干旱区生态环境知识资源中心(Arid): 水稻PSM标记的发展及抗虫基因的分子定位

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	水稻PSM标记的发展及抗虫基因的分子定位
其他题名	Development of Position-specific Microsatellite Markers and Molecular Mapping of Insect Resistant Genes in Rice (Oryza sativa L.)
	黄朝锋; 张桂权
来源期刊	分子植物育种
ISSN	1672-416X
出版年	2003
卷号	1 期号:4 页码:572-574
中文摘要	水稻是世界上最重要的粮食作物之一,为世界近一半人口提供食物来源.水稻微卫星图谱的构建有利于遗传基础的研究及分子育种.本研究通过发展位置特异性微卫星标记,对微卫星标记进行了图谱的整合,并利用微卫星等标记对水稻抗稻瘿蚊基因Gm6和抗褐飞虱基因Bph3进行了分子定位.主要研究结果如下:1、利用网上公布的序列信息进行位置特异性微卫星引物的设计和微卫星图谱的整合.共发展了198个PSM标记,有105个标记的基序为GA/CT重复,占53.3%,绝大多数标记(98.0%)的重复序列长度在20bp以上.将原有微卫星标记和本实验发展的PSM标记整合到RGP遗传图谱上,整合后总的SSR标记数为718个,新图谱的遗传距离总长度为1527.2cM,平均标记密度为每2.13cM有一个SSR标记,其中第1染色体的标记最密(1.73cM/个),而第11染色体的标记密度最低(3.32cM/个).将本研究发展的微卫星图谱与IRMI发表的高密度微卫星进行了比较,结果显示本研究发展的微卫星图谱标记分布比较均匀,只有3个区域标记间遗传间距在10cM以上,5个区域在8-10cM,而IRMI微卫星图谱分别有17和12个.2、采用G2417-2-1抗蚊青占(239株)和G3004-4抗蚊青占(243株)两个F2作图群体对水稻抗稻瘿蚊进行了分子定位,并对该基因区域进行物理作图.利用两个SSR标记(RM348和RM255)和一个STS标记(RG476/ ScaⅠ)将Gm6基因初定位在第4染色体的长臂上,然后利用网上公布的序列信息在附近设计高密度的位置特异微卫星标记,对Gm6基因作精细定位,将其定位在PSM112和PSM114两标记之间,遗传距离为0.4 cM的区域内,另有PSM101、PSM106和PSM115三个标记与Gm6基因完全连锁.筛选出两个BAC重叠克隆AL606645和AL606660用于Gm6基因的物理作图,利用这两个克隆上有多态的位置特异微卫星标记将该基因限定在大约158kb的物理区域内.3、利用RH *七丝软占F2作图群体对水稻抗褐飞虱基因Bph3进行分子定位.根据F3株系表型鉴定结果,从F2作图群体的237株中选出27株抗感极端株用于Bph3基因的分子定位.选择第4染色体上具有多态性的微卫星标记对该基因进行连锁分析,发现两个标记RM261和RM119与Bph3的遗传距离均为8.0cM.在这两标记之间的区域内发展位置特异微卫星标记,两个新发展的微卫星标记PSM323和PSM194与Bph3基因的遗传距离分别为8.0cM和3.8cM.因而Bph3基因被定位在第4染色体长臂上PSM323和PSM194两标记之间.
英文摘要	Rice is the important crop for human consumption, providing staple food for about half the world’ s popu-lation. The construction of rice microsatellite map will contribute greatly to genetic research and molecular breeding. In this study, PSM (position-specific microsatellite) markers were developed in rice, and then an in-tegrated microsatellite map was constructed by using these PSM markers and also some other published RM markers. Two insect resistant genes viz. Gm6 for gall midge and Bph3 for brown planthopper were mapped using some SSR markers and one STS marker in rice. The main results were as follows: Based on published sequences on the websites, position-specific microsatellite primers were designed and an integrated microsatellite map was constructed in silico. A total of 198 primer pairs were designed, among which 105 primer pairs (53.3%) whose motif types were poly (GA/CT) n. The majority (98%) of developed markers had SSRs2￡20bp in length. Previously mapped SSR markers and newly developed PSM markerswere integrated into RGP map and finally a 718-SSR-marker genetic map covering 1527.2 cM was constructed. The integrated map was estimated to have one SSR marker at every 2.13 cM on an average. Chromosome 1 has a highest average density of SSR markers i.e. one at every 1.73 cM, whereas chromosome 11 has a low-est average density of SSR markers i.e. one at 3.32 cM. Compared with newly developed IRMI map with high density of SSR markers, our developed map has more uniform distribution of makers with 3 genetic gaps5:10 cM and 5 genetic gaps between 8 cM and 10 cM, whereas there are 17 and 12 gaps on IRMI map, respectively. Two segregating populations derived from two crosses between two susceptible varieties, ’ G2417 - 2 -1’ and ’ G3004 - 4’, and the resistant variety ’ Kangwenqingzhan’ were used for maping and physical mapping of a gall midge resistance gene Gm6 in rice. Based on linkage analysis of these two populations consisting of 239 and 243 F2 individuals, respectively, Gm6 was mapped on the long arm of chromosome 4 by using two SSR markers (RM348 and RM255) and one STS marker (RG476/Sca I ). Position-specific microsatellite markers were developed for fine mapping of Gm6 gene on the basis of published sequences on the websites. The results showed that Gm6 was mapped between PSM112 and PSM114 at the genetic interval of 0.4 cM, whereas three additional markers PSM101, PSM106, and PSM115 showed no recombination with Gm6, Two overlapping BAC clones AL606660 and AL606645 covering Gm6 gene were used for physical mapping. With the known physical information of all the markers in the two overlapping BAC clones, a sequence of - 158kb was defined as the Gm6 containing region. An F2 mapping population derived from a cross between the resistant variety ’RH’ and the susceptible variety ’ Qishiruanzhan’ was used to map Bph3 gene that confers resistance to brown planthopper of rice. Based on the phenotypic results of F3 lines, 27 F2 individuals selected from the population consisting of 237 F2individuals were used for molecular mapping of Bph3 gene. Through linkage analysis of polymorphic SSR markers of chromosome 4 with Bph3 gene, two SSR markers (RM261 and RM119) were found to have dis-tances of 8.0 cM and 8.0 cM with Bph3, respectively. For further analysis, PSM (position-specific mi-crosatellite) markers were developed in the region between RM261 and RM119. Two polymorphic PSM mark-ers (PSM323 and PSM194) were mapped with Bph3 gene of 8.0cM arid 3.8cM distances, respectively. Therefore, Bph3 gene was mapped between PSM323 and PSM194 on the long arm of chromosome 4.
中文关键词	水稻 ; 微卫星标记 ; 稻瘿蚊 ; 褐飞虱 ; 基因定位 ; 物理作图
英文关键词	Rice Microsatellite Gall midge Brown planthopper Gene mapping Physical mapping
语种	中文
国家	中国
收录类别	CSCD
WOS类目	AGRONOMY
WOS研究方向	Agriculture
CSCD记录号	CSCD:1369253
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/203974
作者单位	华南农业大学, 广州, 广东 510642, 中国
推荐引用方式 GB/T 7714	黄朝锋,张桂权. 水稻PSM标记的发展及抗虫基因的分子定位[J],2003,1(4):572-574.
APA	黄朝锋,&张桂权.(2003).水稻PSM标记的发展及抗虫基因的分子定位.分子植物育种,1(4),572-574.
MLA	黄朝锋,et al."水稻PSM标记的发展及抗虫基因的分子定位".分子植物育种 1.4(2003):572-574.