OsBRCA1  

Mapping of Early Senescence-related OsBRCA1 in Rice by BSA-seq Technique

作　　者：蒋家焕[1] 朱永生[1] 陈丽萍[1] 郑燕梅[1] 蔡秋华[1] 谢华安[1] 王爱荣[2] 张建福[1,2] JIANG Jiahuan;ZHU Yongsheng;CHEN Liping;ZHENG Yanmei;CAI Qiuhua;XIE Huaan;WANG Airong;ZHANG Jianfu(Rice Research Institute,Fujian Academy of Agricultural Sciences/Key Laboratory of Hybrid Rice Germplasm Enhancement and Molecular Breeding in South China,Ministry of Agriculture,P.R.China/Fuzhou branch,National Rice Improvement Center of China/Fujian Engineering Laboratory of Crop Molecular Breeding/Fujian Key Laboratory of Rice Molecular Breeding,Fuzhou,Fujian 350003,China;College of Plant Protection,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China)

机构地区：[1]福建省农业科学院水稻研究所/农业农村部华南杂交水稻种质创新与分子育种重点实验室/福州(国家)水稻改良分中心/福建省作物分子育种工程实验室/福建省水稻分子育种重点实验室,福建福州350003 [2]福建农林大学植物保护学院,福建福州350002

出　　处：《福建农业学报》2022年第2期131-137,共7页Fujian Journal of Agricultural Sciences

基　　金：福建省自然科学基金项目(2018J01039、2020J011352)。

摘　　要：【目的】水稻早衰突变体是研究水稻衰老机制的良好载体。定位和克隆水稻早衰相关基因,有助于理解水稻早衰的遗传规律和分子机制,为相关基因的作用机制研究奠定基础。【方法】用EMS化学诱变处理粳稻云引,获得一个稳定遗传的早衰突变体w14,与日本晴杂交构建F;分离群体,并在群体中各选择100个隐性单株和显性单株的DNA等量混合,利用BSA-seq分析两个DNA池之间的差异位点,定位水稻早衰相关基因。【结果】突变体w14的早衰表型受单隐性基因控制,对两个DNA池测序结果进行单基因定位和突变基因的鉴定,发现单基因主峰位于Chr3,候选区间为Chr3:27.5~29.5 Mb,进一步在目标区域内找到2个符合条件的候选因果变异。【结论】候选基因LOC_Os03g49210突变位点是由野生型的C突变为T,且该突变位于候选基因的第2外显子上,属于错义突变,造成了该基因编码的第20个氨基酸由T(苏氨酸)变成I(异亮氨酸),从而可能导致了基因功能的改变,因此,将该基因定为本研究的候选基因,因与人类BRCA1同源,命名为OsBRCA1。【Objective】Molecular mechanism of rice senescence was studied by locating and cloning the specific gene from a mutant artificially induced with early senescence.【Methods】Japonica rice Yunyin was treated with EMS to artificially induce a genetically stable early senescence w14.The mutant was crossed with Nipponbare to construct an isolated F;population.Subsequently,equal parts of the DNA of 100 recessive and dominant plants in the population were mixed.The differential loci between these two DNA pools were analyzed by BSA-seq to locate the target early senescence-related gene.【Results】The early senescence phenotype of w14 was found to be controlled by a single recessive gene.The mutagenic gene mapped and identified by the specific sequence showed the main peak located in Chr 3 and the candidate interval Chr 3:27.5–29.5 Mb.In the region,two qualified candidate causal variants were found.【Conclusion】The mutation site of the candidate gene Loc_Os03g49210 located in the 2^(nd) exon was determined to have changed in the wild-type from C to T.It was a non-synonymous mutation that resulted in the 20;amino acid encoded by the gene changed from T(threonine)to I(isoleucine)and led to the functional alternation.Being homologous with human BRCA1,the target gene was named OsBRCA1.

关 键 词：水稻 突变体 早衰 基因克隆 BSA-seq 

分 类 号：S511[农业科学—作物学]