遗传 ›› 2006, Vol. 28 ›› Issue (1): 85-91.

• 技术与方法 • 上一篇    下一篇

快速检测干旱和脱水可诱导植物启动子瞬间表达特性的方法

萧凤回1;2;段承俐2;翟虎渠1;3 ; 薛刚平4; 张红生1   

  1. (1. 南京农业大学作物遗传与种质创新国家重点实验室,南京 210095;2. 云南农业大学中药材研究所,云南省中药材规范化技术指导中心,云南省生物技术创新人才培养基地,云南 昆明650201;3. 中国农业科学院,北京 100094; 4. CSIRO Plant Industry, Qld. 4068, Australia)
  • 收稿日期:2005-02-21 修回日期:2005-05-18 出版日期:2006-01-10 发布日期:2006-01-10
  • 通讯作者: 翟虎渠

Rapid Determination of Transient Expression Characteristics of Drought- and Dehydration-Inducible Promoters from Plants

XIAO Feng-Hui1, 2, DUAN Cheng-Li 2 , ZHAI Hu-Qu1, 3, XUE Gang-Ping4 , ZHANG Hong-Sheng1   

  1. (1. National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China; 2. Chinese Medicinal Material Research Institute of Yunnan Agricultural University, Yunnan Provincial Center for Chinese Medicinal Material GAP Technology, Kunming 650201, China; 3. China Academy of Agricultural Science, Beijing 100094, China; 4. CSIRO Plant Industry, Qld 4068, Australia)
  • Received:2005-02-21 Revised:2005-05-18 Online:2006-01-10 Published:2006-01-10
  • Contact: ZHAI Hu-Qu

摘要: 选择合适的诱导表达启动子是开展植物耐干旱和脱水等非生物逆境转基因研究的重要环节。我们通过几年的研究,已建立了一套以大麦幼苗完整活体和植物离体叶片为主要材料通过瞬间表达鉴定来快速检测干旱和脱水可诱导基因启动子表达特性的方法。来自大麦和水稻的启动子Dhn4s、Dhn8s、HVA1s、Rab16Bj、wsi18j在大麦、小麦、水稻、高粱和蕨类植物的离体叶片中经干燥诱导可以瞬间表达GFP,在绿豆、番茄叶片中不表达。鉴定了HVA1s和wsi18j在大麦不同器官或组织中启动子的定性表达情况。进一步建立了GFP荧光点/GUS染色点计数分析和GUS活性/XYN活性测定分析的启动子表达的定量分析方法,并讨论该方法在环境可诱导植物启动子功能分析中的应用价值和前景。

关键词: 检测, 植物启动子, 干旱, 瞬间表达, 脱水

Abstract: The selection of suitable inducible-promoters is one of the most important chains in transgenic researches for plant tolerances to drought, dehydration and other abiotic stresses. Based on laboratory researches in the past years, it was set up a set of methods that could be used to rapidly determine the transient expression characteristics of drought- and dehydration-inducible promoters by using whole barley seedling and plant leaves in vitro as bombardment objects. Promoters Dhn4s, Dhn8s, HVA1s, Rab16Bj and wsi18j isolated from barley and rice were able to express GFP in the leaves of barley, wheat, rice, sorghum and fern after drying treatment, but not in laves of mungbean and tomato. The qualitative expression of HVA1s and wsi18j in different organs and tissues of barley was identified. A method for quantitative analysis of promoter transient expression was established by means of GFP foci/GUS foci counting or GUS activity/XYN activity test. Finally, the value and prospect of the methodology were discussed in terms of its application to the analysis of plant promoters inducible by environmental factors.

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