联系我们 加入收藏
 
年期检索 高级检索
33
2025年4月5日 星期六
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
体外改组强耐盐基因NHXFS1在烟草中的表达及耐盐性分析
黄继斌1,张辉2,郭力榕2,徐源2,夏涛2
1. 华东师范大学
2. 华东师范大学生命科学学院
Expression and Salt Tolerance Analysis of a Novel Na+/H+ Antiporter Gene NHXFS1 in Tobacco (Nicotiana benthamiana L.)
全文: PDF (808 KB)   HTML (1 KB) 
输出: BibTeX | EndNote (RIS)      
摘要 位于植物液泡膜的Na+/H+逆向转运蛋白(NHX1)是将Na+区隔化到液泡中的主要蛋白,对植物耐盐性起着极其重要的作用。NHXFS1基因是以拟南芥(Arabidopsis thaliana)、水稻(Oryza sativa)和菊花(Flos chrysanthemi)的液泡膜Na+/H+逆向转运蛋白基因AtNHX1、OsNHX1和DmNHX1为模板,通过DNA家族改组技术,对其进行体外定向分子进化获得的一个功能显著增强的新基因。本研究利用根癌农杆菌(Agrobacterium tumerfaciens)介导的叶盘法将NHXFS1基因转入烟草 (Nicotiana benthamiana)中,通过潮霉素抗性筛选出再生植株,经PCR和Southern blot鉴定证实NHXFS1基因成功地整合到烟草基因组中。耐盐性测试结果显示,高盐胁迫条件下,萌发期和幼苗期的转基因烟草长势明显优于对照组,与对照组相比,转基因烟草叶片中积累了更多的脯氨酸和Na+。 RT-PCR和Real-time PCR结果表明,NHXFS1基因在烟草中正常表达,盐胁迫条件下NHXFS1基因在根、茎、叶中的表达量分别上调了1.01、0.60和1.79倍,跟对照组相比有明显提高(P<0.05)。研究结果提示,体外改组获得的新基因NHXFS1在植物耐盐基因工程和分子育种中具有较大的应用价值,同时也证明通过基因改组方法开发新基因用于改良植物耐盐性是可行的
服务
把本文推荐给朋友
加入我的书架
加入引用管理器
E-mail Alert
RSS
作者相关文章
黄继斌
张辉
郭力榕
徐源
夏涛
关键词 NHXFS1基因烟草Na+/H+逆向转运蛋白耐盐性DNA家族改组    
Abstract:Plant tonoplast Na+/H+ antiporter comparting sodium into vacuolar plays an important role in salinity tolerance. NHXFS1 is a novel tonoplast Na+/H+ antiporter gene which is obtained from AtNHX1 of Arabidopsis thaliana, OsNHX1 of Oryza sativa and DmNHX1 of Flos chrysanthemi by DNA family shuffling in vitro. To study the expression pattern and salt tolerance ability of NHXFS1 in plant, we transformed NHXFS1 into tobacco (Nicotiana benthamiana) by Agrobacterium-mediated leaf disc method, the transgenic plants were screened with hygromycin B and identified by PCR and Southern blot. 15 NHXFS1 positive plants were identified from 43 hygromycin-resistant tobacco plants that we had gotten. Southern blotting analysis with NHXFS1 probe indicated that the NHXFS1 gene was integrated into tobacco genome. According to segregate ratio, transgenic tobacco line with single copy was inbreeded to homozygote. The result of salinity tolerance test demonstrated that the growth vigor of transgenic plants under high salt stress was better than that of the control ones. The transgenic seeds could still germinate in MS plate with 400 mmol/L NaCl, but the controls could not. Also, the growth statuses of transgenic plants were better than that of controls in soil contained high NaCl concentration. When NaCl concentration reached 300 mmol/L, the wild-type plants were nearly wilting, while the transgenic ones were still vital. Results showed that the transgenic plants could accumulate more proline and Na+ ion. RT-PCR analysis showed that the NHXFS1 gene was expressed in tobacco. The result of Real-time PCR showed that the transcript level of NHXFS1 was significantly higher in different tissues after 200 mmol/L NaCl treatment, which increased 1.01, 0.60 and 1.79 fold in root, stem and leaf, respectively. The data suggest that this novel gene NHXFS1 can be applied in plant salt tolerance engineering and molecular breeding. And it also manifest that it's available to develop novel genes improving plant salt tolerance by DNA family shuffling method
Key wordsNHXFS1 gene    Tobacco    Na+/H+ antiporter    Salt tolerance    DNA family shuffling
收稿日期: 2012-01-06     
通讯作者: 夏涛   
引用本文:   
黄继斌1,张辉2,郭力榕2,徐源2,夏涛2. 体外改组强耐盐基因NHXFS1在烟草中的表达及耐盐性分析[J]. , 2012, 20(7): 774-781.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2012/V20/I7/774
 
版权所有 © 2014 《农业生物技术学报》编辑部   京ICP备11035905号-3
地址:北京市海淀区圆明园西路2号中国农业大学生命科学楼1053室 邮编:100193
电话:010-62733684 传真:010-62731615 E-mail: nsjxb@cau.edu.cn