TANG Qianying,LÜ,Feng,et al.Map-based cloning of a female and male sterile mutant in rice[J].Journal of Nanjing Agricultural University,2018,41(6):983-993.[doi:10.7685/jnau.201803012]





Map-based cloning of a female and male sterile mutant in rice
唐倩莹1 吕凤1 王启明1 王倩1 尤世民1 柏文婷1 郑海1 王沛然1 赵志刚1 万建民12
1. 南京农业大学作物遗传与种质创新国家重点实验室, 江苏 南京 210095;
2. 中国农业科学院作物科学研究所, 北京 100081
TANG Qianying1 LÜ Feng1 WANG Qiming1 WANG Qian1 YOU Shimin1 BAI Wenting1 ZHENG Hai1 WANG Peiran1 ZHAO Zhigang1 WAN Jianmin12
1. State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China;
2. Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China
ricesterilemutantgene cloning
[目的]水稻的育性对产量有非常重要的影响,对水稻雌、雄不育(female and male sterility)突变体fms进行表型研究和基因图位克隆以丰富水稻生殖发育的分子机制。[方法]用甲磺酸乙酯(ethyl methanesulfonate,EMS)化学诱变粳稻品种‘宁粳4号’,获得1个稳定的雌、雄不育突变体fms,利用醋酸洋红染色、半薄切片和激光共聚焦显微技术对野生型和突变体的花药和胚囊进行细胞学观察和比较;利用FMS/fms杂合子和‘N22’杂交构建的F2群体进行遗传分析和基因图位克隆。[结果]细胞学观察发现:fms的花药壁中绒毡层和中间层缺失,花粉母细胞发育停滞,导致雄配子完全不育;突变体胚囊发育早期孢母细胞增多,成熟胚囊完全退化或出现多于8个的细胞核,导致雌配子不育。遗传分析表明,该不育性状由1对隐性核基因控制。通过图位克隆将FMS基因初步定位在第12号染色体长臂上的分子标记RM27877和RM28404之间。进一步扩大群体并加密分子标记,最终将候选基因定位在标记Ta-4和Ta-7之间,物理距离818 kb,该区间内预测有13个开放阅读框。基因组测序结果显示,fmsOs12g0472500的第1个外显子上有1个碱基突变,使色氨酸变成终止密码子,造成翻译提前终止。表达分析显示FMS在花药中特异性高表达,在胚囊中表达量较高。[结论]水稻育性突变体fms突变性状由Os12g0472500基因的隐性突变引起,该基因编码的蛋白影响水稻花粉和胚囊的发育。
[Objectives] Given the great importance of fertility in rice population, we try to study the sterile mutant fms (female and male sterility)clearly so that much more genes and finer gene regulatory network about rice sterility can be found. [Methods] Cytological observation was made on the stable mutant fms, which was obtained by ethyl methanesulfonate (EMS)-induced transformation of japonica rice variety ‘Ningjing 4’. Besides, we crossed the sterile mutant fms with ‘N22’ to construct a F2 population for genetic analysis and gene mapping. [Results] Both the male and female gametes of the fms mutant were sterile. Through the transverse section, laser scanning confocal microscopy and magenta stain, we observed the absence of the tapetum and the middle layer of the anther cell and the developmental arrest of the pollen mother cell. Moreover, in the embryo sac of the mutant, the number of early sporocyte increased, and the mature embryo sac completely degenerated or displayed more than 8 nucleus, causing the female gametes to be sterile. Genetic analysis indicated that the sterility trait of mutant fms was controlled by a recessive nuclear gene and finally the locus was mapped to a 818 kb region between the markers Ta-4 and Ta-7 on the long arm of rice chromosome 12. There were 13 ORF (open reading frame)in this region. Based on the genome sequencing result, a nucleotide substitution was found on the gene Os12g0472500, which turned tryptophan into an end codon, resulting in the premature termination of translation. The expression of FMS was both higher and specific in anther, and then in the embryo sac. [Conclusions] The fertility of fms mutant is controlled by a recessive nuclear gene (Os12g0472500)which affects the development of rice pollen and embryo sac.


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更新日期/Last Update: 2018-11-23