System optimization for tomato Micro-Tom transgenic regeneration
Received date: 2016-04-25
Online published: 2018-05-07
转基因植株的再生频率较低一直是植物基因工程研究的瓶颈之一. 以番茄的 Micro-Tom 突变体为材料, 探索了外植体植物激素配比、预培养时间、共培养时间、筛选剂对愈伤组织发生和不定芽生长的影响. 结果表明: 子叶比下胚轴更适合作为愈伤组织和不定芽诱导的外植体; 当玉米素 (zeatin, ZT) 质量浓度为 0.5 mg/L, 吲哚乙酸 (indole acetic acid, IAA) 质量浓度为 1.0 mg/L 时最有利于不定芽诱导; 外植体预培养的适宜时间为 3 d, 农杆菌和外植体共培养的适宜时间为 2 d; 外植体对卡那霉素的耐受上限为 40 mg/L, 替门汀的适宜质量浓度为 150 mg/L. 实验所建立的转基因再生体系为 Micro-Tom 番茄的遗传工程应用奠定了基础.
关键词： Micro-Tom 番茄; 玉米素; 替门汀; 遗传转化; 农杆菌
邱实, 张文举, 许馥慧, 邓志瑞 . Micro-Tom番茄转基因植株再生体系的优化[J]. 上海大学学报(自然科学版), 2018 , 24(2) : 322 -330 . DOI: 10.12066/j.issn.1007-2861.1773
Low redifferentiation frequency of transgenic plant is a major barrier in plant gene engineering. Tomato species Micro-Tom was used as the experimental material. Effects of plant hormone combination, preculture time, and co-culture time of kanamycin and timentin on callusogenesis and adventitious bud growth were explored. The results show that cotyledon is more suitable than hypocotyledon as explants for callusogenesis and adventitous bud induction. Combination of 0.5 mg/L zeatin (ZT) with 1.0 mg/L indole acetic acid (IAA) brings about the highest adventitous bud induction. Suitable preculture time for explants is 3 d, and that for co-culture with Agrobacterium tumefaciens is 2 d. The upper tolerance limit of explant to kanamycin for selection is 40 mg/L and suitable timentin concentration for Agrobacterium tumefaciens inhibition is 150 mg/L. The established system may provide a good base for genetic engineering for Micro-Tom tomato.
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