水稻雄性不育突变体tpa1的表型鉴定与精细定位  

Phenotypic identification and fine mapping of male sterile mutant tpa1 in rice

作　　者：万应春 班义结 蒋钰东[2] 王亚欣 刘晶晶 刘晓晴 程育林 王楠[1] 冯萍[1] WAN Ying-Chun;BAN Yi-Jie;JIANG Yu-Dong;WANG Ya-Xin;LIU Jing-Jing;LIU Xiao-Qing;CHENG Yu-Lin;WANG Nan;FENG Ping(Rice Research Institute of Southwest University/Chongqing Key Laboratory of Application and Safety Control of Genetically Modified Crops/Engineering Research Center of South Upland Agriculture,Ministry of Education,Chongqing 400716,China;Key Laboratory of Southwest Rice Biology and Genetic Breeding,Ministry of Agriculture and Rural Affairs/Luzhou Branch of National Rice Improvement Center,Rice and Sorghum Research Institute,Sichuan Academy of Agricultural Sciences(Deyang Branch of Sichuan Academy of Agricultural Sciences),Deyang 618000,Sichuan,China)

机构地区：[1]西南大学水稻研究所/转基因植物与安全控制重庆市市级重点实验室/南方山地农业教育部工程研究中心,重庆400716 [2]四川省农业科学院水稻高粱研究所(四川省农业科学院德阳分院),国家水稻改良中心四川泸州分中心/农业农村部西南水稻生物学与遗传育种重点实验室,四川德阳618000

出　　处：《作物学报》2024年第5期1104-1114,共11页Acta Agronomica Sinica

基　　金：重庆市研究生科研创新项目(CYS22224);重庆市技术创新与应用发展专项重点项目(CSTB2022TIAD-KPX0014)资助。

摘　　要：雄性不育材料是杂交水稻育种的关键。本研究通过甲基磺酸乙酯(ethyl methane sulfonate,EMS)诱变优良籼稻保持系西农1B,筛选得到一个雄性不育突变体tpa1。tpa1在营养生长阶段与野生型无差异,生殖生长阶段表现雄配子不育,雌配子发育正常。表型观察发现,tpa1花粉完全破碎消失,花药外壁角质层异常,花药内壁乌式体排列异常,胼胝质合成异常,绒毡层凋亡异常,且花粉外壁缺失柱状层。遗传分析表明该突变性状受1对隐性核基因控制,利用突变体tpa1与缙恢10号构建遗传群体,最终将TPA1基因定位于4号染色体引物标记N9和N11之间,物理距离为74 kb,该区间共15个预测基因,通过重测序仅发现在LOC_Os04g53380的外显子上发生了单碱基替换,导致了翻译的提前终止,随后对野生型和突变体tpa1该位点进行测序证实了这一突变,因此将该基因确定为TPA1的候选基因,TPA1是一个未被报道过的新的雄性不育基因。本研究将为TPA1基因的功能研究奠定基础。Male sterile material is the key to hybrid rice breeding.In this study,a male sterile mutant tpa1 was screened by ethyl methane sulfonate(EMS)mutagenesis of excellent indica cultivar Xinong 1B.There was no difference between tpa1 and wild type at vegetative growth stage.At reproductive stage,male gametes were sterile and female gametes developed normally.Phenotypic observation showed that the pollen of tpa1 was completely broken and disappeared,the stratum corneum of the outer wall of the anther was abnormal,the Ubisch body of the inner wall of the anther was abnormal,the callose synthesis was abnormal,the tapetum apoptosis was abnormal,and the outer wall of the pollen was missing bacula layer.Genetic analysis showed that the mutant trait was controlled by a pair of recessive nuclear genes.A genetic population was constructed using the tpa1 mutant and Jinhui 10.Finally,TPA1 gene was located between the markers N9 and N11 on chromosome 4,with a physical distance of 74 kb.There were 15 predicted genes in this interval.Resequencing identified that only a single base substitution occurred in the exon of LOC_Os04g53380,leading to an early termination of translation.Subsequent sequencing of the wild-type and mutant tpa1 confirmed this mutation,thus identified the gene as a candidate gene for TPA1,indicating that TPA1 was a new male sterile gene.This study will lay a foundation for the functional study of TPA1 gene.

关 键 词：水稻(Oryza sativa L.) 雄性隐性核不育 tpa1 精细定位 

分 类 号：S511[农业科学—作物学]