【作者】 李红霞；

【导师】 张改生；

【作者基本信息】 西北农林科技大学 ， 遗传学， 2005， 硕士

【摘要】 近年,多种基于DNA 多态性的分子标记定位,已被广泛应用于遗传学研究的各个领域。小麦雄性不育研究亦不例外,将其作为育性基因定位的重要方法来研究。为了进一步探讨粘类小麦雄性不育系育性恢复的遗传规律,本试验专门对粘类小麦雄性不育恢复基因进行了遗传分析,选取具有高恢复力的恢复系亲本材料Rk5451 和Rk5253 为父本与ms（Kots）-90-110 杂交,调查F₁代自交结实率。F₁代再与保持系90-110 回交,统计BC1F1的育性分布范围;利用RAPD 分子标记对粘类小麦雄性不育系ms（Kots）-90-110 的恢复系Rk5451 的恢复基因进行了标记定位。以90-110//ms（Kots）-90-110/RK5451 的BC1F1分离群体为研究对象,利用分离群体分组分析法（Bulked　Segregant　Analysis,　BSA）,以350 个随机引物对Rk5451 的主效恢复基因进行RAPD 分析,获得了下述重要结果:　1.系统考察了粘、易、偏和二角型4 种非1BL/1RS 小麦雄性不育系的育性恢复性,结果表明供试的4 种非1BL/1RS 小麦雄性不育系均属易恢复、易保持不育类型;粘型1BL/1RS 小麦雄性不育系优于其余几种,具最高恢复力、且稳定性最好;对四种异质非1BL/1RS 小麦不育系杂种F₁ 恢复度（即平均自交结实率）进行了比较,K 型和V型细胞质的恢复度相近,且恢复度高,变异小;而Ven型和B 型细胞质的恢复度相对较低。　2. 同一不育系与不同恢复系测交,F₁ 恢复度差异显著,表明粘类非1BL/1RS 不育系的易恢复性与恢复系核基因组成有关;核背景相同而质背景不同的4 种不育系间平均自交结实率无显著差异,表明本试验4 种异质雄性不育细胞质源对恢复度影响差异不显著;对平均自交结实率和育性变异系数进行相关分析,4 种不育系平均自交结实率与变异系数呈明显负相关;4 种非1BL/1RS 小麦雄性不育系和恢复系基因除主效育性基因外,在不同核型中存在育性微效基因和抑制基因。父本为972376 的杂交组合F₁ 平均自交结实率显著低于其他组合,表明恢复系972376 核基因组成为主效恢复基因+育性抑制基因类型。　3.对粘类非1BL/1RS 小麦雄性不育系及杂种F₁ 配子传递方式分析,发现粘类1BL/1RS 小麦雄性不育系具有配子体不育的特征。供试90-110//ms（Kots）-90-110/RK5253及90-110//ms（Kots）-90-110/RK5451的BC1F1分离群体明显偏离1:1 的分离比,ms（Kots）-90-110 与RK5451 自交F2后代全都表现可育,无不育株,且自交结实率F2明显高于F1,进一步表明粘类1BL/1RS 小麦雄性不育系杂种F₁是以配子体不育方式传递的。　4.对供试90-110//ms（Kots）-90-110/RK5253及90-110//ms（Kots）-90-110/RK5451的BC1F1育性分布分析认为,粘型非1BL/1RS 小麦雄性不育系育性恢复是由一对主效恢更多还原

【Abstract】 using DNA markers is very important in hybridized wheat male sterile research. In order to probe into the fertility characteristics of male sterile lines of Nian type, Genetic behavior of fertility restorer gene for male sterile line of Nian type was analyzed. Rk5451 and Rk5253 were found to be good fertility restorer lines in hybrid wheat breeding. The sterile line ms（Kots）-90-110 were used as female to cross with Rk5451 and Rk5253. The analysis of the 2F₁s indicated that the restoring ability of Rk5451 and Rk5253 have high restoring degree. Distribution of fertility restoring degree of Bc₁F₁ from 90-110//（Kots）-90-110/ Rk5451 was counted. Bulked Segregant Analysis was employed to identify Random Amplified Polymorphic DNAs（RAPD） makers linked to the restorer gene in the BC1F1 population of 90-110//（Kots）-90-110/ Rk5451. The results were as follows: 1.the fertility restoration performance of the 4 male sterile lines of alloplasmic non-1BL/1RS wheat with Aegilops Kotschyi, Ae.Variabilis,Ae. Ventricosa and Ae. Bicornis were systematically investigated in this paper.The 4 male sterile lines of alloplasmic non-1BL/1RS wheat tested are all belonged to the sterile type of both good restoration and fine maintenance performance,especially Aegilops Kotschyi male line is the best one among them;Average seed sets of F₁ are different, Wheat with Aegilops Kotschyi and Ae.Variabilis are similar with high restoring degree and stable fertility.On the contrary, Wheat with Ae. Ventricosa and Ae. Bicornis have low seed sets of F₁ and high variation coefficient. 2.When the 4 male sterile lines of alloplasmic non-1BL/1RS wheat,as female parents,were test crossed with any different male parents,their average seed set of F1 was influenced by its alloplasm to some extent. there is not significance of different sterile cytoplasms;there is significant negative correlation between the average seed set of F1 and the C.V. of the seed set among the single plant;The same minor enhance gene and inhibiting gene of fertility,besides major fertility gene, exist in the different Karyotype of the male sterile lines and the restorer lines of 4 alloplasmic non-1BL/1RS wheat. 3.The manner of gametes transmission of non-1BL/1RS male sterile lines with Ae. Kotschyi Ae.Variabilis,Ae. Ventricosa and Ae. Bicornis cytoplasms in wheat and seed set of F₁ is analyzed in this paper. Obviously, non-1BL/1RS male sterile lines of Nian type have a Characteristic of gametophte sterility;Fertility Separation ratio of 90-110//ms（Kots）-90 -110/RK5451　depatured　seriously 1:1.In F₂ population of ms（Kots）-90-110/　RK5451 completely appear fertile without sterile.Test on hybridization F1 and F2 indicates that average seed set of F2 is higher than F1 These results prove again that the hybridization F1 of non-1BL/1RS male sterile lines with Ae. Kotschyi Ae.Variabilis,Ae.Ventricosa and Ae.Bicornis cytoplasms inherits in mode of gametophyte sterility. 4. Distributions of fertility restoring degree of BC1F1 of 90-110//ms（Kots）-90-110/ RK5253 and 90-110//ms（Kots）-90-110/ RK5451 probe that restoration fertility of male sterile lines of alloplasmic non-1BL/1RS wheat with Aegilops Kotschyi was influenced by major fertility restorter gene and some minor enhance gene. 5.Bulked Segregant Analysis was employed to identify Random Amplified Polymorphic DNAs（RAPD） makers linked to the restorer gene in the B1C1F population of 90-110//（Kots）- 90-110/ Rk5451, Among the total 350 arbitrary 10-mer oligonucleotide primers screened ,27 produced polymorphisms between the parents. S120,which of them ,was identified to be associated with restorer gene. The polymorphic band was S120-1745. RAPD markers amplified with primer S120 in BC1F1 population have some polymorphisms.更多还原