大豆疫霉根腐病抗病基因的RAPD标记研究-微生物与免疫学专业论文.docx

东北农业大学硕士学位论文 摘要 摘要 、 (旭疫斟躏病是一种毁灭性的大吐传病害，由 Phytophthora sojae 引起。阴1 ..年首次在我碍事龙江省发现糊害， j脯，铺害接制品为盼，对黑龙江做胜产构 .. 成严重威胁电 本文以大豆抗疫霉根腐病 2对活等基因系 (Williarns与Williams79 、Williams 与 Wìlliams82) 为材料，从大豆叶片中提取圳，对:ìlí:等基因系基因组INA.进行队阳分析， 结果显示:经 1创个 10-核育唆随机引你才 2 对远等基因系基因组臼峭的扩增，有 3 个引物 (0附-{)S ..OPA-02和 α呛-ü4 )在 Williams79 中各扩增出 1号条特异带，有 4个引物((FF-16、 OPB-{)5 、αP侃和创刊5)在Wí11iams82中各扩增出了 1-2条特异带，部归亲本 (Williams) 中未见有上述的片段被扩增，多次重复 αl(J-04、创刊5 均获得较一致的结果。当以这两个 随机引切才 4 个已知含 .Rpsl-c、3 个己知含 Rpsl-k的加商材料和 2 个不含 Hps1-c、Hpsl-k 基因的尉商材料进行检测时，它们分脑如商材料中出现而在感病柳斗中均未出现。 !因比， 推波!路些持异性条带〈以 A 片段〉分别与大豆抗疫霉根腐病抗性基因 .Rpsl-c、励Is1吹连锁， 得到显性阴阳 OPQ--O知、 OP刊JIUJe叫 J 关键词:大豆疫霉根腐病:近等基因系:抗病基因:队 PD 分析. ? 东北农业大学 i t.学位论文 摘萤 Abstract ? Phytophthora r∞川、ot of soybean caused by Phylophthora .sojae is one of the w 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. 1t is forming great potential threat to soybean production of this province. For identíficatíon of molecular markers linked to the resistance genes Rpsl-c and Rpsl吮 two near- isogenic lines(NILs). Williams79 and Wi 11 iams82. and a recurγent parent of Wi 11 iarns were screened by 队PD. GenαDic DNAs extracted fr 侃1 soybean leaves of the two ILs were analyzed by 队PD using 160 different l D-mer rand 佣) primers. Polymorphic fragments were detected in the two NILs with 7 of the 160 primers. 1 to 2 specific DNA fragments were amplified fr ，侃I Williams79 which contaíns Hps]-c with primer OP怜05 、OPA-02 andαlQ--04. 1-2 specific DNA fra 阴阳t5 were amplified f 创1 Williams82 which contains HpsJ-k wi th primer OPF-16 、OPB-D5、OPD-06 and OPH-Q5. All these polymorphic D\A fragments were not detected in the recurrent parent Williams. The experiment of α寺。4ωd OPH-{)5 was repeated severa] times and the resul ts were the sarne. Using these two polymorphic primers to detec t 4 resistance cu1t í vars