糜子GBSSI基因功能标记的开发与验证  

Development and validation of functional markers of GBSSI gene in proso millet

作　　者：丁敏 段政勇 王宇卓 薛亚鹏 王海岗[2] 陈凌[2] 王瑞云[1,2] 乔治军 DING Min;DUAN Zheng-Yong;WANG Yu-Zhuo;XUE Ya-Peng;WANG Hai-Gang;CHEN Ling;WANG Rui-Yun;QIAO Zhi-Jun(College of Agronomy,Shanxi Agricultural University,Taigu 030801,Shanxi,China;Center for Agricultural Genetic Resources Research,Shanxi Agricultural University/Key Laboratory of Crop Gene Resources and Germplasm Enhancement on Loess Plateau,Ministry of Agriculture and Rural Affairs/Shanxi Key Laboratory of Genetic Resources and Genetic Improvement of Minor Crops,Taiyuan 030031,Shanxi,China)

机构地区：[1]山西农业大学农学院,山西太谷030801 [2]山西农业大学农业基因资源研究中心/农业农村部黄土高原作物基因资源与种质创制重点实验室/杂粮种质资源发掘与遗传改良山西省重点实验室,山西太原030031

出　　处：《作物学报》2023年第3期703-718,共16页Acta Agronomica Sinica

基　　金：财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-06-14.5-A16);山西省现代农业产业技术体系建设(杂粮)项目(2022-03);国家自然科学基金项目(31271791);山西省自然科学基金项目(201901D11126)资助。

摘　　要：根据糜子胚乳直链淀粉合成调控基因Waxy(GBSSI)上已知功能位点的核苷酸差异设计功能分子标记,并对山西省270份糜子资源进行基因型检测和基因分型。通过对粳糯性状的鉴定,将表型与基因型结合对分子标记进行验证。对46份不同基因型糜子资源进行直链淀粉含量测定,分析该基因的遗传效应,评估其育种利用价值。GBSSI基因在糜子中以2种形式(L和S)存在,针对S型基因15 bp的InDel位点设计了一个功能标记(RYW214),该标记能有效区分126 bp、141 bp和杂合(126/141 bp)3种带型,粳糯性鉴定显示该标记结果与表型鉴定结果吻合度高,达93.30%,Pearson相关性指数r为0.745。针对L型基因上的一个单核苷酸插入位点和一个SNP位点,设计了2个CAPS标记(RYW215、RYW216),该标记可对L基因准确分型。270份资源中,共有11种基因型。其中,基因型S_(-15)/L_(F)最多,有84份(31.10%),其次是S_(0)/S_(-15)/L_(F)(22.96%),S_(-15)/L_(Y)/L_(F)最少(0.74%),未发现S_(-15)/L_(C)。46份材料中,直链淀粉含量较高的基因型为S_(0)/L_(Y)/L_(F)(15.50%),最低为S_(-15)/L_(F)(0.58%)。突变基因型S_(-15)会导致直链淀粉含量出现质的下降,杂合基因型S0S_(-15)直链淀粉含量稍高于纯合基因型S_(-15)。Functional molecular markers were designed based on nucleotide differences of known functional loci on Waxy(GBSSI),a regulation gene for amylose synthesis in proso millet.Genotypes and phenotypes of 270 proso millet accessions from Shanxi province were tested and verified by combining phenotypes and genotypes.The amylose content of 46 proso millet acces-sions with different genotypes was determined to explore the genetic effect of this gene and evaluate its breeding value.Waxy gene of proso millet existed in two forms(L and S),targeting 15 bp on S type gene.A functional marker was designed for InDel(RYW214),which could effectively distinguish 126 bp,141 bp,and heterozygous(126/141 bp)types.The consistency between the marker and phenotypic identification was 93.30%,and the Pearson correlation index(r)was 0.745.Two CAPS markers(RYW215,RYW216)were designed for a single nucleotide insertion site and a SNP site in the L-type gene.Among 270 acces-sions,genotype S_(-15)/L_(F)accounted for the most(31.10%),S_(0)/S_(-15)/L_(F)was the second(22.96%),and S_(-15)/L_(Y)/L_(F)was the least(0.74%).Genotype S_(-15)/LC was absent.Among 46 proso millet accessions,the genotypes with the highest mean amylose content were S_(0)/L_(Y)/L_(F)(15.50%)and S_(-15)/L_(F)(0.58%).The mutant alleles of L type gene had little effect on amylose content,and the existence of wild type S_(0)was enough to guarantee amylose content in endosperm.The existence of mutant S_(-15)would lead to a qualitative decline in amylose content,and the amylose content of heterozygous genotype S_(0)S_(-15)was slightly higher than that of homozygous genotype S_(-15).

关 键 词：糜子 GBSSI基因 功能分子标记 直链淀粉含量 

分 类 号：S516[农业科学—作物学]