Gaube Paul, Kaltenpoth Martin, Flórez Laura V
Insect Symbiosis Research Group, Max Planck Institute for Chemical Ecology, Hans-Knöll-Straße 8, Jena, Germany.
Molecular Biodiversity Group, Department of Animal Ecology and Tropical Biology (Zoology III), Theodor Boveri-Institute, Julius Maximilian University, Am Hubland, Würzburg, Germany.
Bio Protoc. 2017 Dec 20;7(24):e2663. doi: 10.21769/BioProtoc.2663.
To investigate the establishment and consequences of host-microbe interactions, it is important to develop controlled infection assays suitable for each system, as well as appropriate methods to evaluate successful infection and its associated effects. Here, we describe a procedure for bacterial inoculation of soybean plants, followed by the assessment of systemic infection and impact on plant fitness. Soybean () seedlings were mechanically wounded using a device that mimics insect herbivory and inoculated with known cell numbers of bacteria previously isolated from an insect host. The impact on the plants was evaluated by monitoring changes in height, time to flowering and chlorophyll content during plant development, and by quantifying seed production in comparison to plants inoculated with sterile water. The presence and proliferation of bacterial infection were examined in tissues from developed plants using quantitative PCR and fluorescence hybridization (FISH).
为了研究宿主与微生物相互作用的建立及其后果,开发适用于每个系统的可控感染试验以及评估成功感染及其相关影响的适当方法非常重要。在此,我们描述了一种对大豆植株进行细菌接种的程序,随后评估全身感染情况以及对植物适应性的影响。大豆()幼苗使用模拟昆虫食草行为的装置进行机械损伤,并用先前从昆虫宿主中分离出的已知细胞数量的细菌进行接种。通过监测植物发育过程中高度、开花时间和叶绿素含量的变化,并与接种无菌水的植物相比定量种子产量,来评估对植物的影响。使用定量PCR和荧光原位杂交(FISH)在发育成熟植株的组织中检测细菌感染的存在和增殖情况。