【作者】 陆光远；

【导师】 杨光圣； 傅廷栋；

【作者基本信息】 华中农业大学 ， 作物遗传育种， 2004， 博士

【摘要】 油菜是我国主要的油料作物，大约有50％的食用植物油来源于油菜。长期以来，选育高产优质油菜一直是育种家所面临的主要任务。实践证明，杂种优势利用是提高油菜产量的主要途径。目前，细胞核雄性不育是甘蓝型油菜杂种优势利用的一条有效途径。 1985年，李树林等报道了一批新型的细胞核雄性不育材料，他们同时提出这些材料的育性是受由两对显性基因互作控制的，即双基因互作控制模式。这一遗传模式的提出，为显性细胞核雄性不育应用于杂交种生产提供了理论依据。由于显性细胞核雄性不育具有败育彻底、不育性稳定、无不良胞质效应、可以实现“三系”配套等优点，因而在杂交油菜育种中具有重要的应用价值。为了使显性细胞核雄性不育得到更加广泛的应用，很有必要对其进行深入的研究。 本研究以甘蓝型油菜显性细胞核雄性不育纯合两型系Rs1046AB为亲本，与不同来源的甘蓝型油菜品种（品系）进行杂交或回交，开展的研究内容包括：显性细胞核雄性不育的遗传分析；显性细胞核雄性不育在不同细胞质、细胞核和环境条件下的遗传表现；显性核不育基因的图谱定位；上位抑制基因的图谱定位；显性细胞核雄性不育“三系”的分子标记辅助选择育种等。取得的主要实验结果如下： 1．经测交分析，从14份甘蓝型油菜品种（品系）中筛选到3份带有抑制基因的材料，它们可以作为显性细胞核雄性不育系的恢复系，其余11份材料不带有抑制基因，可能是双隐性类型，可以作为显性细胞核雄性不育系的临时保持系。 2．以Rs1046B为共同母本，与不同来源的品种（品系）组配了11个同质异核的杂交组合。育性调查发现，不同杂交组合间的不育株率相差很大（最低为22.7％，最高为55.3％），个别组合还出现了部分可育现象。这似乎说明，某些核背景对显性细胞核雄性不育的育性表现存在一定影响。 3．以Rs1046B为亲本之一，与2份双隐性材料一共配制了6对正反交组合，运用最小差数法和成对数据法对各组合的不育株率进行统计分析，结果发现正反交组合的育性差异不显著。由此说明，显性细胞核雄性不育的育性表达不受细胞质的影响。 4．将7个杂交组合种植于兰州（5月至10月）和武昌（10月至次年5月）两地并进行育性对比观察，结果发现部分杂交组合后代的不育株率在两地相差较大，在兰州的生态条件下不育株率接近正常的50％，而在武昌的生态条件下不育株率大为降低。这说明，不同的生态环境条件对显性细胞核雄性不育的育性表现存在一定的影响。 5．在个别组合F₁不育株（Msmsrfrf）上发现有少量的部分可育花，将其自交，后代发生1∶1的育性分离，继续兄妹交可以选育到杂合两型系。遗传分析表明，部华中农业大学博士学位论文2003 分可育株自交后代发生1二1分离的原因可能是部分可育株产生了Msrf和msrf 两种雌配子而仅仅产生msrf一种雄配子的缘故。6.在回交分离群体[（Rs 1 o46AX“samourai”）X“samourai”]中，运用BSA法构 建了不育集团和可育集团，在两个集团之间一共筛选了480对AFLP引物，找到 了5个与不育基因紧密连锁的AFLP标记，其中4个标记（E3M151。。、E7M1230、 Pl3M8’00和P10M1335。）位于不育基因的同一侧，与不育基因的遗传图距分别为 3.7。M、5.2 cM、5.9 cM和8.9 eM，剩下的1个标记（P6M641。）位于不育基因 的另一侧，与不育基因的遗传图距为5.9 cM。将最有应用价值的标记E3M15100 和P6M6引。的差异片段进行回收、克隆和侧序，结合PCR步行方法，成功地将 这两个AFLP标记转化为便于在育种应用的SCAR标记。7.在兄妹交群体（Rs 1 046A x Rs1046B）中运用BSA法构建了不育集团和可育集团， 在2个集团间一共筛选了640条RAPD引物和1 024对AFLP引物，最后找到了 一个与抑制基因紧密连锁的AFLP标记EI 6MG4205，该标记与抑制基因之间的交 换率仅为6.1%。将标记差异片段进行了回收、克隆和测序，序列分析和BLAST 搜索结果表明，标记片段包含有一个O盯结构区，该ORF区编码的34个氨基 酸序列与4个来源于拟南芥的蛋白质高度相似，但其具体功能不详。8.利用BCI群体[（Rslo46AX“Samourai”）X“samourai”]构建了一个包含15个 主要连锁群、2个三联体和1个连锁对的甘蓝型油菜分子标记遗传图谱，图谱总 长2667.1 cM，整合了138个AFLP标记、83个SSR标记和1个形态标记，标记 间平均距离为13.27 cM。不育基因入介定位于第十连锁群（LG10）上，第八、第 十六连锁群的末端各有一个明显的偏分离标记密集区。9.文中还讨论了建立显性细胞核雄性不育三系选育的分子标记辅助选择体系的可 能性。更多还原

【Abstract】 Rapeseed is one of the major oilseed crops in China. It contributes to appropriately 50% of the vegetable oil consumption. In the past and the near future, canola breeders still face the common and major goal of improving the productivity level of rapeseed. To achieve this, heterosis utilization is proved to be a very effective way, and now genie male sterility (GMS) is becoming a practicable approach to improve yield performance in Brassica napus.Li et al (1985) had reported several new GMS mutants in Brassica napus, also they proposed that the sterility of these mutants followed a digenic mode of inheritance with epistasic interaction. This hypothesis hence provides a theoretical base for employing the GMS in hybrid seed production. This GMS system is a promising approach to be use in hybrid breeding, because its sterility is stable and complete, no negative cytoplasmic effects are involved and, it can be used in a "three-line" system. To make this GMS system a wider application, it is necessary to initiate further study on it.In the present study, Rsl046AB, a homozygous GMS two-type line in Brassica napus, is used as one of the parent to cross with several Brassica napus accessions to study: genetic analysis of the dominant GMS; fertility performance of the dominant GMS under various genetic background, cytoplasm and environments; genetic mapping of the dominant GMS gene (My); genetic mapping of the suppressor gene (Rf) and the establishment of a MAS system for breeding dominant GMS three lines. Main results of the present studies are as follow:1. Three out of the fourteen accessions carrying Rfallele were identified by test-crossing, which could be served as restorers. The other eleven accessions did not carry Rfallele and were possibly in recessive nature of Rf and Ms loci, which could be served as temporary maintainers.2. Using Rsl046B as the mother parent, eleven crosses were made with eleven accessions having different genetic background. Fertility investigation showed that the percentage of male sterile plants varied among the eleven F1populations (from minimum of 22.7% to maximum of 55.3%), and a few partial fertile plants were observed in some FI populations, it seemed that a special genetic background had influence on the expression of the genie male sterility gene.3. Six pairs of reciprocal crosses were made by crossing Rsl046B with two cultivars in recessive nature of Rf and Ms loci. Statistic analysis revealed that the percentage of male sterile plants in the reciprocal crosses were not significantly differrents, which indicated that the expression of the male sterility gene is not influenced by cytoplasm.4. Seven crosses were grown in Lanzhou (from May to October) and Wuchang (from October to May in the next year). Fertility investigation showed that the percentage of male sterile plants in seven crosses grown in Lanzhou were near the expected 50%, but this values decrease sharply in five of the seven crosses grown in Wuchang, thisindicated that fertility performance could be affected by environments.5. A few partial fertile flowers could be observed in the FI male sterile plants (Msmsrfrf). When selfmg such plant, an unexpected 1 : 1 fertility segregating ratio could be observed in the resulting progeny, in which when sibmating sterile plants with fertile plants, the heterozygous GMS two-type line could be bred. Furthur genetic analysis revealed that the partial fertile plants generated two kinds of female gametes (Msrf and msrf) but only one kind of male gamete (msrf). This might partially account for the 1 : 1 fertility segregation in the progeny derived from partial fertile plant.6. In the BCi population of [(Rsl046AX "Samourai" ) X "Samourai" ], AFLP technology in conjunction with bulked segregant analysis were perform to identify genetic markers for the male sterility gene. From the survey of 480 pair of AFLP primers, five markers closely linked to the male sterility were obtained. Four of them (E3M15100. E7Ml230. P13M84oo and PlOMl3 350) flanking the male steri更多还原