Chaudhary Dhiraj Kumar, Kim Jaisoo
Ecology Laboratory, Department of Life Science, Kyonggi University, Suwon, South Korea.
Bio Protoc. 2019 Oct 5;9(19):e3388. doi: 10.21769/BioProtoc.3388.
Unculturable bacteria are those bacteria which proliferate in their native habitat but unable to grow or thrive in the normal laboratory media and conditions. The molecular techniques have revealed the significance of these uncultured bacteria in terms of their functional diversity and potential to produce secondary metabolites. To achieve these benefits, scientists have attempted to isolate and cultivate unculturable bacteria in the laboratory using transwell plates, optical tweezers, laser microdissection, microbioreactors, and diffusions bioreactors. However, these techniques are still inadequate to resolve the difficulties of cultivating unculturable bacteria. Therefore, it is essential to develop new cultivation method that enables growth of diverse range of bacteria in the laboratory conditions. Diffusion bioreactor is a membrane bound chamber which allows microbes to proliferate in their native environment by providing the excess to naturally occurring nutrients and signaling compounds. This paper presents efficient and reliable protocol to construct a diffusion bioreactor and its utilization to isolate and cultivate unculturable soil bacteria in laboratory.
不可培养细菌是指那些在其原生栖息地中能够增殖，但在正常实验室培养基和条件下无法生长或繁衍的细菌。分子技术已经揭示了这些未培养细菌在功能多样性和产生次生代谢产物潜力方面的重要性。为了获得这些益处，科学家们尝试在实验室中使用Transwell板、光镊、激光显微切割、微生物反应器和扩散生物反应器来分离和培养不可培养细菌。然而，这些技术仍不足以解决培养不可培养细菌的难题。因此，开发一种新的培养方法以使各种细菌能够在实验室条件下生长至关重要。扩散生物反应器是一种膜结合腔室，它通过提供过量的天然存在的营养物质和信号化合物，使微生物能够在其原生环境中增殖。本文介绍了构建扩散生物反应器的高效可靠方案及其在实验室中分离和培养不可培养土壤细菌的应用。
