基于不定芽原位诱导的大豆转基因方法

A New Soybean Transformation Method Based on Adventitious Shoots Induction

建立了一种新型农杆菌介导的大豆转基因方法(专利公开号:102181479A),以萌发大豆为外植体,最大限度地保持了大豆植株的完整性,在非无菌环境下进行转化,利用草丁膦筛选阳性植株。该方法不需要组织培养,脱离了传统组培的限制。对16个大豆基因型进行转化,经PCR、RT-PCR、qPCR和GUS染色鉴定转基因株系,平均转化率为23%。获得当代转基因植株接近实生苗,收获种子仅需3～4个月。该方法将有利于促进大豆基因功能研究和规模化转化。

In this work,a novel Agrobacterium-mediated transformation methodology,independent of organ culture,was established.The germinated entire seedlings of soybean were used as explants.The transformation system maximum to maintain the integrity of soybean itself,keep off the limitations of traditional tissue culture.The transformation was under non-sterile environment.The transgenic regeneration plants were selected with herbicide glufosinate.The transgenic lines were confirmed by PCR,RT-PCR,qPCR and GUS stainig.Sixteen genotypes of soybean were transformed,and the average transgenic frequency was 23%.Transgenic soybean plants were usually produced in less than 2 month after Agrobacterium inoculation and seeds could be harvested in 3-4 months.The growth characteristics and morphology of transgenic plants were identical to the untransformed wild type plants.This method would be beneficial to facilitate the characteristic of gene functions and boosted the manipulation of soybean transformation for commercial purpose.