拟南芥AtTIP5;1基因转化非洲紫罗兰的研究
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Agrobacterium-Mediated Genetic Transformation of AtTIP5;1 Gene into Saintpaulia ionantha
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    摘要:

    为研究AtTIP5;1基因的功能,以非洲紫罗兰叶片为受体材料,进行AtTIP5;1基因遗传转化条件的研究,并对转基因植株在高浓度硼酸胁迫下的表现进行分析,以明确利用AtTIP5;1基因提高非洲紫罗兰抗逆性的可行性。结果表明:(1)以非洲紫罗兰叶片为受体的最适转化条件为:叶片预培养2 d,农杆菌共培养2 d,共培养时添加100 μmol/L乙酰丁香酮;最适潮霉素筛选浓度为40 mg/L。(2)转化植株经PCR和RT-PCR检测显示,AtTIP5;1基因成功转入了非洲紫罗兰,获得了17个稳定的转基因植株。(3)对转基因植株和对照植株进行3 mmol/L硼酸胁迫处理结果表明,AtTIP5;1基因表达受高浓度硼酸诱导,且AtTIP5;1基因表达可以减缓植株受高浓度硼酸胁迫导致的组织褐化、叶片卷曲变形症状;转AtTIP5;1基因植株干物质率、可溶性糖、可溶性蛋白质含量下降,而POD、SOD活性上升,证明转AtTIP5;1基因的非洲紫罗兰植株提高了对高浓度硼酸的耐受能力。研究结果为深入探讨AtTIP5;1基因的功能以及非洲紫罗兰基因工程育种奠定了基础。

    Abstract:

    In order to study the function of AtTIP5;1 gene,we employed Saintpaulia ionantha leaves as receptor materials and optimized the genetic transformation conditions.The transgenic plants were exposed to high concentration of boric acid and the resistant ability was evaluated.The results showed:(1)The receptor leaves pre-cultured for 2 d,infected for 15 min,co-cultured of 2 d with 100 μmol/L AS,and selected with 40 mg/L hygromycin were the optimum transformation conditions.(2)After PCR and RT-PCR screening,17 transformed plants containing AtTIP5;1 transgene were obtained.(3)Transformed and control plants were treated by high concentration boric acid (3 mmol/L) and physiological indexes of these plants were evaluated.The results indicated that the AtTIP5;1 expression could be induced by boron.Transgenic plants showed a lower degree of plant tissue browning,leaves curling,and a higher rate of dry matter,soluble sugar and soluble protein content.Meanwhile,the increase of POD and SOD activities were weaken.Our results demonstrate that AtTIP5;1 trasformation can improve the tolerance of S.ionantha to the high boron stress.Our study is helpful for further investigation on AtTIP5;1 gene function and gene engineering in breeding of S.ionantha.

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叶志琴,李佳慧,王广东.拟南芥AtTIP5;1基因转化非洲紫罗兰的研究[J].西北植物学报,2014,34(12):2412-2417

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  • 在线发布日期: 2015-01-16
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