【作者】 夏雪岩；

【导师】 罗耀武； 常金华；

【作者基本信息】 河北农业大学 ， 作物遗传育种， 2004， 硕士

【摘要】 高粱蚜是高粱上的一种重要的害虫，每年对全球的高粱产量和品质造成严重损失。抗蚜品种是防止高梁蚜的最经济、安全、有效的方法。近年来，分子标记技术的发展为加快作物育种进程提供了强有力的武器。但应用于高粱上的分子标记研究起步较晚，从而影响了高粱蚜抗性育种的进程。 本研究利用河农16×千三的杂交组合，对高粱蚜抗性鉴定方法，抗性遗传和SSR分子标记等进行了深入的研究。其目的在于：探索高粱蚜抗性遗传鉴定方法，充分了解高粱蚜抗性遗传的特点，建立一套适合高粱蚜抗性的SSR体系，找到与高粱蚜抗性基因连锁的分子标记，为最终掌握高粱蚜抗性遗传规律，克隆高粱蚜抗性基因奠定基础。主要研究结果如下： 1．河农16×千三杂交组合的亲本、F₁、F₂田间鉴定及遗传分析，结果表明：河农16具有对高粱蚜免疫的抗性，千三为高感高粱蚜品种，所得F₁为高抗，F₂抗、感分离比例符合3∶1，因此河农16的抗性是由一对基因控制的，抗虫性为显性，F₁表现完全显性。 2．选取河农16作为材料，采用单因素筛选和完全随机组合相结合的方法，建立了一套适合高粱抗高粱蚜的SSR体系。其中TaqDNA聚合酶0.3μL（5u／μL），10×PCR Buffer 2.0μL，引物1.5μL（10μmol／L），模板DNA 60 ng。利用该体系进行扩增，所得谱带清晰、稳定、非特异性带少。 3．用99对SSR引物对杂交组合河农16×千三的亲本、抗感基因池进行扩增，结果表明：在两亲本之间，有17对引物表现明显而稳定的多态性，散布于8个连锁群上，多态性频率为17.8％；但在抗感基因池间只有一对差异引物。这说明河农16和千三间遗传背景的差异是比较大的，因此分离群体多样性越大，越适宜构建作图群体。 4．分别用亲本间有多态性的17对SSR引物对河农16×千三的F₂隐性单株进行扩增，并对扩增结果进行分析，表明：与高粱蚜感虫基因相关的只有1个SSR位点：Xtxp6。采用MAPMAKER软件，对Xtxp6的所有F₂单株的SSR扩增结果进行分析，初步确定了高粱蚜抗性标记基因与SSR标记位点Xtxp6连锁，Xtxp6位于该连锁图的第9个连锁群的零位点上，距离为11.7cM。 本研究创新点在于利用一个对高粱蚜免疫的品种—河农16作为材料，用单因素筛选和完全随机组合相结合的方法，建立了一套适合高粱抗高粱蚜的SSR体系，利用该技术找到了与高粱蚜抗性基因相关的分子标记，为今后进一步克隆该基因奠定基础。更多还原

【Abstract】 Aphis sacchari. Zehntner was one of important pests in sorghum. This leaded to serious loss of yields and quality in sorghum of the world. Breeding resistance cultivars was the most economical, preferable, and effective method to control the Aphis sacchari. Zehntner. In the near years, development melocular marker technology fasten the progress of crop breeding. Moreover, the study of molecular biology of Aphis sacchari. Zehntner in sorghum lagged to other crops and further affected the progress for breeding Aphis sacchari. Zehntner resistant varieties.The research on the identification method resistant inheritance and SSR molecular markers of Aphis sacchari. Zehntner was carried out by using the cross of He nong 16xQian san.The intention was to explore a identification method of Aphis sacchari. Zehntner resistant, understand the characteristics of resistant inheritance of Aphis sacchari. Zehntner, establish suited SSR technique system and search the molecular marker linked with the resistant gene. It would build a basis for mastering regulation of Aphis sacchari. Zehntner resistant inheritance and cloning the resistant gene. The main results as follows:1 Identification and genetic analysis of resistance of Aphis sacchari. Zehntner by parents F1 and F2 population of He nong 16xQian san indicated that He nong 16 was immune to Aphis sacchari. Zehntner, Qian san was impressible, Fi were high resistant to Aphis sacchari. Zehntner, the segregation rate of resistant and susceptible was 3:1 in F2 population. So this resistance were controlled by one gene, Fi showed complete dominance.2 He nong 16 which was a hard-won variety immuned to Aphis sacchari. Zehntner was used in this expirement. Single factor selection was made to investigate better concentration in SSR technique system.Then, random thorough experiment was made to seek the best combination among factors.At last, the best SSR technique system was established. Taq DNA polymerase 0.3uL(5u /uL ), 10XPCR Buffer 2.0uL, primer 2.0uL (10umol/L), template DNA 60 ng.Using this system, amplification bands were clear, steady and non-especial bands were shorter.3, 99 pairs of SSR primers were amplified among DNA of P1 P2 and theirsegregated bulk of the cross of He nong 16xQian san. The results indicated that 17 pairs of SSR primers had visible and steady polymorphism between parents, distributed in 8 linkages, the rate of polymorphism was 17.8%. One pair of SSR primers had polymorphism between gene pools. It indicated the inheritance background of He nong 16 was very different from Qian san. So the higher genetic diversity in population ,the better for construct a linkage map. 4 The PCR amplification products of F2 recessive individuals from the cross He nong 16xQian san by 17 pairs of primers were analyzed. There was one SSR loci ?Xtxp6 related to the susceptible genes of Aphis sacchari. Zehntner. The PCR amplification products of all of F2 individuals were analyzed by using MAPMAKER soft program. It was assured elementarily that Aphis sacchari. Zehntner resistant gene linked to Xtxp6. It located at zero loci of 9 linkage, with a distance of 11.7 cM.The innovation of this research was that He nong 16 which was a hard-won variety immuned to Aphis sacchari. Zehntner was used, single factor selection and random thorough experiment were made to establish a suited SSR technique system, SSR molecular marking technology was introduced to find genes relevant to Aphis sacchari. Zehntner resistance. This would build a basis for isolating and cloning the Aphis sacchari. Zehntner resistant genes in the future.更多还原