联系我们 加入收藏
 
年期检索 高级检索
33
2025年4月3日 星期四
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
甘蔗尿苷二磷酸葡萄糖焦磷酸化酶基因(UGPase)转化拟南芥及转基因植株的生理特性分析
连玲1,叶冰莹2,6,陈如凯3,谢华安4,张建福5,陈由强2,6
1. 福建省农业科学院
2. 福建师范大学生命科学学院,福建 福州 350108
3. 福建农林大学甘蔗综合研究所
4. 福建省农业科学院国家水稻改良中心福州分中心
5. 农业部闽台农作物种质资源利用重点开放实验室;福州国家水稻改良分中心;福建省杂交水稻育种工程技术研究中心;福建省农业科学院水稻研究所
6. 福建师范大学生命科学学院,福建 福州 350108
Transformation of UDP-glucose Pyrophosphorylase Gene (UGPase) from Saccharum officinarum into Arabidopsis thaliana and Analysis of Physiological Characteristics of Transgenic Plants
全文: PDF (1131 KB)   HTML (1 KB) 
输出: BibTeX | EndNote (RIS)      
摘要 尿苷二磷酸葡萄糖焦磷酸化酶(UDP-glucose pyrophosphorylase, UGPase)是植物糖代谢的主要参与酶之一,在植物的生长发育过程中起着重要作用。本研究将甘蔗(Saccharum officinarum)UGPase基因cDNA片段连接至载体pBI121,通过BamHⅠ和SacⅠ酶切鉴定及测序验证,结果表明,植物表达载体成功构建;通过农杆菌(Agrobacterium tumefaciens)的介导,采用浸花法转化拟南芥(Arabidopsis thaliana)。结合卡那霉素抗性筛选和PCR检测,获得了5株T0代转基因植株。对T1代转基因植株进行PCR及Southern blot分析,结果表明,目的基因已成功转入拟南芥中,并且不同的转化植株含有目的基因的拷贝数不同。对T2代转基因植株进行PCR和RT-PCR检测,结果表明,目的基因不仅能在自交系后代中稳定遗传,而且在RNA水平也有表达。同时,对T2代转基因植株的可溶性总糖、蔗糖及淀粉含量进行测定,结果表明,与野生型相比,转基因植株中可溶性总糖含量没有明显的变化,但蔗糖含量有所提高,并且差异明显,比野生型植株提高了50.85%~96.99%,而淀粉含量都较野生型植株的低,降低了9.69%~36.76%。说明UGPase在蔗糖与淀粉的转换过程中起着较为重要的作用,其催化的反应方向影响着组织中这两种产物(蔗糖和淀粉)的分配。
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
连玲
叶冰莹
陈如凯
谢华安
张建福5
陈由强
关键词 UGPase载体构建转化糖含量测定    
Abstract:UGPase(UDP-glucose pyrophosphorylase) is one of the enzymes which mainly involves in plant carbohydrate metabolism, and plays an important role in the process of plant growth and development. The plant expression vector was constructed by ligating the cDNA fragment of UGPase from Saccharum Officinarum into expression vector pBI121, then the recombinant plasmid pBI-UGP was confirmed by restriction enzyme digestion analysis with BamHⅠ/SacⅠ, and further verified by DNA sequencing. The expression vector was introduced into Arabidopsis thaliana by floral-dip method, with Agrobacterium tumefaciens as media. Five T0 transgenic plants were selected by combining kanamycin screening with PCR detection. After selecting by kanamycin screening, T1 transgenic plants were identified by PCR and Southern blot, the results showed that the target gene was transformed into Arabidopsis thaliana successfully, and different transgenic plants contained various copy numbers of UGPase from Saccharum officinarum. T2 transgenic plants were identified by PCR and RT-PCR, the results indicated that the gene not only inherited stably in self-progeny, but also expressed at the RNA level. Meanwhile, content of soluble sugar, sucrose and starch were determinated in T2 transgenic plants and wild plants. Compared to wild plants, the soluble sugar content of transgenic plants had little change, however, sucrose content of transgenic plants improved obviously, increased by 50.85%~96.99%. Starch content of transgenic plants was lower than that of wild plants, decreased by 9.69%~36.76%. The research proved that UGPase plays a critical role in the process of converting sugar and starch, and the direction of its catalysis influneces distribution of these products in plant organization. It will lay foundation for further study of UGPase.
Key wordsUGPase    vector construction    transformation    sugar analysis
收稿日期: 2011-10-10     
通讯作者: 陈由强   
引用本文:   
连玲1,叶冰莹2,6,陈如凯3,谢华安4,张建福5,陈由强2,6. 甘蔗尿苷二磷酸葡萄糖焦磷酸化酶基因(UGPase)转化拟南芥及转基因植株的生理特性分析[J]. , 2012, 20(5): 481-488.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2012/V20/I5/481
 
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