【作者】 张俊杰；

【导师】 孙万仓；

【作者基本信息】 甘肃农业大学 ， 作物遗传育种， 2011， 硕士

【摘要】 白菜型冬油菜（Brassica.rapa, L.,var.biennis,f.winterness）具有优异的抗寒性,既是重要的油料作物,又是冬、春季覆盖作物,在北方地区的农业生产中具有重大的经济意义和生态意义。本研究利用白菜型冬油菜超强抗寒性品种陇油6号和抗寒性较弱品种天油8号进行正反交得到F₁代以及F₂代。以F₂代两群体为基础研究群体,对白菜型冬油菜的抗寒性的遗传、抗寒性相关的SSR分子标记以及不同抗寒品种的蛋白质表达等进行了分析研究,主要研究结果如下:1.通过陇油6号和天油8号F₂代植株的抗寒与不抗寒分离比例进行统计,实测值与理论值的比率用卡方适合性进行测验,确定分离比率。结果表明,白菜型冬油菜抗寒性是受两对主基因所控制的,用R1和R2表示,这两对基因作用于同一性状,其效应是累加的,等位基因间无显隐性关系;用抗寒性弱的亲本做母本,分析比例有一定偏离。正反交分离比不同,说明白菜型油菜的抗寒性遗传除受细胞核基因控制外,还与细胞质遗传有关。2.利用SSR分子标记引物对陇油6号和天油8号的508个F₂代植株分析结果,获得了7个与白菜型冬油菜抗寒性Rf紧密连锁的分子标记,应用Mapdraw软件构建了遗传图谱。7个标记与抗寒连锁程度由近及远分别是:BRMS-050、Ol12-A04、BRMS-042-2、BRMS-036、Ra3-D02b、Na10-C06、Ra3-D04,连锁距离分别为:4.09 cM、5.83 cM、9.49 cM、12.73 cM、12.96 cM、16.88 cM、24.61 cM,其中,标记BRMS-050、Ol12-A04距抗寒性Rf的遗传距离较近,这些标记可以作为白菜型冬油菜分子标记辅助选择育种的参考依据。3.对冬油菜亲本材料陇油6号、天油8号进行蛋白质双向电泳（2-DE）分析,在1000μg/IPG胶条的上样量条件下,超强抗寒性品种陇油6号和弱抗寒性品种天油8号分别检测到有效蛋白点1207和1053个。以天油8号蛋白质胶为参照胶,陇油6号与之比较,差异蛋白点有76个,其中49个蛋白点表达量上调,21个蛋白点表达量下调,新增的蛋白点5个,减少的蛋白点1个。结果表明,白菜型冬油菜陇油6号的抗寒性与某些蛋白质的表达量有关,抗寒性强的品种蛋白质表达量高于抗寒性弱得品种,陇油6号的强抗寒性还与特异蛋白的表达有关。更多还原

【Abstract】 Winter rapeseed （Brassica campestris） has excellent cold hardiness, it is not only major oil crop, but it is winter and spring cover crop, and it causes great economic and ecological significance on agricultural production in the north areas.This study used two varieties of winter Brassica.Rapeseeds: super-cold Longyou6 and weaker-cold Tianyou8, obtained F₁ and F₂ generation through reciprocal cross. The two groups of F₂ generation were used for Basic research. Analyzed and research on genetic of cold hardiness in winter Brassica Rapa, SSR marker be relared to cold hardiness and protein expression of different cold hardiness varieties. The main results are as follows:1. Statistical separation ratio about the number of F₂ generation plants of Longyou6 and Tianyou8 by cold hardiness or not. The ratio of measurement values and the theoretical value used chi-square Suitability for testing, to checking separation ratio. The results showed that cold resistance of winter Brassica rapeseed was controlled by two the main genes, which expressed with R1 and R2. The two pairs of genes acted on the same characters, the effect was cumulative, and there was no recessive relationship between the allele. Separation ratio has deviation when weaker-cold varieties as female parent.Reciprocal cross ratio was different, it declared that the cold resistance heredity of winter Brassica rapeseed not only controlled by nuclear genes, but it connected with the cytoplasmic heredity.2. Using the primer of SSR molecular marker analysis of the 508 per plant of F₂ generation of Longyou6 and Tianyou8, and the results following: which totally got 7 molecular markers closely linked with cold hardiness Rf in winter Brassica rapa. Applying software of Mapdraw builds genetic map. 7 markers and linkage degree of cold hardiness from near to far were as follows: BRMS-050、Ol12-A04、BRMS-042-2、BRMS-036、Ra3-D02b、Na10-C06、Ra3-D04, Linkage distance was that: 4.09、5.83、9.49、12.73、12.96、16.88、24.61, Among them, the marks BRMS-050, Ol12-A04 was nearer to the cold hardiness gene Rf genes from the genetic distance, These marks can be used as reference of molecule marker assisted selection breeding in winter Brassica rapa.3. Analyzed on parental materials Longyou6 and Tianyou8 of winter rapeseed with two-dimensional electrophoresis of proteins （2-DE）. When loading quantities were at 1000μg/IPG strips. Super cold varieties Longyou6 and weaker-cold varieties Tianyou8 were detected effective protein spots 1207 and 1053. Using Tianyou8 protein gel as the reference gel, Longyou6 in contrast, there were 76 differential protein spots, and in them, the expression of 49 protein level spots increased, 21 protein points decreased, increased 5 new protein spots and reduced 1 protein spots. The results showed that cold resistance of winter Brassica rapa Longyou6 related to the expression level of certain proteins, the varieties of strong cold hardiness had a high expression level of protein than weaker-cold varieties. Strong cold hardiness of Longyou6 also related to the specific protein expression.更多还原