水稻光叶性状基因gl1的精细定位与候选基因分析  

FINE MAPPING AND CANDIDATE GENE ANALYSIS OF GLABROUS LEAF AND HULL GENE(gl1) IN RICE(Oryza sativa L.)

作　　者：洪隽[1] 王启钊[1] 富昊伟 吴殿星[1] 舒庆尧[1] 

机构地区：[1]浙江大学原子核农业科学研究所/农业部浙江省核农学重点实验室,浙江杭州310029 [2]浙江嘉兴市农业科学院,浙江嘉兴314016

出　　处：《核农学报》2011年第6期1088-1093,1190,共7页Journal of Nuclear Agricultural Sciences

基　　金：国家科技支撑项目(2009BAA24B05)

摘　　要：光叶水稻是一类在叶片和谷粒等表面都光滑无毛的水稻,是美国南部和非洲一些国家的主栽品种。已有研究表明水稻的光叶性状受一对位于第5染色体上的隐性基因gl1控制。本试验旨在对gl1进行精细定位,分析候选基因,为最终克隆gl1基因、研究毛状体形态建成的调控奠定基础。首先根据与gl1连锁的RFLP标记在第5染色体上的大致位置,有选择地用第5染色体上的部分微卫星(SSR)标记研究热带光叶粳稻(AP9)与毛叶籼稻(浙恢7954)的杂交分离群体中与光叶性状的共分离,获得了与gl1连锁的SSR标记RM17990。在此基础上,采用与RM17990相邻的更多SSR标记以及插入/缺失(InDel)标记将gl1定位在RM17759～RM17831之间(物理区间为538kb～1.649Mb)。为进一步缩小范围,发展了毛细管电泳方法和能够识别单碱基多态性(SNP)的四引物扩增受阻突变体系-PCR(Tetra-primer amplification refractory mutation system PCR,ASPCR)标记,将gl1的区间缩至770.5～977.1kb之间。结合其他实验室有关gl1定位的最新结果,本研究将gl1定位在971～977.1kb之间,并对其中的全部4个注释基因进行测序分析,发现只有LOC_Os05g02750的5’端非翻译区在光叶水稻和毛叶水稻之间存在一个碱基差异,该差异可能是造成光叶突变的原因。Glabrous rice is a kind of rice that is hairless on the surface of its leaves and grains.It is the main grown cultivar in the southern region of the United States and in many African countries.It has been proved that the glabrous trait is controlled by the recessive gene gl1 on chromosome 5.This study aimed to fine map and identify candidate genes for the eventual cloning of gl1 and for further analysis of trichome morphogenesis and regulation.One segregating population was established by crossing a glabrous tropic Japonica rice cultivar（AP9） and non-glabrous indica cultivar（Zhehui 7954）,and microsatellite（SSR） markers were identified around the region of RFLP markers which were previously identified to be linked with gl1.These SSR markers were used for linkage analysis of the AP9×Zhehui 7952 population and proved that gl1 was linked to SSR marker RM17990.Further analysis using more new SSR markers and insertion/deletion（InDel） markers mapped gl1 to the region between RM17759 and RM19831（physical interval 538kb^1.649Mb）.To further delimit gl1,capillary electrophoresis and tetra-primer amplification refractory mutation system PCR markers（referred as ASPCR markers hereafter） were used for SNP analysis and bracketed gl1 to 770.5￣977.1kb.This region was further narrowed down to a 6.1kb segment between 971~977.1kb by combining mapping results of this experiment and a newly published mapping report.After sequencing all four annotated genes within the 6.1kb fragment,only one SNP was discovered between glabrous rice and pubescent rice in the 5＇-end untranslated region of LOC_Os05g02750,hence LOC_Os05g02750 is very likely to be the gene controlling glabrous traits in rice.

关 键 词：水稻 光叶 gl1 毛状体 精细定位 

分 类 号：S511[农业科学—作物学]