Yu Wenjia, Macho Alberto P
Shanghai Center for Plant Stress Biology, CAS Center for Excellence in Molecular Plant Sciences; Chinese Academy of Sciences, Shanghai 201602, China.
University of Chinese Academy of Sciences, Beijing, China.
Bio Protoc. 2021 Aug 5;11(15):e4116. doi: 10.21769/BioProtoc.4116.
is a devastating soil-borne bacterial pathogen that causes disease in multiple host plants worldwide. Typical assays to measure virulence of in laboratory conditions rely on soil-drenching inoculation followed by observation and scoring of disease symptoms. Here, we describe a novel inoculation protocol to analyze the replication of upon infiltration into the leaves of , in which gene expression has been altered using . The protocol includes five major steps: 1) growth of plants; 2) infiltration of ; 3) inoculation; 4) sample collection and bacterial quantitation; 5) data analysis and representation. The transient gene expression or gene silencing prior to inoculation provides a straightforward way to perform genetic analysis of plant functions involved in the interaction between pathogen and host, using the appropriate combination of and strains, with high sensitivity and accuracy provided by the quantitation of bacterial numbers in plant tissues.
是一种极具破坏力的土传细菌病原体，在全球多种寄主植物中引发病害。在实验室条件下测量其毒力的典型测定方法依赖于土壤浇灌接种，随后观察并对病害症状进行评分。在此，我们描述了一种新颖的接种方案，用于分析其渗入经[某种技术]改变了基因表达的[植物名称]叶片后的增殖情况。该方案包括五个主要步骤：1）[植物名称]植株的培养；2）[物质名称]的渗入；3）[病原体名称]接种；4）样本采集与细菌定量；5）数据分析与呈现。在[病原体名称]接种前进行的瞬时基因表达或基因沉默，提供了一种直接的方法，通过使用[病原体名称]和[植物名称]菌株的适当组合，利用植物组织中细菌数量的定量所提供的高灵敏度和准确性，对参与病原体与寄主相互作用的植物功能进行遗传分析。
