| 草莓叶盘再生和遗传转化体系的优化 |
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| 引用本文： | 柏锡,狄少康,张秀芹,朱延明.草莓叶盘再生和遗传转化体系的优化[J].东北农业大学学报,2012(10):141-150. |
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| 作者姓名： | 柏锡 狄少康 张秀芹 朱延明 |
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| 作者单位： | 东北农业大学生命科学学院,哈尔滨 150030 |
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| 基金项目： | 黑龙江省自然科学基金(C2005-25);国家自然科学基金(30270916); |
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| 摘 要： | 农杆菌介导的叶盘法是目前草莓遗传转化的主要方法,建立高效稳定的叶盘再生体系是获得转基因草莓的必要条件.文章探讨了影响草莓离体叶片不定芽诱导的主要因素,建立了一个稳定的草莓植株再生体系,确定草莓品种弗吉尼亚不定芽诱导的最佳培养基组分为:MS+TDZ 2 mg·L-1+IBA 0.1 mg·L-1+2,4-D 0.2 mg·L-1,pH 5.8.发现叶盘近轴面向下放置能显著提高再生频率,且叶片幼嫩程度与草莓不定芽分化率呈正相关.并以此体系对草莓叶盘遗传转化进行优化,发现草莓品种弗吉尼亚遗传转化最佳预培养时间为2 d,最佳侵染时间为7 min.同时,利用该体系,将由不同启动子诱导的组织型纤溶酶原激活剂(Tissue-type plasminogen activator, t-PA)基因对草莓进行遗传转化.
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| 关 键 词： | 草莓 遗传转化 组织型纤溶酶原激活剂 |
Establishment of strawberry regeneration and transformation system through leaf discs |
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| BAI Xi,DI Shaokang,ZHANG Xiuqin,ZHU Yanming.Establishment of strawberry regeneration and transformation system through leaf discs[J].Journal of Northeast Agricultural University,2012(10):141-150. |
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| Authors: | BAI Xi DI Shaokang ZHANG Xiuqin ZHU Yanming |
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| Institution: | (School of Life Sciences,Northeast Agricultural University,Harbin 150030,China) |
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| Abstract: | The Agrobacterium-mediated transformation method through leaf discs was mainly adopted in strawberries.Therefore,it is necessary to establish an ideal regeneration system of strawberry.In this study,it investigated some key factors affecting the shoot regeneration of strawberry leaf.The optimum strawberry shoot induction medium of c.v Fugilia was confirmed as the following: MS+ TDZ 2 mg·L-1+IBA 0.1 mg·L-1+2,4-D 0.2 mg·L-1,pH 5.8.The results demonstrated that adaxial surface towards medium could obviously increase the strawberry regeneration frequency,and the positive correlation was observed between the age of strawberry leaf discs and shoot regeneration frequency.Some key factors affecting Agrobacterium-mediated transformation system were also investigated,and it found that the optimum pre-culture time was 2 d and the optimum infection time was 7 min.In addition,tissue-type plasminogen activator gene was also transgened with three different promoters into strawberry with this system. |
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| Keywords: | strawberry transformation tissue-type plasminogen activator |
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