Micro-Tom番茄转基因植株再生体系的优化

System optimization for tomato Micro-Tom transgenic regeneration

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MicroTom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.

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.