利用小孢子胚离体培养筛选抗除草剂的油菜  

In vitro Selection of Herbicide Resistance in Microspore-derived Embryos of Oilseed Rape

作　　者：许玲[1] 张国庆[1] 顾宏辉[1,2] 周伟军[1] 

机构地区：[1]浙江大学农业与生物技术学院 [2]浙江省农业科学院作物研究所,杭州310021

出　　处：《农业生物技术学报》2005年第4期411-415,共5页Journal of Agricultural Biotechnology

基　　金：浙江省重点科技项目(No.021102109;2004C22013);浙江省自然科学基金(Y304162);杭州市科技项目(No.2003132E32)资助。

摘　　要：选用4个甘蓝型油菜(BrassicanapusL.)F1(7039,7040,282和5102)为小孢子培养供体材料,分别对小孢子胚在含草甘膦和盖草能的培养基上进行培养,以筛选出抗草甘膦和抗盖草能的胚状体,进而获得再生植株。其中,基因型7039和7040用于草甘膦筛选,282和5102用于盖草能的筛选。选取子叶期胚状体,在含0.006%的草甘膦及0.01%和0.02%盖草能的MS-2培养基上培养2周,不抗草甘膦和不抗盖草能的胚状体2周内变褐死去,抗草甘膦和抗盖草能的胚状体转绿。随后转绿胚状体转移至正常MS-2培养基中继续培养,直至获得再生植株。再生植株喷施0.25%的草甘膦液表明均抗草甘膦;而对植株喷施0.05%的盖草能液时,0.02%筛选出的植株大部分为抗盖草能,而0.01%筛选出的植株却大部分死去,表明用0.02%浓度筛选抗盖草能植株的方法更有效。用170mg/L秋水仙碱直接处理再生植株20和30h,染色体加倍率分别达到了34%和52%。Using 4 oilseed rape （Brassica napus L.） F1 hybrids （7039, 7040, 282 and 5102） as donor plants for microspore culture, the experiment was conducted to select glyphosate and haloxyfop resistant embryos through glyphosate- and haloxyfop-contained cultural media with microspore-derived embryos in vitro respectively. The genotypes 7039 and 7040 were used to select regenerated plants which showed glyphosate resistance and the other two genotypes were used for haloxyfop resistance selection. The embryos at cotyledonary stage were taken to grow on glyphosate- and haloxyfop-contained MS-2 medium for two weeks. The embryo which was not resistant to glyphosate and haloxyfop collapsed in short time, while those which were resistant to glyphosate and haloxyfop turned green and survived within two weeks. The green embryos were then transferred into the normal MS-2 medium for further plant regeneration. All regenerated plants showed tolerance to glyphosate after tested by spraying 0.25% glyphosate herbicide. However, when the regenerated plants selected from 0.02% haloxyfop were sprayed with 0.05% haloxyfop, most of these regenerated plants grew well, while the survival rate of the regenerated plants from 0.01% haloxyfop-contained medium were low. The present experiment indicated that the method of using 0.02% haloxyfop to select haloxyfop resistance plants was better than that using 0.01% haloxyfop. The chromosome doubling efficiency of regenerated plants reached 34% and 52% after directly treated with 170 mg/L colchicine for 20 and 30 h, respectively.

关 键 词：甘蓝型油菜 小孢子培养 胚胎 离体筛选 抗草甘膦 抗盖草能 植株再生 胚状体 筛选 抗除草剂 

分 类 号：S565.4[农业科学—作物学] S565.403.5