谷子条纹叶突变体wsl2的鉴定及候选基因分析  

Identified and Candidate Gene Analysis of a White Stripe Leaf Mutant wsl2 in Foxtail Millet

作　　者：王秋兰[1] 王智兰[1] 韩芳 杜晓芬[1] 连世超 韩康妮 周雪 李慧娟 张林义[1] 王军[1] 郭二虎[1] WANG Qiulan;WANG Zhilan;HAN Fang;DU Xiaofen;LIAN Shichao;HAN Kangni;ZHOU Xue;LI Huijuan;ZHANG Linyi;WANG Jun;GUO Erhu(Millet Research Institute,Shanxi Academy of Agricultural Science,Shanxi Key Laboratory of Genetic Resources and Breeding in Minor Crops,Changzhi 046011,China;Institute of Agricultural Sciences of Yan′an,Yan′an 716000,China)

机构地区：[1]山西省农业科学院谷子研究所,杂粮种质资源发掘与遗传改良山西省重点实验室,山西长治046011 [2]延安市农业科学研究所,陕西延安716000

出　　处：《华北农学报》2020年第1期214-221,共8页Acta Agriculturae Boreali-Sinica

基　　金：山西省青年基金项目(201701D221199);山西省“农谷”研发专项(YCX2017D2201);山西农业科学院国家自然科学基金培育项目(YGJPY1906);山西省重点研发计划重点项目(201703D211008);国家重点研发计划项目(2018YFD1000706;2018YFD1000700);山西省农业科学院特色产业重点研发专项(YCX2019T05);国家现代农业产业技术体系建设专项(CARS-06-13.5-A21)

摘　　要：谷子叶色突变体是探究叶绿体发育机制和C4光能利用率的理想材料之一。为了研究谷子叶色突变的分子机制,从谷子主推品种长农35号的甲基磺酸乙酯(EMS)诱变库中筛选鉴定到一个苗期条纹叶突变体wsl2,通过表型鉴定、遗传背景检测和遗传分析,同时利用MutMap方法快速精细定位突变基因,并根据突变位点开发共分离分子标记等方面对该突变体进行研究。结果表明,wsl2在苗期表现为条纹叶表型,但从拔节期开始恢复为正常叶片表型;wsl2与野生型遗传背景相同且wsl2受一个隐性核基因控制;MutMap精细定位的关联区间内包含9个非同义突变基因,其中,Seita.9G561800是一个编码与叶绿体相关的PsbP蛋白基因,含有6个外显子和5个内含子,在第1外显子77 bp处发生G/T碱基突变,导致一个精氨酸(R)突变成亮氨酸(L)。根据突变碱基设计了dCAPS标记MRI498-1(Cac8Ⅰ)和共分离标记MRI501-3,进一步验证了wsl2突变体的候选基因突变位点。研究鉴定了一个新的条纹叶突变体wsl2,对PsbP基因光合作用反应机制的进一步研究奠定了理论基础,丰富了谷子叶色突变体资源,同时验证了MutMap方法克隆谷子突变基因的有效性。Leaf color mutant is one of the ideal materials to explore the development mechanism of chloroplast and the utilization of C4 light energy in foxtail millet.To study the molecular mechanism of leaf color mutant in foxtail millet,we screened and identified a stripe leaf mutant wsl2 from the EMS mutagenesis library of the main variety Changnong 35.The mutant was studied through phenotype identification,genetic background detection and genetic analysis,location of the mutant gene rapidly and precisely with MutMap method and developing co-isolated molecular markers according to the mutation site.The results showed that wsl2 showed stripe leaf phenotype at seedling stage,but returned to normal leaf phenotype from jointing stage.wsl2 had the same genetic background as the wild type by detecting the genetic background.And wsl2 was controlled by a recessive nuclear gene with genetic analysis.The association interval contained 9 non-synonymous mutant genes by MutMap method.Seita.9G561800 encoded a PsbP-like protein associated with chloroplast,containing 6 exons and 5 introns.A G/T base mutation occured at 77 bp in the first exon,resulting in an arginine(R)becoming leucine(L).The mutation site of candidate gene in wsl2 mutant was further verified by the dCAPS marker MRI498-1(Cac8Ⅰ)and the co-isolation marker MRI501-3.This study identified a new stripe leaf mutant wsl2,which laid a theoretical foundation for further study on the mechanism of photosynthesis reaction of PsbP gene,enriched the resources of leaf color mutant,and verified the effectiveness of MutMap method in cloning mutant gene in foxtail millet.

关 键 词：谷子 条纹叶突变体wsl2 MutMap 候选基因分析 

分 类 号：S515[农业科学—作物学] Q78[生物学—分子生物学]