MsNHX1基因转化紫花苜蓿及转基因植株的鉴定

Agrobacterium-mediated Transformation of Alfalfa by MsNHX1 Gene and Molecular Detection for Transgenic Alfalfa

以紫花苜蓿"中苜1号"(Medicago sativa L.cv.Zhongmu No.1)7日龄无菌苗子叶为外植体,建立适用于农杆菌介导的转基因组织再生体系,并进行MsNHX1基因转化试验。MsNHX1基因编码"中苜1号"液泡膜Na+/H+逆向转运蛋白,将其转化到"中苜1号"中,以期获得耐盐性更好的苜蓿材料。利用PCR技术、RT-PCR技术及Dot Blotting技术对转基因植株进行鉴定,结果显示:MsNHX1基因已经整合到"中苜1号"基因组内,并且可以转录为mRNA。

In order to obtain the materials with better performance of salt tolerance, the plant regeneration system mediated by Agrobacterium tumefaciens was established using the cotyledons as the explants from the sterile 7-day old seedlings of Medicago sativa L. cv Zhongmu No. 1 and the genetic transformation of MsNHX1 gene was conducted. The vacuolar Na^＋/H^＋ antiporter of Zhongmu No. 1 was coded by gene MsNHX1 and then transformed back into Zhongmu No. 1. PCR, RT-PCR, and Dot Blotting analysis were used to verify the transgenetic plants and the results indicate that MsNHX1 gene had been inserted into the genome of the transgenic plants and the extrinsic gene had been transcripted to mRNA.