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

doi:10.11733/j.issn.1007-0435.2008.02.003

›› 2008, Vol. 16 ›› Issue (2): 115-120.DOI: 10.11733/j.issn.1007-0435.2008.02.003

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

LIU Xiao-lin¹, KANG Jun-mei¹, SUN Yan², WANG Ji-feng¹, HU Xiao-yan^1,3, YANG Qing-chuan¹

1. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100094, China;
2. China Agricultural University, Beijing 100094, China;
3. College of Resources and Environment, Northwest A&F University, Yangling, Shaanxi Province 712100, China

Received:2007-06-07 Revised:2007-09-14 Online:2008-04-15 Published:2008-04-15

MsNHX1基因转化紫花苜蓿及转基因植株的鉴定

刘小琳¹, 康俊梅¹, 孙彦², 王继峰¹, 胡晓艳^1,3, 杨青川¹

1. 中国农业科学院北京畜牧兽医研究所, 北京, 100094;
2. 中国农业大学草地研究所, 北京, 100094;
3. 西北农林科技大学资源环境学院, 陕西, 712100

通讯作者: 杨青川,E-mail:qchyang66@yahoo.com.cn
作者简介:刘小琳(1982- ),女,内蒙古赤峰市人,硕士研究生,研究方向为牧草生物技术
基金资助:
“十五”国家高技术发展计划(863计划)项目(2002AA241101);国家自然科学基金项目(30471110)

Abstract

Abstract: 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.

Key words: Medicago sativa L.cv.Zhongmu No.1, Alfalfa, MsNHX1 gene, Transformation, Transgenic plants identification

摘要： 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。

关键词: 中苜1号, 紫花苜蓿, MsNHX1基因, 转化, 转基因植株鉴定

CLC Number:

Q785

LIU Xiao-lin, KANG Jun-mei, SUN Yan, WANG Ji-feng, HU Xiao-yan, YANG Qing-chuan. Agrobacterium-mediated Transformation of Alfalfa by MsNHX1 Gene and Molecular Detection for Transgenic Alfalfa[J]. , 2008, 16(2): 115-120.

刘小琳, 康俊梅, 孙彦, 王继峰, 胡晓艳, 杨青川. MsNHX1基因转化紫花苜蓿及转基因植株的鉴定[J]. , 2008, 16(2): 115-120.

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

MsNHX1基因转化紫花苜蓿及转基因植株的鉴定

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