黄瓜再生体系建立及农杆菌介导遗传转化-蔬菜学专业毕业论文.docxVIP

黄瓜再生体系的建立及农杆菌介导的遗传转化 period (9-15 d), IAA and GA contents first increased and then decreased, ZR content was reduced, while ABA content first decreased and then increased. It indicated that it was important of endogenous hormone to regeneration of cucumber. The endogenous hormones contents changed with the different stage of cucumber regeneration. The types, concentration and coordination balance of endogenous hormones were pivotal in bud formation and differentiation. 3.Based on the regeneration of cucumber, we established efficiency Agrobacterium-mediated transformation system, mainly including preculture, coincubate, differentiation culture, and rooting culture. The production of non-transformed adventitious buds was inhibited on 150 mg/L Kanamycine. 250 mg/L carbenicilin can effectively restrain the growth of Agrobacterium. 20 transformed plants were obtained from 250 cotyledonary node explants and 12 transgenic plants were confirmed using PCR specific analysis, and the transgenic rate was approximately 4.8 %. The expression level of cuPLDα gene in transgenic plants were markedly lower than in wildtype plants, indicating that the entire foreign gene has been successfully inserted in cucumber genome and inhibited the cuPLDα gene expression Key words: Cucumber; Cotyledonary node; Agrobacterium tumefaciens; Transformation; Phospholipase Dα gene 4 英文缩略词表 英文缩写 英文名称 中文名称 ABA AS Abscisic acid Acetosyringone 脱落酸 乙酰丁香酮 APX Ascorbate peroxidase 6-苄氨基嘌呤 6-BA 6-Benzylaminopurine 抗坏血酸过氧化物酶 Cb Carbenicillin 羧苄青霉素 CAT Catalase 过氧化氢酶 cuPLDα PLDα gene of cucumber 黄瓜磷脂酶 Dα 基因 2,4-D 2,4-dichlorophenoxyacetic acid 2,4-二氯苯氧乙酸 FW Fresh weight 鲜重 GA Gibberellic acid 赤霉素 GUS β-glucuronidase β-葡萄糖醛糖苷酶 Hpt-Ⅱ Hygromycin phosphotransferaseⅡ 潮霉素磷酸转移酶Ⅱ IAA Indole-3-Acetic Acid 吲哚乙酸 IBA Indole-3-Butyric Acid 吲哚丁酸 Kan Kanamycine 卡那霉素 NAA Naphthylacetic acid 萘乙酸 Npt-Ⅱ Neomycin phosphotransferase Ⅱ 新霉素磷酸转移酶Ⅱ PCR Polymerase Chain Reaction 聚合酶链式反应 PLD Phospholipase D 磷脂酶 D POD Peroxidase 过氧化物酶 Rif