Plant Chemetics Lab, Department of Plant Sciences, University of Oxford, Oxford, OX1 3RB, UK.
Department of Plant Sciences, University of Oxford, Oxford, OX1 3RB, UK.
Plant J. 2021 Feb;105(3):831-840. doi: 10.1111/tpj.15056. Epub 2021 Jan 12.
The lengthy process to generate transformed plants is a limitation in current research on the interactions of the model plant pathogen Pseudomonas syringae with plant hosts. Here we present an easy method called agromonas, where we quantify P. syringae growth in agroinfiltrated leaves of Nicotiana benthamiana using a cocktail of antibiotics to select P. syringae on plates. As a proof of concept, we demonstrate that transient expression of PAMP receptors reduces bacterial growth, and that transient depletion of a host immune gene and transient expression of a type-III effector increase P. syringae growth in agromonas assays. We show that we can rapidly achieve structure-function analysis of immune components and test the function of immune hydrolases. The agromonas method is easy, fast and robust for routine disease assays with various Pseudomonas strains without transforming plants or bacteria. The agromonas assay offers a reliable approach for further comprehensive analysis of plant immunity.
生成转化植物的漫长过程是目前研究模式植物病原体丁香假单胞菌与植物宿主相互作用的一个限制因素。在这里，我们提出了一种简单的方法，称为 agroinfiltration，我们使用抗生素混合物在侵染的烟草原生质体叶片中定量检测丁香假单胞菌在平板上的生长。作为概念验证，我们证明了 PAMP 受体的瞬时表达减少了细菌的生长，而宿主免疫基因的瞬时耗竭和 III 型效应物的瞬时表达增加了 agroinfiltration 测定中丁香假单胞菌的生长。我们表明，我们可以快速实现免疫成分的结构-功能分析，并测试免疫水解酶的功能。该 agroinfiltration 方法简单、快速且稳健，可用于各种 Pseudomonas 菌株的常规疾病测定，而无需转化植物或细菌。该 agroinfiltration 测定为进一步全面分析植物免疫提供了一种可靠的方法。
