秋水仙碱处理油菜离体小孢子的染色体加倍效应  

Chromosome Doubling of Brassica napus L. Haploid by Treating in vitro Microspores with Colchicine

作　　者：石淑稳[1] 周永明[2] )吴江生[2] 

机构地区：[1]华中农业大学植物科学与技术学院 [2]华中农业大学作物遗传改良国家重点实验室

出　　处：《华中农业大学学报》2002年第4期329-333,共5页Journal of Huazhong Agricultural University

基　　金：8 63课题 (2 0 0 1AA2 4110 1) ;国家生物技术育种 (96-C0 1-0 1-0 4);国家自然科学基金 (3 9670 467)资助项目

摘　　要：用秋水仙碱对 5 2份甘蓝型油菜品系 (种 )和它们的杂种离体小孢子的单倍体二倍化技术进行了研究。从花药中分离出的单核晚期小孢子接种在含 10～ 80 0mg/L秋水仙碱的NLN液体培养基中处理 16～ 90h后转入无秋水仙碱的相同培养基诱导胚状体。结果表明 ,用 10mg/L秋水仙碱处理小孢子 4 8h ,10份材料的双单倍体植株变幅为 37.10 %～ 90 .12 % ,平均为 6 5 .4 4 %。 5 0mg/L处理 4 8h的 8份材料是 4 8.72 %～ 97.81% ,平均88.86 %。 10 0～ 80 0mg/L处理 16～ 4 8h的试验 ,多数加倍率在 90 %～ 10 0 %之间。但该处理药用量大 ,费用高 ,而且对小孢子毒性大 ,胚状体再生率低。试验还表明 ,秋水仙碱处理小孢子诱导的二倍体植株所产生的花一般均能结籽 ,带有不孕花的嵌合植株极少。用秋水碱处理小孢子再生的植株的根或芽所产生的二倍体植株多是可孕和不孕花共生的嵌合植株 ,自交后产生的单株种子很少 ,难以达到遗传和育种所需的群体量。Since 1996, the chromosome doubling of Brassica napus L. haplioid has been studied by treating its in vitro microspores with colchicine. Isolated microspores were treated in NLN media with 10～800 mg/L colchicine for 16～90 h followed by a reculture in the same kind of medium but without colchicine to induce embryos formation. The results show that the ratio range of diploid plants from the 10 crosses is 37.10%～90.12% with the mean of 65.44% by treating their microspores for 48 h with 10 mg/L colchicine. After the treatments of 50mg/L for 48h, the doubling ratio of 8 crosses is 48.72%～ 97.81 % with the mean of 88.86%. However, the doubling rate reaches 90%～100% in majority of experiments using 100～800 mg/L for 16～48 h. But the embryos yield is very low due to poison to the microspores growth when using colchicine with the concentration of more than 200 mg/L. The results also indicate that all flowers born in diploid plants induced by colchicine treatment of microspores are fertile and may produce a great number of seeds in each plant. Whereas fewer seeds can be harvested from chimeric plants born both fertile and sterile flowers by the method of treatment of microspore derived plants.

关 键 词：秋水仙碱处理 油菜 离体小孢子 染色体加倍效应 

分 类 号：S565.4[农业科学—作物学]