摘要
含双元载体的非致瘤根癌农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍ ｅｆａｃｉｅｎｓ） ＬＢＡ４４０４经乙酰丁香酮和复合酚类化合物活化预处理后，感染胡萝卜（Ｄａｕｃｕｓ ｃａｒｏｔａ）悬浮培养细胞，在含５０ ｍ ｇ／ＬＫｍ 的选择培养基上筛选Ｋｍ ｒ 克隆。实验表明：酚类化合物活化预处理组的转化率分别为０．２４％ 和０．１７％ ，而未经酚类化合物活化处理组的转化率仅为０．０２％ 。Ｋｍ ｒ 克隆在含５０ ｍ ｇ／Ｌ Ｋｍ 的无激素培养基上通过体细胞胚胎发生形成完整植株。组织化学分析表明：报告基因ＧＵＳ的活性可出现于Ｋｍ ｒ 克隆细胞、再生植株的根、茎、叶、叶柄维管束薄壁细胞、叶片的气孔保卫细胞、皮层细胞、叶肉细胞和毛状体。
Non tumorigenic Agrobacterium tumefaciens strain LBA4404 harboring binary vector pretreated with acetosyringone and complex phenolic compounds significantly increased the rate of carrot (Daucus carota) suspension cell transformation and plant regeneration. Kanamycin resistant colones were selected on media containing 50 mg/L kanamycin.The author's experimental data indicated that transformation frequencies in the group activated by acetosyringone and complex phenolic compounds pretreatment were 0 24% and 0 17% respectively while that in the phenolic compound non pretreated was only 0.02%.Transgenic carrot plants were obtained from the resistant colones via somatic embryogenesis on hormone free media containing 50 mg/L kanamycin.Histochemical analysis showed that high levels of GUS activity appeared in the parenchymatous cells of vascular bundles of roots,stems,leaves and petioles as well as stomatic guard cells,cortical cells,mesophyllous cells and trichome of leaves,also epidermal cells also parenchymatous cells under the epiderm of petiols.
关键词
胡萝卜
悬浮培养
细胞
根癌农杆菌
遗传转化
Daucus carota
Suspension culture cells
Agrobacterium tumefaciens
Genetic transformation
Phenolic compounds
Binary vector
GUS
