Visualization of Gene Reciprocity among Lactic Acid Bacteria in Yogurt by RNase H-Assisted Rolling Circle Amplification-Fluorescence In Situ Hybridization

Microorganisms

Toshiro Kobori¹  Tsunehiro Aki²  Kyohei Horio²  Hirokazu Takahashi²  Yoshiko Okamura²  Kenshi Watanabe²  Yutaka Nakashimada² 

[1] Division of Food Biotechnology, Food Research Institute, National Agriculture and Food Research Organization, Tsukuba, Ibaraki 305-8642, Japan;Graduate School of Integrated Sciences for Life, Hiroshima University, Higashihiroshima 739-8530, Japan;

关键词: mRNA;    fluorescence in situ hybridization;    lactic acid bacteria;    single cell;    symbioses;   

DOI  :  10.3390/microorganisms9061208

来源: DOAJ

【 摘 要 】

Recently, we developed an in situ mRNA detection method termed RNase H-assisted rolling circle amplification-fluorescence in situ hybridization (RHa-RCA-FISH), which can detect even short mRNA in a bacterial cell. However, because this FISH method is sensitive to the sample condition, it is necessary to find a suitable cell permeabilization and collection protocol. Here, we demonstrate its further applicability for detecting intrinsic mRNA expression using lactic acid bacteria (LAB) as a model consortium. Our results show that this method can visualize functional gene expression in LAB cells and can be used for monitoring the temporal transition of gene expression. In addition, we also confirmed that data obtained from bulk analyses such as RNA-seq or microarray do not always correspond to gene expression in individual cells. RHa-RCA-FISH will be a powerful tool to compensate for insufficient data from metatranscriptome analyses while clarifying the carriers of function in microbial consortia. By extending this technique to capture spatiotemporal microbial gene expression at the single-cell level, it will be able to characterize microbial interactions in phytoplankton–bacteria interactions.

【 授权许可】

Unknown