摘要
为创建大白菜—结球甘蓝二体异附加系,以部分大白菜—结球甘蓝单体异附加系为试材,采用常规压片、核型分析以及SSR标记从其自交后代中鉴定出3个二体异附加系,并对单体异附加系的雌雄配子传递率和结籽率进行了测定,为大白菜—结球甘蓝异代换系、易位系的创建提供了关键材料,并为大白菜遗传改良以及研究A、C染色体组间的关系及基因的定位和表达奠定了基础。主要研究结果如下:
1.以二倍体大白菜为对照,对7个单体异附加系自交后代结籽率进行了统计,结果表明,单体异附加系自交后代结籽率均低于二倍体大白菜,其中,6号单体异附加系结籽率最高,可达到65.48%,9号单体异附加系最低,结籽率仅为14.40%。
2.部分大白菜—结球甘蓝单体异附加系分别与二倍体大白菜亲本进行正反交,测定单体异附加系中外源染色体的雌雄配子传递率。结果表明,附加结球甘蓝2号、3号、4号、5号和6号染色体的单体异附加系n+1雌配子的传递率分别为25.64%、48.72%、28.95%、32.35%和33.33%;n+1雄配子的传递率分别为19.05%、28.57%、34.09%、19.51%和29.79%。
3.采用根尖细胞染色体中期计数法,对3个大白菜—结球甘蓝单体异附加系自交后代102个植株染色体数目进行了鉴定,从中鉴定出2号、4号和5号二体异附加系,其所占比例分别为12.5%、8.33%、10.81%。
4.对3个大白菜—结球甘蓝二体异附加系植株花粉母细胞减数分裂进行了观察,结果表明,终变期以11Ⅱ联会形式为主,中期Ⅰ有游离染色体,后期Ⅰ基本以11-11形式分离,后期Ⅱ的分离方式多样,但以11-11-11-11为主。
5.大白菜—结球甘蓝二体异附加系植株从叶形、叶色、叶表面特征、花器多方面表现出类似于甘蓝的一些特征。但是由于附加的甘蓝染色体不同,3个二体异附加系形态特征各异。
6. 3个大白菜—结球甘蓝二体异附加系植株的花粉特征和结籽率与二倍体大白菜具有显著差异。花粉整齐度、花粉生活力和结籽率均存在显著的正相关关系。
7.选取结球甘蓝07连锁群特异SSR引物BN72a和Ol10B01,分别对异源三倍体杂交种及其亲本大白菜、结球甘蓝和5号单体及二体异附加系进行SSR扩增,扩增结果表明,引物BN72a和Ol10B01在大白菜—甘蓝5号单体及二体附加系扩增结果与结球甘蓝及异源三倍体杂交种一致,而二倍体大白菜上没有相应的扩增产物。说明筛选的大白菜—结球甘蓝5号二体异附加系外源染色体与甘蓝07连锁群相对应。
In order to creating Chinese cabbage-cabbage disomic alien addition lines,part Chinese cabbage-cabbage monosomic addition line was used, and its selfing F1 generation was tested by means of identification of cytology and SSR analysis,Three disomic chinese cabbage-cabbage chromosome alien addition lines were identified. Transmission rate of gametes and seeding set of Chinese cabbage-cabbage monosomic alien addition line were counted. This study provided basic material for creating alien substitution lines and translation lines of Chinese cabbage and cabbage, and helped to carry out Chinese cabbage genetic improvement, explore the relation between A and C genomes, study gene intactions, location and expression. The results were as followed:
1.With Chinese cabbage(2n=2x=20) compared, seven monosomic aline addition lines seeding set by self were counted. Result indicate that monosomic aline addition lines seeding set by self were lower than Chinese cabbage. 6# Chinese cabbage-cabbage disomic alien addition lines seeding set was the highest,the rate was 65.48%,9# disomic Chinese cabbage-cabbage alien addition lines seeding set was the lowest,the rate was 14.40%。
2.Reciprocal crosses were made between the part Chinese cabbage-cabbage monosomic aline addition lines(2n+1) and Chinese cabbage.The transmission rate of gametes the monosomic aline addition lines from alien addition lines were determined. Result indicate that the n+1 female gamete transmission rates of 2#、3#、4#、5#、6# Chinese cabbage-cabbage monosomic alien addition lines were respectively 25.64%、48.72%、28.95%、32.35% and 33.33%; The n+1 male gamete transmission rates were respectively 19.05%、28.57%、34.09%、19.51% and 29.79%。
3. The F1 generation of three Chinese cabbage-cabbage monosomic aline addition lines selfing were indentified by taking account of the chromosome number in mitosis metaphase.The 2#、3# and 5# chinese cabbage-cabbage disomic alien addition lines were obtained.The rate is 10.09%
4.PMC meiosis behavior of three Chinese cabbage-cabbage disomic alien addition lines showed that: the chromosome configurations at diakinesis were mainly showed 11II ,two chromosomes free move to cytoplasm at metaphaseⅠ, the chromosome configurations at anaphaseⅠwere mainly showed 11-11,it has many chromosome separation at MetaphaseⅡ,but chromosomes mainly distribute with 11/11/11/11.
5.The field character of the Chinese cabbage-cabbage disomic alien addition lines, such as leaf surface character, leaf color, waxy of leaf,floral organs and so on, were similar to cabbage.The different cabbage chromosome caused different morphological features.
6.The characters of pollen characteristics and seeding set were counted. The results showed that there was significant difference between three Chinese cabbage-cabbage disomic alien addition lines and Chinese cabbage. Significant positive correlation was pollen regularity,pollen viability and seeding set.
7.The primes of BN72a and Ol10B01 were amplified on cabbage, allotriploid hybrids,5# Chinese cabbage-cabbage disomic alien addition lines and 5# Chinese cabbage-cabbage monosomic alien addition lines. The amplified result that 5# Chinese cabbage-cabbage disomic alien addition lines and 5# Chinese cabbage-cabbage monosomic alien addition lines were similar to cabbage,but Chinese-cabbage didn’t have.It make out the alien chromosome of 5# Chinese cabbage-cabbage disomic alien addition lines corresponded with 07 linkage group of cabbage.
引文
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