MsNHX1基因转化紫花苜蓿及转基因植株的鉴定

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

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

收稿日期:2007-06-07 修回日期:2007-09-14 出版日期:2008-04-15 发布日期:2008-04-15
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)

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

摘要/Abstract

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

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

中图分类号:

Q785

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.

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.

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参考文献

[1] Rhoades J D,Kandiah A,Mashali A M.The use of saline wa-ters for crop production[J].FAO,Irrigation and Drainage Pa-per,1992,80-92
[2] Jacoby.B.Mechanisms involved in salt tolerance by plants.M.Pessarakli (ed.),Handbook of Plant and Crop Stress.Marcel Dekker[M].New York,1993.97-123
[3] 牛东玲,彭宏春,王启基,等.柴达木盆地弃耕地盐渍状况的主分量分析[J].草业学报,2001,10(2):39-46
[4] HINCHEE Maw,DVCONNER Ward,NEWELL C A.Pro-duction of transgenic soybean plants using Agrobacterium me-diated DNA transfer[J].Bio Technology,1988(6),915-922
[5] Hamada A,ibino T,amura T,et al.Na⁺/H⁺ antiporter from Synechocystis species PCC 6803,homologous to SOS1,contains an aspartic residue and long C-terminal tail important for the carrier activity[J].Plant Physiology,2001a,125:437-446
[6] Laura Heggie and Marcel A.K.Jansen.Transgenic tobacco (Nicotiana tabacum L.cv.Samsun-NN) plants over-express-ing a synthetic HRP-C gene are altered in growth,develop-ment and susceptibility to abiotic stress[J].Plant Physiology and Biochemistry,2005,(43),1067-1078
[7] Takayuki Tobge,Yasutaka Nishiyama.Functional genomics by integrated analysis of metabolome and transcriptome of Arabi-dopsis plants over-expressing an MYB transcription factor[J].The Plant Journal.2005,(42),218-235
[8] 萨姆布鲁克J.,拉塞尔D.W.分子克隆实验指南[M].北京:科学出版社,2002
[9] Sambrook J.Molecular Cloning,A Laboratory Manual,2nd ed[M].New York:Cold Spring Harbor Laboratory Press,1989.475-490
[10] Gallego P,Hita O,Villalobos N,et al.Somatic embryogene-sis and plant regeneration in Medicago arborea L.[J].Vitro Cell.Dev.Biol.Plant,2001,37(2):199-203
[11] Cuadrado Y,Guerra H,Martin A B,et al.Differences in in-vertase activityin embryogenic and non-embryogenic calli from Medicagoarborea[J].Plant Cell Tiss.Org.Cult.,2001,67:145-151
[12] Iantcheva A,Slavov S,Prinsen E,et al.Embryo induction and regenerationfrom root explants of Medicago truncatula af-ter osmoticpre-treatment[J].Plant Cell Tiss.Org.Cult.,2005,81,37-43
[13] 金淑梅,管清杰,罗秋香,等.苜蓿愈伤组织高频再生遗传和转化体系的建立[J].分子植物育种,2006,4(4):571-578
[14] 陈晨,曹致中,贺顺姬,等.农杆菌介导的紫花苜蓿遗传转化体系的建立与优化[J].甘肃农业大学学报,2004,39(5):516-519
[15] 金芜军,郝旸,程红梅,等.用复合PCR方法检测6种转基因玉米外源DNA中的特异性[J].农业生物技术学报,2005,11(5):562-567
[16] 毛培胜,张涛,杨青川.紫花苜蓿品种鉴定的RAPD分子标记技术研究[J].草地学报,2007,15(2):124-128

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

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