普通野生稻早抽穗基因OsEHD8的鉴定与进化分析  

Identification and Evolutionary Analysis of the Early Heading Gene OsEHD8 in Common Wild Rice(Oryza rufipogon Giff.)

作　　者：鄢柳慧 钟琦 马增凤[1] 韦敏益 刘驰[1] 秦媛媛[3] 周小龙 黄大辉[1] 卢颖萍 秦钢[1] 张月雄[1] YAN LiuHui;ZHONG Qi;MA ZengFeng;WEI MinYi;LIU Chi;QIN YuanYuan;ZHOU XiaoLong;HUANG DaHui;LU YingPing;QIN Gang;ZHANG YueXiong(Rice Research Institute,Guangxi Academy of Agricultural Sciences/Guangxi Key Laboratory of Rice Genetics and Breeding/State Key Laboratory for Conservation and Utillzation of Subtropical Agro-Bioresources,Nanning 530007;Liuzhou Branch,Guangxi Academy of Agricultural Sciences/Liuzhou Research Center of Agricultural Sciences,Liuzhou 545000,Guangxi;Agricultural Science and Technology Information Research Institute,Guangxi Academy of Agricultural Sciences,Nanning 530007)

机构地区：[1]广西农业科学院水稻研究所/广西水稻遗传育种重点实验室/亚热带农业生物资源保护与利用国家重点实验室,南宁530007 [2]广西农业科学院柳州分院/柳州市农业科学研究中心,广西柳州545000 [3]广西农业科学院农业科技信息研究所,南宁530007

出　　处：《中国农业科学》2024年第14期2703-2716,I0001-I0010,共24页Scientia Agricultura Sinica

基　　金：国家自然科学基金(32360490);广西自然科学基金(2013GXNSFBA019067)。

摘　　要：【目的】抽穗期是影响水稻(Oryza sativa L.)区域适应性和产量的关键性状。鉴定早抽穗期基因对于丰富和完善水稻抽穗期调控网络,为品种适应性改良提供重要信息。【方法】以早抽穗的染色体片段代换系CL33和晚抽穗的受体亲本93-11为研究材料,对其主要农艺性状进行调查分析;构建2个极早抽穗和极晚抽穗的DNA混合池;对其进行全基因组重测序和BSA-Seq分析定位抽穗期关联的区域,开发该区域内InDel标记进行精细定位,对定位区间进行基因预测和候选基因分析,确定LOC_Os08g07740为目标候选基因;随后对该基因进行克隆和等位基因分析。此外,还利用生物信息学软件对LOC_Os08g07740上下游约40 kb范围内的基因组数据进行分析,确定该基因在Indica、Japonica和O.rufipogon 3个水稻亚群间的遗传和系统进化关系。【结果】在南宁自然长日照和自然短日照条件下,CL33的抽穗期均比受体亲本93-11短20—25 d;此外,在自然长日照条件下,CL33植株除了穗长变短和每穗粒数减少之外,其他农艺性状与93-11无显著差异。遗传分析表明,CL33早抽穗性状受1对隐性基因控制。该基因被精细定位于水稻第8染色体短臂PSM8-6与PSM8-8标记间约100 kb区间内,该区域内有15个预测的候选基因。其中,ORF13(LOC_Os08g07740)与已知抽穗期基因DTH8/Ghd8等位;通过对ORF13基因的测序及序列比对,该基因全长888 bp,无内含子,编码295个氨基酸;与受体亲本93-11相比,CL33中的LOC_Os08g07740在第535—536位碱基和第820—821位碱基之间分别有6 bp的插入和9 bp的缺失,导致编码的氨基酸序列改变。因此,确定其为目标候选基因,暂命名为Os EHD8。进化分析表明,LOC_Os08g07740内,相较于O.rufipogon,Indica和Japonica的遗传多样性显著下降,其中,Indica减少了62.53%,Japonica减少了53.76%,而Indica和Japonica之间的差异不显著。该基因共有13个单倍型,其中Hap_2单倍型可能是3个亚群【Objective】The heading date plays a crucial role in influencing the regional adaptation and yield of rice(Oryza sativa L.).The identification of early heading genes contributes significantly to enhancing and fine-tuning the regulatory network that controls rice heading,which provides valuable genetic resources for molecular breeding with the goal of achieving early maturity and high yield in rice.【Method】CL33,a chromosome segment substitution line with early heading,and 93-11,its recipient parent with late heading,were used as research materials to investigate and analyze their major agronomic traits.Two DNA pools were constructed,comprising plants exhibiting extremely early and late heading.Whole-genome resequencing and BSA-Seq analyses were then conducted to locate the genomic region associated with the heading date.In the subsequent steps,InDel markers within this identified region were developed for fine mapping.The gene LOC-Os08g07740 emerged as the primary candidate gene within localization intervals,determined through gene prediction,candidate gene analyses,as well as references to relevant literatures.This candidate gene was subsequently cloned and analyzed for allelic variations.Moreover,we explored the genetic and phylogenetic relationships of the LOC_Os08g07740 gene within the three rice subgroups,Indica,Japonica and O.rufipogon.This analysis involved studying genomic data within approximately 40 kb upstream and downstream of the gene utilizing bioinformatics software.【Result】Under both natural long-day and short-day conditions in Nanning,Guangxi,CL33 exhibited a 20-25 days shorter than its recipient parent 93-11.Moreover,under natural long-day conditions,the agronomic traits of CL33 were largely similar to those of 93-11,with the exception of a shortened spike length and a reduced number of grains per spike.Genetic analysis revealed that the early heading trait in CL33 was controlled by a recessive gene.This gene was finely localized within a 100 kb region between the markers PSM8-6 and PS

关 键 词：水稻 抽穗期 DTH8/Ghd8 BSA-seq 进化 

分 类 号：S511[农业科学—作物学]