【作者】 陈庆山；

【导师】 杨庆凯；

【作者基本信息】 东北农业大学 ， 作物遗传育种， 2001， 博士

【摘要】 大豆灰斑病（Cercospora sojina Hara）是世界性病害，也是我国东北大豆主要病害之一，危害非常严重。本研究收集了黑龙江省94份大豆灰斑病种质资源，利用RAPD和SSR技术对种质资源间的遗传关系、大豆灰斑病10个生理小种的专化抗性进行了研究。主要研究结果如下： 1.对94份大豆灰斑病种质资源进行了10个生理小种的抗性鉴定，并用RAPD技术研究了种质资源间的遗传关系。其中，抗8个生理小种以上的材料共有17份，感8个生理小种以上的材料共有15份。41条引物共扩出485条带，多态条带总数为145条。材料间的相似系数介于0.43-0.96。用SSR技术对其中48份进一步进行了研究。61个SSR引物共在48个大豆品种或品系中得到197个等位基因，材料间的相似系数介于0.17-0.92。 2.根据大豆灰斑病种质资源10个生理小种的抗感鉴定结果与RAPD和SSR的分子数据，得到了许多与各生理小种专化抗性有关的分子标记。相似度大于0.7，Fisher准确性测验达到显著相关的有RAPD引物C07-2、F02-3、Q12-3和SSR引物Sat₁08-b、Sat₁21-b、Satt049-c、Satt199-b、Satt239-c、Satt247-d、Satt303-b、Satt371-a、Satt453-a、Satt516-c、Satt543-b、Satt587-a、Sct₁86-a、Sct₁86-e、Sct₁86-f、Sct₁89-b、SOYPRP1-a，可以建立各生理小种的分离群体进一步确定连锁距离。 3.对RAPD、琼脂糖分离的SSR、聚丙烯酰胺分离的SSR在揭示大豆灰斑病种质资源平均等位基因数、预期异质结合度和相似系数进行了比较。结果表明：聚丙烯酰胺分离的SSR得到的平均等位基因数最多，预期异质结合度最大，相似系数最低；琼脂糖分离的SSR次之；RAPD再次。说明聚丙烯酰胺分离的SSR揭示种质资源多态能力最强。SSR聚类结果与RAPD技术有差异，但两种SSR技术聚类结果吻合得很好，相关系数达到显著水平。 4.本实验结果表明：40566、40567、91-135、东农39；9674；1574、1572三组抗病材料在聚类树状图上分组明显，说明这三类抗病材料抗性来源有可能不同，可作为亲本相互杂交以富集不同抗性，拓宽今后育成品种的遗传基础。 5.RAPD和SSR数据聚类结果表明：大豆灰斑病种质资源的部分材料是同一育种单位内的聚到一起，而大部分材料是各育种单位间相互交织在一起。说明了各育种单位材料间亲缘关系相近，抗病育种选择的亲本间亲缘关系相近。黑龙江省大豆灰斑病种质资源的遗传基础狭窄，急需拓宽，以育出抗性更强、抗谱更广、抗病更持久的品种。更多还原

【Abstract】 Ce^cospora sojina Hara occurs worldwide and is one of major soybean disease in the northeast of china, harming soybean seriously. Ninety-four soybean germplasm were collected in this study. Genetic relation and specific resistance to ten races were analyzed with RAPD and SSR. The main results are summarized as the followings:1.Filly of the Ninety-four soybean germplasm of frog-eye leaf spot were evaluated for resistance to 10 races and their genetic relations were analyzed with RAPD. Seventeen cultivars or lines were resistant to over 8 races, and fifteen were susceptible to over 8 races. Among the total 485 bands amplified with 41 primers, 145 were polymorphic. The genetic similarities of soybean germplasm were between 0.43 0.96. SSR was used to study more on forty-eight ones from the Ninety-four soybean germplasm. 197 alleles were detected with 61 primers. The genetic similarities of soybean gemiplasm were between 0.17-0.92.2.Based on the results identified with 10 races of soybean gerrnplasm and data from RAPD and SSR, some markers with speaflc resistance to ten races were got With the scale of similarity over 0.7 and significant correlation of Fisher Exact Test. there are 007-2.. F02-3.. Q12-3 from RAPD and Sat₁08-b.. Sat₁21-b.. Satff)49-c.. Sattl99-b.. 5att239-c.. Satt247-d.. Satt3O3-b.. Satt37l-a.. Satt453-a.. SattSl6-c. Satt543-b. SattSS7-a.. Sct₁86-a.. Sct₁86-e.. Sct₁86-f.. Sct₁89-b.. SOYPRPI-a from SSR with different races. For the further studying, the segregate populations on different races were needed to learn the linkage distance.3.The differences among RAPD, SSR （detected with agarose gel）, and SSR （detected with polyacrylamide gel） was determined by evaluating number of alleles analyzed per lod, expected heterogeneity and genetic similarity between soybean gerrnplasm. SSR markers, detected with polyacryiamide gel, had the highest mean number of alleles, expected heterogeneity value and lowest similarity coefflaent SSR detected with agarose gel took the second place. RAPD took the last The difference existed between the results of similarity and duster analysis revealed by RAPD and SSR. The results between SSR （detected with agarose gel） and SSR （detected with polyacrylamide gel） came the same.4. Our^expertments showed that 40566.. 40567.. 91-135.. Dongnong39; 9674;1574.. 1572 were highly resistant to Cemospom sojina Ham, but they cut to three groups with duster analysis. The reason is that they maybe carried different resistance genes. If they were used as parents to make a cross, the different resistance genes were nched. It is good for the varieties to keep resistance for a long time.5.The duster analysis by RAPD and SSR showed: Some soybean germplasm from the same breeding unit were clustered in a group, but most soybean germplasm were clustered into different groups with germplasm from different breeding units. It means that the genetic relation among the germplasm from different breeding unit is very high. So, the conclusion is that the genetic base of the soybean germplasm resistant to Cetvospo sojina Ham in Province Hei Longjiang is very narrow. It must be done to broaden the genetic base to cultivate varieties with long-time more-races resistance to Cercospora sojina Ham.更多还原