Agrobacterium-mediated transient expression in plant organs is a quick and reliable method for studying gene functions. Due to the significance of transient transformation, substantial efforts have been dedicated to developing such protocols in various plants including the model Arabidopsis thaliana. Despite the importance, a reliable protocol is still lacking in Brassicaceae due to their recalcitrance towards Agrobacterium-mediated transient transformation. We have developed protocols for transient expression in Brassica juncea (PI 211000) and tested three other Brassica sp. for the suitability of the protocol. Co-infiltration of a bacteria-derived avirulence protein AvrPto1 significantly improved expression in B. juncea cotyledonary leaves. The protocol was used successfully in studying protein localization, protein–protein interaction by co-immunoprecipitation assay and transient silencing in B. juncea indicating it to be an excellent model system for transient expression. The efficiency of the protocol varied between Brassica sp. and depended highly on the Agrobacterium strain used. The protocol would be useful in designing functional analyses of genes using transient expression in Brassicaceae, including Arabidopsis and enable inclusion of mutant lines for such studies.

中文翻译：

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农杆菌介导的瞬时表达对芥菜基因功能研究的规程：在其他芸苔科中的适用性

农杆菌介导的植物器官瞬时表达是研究基因功能的一种快速可靠的方法。由于瞬时转化的重要性，已经在包括模型拟南芥（Arabidopsis thaliana）模型在内的各种植物中致力于开发这样的方案。尽管很重要，但是由于十字花科对农杆菌介导的瞬时转化不服从，因此在十字花科中仍然缺乏可靠的方法。我们已经开发了在芥菜型油菜中瞬时表达的方案（PI 211000），并测试了其他三个甘蓝型油菜。协议的适用性。细菌衍生的无毒蛋白AvrPto1的共浸润显着改善了其表达芥菜子叶。该协议已成功用于研究蛋白定位，蛋白-蛋白相互作用（通过共免疫沉淀测定法）和芥菜中的瞬时沉默，表明它是瞬时表达的出色模型系统。该协议的效率在甘蓝型油菜之间有所不同。并且高度依赖于所用农杆菌菌株。该协议可用于在十字花科（包括拟南芥）中使用瞬时表达设计基因的功能分析，并使此类研究包含突变株系。