甘蓝型油菜小孢子秋水仙碱处理提高双单倍体频率研究  

Studies on Efficient Production of Doubled Haploid Plants by Colchicine Treatments in Microspore Culture of Brassica napus

作　　者：周伟军[1] 唐桂香[1] 张国庆[1] Hagberg P 

机构地区：[1]浙江大学农业与生物技术学院,杭州310029 [2]瑞典Svalof Weibull AB,Nilsson-Ehle Laboratory,svalov26881

出　　处：《中国农业科学》2002年第4期410-414,T001,共6页Scientia Agricultura Sinica

基　　金：教育部留学回国人员科研启动基金;浙江省重点科技项目 (99110 2 3 3 3 )

摘　　要：以 2个对小孢子培养反应较好的甘蓝型冬油菜F1杂种为供体材料 ,研究了 3种不同时期秋水仙碱处理对小孢子胚胎发生、成苗与加倍率的影响。对分离小孢子直接进行 15h以及 0、5 0、5 0 0和 10 0 0mg/L秋水仙碱处理 ,在液体NLN培养基中胚胎发生好 ,培养 5周后获得大量正常胚体。将这些发育优异的胚体移入固体MS培养基、经过 10d低温 (2℃ )诱发 ,常温 (2 4℃ )下培养能很好地萌发 ,并迅速长成完全正常的再生植株。 5 0 0mg/L秋水仙碱处理 15h获得了 83%～ 92 %的高染色体加倍率 ,并且嵌合体和多倍体数很少。对正常胚体在移入固体培养基前用 0、2 5、5 0、2 5 0、5 0 0和 10 0 0mg/L秋水仙碱 15和 30h进行染色体加倍处理 ,或对再生植株在幼苗期进行 12 5mg/L秋水仙碱 2 0h浸根处理 ,其加倍效果均不及采用分离小孢子直接进行秋水仙碱处理 ,且其嵌合体的发生频率也较高 ;后者还产生大量待特殊处置的秋水仙碱废液 ,并推迟再生株的生育进程。因此 ,利用分离小孢子直接进行秋水仙碱加倍染色体更为安全。Using two winter rape (Brassica napus) F 1 hybrids that had good responses to microspore culture as the donor plant in the experiment, the effects of colchicine treatments in three different periods on embryogenesis, shoot production and doubling efficiency were investigated. Immediate colchicine treatment of isolated microspores with 0, 50, 500 and 1 000 mg/L concentrations for 15 h, large amounts of normal embryos were produced when cultured in NLN medium for 5 weeks. After transferred to solid MS medium and induced at low temperature (2℃) for 10 days, the embryos germinated well at 24℃ and quickly generated shoots. High doubling efficiency of 83%～92% was obtained from 500 mg/L colchicine treatment for 15 h with less chimeric and polyploid plants. Colchicine treatments of normal embryos with 0, 25, 50, 250, 500 and 1 000 mg/L concentrations for 15 and 30 h before transferring to solid medium, or regenerated plants treated with 125 mg/L concentration for 20 h at the seedling stage had low doubling efficiencies but high chimeric percentages. In addition, plant treatment for chromosome doubling produced large volumes of spent colchicine and delayed plant growth and development. Therefore, immediate chromosome doubling of isolated microspores by colchicine treatment is safer, quicker and more efficient.

关 键 词：甘蓝型油菜 小孢子 秋水仙碱处理 双单倍体频率 

分 类 号：S565.4[农业科学—作物学]