Measuring Bacterial Colonization on Arabidopsis thaliana Roots in Hydroponic Condition

文献类型： 外文期刊

第一作者： Shu, Xia

作者： Shu, Xia;Li, Huatai;Liu, Yunpeng;Zhang, Ruifu;Shu, Xia;Wang, Jing;Zhang, Ruifu;Wang, Shan

作者机构：

期刊名称：JOVE-JOURNAL OF VISUALIZED EXPERIMENTS （ 影响因子：1.2； 五年影响因子：1.6 ）

ISSN： 1940-087X

年卷期： 2024 年 205 期

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收录情况： SCI

摘要： Measuring bacterial colonization on Arabidopsis thaliana root is one of the most frequent experiments in plant-microbe interaction studies. A standardized method for measuring bacterial colonization in the rhizosphere is necessary to improve reproducibility. We first cultured sterile A.thaliana in hydroponic conditions and then inoculated the bacterial cells in the rhizosphere at a final concentration of OD600 of 0.01. At 2 days post-inoculation, the root tissue was harvested and washed three times in sterile water to remove the uncolonized bacterial cells. The roots were then weighed, and the bacterial cells colonized on the root were collected by vortex. The cell suspension was diluted in a gradient with a phosphate-buffered saline (PBS) buffer, followed by plating onto a Luria-Bertani (LB) agar medium. The plates were incubated at 37 degrees C for 10 h, and then, the single colonies on LB plates were counted and normalized to indicate the bacterial cells colonized on roots. This method is used to detect bacterial colonization in the rhizosphere in mono-interaction conditions, with good reproducibility.

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