Establishment of cell-specifically inducible expression systems in transgenic barley and maize

转基因大麦和玉米细胞特异性诱导表达系统的建立

• 批准号: 19985040
• 负责人: Dr. Jochen Kumlehn
• 金额: --
• 依托单位: Leibniz-Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK)
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2006
• 资助国家: 德国
• 起止时间: 2005-12-31 至 2009-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/19985040?language=en
• 关键词: Establishment; cell; specifically; inducible; expression

Experimental approaches to over-expression or knock-down of genes implicated in plant pathosystems using constitutive or tissue-specific promoters often result in pleiotrophic effects such as developmental aberrations or reduced viability. Hence, the immediate gene function confined to the context of plant-pathogen interaction cannot be studied properly using these conventional expression systems in transgenic plants. Thus, this project aims at establishing gene expression systems in barley and maize based on the ethanol-inducible AlcRlalcA promoter system derived from Aspergillus nidulans. This two-component system provides control of transgene expression at three levels: (1) the transactivator gene AlcR can be driven by an appropriate cell-specific promoter, (2) the inducing agent can be applied at any developmental stage of choice, and (3) locally confined to only a part of the plant investigated. Two candidate promoters specific for highly relevant tissues in plant-pathogen interactions will be functionally verified first by Agrobacterium-mediated transformation of barley using conventional reporter gene constructs. These promoters will then be used in conjunction with the transactivator AlcR to develop cell-specifically inducible expression systems. In parallel we will establish genetic transformation of maize via infection of either immature embryos or androgenetic pollen cultures with Agrobacteria. Eventually, it is envisaged to provide a proof of concept by transformation of barley and maize using the cell-specifically inducible expression systems with the alcA promoter driving the GFP gene. In various follow-up approaches, the established systems will be highly valuable for functional characterisation of genes putatively implicated in the compatible interaction of barley or maize with various pathogenic or mutualistic fungi.

使用组成型或组织特异性启动子过量表达或敲低植物病理系统中涉及的基因的实验方法通常导致多效性效应，例如发育畸变或活力降低。因此，直接的基因功能局限于植物-病原体相互作用的背景下，不能正确地使用这些常规的表达系统在转基因植物中进行研究。因此，本项目旨在建立大麦和玉米基因表达系统的基础上，乙醇诱导AlcRlalcA启动子系统来自构巢曲霉。该双组分系统在三个水平上提供转基因表达的控制：（1）反式激活因子基因AlcR可以由适当的细胞特异性启动子驱动，（2）诱导剂可以在选择的任何发育阶段施用，和（3）局部地仅限于所研究的植物的一部分。两个候选启动子特异性的高度相关的组织在植物病原体的相互作用将功能验证首先通过农杆菌介导的转化大麦使用常规的报告基因构建体。然后将这些启动子与反式激活因子AlcR结合使用，以开发细胞特异性诱导型表达系统。同时，我们将通过用农杆菌感染未成熟胚或雄核发育花粉培养物来建立玉米的遗传转化。最后，设想通过使用具有驱动GFP基因的alcA启动子的细胞特异性诱导型表达系统转化大麦和玉米来提供概念验证。在各种后续方法中，所建立的系统将是非常有价值的功能特性的基因pupilli牵连在大麦或玉米与各种致病性或互利共生真菌的相容性相互作用。