J Integr Plant Biol. ›› 2003, Vol. 45 ›› Issue (7): 871-877.

• Research Articles • Previous Articles     Next Articles

Fine Mapping of the Blast Resistance Gene Pi15, Linked to Pii,

PAN Qing-Hua, HU Zhen-Di, Tanisaka TAKATOSHI, WANG Ling   

Abstract:

The gene Pi15 for resistance of rice to Magnaporthe grisea was previously identified as being linked to the gene Pii. However, there is a debate on the chromosomal position of the Pii gene, because it was originally mapped on chromosome 6, but recent work showed it might be located on chromosome 9. To determine the chromosomal location of the Pi15 gene, a linkage analysis using molecular markers was performed in a F2 mapping population consisting of 15 resistant and 141 susceptible plants through bulked-segregant analysis (BSA) in combination with recessive-class analysis (RCA). Out of 20 microsatellite markers mapped on chromosomes 6 and 9 tested, only one marker, RM316 on chromosome 9, was found to have a linkage with the Pi15 gene with a recombination frequency of (19.1 ± 3.7)%. To confirm this finding, four sequence-tagged site (STS) markers mapped on chromosome 9 were tested. The results suggested that marker G103 was linked to the Pi15 gene with a recombination frequency of (5.7 ± 2.1)%. To find marker(s) more closely linked to the Pi15 gene, random amplified polymorphic DNA (RAPD) analysis was performed. Out of 1 000 primers tested, three RAPD markers, BAPi15486, BAPi15782 and BAPi15844 were found to tightly flank the Pi15 gene with recombination frequencies of 0.35%, 0.35% and 1.1%, respectively. These three RAPD markers should be viewed as the starting points for marker-aided gene pyramiding and cloning. A new gene cluster of rice blast resistance on chromosome 9 was also discussed.

稻瘟病抗病基因Pi15 的精细定位
潘庆华 胡珍娣 谷坂隆俊 王 玲
(1. 华南农业大学资源环境学院植物抗病遗传学研究室,广州 5 1 0 6 4 2;
2. Laboratory of Plant Breeding, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan)

摘要: 稻瘟病抗病基因Pi15曾被作者鉴定为与已知抗病基因Pi15具有连锁关系,但是,pii基因究竟位于染色体6还是9上存在争议。为了确定Pi15基因的染色体位置,利用分子标记在由15个抗病个体和141个感病个体组成的F2群体中,通过混合群体分离法(BSA)与隐性群体分析法(RCA)相结合的手段,对目标基因进行了连锁分析。首先,从染色体6和9分别选择lO个微卫星标记进行了分析,结果表明,只有位于染色体9的RM316与目标基因连锁,重组率为(19.1±3.7)%。为了进一步确定这种连锁关系,从染色体9选择了4个序列标定位点(STS)标记进行分析,结果表明,只有G103与目标基因连锁,重组率为(5.7±2.1)%。为了获得与目标基囚更加紧密连锁的分子标记,对目标基因进行了RAPD分析。在筛选、分析了l000个随机引物之后,从中获得了3个目标基因紧密连锁的分了标记BAPi15486、BAPi15782、BAPi15844。它们与目标基因的重绀率分刖为O.35%、O.35%和1.1%。这些紧密连锁的分子标记可作为分了标记辅助基因聚合和克隆的出发点。

关键词: 混合群体分离法( BSA);微卫星;水稻;稻瘟病;RAPD;隐性群体分析法( RCA);序列标定位点( STS)
通讯作者。 E-mail: <panqh@scau.edu.cn>。

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