联系我们 加入收藏
 
年期检索 高级检索
33
2025年8月19日 星期二
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
转Barnase基因无籽植物的研究
陈桂敏1,吴刚2,武玉花2,卢长明3
1. 淮阴师范学院
2.
3. 中国农业科学院油料作物研究所
Developing Transgenic Plants with No Seeds by Transforming Barnase Gene
全文: PDF (443 KB)   HTML (1 KB) 
输出: BibTeX | EndNote (RIS)      
摘要 种子败育的无籽瓜果具有较强的市场竞争力和可观的经济价值。来源于解淀粉芽胞杆菌(Bacillus Amyloliquefaciens)的Barnase基因编码12 kD的胞外小分子核糖核酸酶(RNAase)。在缺乏其天然抑制物Barstar的情况下,单独表达Barnase通常会造成宿主细胞的死亡。本研究用油菜(Brassica napus)种子特异启动子Napin调控Barnase基因,构建T-DNA表达载体,并用叶盘转化法转化烟草(Nicotiana benthamiana),获得55株转基因植株。随机抽取10株对其花器官进行形态学分析,并比较转基因植株和非转基因植株在不同发育时期花器官的形态学变化,发现转基因植株和非转基因植株在授粉前的花器官形态没有明显差异,但是受精以后转基因植株花瓣逐渐枯萎,不能正常发育形成种子,而非转基因植株能够正常形成种子。为了进一步了解转基因烟草种子败育的原因,将转基因植株和非转基因植株正反杂交,均不能正常结籽,表明转基因烟草的种子败育不是由自交不亲和性引起的。将转基因植株和非转基因植株花粉分别使用亚历山大染液染色,显微观察花粉的大小、形状和色泽,发现转基因花粉和非转基因花粉无明显差别,转基因植株和非转基因植株均能产生具有正常授粉活性的花粉。以上研究表明,使用种子特异性启动子驱动Barnase基因表达,外源基因不会通过花粉渗漏表达。本研究成功获得了Barnase基因在种子中特异表达的无籽转基因烟草,该研究为无籽植物的研制提供了一种全新的模式和思路。
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
陈桂敏
吴刚
武玉花
卢长明
关键词 Napin启动子Barnase基因转基因烟草无籽    
Abstract:Seedless fruit has strong market competitiveness and the considerable economic value, seedless trees can effectively reduce the pollution of the environment, seedless transgenic plants can help prevent foreign gene dispersal. Barnase gene comes from Bacillus amyloliquefaciens, encoding a 12 kD small molecule extracellular ribonuclease (RNAase). In the absence of its natural inhibitor Barstar, the expression of Barnase gene usually causes the death of its host cell. The T-DNA vector i. e. pBIloxP2 PNap TAibarnase carrying Barnase gene driven by Brassica napus seed-specific promoter Napin was constructed and transformed into tobacco(Nicotiana benthamiana) with the leaf disc transformation method, and 55 tobacco transformants were obtained. In which, 10 transformants were randomly selected for floral morphology analysis. Comparison of floral organs indicated that no obvious morphological difference was observed at different developmental stages between the transgenic and non-transgenic plants before pollination. Whereas, the floral organs of transgenic plants were gradually shrinking and dry, and failing to produce seeds after pollination; the petals of non-transgenic tobaccos were normally withered and can produce seeds within torus after pollination. In order to understand the mechanism of seed abortion of transgenic tobaccos, the cross between transgenic and non-transgenic plants were performed. However, no seed was obtained with the transgenic plants either as female parent or male parent. The hybrid results revealed that the seed abortion of transgenic tobacco was not caused by the self-incompatibility. To analyze the pollen activity, Alexander solution was used to stain the pollens of transgenic and non-transgenic tobaccos, then we observed the size, shape and color of pollens from transgenic and non-transgenic tobaccos with microscope, no difference was found. Therefore both the transgenic and non-transgenic tobaccos could generate active pollens. It is concluded that the seed specific promoter Napin used in this study only drives the Barnase gene expression in the seed, and no leakage expression takes place in pollens. We successfully obtain seedless transgenic tobaccos through expressing the Barnase gene in the seed tissue. The present study creates a novel model of producing seedless plants via genetic transformation, especially for vegetatively propagated crops.
Key wordsNapin promoter    Barnase gene    Transgenic tobacco    No seeds
收稿日期: 2011-10-31     
通讯作者: 卢长明   
引用本文:   
陈桂敏1,吴刚2,武玉花2,卢长明3. 转Barnase基因无籽植物的研究[J]. , 2012, 20(2): 113-120.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2012/V20/I2/113
 
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