黄瓜子叶高效再生体系的建立与遗传转化  

Establishment of High Effective Regeneration System in Cucumber(Cucumis sativus L.)and Agrobacterium tumefaciens Mediated Genetic Transformation

作　　者：王艳蓉[1] 陈丽梅[1] 潘俊松[1] 何欢乐[1] 蔡润[1] 

机构地区：[1]上海交通大学植物科学系,上海201101

出　　处：《上海交通大学学报（农业科学版）》2006年第2期152-156,164,共6页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：上海市科委攻关项目(043919317)

摘　　要：实验以S05、S062个黄瓜品系的子叶为外植体,以MS为基本培养基,添加不同浓度的BA、ABA、AgNO3进行再生培养。结果表明,在MS培养基上,ABA与BA组合能有效地抑制愈伤组织的形成,促进芽的发生,其再生率超过90%,每个外植体的出芽数为1～3个。S05、S06的最佳芽诱导培养基分别为:MS+BA3.0mg·L-1+ABA1.0mg·L-1+AgNO32.0mg·L-1和MS+BA1.5mg·L-1+ABA0.5mg·L-1+AgNO32.0mg·L-1。在此基础上,以S06子叶为受体材料,建立了pBI121-ATT1为中间载体,根癌农杆菌LBA4404介导的遗传转化体系,最终获得了3株抗性植株,经过PCR检测,初步确定均为阳性转化植株。In this experiment, we chose cotyledons of S05 and S06 that are two lines of cucumber （Cucumis sativus L） as explants, and researched MS medium containing different concentration BA.ABA and AgNOs how to influence the regeneration of these explants. The results showed that the combination of BA and ABA efficiently restrict the callus inducing and promote the growth of shoots on the MS medium. The regeneration rate reaches over 90% and the number of shoots per explants is from 1 to 3. The best shoot induction medium for S05 and S06 arc MS＋BA 3.0 mg·L^-1 ＋ABA 1.0 mg·L^-1＋AgNO3 2.0 mg·L^-1 and MS＋BA 1.5 mg·L^-1 ＋ABA 0.5 rag·L^-1 ＋AgNO3 2.0 mg·L^-1. On this base, we chose cotyledons of S06 as the plant material. They were transformed mediated with Agrobacterium tumefaciens strain LBA4404 that carries the binary vector pBI121 containing an ATT1 gene. And finally, we got three putative transgenic plants, they were all showed the presence of the expected DNA fragments in the PCR analysis.

关 键 词：黄瓜 子叶 不定芽 植株再生 根癌农杆菌 

分 类 号：S642.2[农业科学—蔬菜学]