谷子SiMS1雄性不育基因的克隆和功能分析

Abstract: Foxtail millet, originated from China, takes great position in dry land agriculture and the structural reforming of crops cultivation. First developed in foxtail millet and put heterosis utility into a reality, partial male sterile lines can reproduce itself by self-fertilization and produce hybrid seeds by crossing with a restoration line, which created a different way of heterosis utility from the known CMS method and the photo and/or temperature sensitive male sterile lines. Cloning and characterization of male sterile genes can provide fundamental information for heterosis utility breeding in crops and so is the situation of CMS and photo sensitive male sterile lines in rice. The purpose of this project is cloning and characterization of the male sterile gene SiMS1 from the widely used line 1066A in foxtail millet breeding, deciphering its gene and molecular net work or pathway which causes the male sterile character. Systematic analysis of haplotypes of SiMS1 from different foxtail millet partial male sterile lines used in hybrid seed production currently and the origin of SiMS1 will also be carried out. With the development of several near-isogenic male sterile lines of SiMS1 in different genetic background, environmental factors, as photo period and temperature, that influencing SiMS1 expression and its pathway will be analyzed. The results of this project will provide theoretic information for foxtail millet herterosis utility breeding and also provide novel knowledge for plant breeding.

摘要：谷子是中国起源的禾谷类粮饲兼用作物，在旱作农业和种植业结构调整中具有重要地位。谷子高度雄性不育系依靠自身少量的自交结实保持不育性和不育系种子，以具有显性抗除草剂特性恢复系实现了杂种优势的生产利用，为作物杂种优势利用方法增添了新途径。这种雄性不育不同于三系法的细胞质不育和光温敏两系法的核不育，是一种不完全败育的特殊核不育类型，但其导致雄性不育的基因尚未克隆，花粉败育的分子和环境影响基础不清。本项目以谷子育种上广泛应用的1066A不育系为材料，克隆其雄性不育基因SiMS1，研究其功能和分子作用网络；以多个该不育基因的近等基因系为材料，分析不同遗传背景和环境对不育系的不育度影响，解析不育基因表达的影响因素；通过单倍型分析和系谱分析了解目前生产应用的不育系不育基因的异同和1066A不育基因的起源，从而为谷子高度雄性不育系的高效繁种和杂交制种提供指导，丰富植物花粉发育分子机理和作物育种理论

目前，依靠高度雄性不育是谷子杂种优势利用的唯一途径。本项目取得了以下进展：（1）首次从谷子高度雄性不育系1066A中克隆了雄性不育基因SiMS1,并通过转基因互补和基因编辑对基因功能进行了验证，证明了所克隆的基因正确。同时，对sims1的系谱和单倍型分析表明sims1和野生型SiMS1基因序列相比存在一个1095 bp的缺失并在谷子杂交育种中得到广泛利用。通过SiMS1基因的表达谱及蛋白互作相关研究，揭示了SiMS1基因的分子功能。这些研究为谷子高度不育系的高效繁种和杂交制种提供指导，同时也丰富了作物杂种优势利用途径。（2）克隆了谷子中第一个细胞核雄性不育基因SiNP1,并通过基因编辑、表达谱分析、转录组及代谢组学分析对SiNP1基因的功能进行系统研究，为谷子杂种优势利用提供了一个重要的候选基因。（3）首次在谷子上建立了基于普通细胞核雄性不育基因的智能不育系统并获得了第一个谷子智能雄性不育系，为谷子杂种优势利用途径增添了新的内容。.得益于该项目的执行，项目负责人在The Crop Journal刊物发表sci论文一篇，申报国家发明专利2项。在人才培养方面，培养了博士研究生1人，硕士研究生2人。