调控玉米株高和穗位高候选基因Zmdle1的定位  

Candidate Gene Localization of ZmDLE1 Gene Regulating Plant Height and Ear Height in Maize

作　　者：周文期 张贺通 何海军[1] 龚佃明 杨彦忠[1] 刘忠祥[1] 李永生 王晓娟[1] 连晓荣[1] 周玉乾[1] 邱法展[2] ZHOU WenQi;ZHANG HeTong;HE HaiJun;GONG DianMing;YANG YanZhong;LIU ZhongXiang;LI YongSheng;WANG XiaoJuan;LIAN XiaoRong;ZHOU YuQian;QIU FaZhan(Crops Research Institute,Gansu Academy of Agricultural Sciences,Lanzhou 730070;National Key Laboratory of Crop Genetic Improvement,Huazhong Agricultural University,Wuhan 430070)

机构地区：[1]甘肃省农业科学院作物研究所,兰州730070 [2]华中农业大学作物遗传改良全国重点实验室,武汉430070

出　　处：《中国农业科学》2023年第5期821-837,共17页Scientia Agricultura Sinica

基　　金：国家自然科学基金(32160490);作物遗传改良全国重点实验室开放课题资助;甘肃省重大专项(21ZD11NA005,21ZD10NF003);甘肃省农业科学院生物育种专项共同资助(2022GAAS04)。

摘　　要：【目的】株高是玉米株型育种的重要目标性状之一,不仅与玉米籽粒的机械化收获及抗倒伏相关,也与玉米产量密切相关。因此,挖掘玉米株高 QTL/基因并解析其功能具有重要的理论和育种价值,定位一个新的玉米矮秆基因 ZmDLE1,阐明其生物学功能,为加速改良玉米的株型提供重要的理论依据和基因资源。【方法】利用化学诱变剂甲基磺酸乙酯(EMS)诱变甘肃省农业科学院作物研究所自育玉米骨干自交系 LY8405,M2后代分离获得一个单基因调控隐性遗传的玉米矮秆低穗位突变体,M3、M4后代能稳定遗传,命名为 dwarf and low ear mutant1(Zmdle1),通过与 Mo17 杂交构建 F2分离群体,借助极端性状混池测序分析法(BSA-seq)及目标区段重组交换鉴定的方法,基于 Mo17 参考基因组对目标区段内的基因进行挖掘和功能注释,定位候选基因。【结果】开展了 Zmdle1 表型鉴定,突变体 Zmdle1 苗期表型与对照 LY8405 无显著性差异,成熟期植株株高和穗位高较 LY8405 分别降低 87.2 和 55.4 cm,为 35.0%和 62.9%,差异极显著。细胞形态学观察表明,穗下节间数减少及节间细胞长度缩短是导致 Zmdle1 的株高和穗位高显著降低的主要原因。利用 F2:3遗传群体,进行突变基因的遗传分析,后代野生型和突变体植株分离比例符合 3:1(χ^(2)=2.854),说明突变基因为细胞核遗传的单隐性基因。因此,根据 BSA-seq 结果,将候选基因 ZmDLE1 初步定位在玉米第 1 染色体 Bin1.09-1.10 区段约 15 Mb 区间内,进一步利用 Mo17 和Zmdle1 重测序结果开发多态性分子标记,通过图位克隆手段精细定位目标基因,最终将候选基因定位到约 600 kb 大小区间,该区间内有 16 个候选基因。比对重测序数据,发现 Zm00001d033231 在第 2062位置碱基 G 变成 A,导致氨基酸由甘氨酸变成丝氨酸,且转录水平表达比 LY8405 极显著降低,Zm00001d033234 第 223 位置碱基由 T变成C,导致第75位�【Objective】 Plant height is one of the important target traits in maize plant type breeding, which is closely related not only to mechanized grain harvesting and lodging resistance, but also to maize yield. Therefore, it is of great theoretical and breeding value to isolate QTL/gene of maize plant height and analyze its function. This study aims to locate a novel maize dwarf gene ZmDLE1, clarify its biological function, and provide important theoretical basis and gene resources for accelerating the improvement of maize plant type. 【Method】 A single recessive mutant was derived in maize inbred line LY8405, from Crop Research Institute of Gansu Academy of Agricultural Sciences, by chemical mutagenic agent Ethyl Methyl Sulfonate (EMS). A maize dwarf and low ear mutant was isolated from the M2progeny, and the M3and M4progeny could stably inherit, which was named dwarf and low ear mutant1 (Zmdle1). The F2population was constructed by hybridization with Mo17 and was identified by bulked segregate analysis-sequencing (BSA-seq) and target segment recombination exchange. Based on the Mo17 reference genome, the genes in the target region were obtained and functionally annotated to locate candidate genes.【Result】Phenotypic identification of Zmdle1 was carried out, and the phenotype of Zmdle1 at seedling stage was not significantly different from that of the control LY8405. The plant height and ear height of Zmdle1 at mature stage were reduced by 87.2 cm and 55.4 cm, respectively, accounting for about 35.0% and62.9%, difference is extremely significant. Morphological observation showed that the decrease of internode number and the shortening of internode cell length were the main reasons for the significant decrease of plant height and ear height of Zmdle1. The genetic analysis of the mutant gene was conducted by using the F2:3genetic populations. The Zmdle1 mutant is inherited in a 3:1(χ^(2)=2.854) ratio and is a single recessive gene. Therefore, according to the results of BSA-seq, the candidate gene ZmDLE1 wa

关 键 词：玉米 株高 穗位高 BSA-seq 基因定位 关联分析 

分 类 号：S513[农业科学—作物学]