| 水稻雌性败育基因FA的图位克隆 |
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| 引用本文： | 吕凤 唐倩莹 王启明 郑海 尤世民 柏文婷 肖晏嘉 赵志刚,万建民.水稻雌性败育基因FA的图位克隆[J].中国水稻科学,2018,32(6):519-528. |
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| 作者姓名： | 吕凤 唐倩莹 王启明 郑海 尤世民 柏文婷 肖晏嘉 赵志刚 万建民 |
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| 作者单位： | 1.南京农业大学 作物遗传与种质创新国家重点实验室,南京 210095;2. 中国农业科学院 作物科学研究所/农作物基因资源与基因改良国家重大科学工程, 北京 100081 |
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| 基金项目： | 国家自然科学基金委与云南省人民政府共同设立联合基金资助项目(U1502265)。 |
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| 摘 要： | 目的 对水稻胚囊突变体的表型观察、遗传定位和基因克隆,将为研究植物生殖发育奠定理论基础。方法 从粳稻品种宁粳4号突变体库中筛选出一个雌性败育突变体female abortion(fa),对野生型和突变体不同发育时期的胚囊进行细胞学观察并统计胚囊类型。以突变体杂合型为母本,N22为父本构建定位群体,对该表型进行遗传分析,采用图位克隆方法精确定位目的基因。结果 表型分析显示,突变体雌蕊在外观上没有表现出差异,但成熟时植株完全不育。与野生型相比, 该突变体胚囊发育异常,不能形成七细胞八核胚囊结构,而且形成多种类型的异常胚囊。遗传分析表明,该突变性状受一对隐性核基因控制。我们将该基因初定位在第1染色体的分子标记L1和L3之间,通过扩大定位群体,最终将基因定位在分子标记L10和L11之间,物理距离为117 kb。测序分析发现该区间内LOC_Os01g68870外显子上有一个单碱基替换,由胸腺嘧啶(T)替换成胞嘧啶(C),氨基酸由亮氨酸变为脯氨酸,从而导致该表型的出现。qRT-PCR结果显示,相对于OsMADS13、OsAPC6、OsTDL1A基因在突变体fa胚囊中表达量而言,OsDEES1基因在突变体fa胚囊中表达量变化最为显著,FA可能是OsDEES1的上游基因。亚细胞定位结果显示,该蛋白定位于质膜。结论 FA是多孢囊基因MULTIPLE SPOROCYTE 1(MSP1)的新等位基因。本研究进一步明确了该基因对于水稻胚囊发育的重要性,可为探明它所在的调控网络体系提供新的线索。
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| 关 键 词： | 雌性败育突变体 遗传分析 图位克隆 水稻 |
| 收稿时间： | 2018-03-09 |
| 修稿时间： | 2018-04-03 |
Map-based Cloning of Female Abortion (FA) Gene in Rice |
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| Lü Feng,TANG Qianying,WANG Qiming,ZHENG Hai,YOU Shimin,BAI Wenting,XIAO Yanjia,ZHAO Zhigang,WAN Jianmin.Map-based Cloning of Female Abortion (FA) Gene in Rice[J].Chinese Journal of Rice Science,2018,32(6):519-528. |
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| Authors: | Lü Feng TANG Qianying WANG Qiming ZHENG Hai YOU Shimin BAI Wenting XIAO Yanjia ZHAO Zhigang WAN Jianmin |
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| Institution: | 1.State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China;2.National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China |
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| Abstract: | 【Objective】Phenotypic observation, genetic mapping and gene cloning of rice embryo sac mutants will lay a theoretical basis for recovering plant reproductive development.【Method】A female abortion (fa) mutant was obtained from ethyl methylsulfonate (EMS)-treated Ningjing 4. The embryo sacs of the wild-type and mutant were observed at different developmental stages, and the types of mutant embryo sacs were also calculated. The heterozygous mutant was used as the female parent and N22 as the male parent to construct the mapping population. The phenotype was analyzed by genetic analysis, and the target gene was precisely mapped by map-based cloning method. 【Result】Phenotypic analysis showed that the mutant’s pistil showed no difference in appearance, but the plants showed complete sterility at maturity. Compared with the wild type, the development of the mutant embryo sacs was abnormal, which could not form seven-cell eight-nuclear structure. Genetic analysis showed that the fa mutant traits were controlled by a single recessive gene. The gene was initially mapped between the L1 and L3 markers on chromosome 1 and finally between markers L10 and L11 by expanding the mapping population, with a physical distance of 117 kb. Sequencing analysis showed that LOC_Os01g68870 had a single base substitution in exon, from thymine(T) to cytosine(C), leading to amino acid change from leucine to proline and this phenotype. The results of qRT-PCR showed that compared with OsMADS13, OsAPC6 and OsTDL1A, the expression level of OsDEES1 in the mutant fa embryo sacs was the most significant, and FA might act at OsDEES1 upstream. Subcellular localization showed that the FA protein is localized to the plasma membrane. 【Conclusion】 According to the present study, FA gene was a new allele of MSP1. Our work further elucidates the importance of this gene in the development of embryo sacs and provides new clues in regulation the network system. |
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| Keywords: | female abortion mutant genetic analysis map-based cloning rice |
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