【作者】 李雪华；

【导师】 喻德跃； 王敏；

【作者基本信息】 南京农业大学 ， 作物遗传育种， 2003， 硕士

【摘要】 分析基因功能最有效的方法之一是利用突变体。通过比较突变体与其对照在特定环境条件下基因表达的差异可以获取基因功能的重要信息。构建大豆的突变体库，集中各种变异性状，选育和创造近等基因系可以为基因功能的分析提供材料。 本研究通过对12个大豆品种进行⁶⁰Co γ射线诱变，主要从品质、耐盐性、农艺性状等方面鉴定和筛选突变体。 1、通过对3000份大豆辐射M₃代家系蛋白质含量和油份含量的测定和分析，筛选出了大豆高蛋白和低蛋白含量、高油分和低油分含量的家系各200份，方差分析表明，这些家系的蛋白质或油份含量与对照相比差异显著。 2、对筛选出的200份蛋白质含量或油份含量发生变异的M₃代家系，扩繁至M₄代，并进行鉴定。结果表明：M₄代家系中的蛋白质含量或油份含量与对照相比差异显著，与对应M₃代家系的蛋白质含量和油份含量均达到显著相关的水平。 3、对五个M₄代辐射材料中的最高蛋白质含量和最低蛋白质含量的家系并结合其相应的对照品种，用200对引物进行SSR分子标记筛选，结果筛选出了两个可能与之相关的标记：Satt031和Satt055。 4、通过SDS-PAGE电泳，从600份M₄代家系中筛选出了一个11S／7S值为2.8的高比值家系，其对照为南农88-31。此家系的凝胶电泳图谱中出现了7S的亚基条带变浅，11S亚基条带加深现象，从而提高了11S／7S比值，为品质育种提供了新的基因型，并且为控制蛋白质亚基含量的基因功能研究提供了新的研究材料。另外，相关分析结果表明，11S和7S之间具有负相关的关系，与文献报道相一致。 5、对鉴定出的11S／7S突变体进行SSR标记分析。结果在200对引物中筛选出了一个可能与之相关的标记：Satt193。 6、在不同浓度NaCl胁迫下，通过不同家系在发芽率、发芽指数、盐害指数、叶片相对电导率、叶绿素等方面的差异筛选出了与耐盐品种Lee68的耐盐性相近的家系12个，耐盐性明显地高于Lee68的家系四个：T30682、T30014、T30935和T4110。 7、在农艺性状方面发现的突变种质主要为：芽黄、花色变异、茸毛色、结荚习性、株高、不育性等几个方面。 总之，本研究初步构建了如下突变体库。 在品质方面的突变有：蛋白质含量变异的家系200份（高蛋白和低蛋白各100份），油份含量变异的家系200份（高、低油份各100份），其中，蛋白质含量超过46％的大豆突变体库的构建及突变类型的鉴定家系32份，油分含蚤超过22%的家系5份.在115/7S值突变方面，其llsns值不低于2.5的家系3份. 在耐盐性方面的突变有:杭盐的家系（较之Lee68）17份.其中，抗盐性比Lee68（国际公认耐盐品种）高的家系4份，耐盐性与Lee68相似的家系12份;盐歌感的家系5份. 在农艺性状方面:叶形突变21株，叶绿素缺失突变加株，育性的突变12株，来自于南农86一4;花色突变2株，来自于南春201;茸毛色突变20株，分别来自于88一31和菜豆1号;种脐色、感病性和结芙习性突变分别为5株、5株和12株，来自于88一3 1.更多还原

【Abstract】 One of the most effective ways in functional genomics research is the application of mutants. By comparing mutants with the control(CK), it is possible to get knowledge of gene functions. Therefore, construction of mutant pools and collection of kinds of elite mutants are the basis fou functional genomics in soybean.In this study, soybean mutants were produced by irration of Y -ray of the isotope 60Co, and some of those were further characterized. The main results are as follows:1. By measuring of protein and oil contents of 3000 accessions of soybean mutants of the M3 lines, we selected 200 accessions respectively with higher and lower protein or oil contents than control. The variance analysis of irradiation effect showed the selected accessions had a significant difference compared to the control.2. Analysis of mutation in protein and oil contents of 400 accessions of M4 lines M3 lines showed that the contents of protein and oil in M4 were significantly different from those in control, and the correlation analysis indicated that the M4 lines significantly correlated with M3 both in protein and oil contents.3. In research for SSR markers linked with mutation for protein content, 15 templates were screened from five kinds of M4 lines with highest protein content, lowest protein and the control. And 200 random primers were applied to this research. Primers Satt031 and Satt055 found polymorphic bands.4. A study was carried on to select elite mutants rich in 11S globulin among 600 accessions of M4. Specific values of seed storage protein component 11S/7S were analyzed using SDS-PAGE. It was found that the range of the specific values was 0.8-2.8 in the mutants. Compared with the control(1.9), the difference is significant. And we obtained one material that had a specific value of 11S/7S with 2.8. And it could be considered as an excellent genotype for quality genetics and breeding.5. 200 SSR primers were applied to reveal genomic difference between the 11S/7S mutants(2.8) and the control(1.09). The result showed that only primer Satt193 revealed polymorphic bands between these two materials.6.Salt tolerance of 400 accessions in phases of seed germination and seedling screenned. The result of analysis showed that some of the accessions (10) had a similar tolerance in salt with Lee68. Even, T30682 and T4110 were more tolerant than Lee68.In conclusion, we constructed main mutant pools as follows:For the quality of soybean, we obtained 400 accessions of higher and lower protein and oil contents. And we also selected 12 salt-tolerant and 5 salt-sensitive accessions of M3 lines compared with Lee68. For agromomic characters, we identified 21 mutants in shape of leaf, 5 with absense of chlorophyl 1 and 12 fertility mutants.更多还原