普通小麦春化基因VRN1 RNA干扰载体构建及转基因小麦获得

RNA Interference Vector Construction of VRN1 Gene and Obtaining Transgenic Wheat

为进一步探究春化基因VRN1在小麦发育进程中的功能,利用荧光定量PCR分析了VRN1基因在不同发育特性小麦品种新春2号、京841中的表达情况。结果表明,随着生育进程的推进,VRN1基因在新春2号叶片和茎尖中表达量均呈上升趋势,VRN1基因在京841叶片中呈波动上升趋势,在茎尖中表达量趋于0;以p FGC5941载体为基础构建含有VRN1反向重复序列的RNA干扰载体,利用农杆菌介导的茎尖转化法转化新春2号获得了再生植株,并通过PCR法检测获得了转基因阳性植株,为从分子水平上实现小麦发育特性遗传改良和创育小麦新种质奠定了基础。

In order to study the biological function of VRN1 in wheat developmental process,VRN1 expression patterns were analyzed using fluorescence quantitative RT-PCR in two vernalization characteristic varieties Xinchun No. 2 and Jing 841. The results showed that the VRN1 transcription levels increased gradually in Xinchun No. 2leaves and shoot meristems. While the VRN1 transcription levels in Jing 841 leaves showed a wavelike rises,but the expression level in shoot meristems was very low and nearly zero. The VRN1 RNA interference sequence was successfully introduced into vector p FGC5941,and the recombined vector was transferred into the shoot apical meristem of Xinchun No. 2 by Agrobacterium-mediated transformation. Transgenic plants were detected by PCR amplification,which could lay a foundation for genetic improvement of wheat developmental characteristics on molecular level and created new wheat germplasm for wheat breeding.