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Measuring Bacterial Colonization on Arabidopsis thaliana Roots in Hydroponic Condition

文献类型： 外文期刊

作者： Shu, Xia 1 ; Li, Huatai 1 ; Wang, Jing 3 ; Wang, Shan 4 ; Liu, Yunpeng 1 ; Zhang, Ruifu 1 ;

作者机构： 1.Chinese Acad Agr Sci, Inst Agr Resources & Reg Planning, State Key Lab Efficient Utilizat Arid & Semiarid A, Beijing, Peoples R China

2.Huazhong Agr Univ, Coll Life Sci & Technol, State Key Lab Agr Microbiol, Wuhan, Peoples R China

3.Nanjing Agr Univ, Natl Engn Res Ctr Organ Based Fertilizers, Jiangsu Collaborat Innovat Ctr Solid Organ Waste R, Jiangsu Prov Key Lab Organ Solid Waste Utilizat, Nanjing, Peoples R China

4.Jiangsu Vocat Coll Agr & Forestry, Jiangsu Engn & Technol Ctr Modern Hort, Nanjing, Peoples R China

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

页码：

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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