利用双向导入系和重组自交系定位和挖掘水稻苗期耐冷QTL和有利等位基因  

QTL identification and favorable allele mining of cold tolerance at seedling stage by reciprocal introgression and recombinant inbred line populations in rice

作　　者：曹慧敏 杨贤莉 王立志[3] 李苹苹 翟来圆 姜树坤[3] 郑天清[2] 邱先进[1] 徐建龙[2,4,5] CAO Hui-Min;YANG Xian-Li;WANG Li-Zhi;LI Ping-Ping;ZHAI Lai-Yuan;JIANG Shu-Kun;ZHENG Tian-Qing;QIU Xian-Jin;XU Jian-Long(Key Laboratory of Sustainable Crop Production in the Middle Researches of the Yangtze River,Ministry of Agriculture and Rural Affairs(Co-construction by Ministry and Province),College of Agriculture,Yangtze University,Jingzhou 434025,Hubei,China;Institute of Crop Sciences,Chinese Academy of Agricultural Sciences,Beijing 100081,China;Institute of Crop Cultivation and Tillage,Heilongjiang Academy of Agricultural Sciences,Harbin 150086,Heilongjiang,China;Shenzhen Branch,Guangdong Laboratory for Lingnan Modern Agriculture,Agricultural Genomics Institute at Shenzhen,Chinese Academy of Agricultural Sciences,Shenzhen 518120,Guangdong,China;Hainan Yazhou Bay Seed Laboratory,Sanya 572024,Hainan,China)

机构地区：[1]农业农村部长江中游作物绿色高效生产重点实验室(省部共建),长江大学农学院,湖北荆州434025 [2]中国农业科学院作物科学研究所,北京100081 [3]黑龙江省农业科学院耕作栽培研究所,黑龙江哈尔滨150086 [4]岭南现代农业科学与技术广东省实验室深圳分中心,中国农业科学院农业基因组研究所,广东深圳518210 [5]海南省崖州湾种子实验室,海南三亚572024

出　　处：《作物学报》2023年第10期2633-2642,共10页Acta Agronomica Sinica

基　　金：深圳市基础研究专项(JCYJ20200109150713553);海南省崖州湾种子实验室揭榜挂帅项目(B21HJ0216)资助。

摘　　要：苗期低温冷害常造成水稻生长发育受阻,影响形态建成最终导致减产。鉴定和挖掘优良的耐冷基因,选育耐冷水稻品种,是减少冷害造成水稻产量损失的有效措施。本研究以籼稻明恢63和粳稻02428为亲本构建的双向导入系及重组自交系群体为研究材料,通过人工气候室进行苗期耐冷性鉴定,考查枯萎度和冷害恢复生长后的存活率,结合重测序构建的Bin基因型数据,进行苗期耐冷性QTL定位,3个群体在1号、4号、5号、7号和12号染色体上共检测到13个与水稻苗期耐冷性相关的QTL,解释0.36%~13.63%的表型变异。未定位到不同遗传背景下稳定表达的QTL,但有5个同时影响枯萎度和存活率的QTL被检出,分别位于1号、5号、7号和12号染色体,其中位于1号染色体12,732,139~13,202,097 bp和14,445,778~14,585,009 bp及5号染色体14,658,891~15,684,510 bp的3个QTL的LOD值都高于25,被认为是可靠的主效QTL。结合基因注释、表达谱数据和3K种质资源的苗期耐冷表型数据分析,推测位于5号染色体区间内影响枯萎度和存活率的基因很可能是已克隆的耐冷基因OsWKRY45,而1号染色体2个区间的2个基因是新基因位点,其候选基因分别是LOC_Os01g25540和LOC_Os01g25560。单倍型分析结果表明,LOC_Os01g25540的Hap2、Hap4和Hap9,LOC_Os01g25560的Hap1、Hap8和Hap10为有利单倍型。研究结果将为水稻品种耐冷分子改良提供宝贵的耐冷资源和有利基因。Cold damage often hinders rice growth,affects its morphogenesis and leads finally to serious yield loss.Identifying favorable genes tolerant to cold,and breeding elite rice varieties with cold tolerance is an effective way to solve this problem.In this study,two sets of reciprocal introgression lines and a set of recombinant inbred line derived from xian variety Minghui 63 and geng variety 02428 were evaluated for cold tolerance in climate chamber using cold tolerance-related traits such as wilting degree and survival rate after recovered growth.Combined with the available Bin genotype data generated by previous re-sequencing,13 QTLs for cold tolerance at seedling stage were identified on chromosomes 1,4,5,7,and 12,which explained 0.36%to 13.63%of phenotypic variation.No QTL was stably detected in different genetic backgrounds,whereas five QTLs were simultaneously de-tected for both wilting degree and survival rate on chromosomes 1,5,7,and 12.Among them,QTLs in the regions of 12,732,139–13,202,097 bp and 14,445,778–14,585,009 bp on chromosome 1,and 14,658,891–15,684,510 bp on chromosome 5 had high LOD values above 25 and were considered as reliable main-effect QTLs.Combined with gene annotation,gene expres-sion profile and phenotyping data of cold tolerance of 3K germplasms,the previously cloned gene OsWKRY45 for cold tolerance was regarded as the responsible gene for the QTL in the region of 14,658,891–15,684,510 bp on chromosome 5,while the two candidate genes,LOC_Os01g25540 and LOC_Os01g25560 in the two regions on chromosome 1,were the new candidate genes for both wilting degree and survival rate.Haplotype analysis indicated that Hap2,Hap4,and Hap9 of LOC_Os01g25540 and Hap1,Hap8,and Hap10 of LOC_Os01g25560 were favorable haplotypes for cold tolerance.The results provide the novel germ-plasms and favorable genes for molecular improvement of cold tolerance in rice.

关 键 词：水稻 苗期耐冷 枯萎度 QTL定位 单倍型 

分 类 号：S511[农业科学—作物学]