| 阔叶猕猴桃遗传转化技术参数的优化 |
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| 引用本文： | 高月,毕静华,刘永立.阔叶猕猴桃遗传转化技术参数的优化[J].果树学报,2007,24(4):553-556,F0003. |
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| 作者姓名： | 高月 毕静华 刘永立 |
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| 作者单位： | 浙江大学农业与生物技术学院,杭州,310029 |
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| 摘 要： | 为了提高阔叶猕猴桃的遗传转化效率,以阔叶猕猴桃叶柄为试材,通过根癌农杆菌介导法进行了遗传转化技术参数的研究。结果表明,叶柄预培养3～4d、用农杆菌悬浮液(D600nm值0.5)感染10min、共培养48h、共培养时在培养基中加入200μmol/L乙酰丁香酮处理可以获得较高的gus基因表达率。在供试的1200个叶柄中获得了49个抗性芽,转化频率为4.1%。对转基因抗性材料进行PCR检测和gus基因组织化学染色,证实了外源基因已整合到阔叶猕猴桃的基因组中,并得到稳定表达。
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| 关 键 词： | 阔叶猕猴桃 遗传转化 根癌农杆菌 gus基因 |
| 文章编号： | 1009-9980（2007）04-553-04 |
| 修稿时间： | 2006-12-222007-05-08 |
Study on the optimum technological parameters for genetic transformation of Actinidia latifolia |
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| GAO Yue,BI Jing-hua,LIU Yong-li.Study on the optimum technological parameters for genetic transformation of Actinidia latifolia[J].Journal of Fruit Science,2007,24(4):553-556,F0003. |
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| Authors: | GAO Yue BI Jing-hua LIU Yong-li |
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| Institution: | College of Agriculture and Biotechnolo~y, Zhejiang University, Hang,.hou, Zhejiang 310029 China |
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| Abstract: | In order to improve the genetic transformation efficiency of Actinidia latifolia Merr.,its petiole explants were transformed by Agrobacterium tumefaciens for studying the optimum technological parameters.Results showed that the petiole explants pre-cultured for 3～4 days,infected for 10 min with Agrobacterium tumefaciens(concentration:D600 nm 0.5 ),and co-cultured for 48 h in the medium with 200 μmol/L acetosyringone could get the highest level of gus gene expression.49 kanamycin-resistant shoots were obtained from 1 200 petiole explants and transgenic frequency was 4.1%.Successful transformation was confirmed by PCR and histochemical analysis of gus activity in leaves of kanamycin-resistant shoots.The results showed that gus gene was integrated into A.latifolia genomes and had stable expression. |
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| Keywords: | A ctinidia latifolia Genetic transformation A grobacterium tumefaciens gus gene |
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