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利用根据RNAi技术构建的干扰黄烷酮-3-羟化酶基因(F3H)的载体pF3Hi330,通过农杆菌介导辅助抽真空的大豆茎尖转化法和花粉管通道法将pF3Hi330载体整合到大豆基因组中,获得转基因阳性植株4棵,抗性植株经PCR检测,初步证明外源目的基因已整合到大豆基因组中。同时,对转基因阳性植株异黄酮含量进行测定,显示异黄酮含量得到一定的提高。结果表明更多的前体流向异黄酮合成的分支途径,达到提高大豆异黄酮含量的目的。
The pF3Hi330 vector was integrated into the genome of soybean by Agrobacterium tumefaciens-mediated soybean apex transformation and pollen tube pathway by using the vector pF3Hi330 which interferes with flavanone-3-hydroxylase gene (F3H) , 4 transgenic plants were obtained, and the resistant plants were detected by PCR. It was initially proved that the exogenous gene has been integrated into the soybean genome. At the same time, the content of isoflavones in transgenic plants was measured, showing that the isoflavone content increased a certain extent. The results show that more precursors flow to the branch of isoflavone synthesis, to achieve the purpose of increasing the content of soy isoflavones.