Leaf-disc assay based on transient over-expression in Nicotiana benthamiana to allow functional screening of candidate effectors from aphids.
Leaf-disc assay based on transient over-expression in Nicotiana benthamiana to allow functional screening of candidate effectors from aphids.
复制标题
基于本塞姆氏烟草中瞬时过度表达的叶盘测定，可对蚜虫候选效应子进行功能筛选。
DOI：
10.1007/978-1-62703-986-4_11
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Rodriguez PA
中科院分区：
文献类型：
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作者：
Rodriguez PA
像植物病原体一样，蚜虫与它们的寄主形成了密切的联系。在探测和取食时，这些昆虫在宿主内部传递效应器，被认为参与抑制宿主防御和/或营养物质的释放。随着蚜虫基因组和转录组测序数据的日益可用，现在可以使用基于生物信息学和蛋白质组学的方法来识别效应器。下一步是应用与植物-蚜虫相互作用相关的功能分析来确定效应器的活动。本章描述了一种有效的、中等吞吐量的筛选方法，用于鉴定影响植物过度表达中的蚜虫繁殖力的效应器。该检测方法可用于鉴定在棉铃虫毒力中起作用的蚜虫效应因子，并可适用于其他易受农业渗透的植物以及其他基于瞬时表达的检测方法，如RNAi。
Aphids, like plant pathogens, are known to form close associations with their host. While probing and feeding, these insects deliver effectors inside the host, which are thought to be involved in suppression of host defenses and/or the release of nutrients. With increasing availability of aphid genome and transcriptome sequencing data, effectors can now be identified using bioinformatics- and proteomics-based approaches. The next step is then to apply functional assays relevant to plant–aphid interactions to identify effector activities. This chapter describes an effective and medium-throughput screen for the identification of effectors that affect aphid fecundity upon in planta over-expression. This assay will allow the identification of aphid effectors with a role in aphid virulence and can be adapted to other plant species amenable to agroinfiltration as well as to other assays based on transient expression, such as RNAi.