利用花粉介导法获得油菜RNAi转基因植株

Recovery of RNAi Plants in Brassica napus with Pollen-mediated Transformation

以甘蓝型油菜7B为试验材料,利用花粉介导法将携带RNA干扰的HKL1基因的pCaMHKL1-RNAi质粒转入7B中,获得转基因油菜植株,通过PCR分别对T_1,T_2植株进行了转基因鉴定,并对转基因植株后代的生长发育进行研究。结果表明,T_2的RNAi植株与野生型植株相比,出真叶时间较晚,株型较矮小;qRT-PCR结果表明,RNAi植株中HKL1基因的相对表达量为野生型植株的24.6%~67.9%;不同的RNAi植株中HKL1基因的表达量各不相同;利用花粉介导法在甘蓝型油菜中进行RNAi质粒转化,外源基因可以在后代植株中稳定遗传。花粉介导法较之农杆菌侵染、基因枪法等基因转化方法操作更加简单、适用性更强。

In this study, the plasmid pCaMHKL1-RNAi contains RNA interferenced HKL1 gene was introduced into rapeseed line 7B plants by pollen-mediated method. T1 and T2 generation plants were identified by PCR, respectively. The growth and development of the offspring of transgenic plants was studied. The results showed that transgenic seedlings grew euphylla later than wild-type plants.During plant growth, the transgenic plants were dwarfer than the wild-type plants. The results of qRT-PCR showed that the relative expression of HKL1 gene in transgenic T2 plants was 24.6% -67.9% of wild-type plants,the expression of HKL1 gene was different in RNAi type transgenic plants. The transformation of RNAi plasmid was carried out by pollen mediated method, exogenous genes could be stable in different generations. The pollen mediated method is more simple and application than that of the infection of Agrobacteriummediated method and particle bombardment method etc.