冬性花椰菜的小孢子胚诱导和植株再生研究  

Embryogenesis and plant regeneration from isolated microspores of winter cauliflower (Brassica oleracea var. botrytis)

作　　者：顾宏辉[1] 唐桂香[1] 张国庆[1] 周伟军[1] 

机构地区：[1]浙江大学农学系

出　　处：《浙江大学学报（农业与生命科学版）》2004年第1期34-38,共5页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：教育部留学回国人员科研启动基金;中瑞政府间合作项目(C08);浙江省重点科技资助项目(021102109).

摘　　要：以2个冬性花椰菜F1杂交种为供体材料,研究了不同热击温度与时间组合、更新培养液和冷击预处理对花椰菜小孢子培养胚发频率的影响.结果表明,最佳热击组合是32℃、24h;先用17%蔗糖浓度的NLN-17培养液热击培养24h后换成10%蔗糖的NLN-10培养液,并转入24℃继续培养,能显著提高出胚产量;更换培养液和冷击预处理能明显提高胚体质量,减少愈伤状胚的形成.分离小孢子在NLN-13培养液中经32℃、24h热击暗培养后转入24℃下培养,产生了大量正常胚体.小孢子分离30d后将正常胚体移入固体MS培养基中萌发、长根,并经2次继代培养后获得了大量正常的再生植株,其多为自发双单倍体.The effects of various combinations of heat shock temperature and duration, medium renovation and cold shock pretreatment on the embryogenesis of isolated microspores were studied in two F1 hybrids of winter cauliflower (Brassica oleracea var. botrytis). The results indicated that the optimal combination of heat shock temperature and duration was 32 ℃ and 24 h. A decrease in sucrose concentration from 17% to 10% in liquid NLN medium following the first 24 h incubation at 32 ℃ significantly enhanced microspore embryogenesis. Both medium renovation and cold shock pretreatment markedly improved the quality of normal embryos and reduced the number of callus-like embryos. After 24 h induction at 32　℃ and continuous culture at 24　℃ in darkness for 30 d, isolated microspores produced normal embryos, which were then transferred to solid MS medium for embryo germination and root development. After two times of subculture, large amounts of normal regenerated plants were produced from generated shoots, and these regenerated plants were mainly spontaneous doubled haploids.

关 键 词：冬性花椰菜 小孢子 植株再生 胚胎发生 倍性检测 

分 类 号：S635.3[农业科学—蔬菜学]