Cotyledonary node pre-wounding with a micro-brush increased frequency of Agrobacterium-mediated transformation in soybean
Cotyledonary node pre-wounding with a micro-brush increased frequency of Agrobacterium-mediated transformation in soybean
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DOI：
10.5511/plantbiotechnology.27.217
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发表时间：
2010-06
影响因子：
1.6
通讯作者：
Tetsuya Yamada;Satoshi Watanabe;M. Arai;K. Harada;K. Kitamura
中科院分区：
文献类型：
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作者：
Tetsuya Yamada;Satoshi Watanabe;M. Arai;K. Harada;K. Kitamura
一个日本大豆品种Kariyutaka，已知从子叶节的部分形成许多不定芽，在常规方案中，其转化频率（T0个转基因植株的相对数量，这些植株稳定地将转基因传递给它们的T1个后代）对受农杆菌感染的外植体的数量)约为1.0%。我们对该方法进行了改进，得到变换频率为4.4%。改良方案为微刷机械损伤外植体，并在农杆菌侵染期间施用0.02%的Silwet L-77。在与农杆菌共培养8 ~ 12周后，共获得10株植株。共培养后5 ~ 6个月迅速获得T1株种子，其中T0株开花早。共培养后12周产生的4株T0植株中有3株（占全部T0植株的75%）具有稳定的转基因，并能传递给T1后代，而共培养后8周产生的22株T0植株中只有7株（占全部T0植株的32%）稳定地传递给T1后代。
A Japanese soybean variety, Kariyutaka, known to form many adventitious shoots from sections of the cotyledonary node, had a transformation frequency (relative number of T0 transgenic plants, which stably transmitted transgenes to their T1 progenies, to number of explants subjected to Agrobacterium-infection) of about 1.0% in the conventional protocol. We modified this method to obtain the transformation frequency of 4.4%. Modified protocol included mechanical wounding of explants with a micro-brush and application of 0.02% of Silwet L-77 during the infection with Agrobacterium. We obtained T0 plants 8 to 12 weeks after co-cultivation with Agrobacterium. T1 seeds were rapidly obtained from 5 to 6 months after the co-cultivation, for T0 plants flowered precociously. Three T0 plants out of 4, i.e. 75% of all T0 plants, which were generated at 12 weeks after co-cultivation, possessed stable transgene(s) that were transmitted to their T1 progenies, although only 7 T0 plants out of 22, i.e. 32% of all T0 plants, which were generated at 8 weeks after the co-cultivation, stably transmitted the Basta-resistance to their T1 progenies.