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Pathogen Phytosensing: Plants to Report Plant Pathogens

机译:病原体植物感测:植物报告植物病原体

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摘要

Real-time systems that provide evidence of pathogen contamination in crops can be an important new line of early defense in agricultural centers. Plants possess defense mechanisms to protect against pathogen attack. Inducible plant defense is controlled by signal transduction pathways, inducible promoters and cis-regulatory elements corresponding to key genes involved in defense, and pathogen-specific responses. Identified inducible promoters and cis-acting elements could be utilized in plant sentinels, or ‘phytosensors’, by fusing these to reporter genes to produce plants with altered phenotypes in response to the presence of pathogens. Here, we have employed cis-acting elements from promoter regions of pathogen inducible genes as well as those responsive to the plant defense signal molecules salicylic acid, jasmonic acid, and ethylene. Synthetic promoters were constructed by combining various regulatory elements supplemented with the enhancer elements from the Cauliflower mosaic virus (CaMV) 35S promoter to increase basal level of the GUS expression. The inducibility of each synthetic promoter was first assessed in transient expression assays using Arabidopsis thaliana protoplasts and then examined for efficacy in stably transgenic Arabidopsis and tobacco plants. Histochemical and fluorometric GUS expression analyses showed that both transgenic Arabidopsis and tobacco plants responded to elicitor and phytohormone treatments with increased GUS expression when compared to untreated plants. Pathogen-inducible phytosensor studies were initiated by analyzing the sensitivity of the synthetic promoters against virus infection. Transgenic tobacco plants infected with Alfalfa mosaic virus showed an increase in GUS expression when compared to mock-inoculated control plants, whereas Tobacco mosaic virus infection caused no changes in GUS expression. Further research, using these transgenic plants against a range of different pathogens with the regulation of detectable reporter gene could provide biological evidence to define the functional differences between pathogens, and provide new technology and applications for transgenic plants as phytosensors.
机译:提供农作物病原体污染证据的实时系统可能是农业中心早期防御的重要新路线。植物具有防御机制来防御病原体的侵袭。可诱导的植物防御受信号转导途径,与防御相关的关键基因相对应的诱导型启动子和顺式调控元件以及病原体特异性反应的控制。鉴定出的诱导型启动子和顺式作用元件可用于植物前哨或“植物传感器”,方法是将这些融合到报告基因上,以响应病原体的存在而产生具有改变的表型的植物。在这里,我们采用了来自病原体诱导基因启动子区域的顺式作用元件,以及对植物防御信号分子水杨酸,茉莉酸和乙烯有反应的基因。通过将各种调节元件与花椰菜花叶病毒(CaMV)35S启动子的增强子相结合来构建合成启动子,以增加GUS表达的基础水平。首先在拟南芥原生质体的瞬时表达测定中评估每个合成启动子的诱导能力,然后检查其在稳定转基因拟南芥和烟草植物中的功效。组织化学和荧光GUS表达分析表明,与未经处理的植物相比,转基因拟南芥和烟草植物对诱导剂和植物激素处理均具有增加的GUS表达。通过分析合成启动子对病毒感染的敏感性来启动病原体诱导的植物传感器研究。与模拟接种的对照植物相比,被苜蓿花叶病毒感染的转基因烟草植物显示GUS表达增加,而烟草花叶病毒感染未引起GUS表达改变。将这些转基因植物针对一系列不同病原体并通过可检测的报道基因进行调控,进一步的研究可为确定病原体之间的功能差异提供生物学依据,并为转基因植物作为植物传感器提供新的技术和应用。

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