摘要
为鉴定出黑青稞粒色形成关键调控基因,本研究基于黑粒青稞品种昆仑17号和白粒青稞资源白91-97-3籽粒形成的三个关键时期的二代转录组测序(RAN-seq)结果,得到显著差异表达基因HvnISC,同时对该基因进行同源克隆、序列比对和表达量分析。结果表明:HvnISC基因的编码区(CDS)在昆仑17号和白91-97-3中无序列差异;蛋白的二三级结构分析表明该蛋白结构稳定。启动子区域在两品种中发生一个碱基差异,造成该碱基位点结合的转录因子在黑粒品种中发生改变,导致HvnISC基因在黑粒和白粒青稞中的表达产生了差异。实时荧光定量(RT-qPCR)结果表明,该基因在粒色形成的中后期,黑粒中表达量极显著高于白粒(P<0.01)。结合前人研究结果我们提出了该基因通过调控内源环境的离子浓度,影响多酚氧化酶(PPO)的活性,间接地调控了青稞黑粒粒色的形成,该研究结论需要进一步验证。
To identify key regulatory genes involved in the black grain coloration of highland barley,this study analyzed RNA-seq data from three critical grain development stages of the black-grained variety Kunlun 17 and the white-grained variety Bai 91-97-3.The significantly differentially expressed gene HvnISC was identified and subjected to homologous cloning,sequence alignment,and expression analysis.Results revealed no coding sequence(CDS)differences in HvnISC between the two varieties,and secondary/tertiary protein structure analyses indicated structural stability.However,a single nucleotide variation in the promoter region altered transcription factor binding in the black-grained variety,leading to differential HvnISC expression between the two varieties.RT-qPCR confirmed that HvnISC expression was significantly higher(P<0.01)in the black-grained variety during the mid-to-late stages of grain coloration.Based on previous studies,we propose that HvnISC indirectly regulates black pigmentation by modulating endogenous ion concentrations to influence polyphenol oxidase(PPO)activity,though further experimental validation is required.
作者
吴泰瑞
姚有华
姚晓华
WU Tairui;YAO Youhua;YAO Xiaohua(Qinghai University,Xining Qinghai 810016,China;Qinghai Academy of Agriculture and Forestry Sciences,Xining Qinghai 810016,China;National Oat Improvement Center Qinghai Highland Barley Branch,Xining Qinghai 810016,China;Qinghai Key Laboratory of Highland Barley Genetics and Breeding,Xining Qinghai 810016,China;Qinghai-Tibet Plateau Germplasm Research and Utilization Laboratory,Xining Qinghai 810016,China)
出处
《青海农林科技》
2025年第2期55-61,共7页
Science and Technology of Qinghai Agriculture and Forestry
基金
科技特派员专项(2025-NK-P01)
国家大麦产业技术体系项目(CARS-05)。
关键词
青稞
调控基因
HvnISC
多酚氧化酶
克隆和表达
Hulless barley
Regulatory gene
HvnISC
Polyphenol oxidase
Cloning and expression
