Biofilm spatiotemporal mapping project

PI: Martin Robert. Laboratory of Microbial Systems Biology, Graduate School of Pharmaceutical Sciences, Kyoto University

Term: April 2021- March 2024 補助事業期間 令和3年度~令和5年度

Summary

Background: Large aggregates of bacteria growing on solid media or at liquid-air interfaces can form biofilms and macro-colonies with unique biological properties. These structures are important in bacterial resistance to drugs and represent a simple but powerful model to study the evolution of multicellularity, but little is known about functional differentiation in these otherwise free-living organisms.

Issues: There is therefore an important need to characterize the differentiation process at the molecular level.

Task and methods: To address this, we will quantify gene expression and metabolic activity in E. coli biofilms using time-lapse fluorescence imaging of reporter gene expression, and direct spatio-temporal mapping of the cellular proteome using liquid chromatography-mass spectrometry. We will focus on specific environmental conditions and gene deletion mutants that affect biofilm properties such as motility, curli protein production and signaling nucleotide.

Impact: The results are expected to reveal molecular mechanisms leading to complex patterns of biofilm development in bacteria, together with fundamental insight into the origin of multicellularity.

Outcomes 

TBA

Event (2024)

To commemorate the closing of this project Martin Robert is organizing a symposium on microbial multicellularity at the 47th annual meeting of the Molecular Biology Society of Japan (November 27-29, 2024).

https://sites.google.com/kyoto-u.ac.jp/msb/symposium-on-microbial-multicellularity-2024?authuser=0

Presentations

2023

1. Fujino. Y and Robert, M. Exploring culture conditions and fractionation methods for spatio-temporal analysis of E. coli biofilms. The 46th Annual Meeting of the Molecular Biology Society of Japan, Kobe, Japan, December 6-8, 2023

2. Zhao, Y and Robert, M. Investigating the role of YccA on E. coli biofilm growth and development. The 46th Annual Meeting of the Molecular Biology Society of Japan, Kobe, Japan, December 6-8, 2023

3. Sakai. R, Zhao, Y, and Robert, M. Development of imaging system to track biofilm morphology and temporal gene expression in growing biofilms. The 46th Annual Meeting of the Molecular Biology Society of Japan, Kobe, Japan, December 6-8, 2023

4. Robert, M. Biofilming beyond B. subtilis. BACELL 2023, Kobe, Japan, November 20-23, 2023

5. Robert M. E. coli biofilms as a developmental model, 第19回 21世紀大腸菌研究会Tsuruoka, Yamagata, June 29-30, 2023

6. 干場 悠介, 藤野 祐紀子, Zhao Yifan, 酒井 隆之介, Robert M.. Effects of glucose on the growth of E. coli biofilms in rich medium, 第19回 21世紀大腸菌研究会, Tsuruoka, Yamagata, June 29-30, 2023

7.     Zhao Yifan, Sakai R., Fujino Y., Hoshiba Y., Robert M. Investigating the role of YccA on E. coli biofilm growth and development, 第19回 21世紀大腸菌研究会 Tsuruoka, Yamagata, June 29-30, 2023

8. 酒井 隆之介, Zhao Yifan, Robert M.明視野および蛍光モードでバイオフィルムの動態を捉える   タイムラプスイメージングシステムの開発, 第19回 21世紀大腸菌研究会Tsuruoka, Yamagata, June 29-30, 2023

9.     Robert M. Exploring patterns of growth and differentiation during E. coli biofilm formation. 日本農芸化学会関西支部　第525回講演会, Kyoto Prefectural University, Kyoto, May 27, 2023

2022

1.     Fujino Y. Sakai, R. and Robert M., Exploring transitions in metabolic state during the development of bacterial biofilms. RIKEN BDR Symposium 2023 “Transitions in Biological Systems”, Kobe, Japan, March 7-9, 2023

2. Robert M. Opening and closing the box of bacterial metabolic function: from enzyme discovery to adaptive evolution and biofilms using E. coli. Annual Meeting of Japan Society for Environmental Biotechnology. University of Tokyo, November 22, 2022

3.     Fujino Y. and Robert M. Exploring developmental processes in growing E. coli biofilms. 16th Society of Genome Microbiology, Japan (online), March 2-4, 2022

Publications (in preparation)

Keywords

E. coli, biofilm, imaging, gene expression, proteome analysis, metaboli