水稻雌雄不育突变体Osfma2的细胞学观察及基因图位克隆
#共同第一作者;
收稿日期: 2021-03-23
修回日期: 2021-04-22
网络出版日期: 2022-01-10
基金资助
国家自然科学基金资助项目(31991224);国家自然科学基金资助项目(31971909);国家自然科学基金资助项目(U2002202)
Cytological Observation of a Female and Male Sterile Osfma2 Mutant in Rice and Its Map-based Cloning
#These authors contributed equally to this work;
Received date: 2021-03-23
Revised date: 2021-04-22
Online published: 2022-01-10
【目的】通过构建分离群体定位并克隆水稻雌雄不育基因OsFMA2,并对其在水稻育性调控中的功能进行探究。【方法】通过EMS诱变水稻宁粳4号得到稳定的不育突变体,对突变体进行表型及细胞学观察,利用精细定位和Mut-map相结合的方法克隆了水稻雌雄不育基因OsFMA2,通过实时荧光定量PCR检测其在各组织表达水平差异,利用水稻原生质体表达系统进行OsFMA2蛋白的亚细胞定位。【结果】细胞学观察发现突变体为雌雄不育。利用极端个体将其精细定位于水稻第6染色体长臂448 kb的区间内。结合全基因组重测序结果,最终确定OsFMA2为目标基因。该基因编码一个复制蛋白,在单子叶植物中高度保守。OsFMA2具有组织特异性,在幼穗中高表达。亚细胞定位结果显示OsFMA2蛋白定位于细胞核中。【结论】OsFMA2在幼穗中高表达,可能参与了水稻雄配子第一次减数分裂前期同源重组过程,同时,OsFMA2的表达对雌配子发育也产生了不利影响。
张涛荟, 王海宇, 万华, 张莉萍, 谢振威, 陈可毅, 何晓栋, 赵志刚, 万建民 . 水稻雌雄不育突变体Osfma2的细胞学观察及基因图位克隆[J]. 中国水稻科学, 2022 , 36(1) : 13 -26 . DOI: 10.16819/j.1001-7216.2022.210312
【Objective】 The work aims to locate and clone the rice female and male sterile gene OsFMA2 by constructing a segregated population and explore its function in the regulation of rice fertility. 【Method】 Stable sterile mutants were obtained by EMS mutagenesis of japonica rice Ningjing 4, and the phenotypic and cytological observation of the mutant was conducted. Using map-based cloning and Mut-map methods, the female and male sterile gene OsFMA2 was fine-mapped and cloned. The expression pattern of OsFMA2 in various tissues was analyzed by the real-time quantitative PCR technique. Subcellular localization of OsFMA2 protein was performed with the rice protoplast expression system. 【Result】 The cytological observation revealed that the Osfma2 mutant was male- and female-sterile, and the extreme individuals were used to locate the gene in a 448-kb interval on the long arm of chromosome 6. The gene OsFMA2 is predicted to encode a replication protein that is highly conserved in monocots. The gene OsFMA2 was tissue-specific and was highly expressed in young panicles. Subcellular localization analysis showed that the OsFMA2 protein was localized on the nucleus. 【Conclusion】The gene OsFMA2 is highly expressed in young panicles, and it may participate in the homologous recombination of the first meiotic division of male gamete. At the same time, the expression of OsFMA2 also has an adverse effect on the development of female gametes.
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