大豆疫霉根腐病抗病基因的RAPD标记研究

【作者】 曲娟娟；

【导师】 李景鹏；

【作者基本信息】 东北农业大学 ， 微生物与免疫学， 2001， 硕士

【摘要】 大豆疫霉根腐病是一种毁灭性的大豆土传病害，由Phytophthora sojae引起。1991年首次在我国黑龙江省发现该病害，此后，该病害呈逐年扩大趋势，对黑龙江省大豆生产构成严重威胁。本文以大豆抗疫霉根腐病2对近等基因系（Williams与Williams79、Williams与Williams82）为材料，从大豆叶片中提取DNA，对近等基因系基因组DNA进行RAPD分析，结果显示：经160个10-核苷酸随机引物对2对近等基因系基因组DNA的扩增，有3个引物（OPN-05、OPA-02和OPQ-04）在Williams79中各扩增出1-2条特异带，有4个引物（OPF-16、OPB-05、OPD-06和OPH-05）在Williams82中各扩增出了1-2条特异带，在轮回亲本（Williams）中未见有上述的片段被扩增，多次重复OPQ-04、OPH-05均获得较一致的结果。当以这两个随机引物对4个已知含Rpsl-c、3个已知含Rpsl-k的抗病材料和2个不含Rpsl-c、Rpsl-k基因的感病材料进行检测时，它们分别在抗病材料中出现而在感病材料中均未出现。因此，推测这些特异性条带（DNA片段）分别与大豆抗疫霉根腐病抗性基因Rpsl-c、Rpsl-k连锁，得到显性RAPD标记OPQ-04₇₀₀、OPH-05_?。更多还原

【Abstract】 Phytophthora root rot of soybean caused by Phytophthora soJie is one of the most dangerous soybean diseases. The disease was first found in Heilongjiang province of China in 1991 and then the area subject to the disease has continued to expand rapidly. It is forming great potential threat to soybean production of this province. For identification of molecular markers linked to the resistance genes Rpsl and Rpsl, two nearsogenic lines(NILs) Williams79 and Williams82, and a recurrent parent of Williams were screened by RAPD. Genomic DNAs extracted from soybean leaves of the two NILs were analyzed by RAPD using 160 different 10ner random primers. Polymorphic fragments were detected in the two NILs with 7 of the 160 primers. 1 to 2 specific DNA fragments were amplified from Williams79 which contains Rpsl with primer OPN?5 , OPA?2 and OPQ?4. 1? specific DNA fragments were amplified from Williams82 which contains Rpsl with primer 0PF6~ 0PB?5~ OPD?6 and OPH?5. All these polymorphic DNA fra~nents were not detected in the recurrent parent Williams. The experiment of OPQ?4 and OPH?5 was repeated several times and the results were the same. Using these two polymorphic primers to detect 4 resistance cultivars with Rps]x3 with Iisland 2 susceptible cultivars without Rpsl, Asl, 0PQ?4~ and 0PH?5~~were amplified in all of the resistance cultivars and absent in all of the susceptible cultivars. So it is inferred that these polymorphic DNA fragments are probably linked to Rpsl and Rps]-k.更多还原