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杂种不育是水稻籼粳亚种间杂种优势利用的主要障碍,开展杂种不育基因的克隆和功能分析对于克服和缓解亚种间杂种的不育性具有重要的意义.本研究在S-b座位初定位的基础上,根据水稻基因组的序列发展微卫星标记,将S-b座位定位在微卫星标记PSM8和PSM202之间.为了精细定位S-b座位,以S-b座位的近等基因系为材料培育了包含有3910株的F2作图群体,同时根据PSM8和PSM202界定范围内的基因组序列发展了2个微卫星标记,2个插入缺失多态性标记和4个CAPS标记.利用作图群体中的重组株进行花粉育性表型与新发展标记基因型的连锁分析,结果表明标记W4与S-b座位完全连锁,而标记A8和A14位于S-b座位的两侧,与S-b座位的遗传距离分别为0.026和0.038cM.将多态性标记与该区域克隆的序列进行整合,结果表明,新发展的多态性标记锚定在了AC093089,AC079021和AC134931三个首尾相连的克隆上.根据各标记在物理图谱上的位置,最终将S-b座位界定在了A8与A14之间27kb的物理距离内.RiceGAAS注释表明该区域有7个预测ORF.该结果为S-b座位进一步的功能分析奠定了基础.
Hybrid sterility is a major obstacle to the utilization of heterosis among indica-japonica subspecies in rice. Cloning and functional analysis of the hybrid sterility gene are important for overcoming and alleviating the sterility of inter-subspecific hybrids.In this study, Based on the sequence of rice genome, microsatellite markers were developed to locate Sb locus between microsatellite markers PSM8 and PSM202.In order to finely locate Sb locus, Sb alleles of Sb locus were used to cultivate plants containing 3910 Strain F2, and two microsatellite markers, two insertion deletion polymorphism markers and four CAPS markers were developed according to the genome sequences defined by PSM8 and PSM202.The recombinant strains in the mapping population were used for pollen The results showed that the marker W4 was completely linked with Sb locus, while the markers A8 and A14 were located on both sides of Sb locus with the genetic distances of 0.026 and 0.038 cM, respectively, from the Sb locus. The results showed that the newly developed polymorphic markers were anchored to the three end-to-end clones of AC093089, AC079021 and AC134931, respectively Marked on the physical map location, eventually S-b seat delimited between A8 and A14 .RiceGAAS annotations within the physical distance of 27kb indicates that the region has seven predicted the ORF. The result is S-b seating further functional analysis of the foundation.