National Centre for Microbial Resource, National Centre for Cell Sciencegrid.419235.8, Pune, Maharashtra, India.
Environmental Biotechnology and Genomics Division, CSIR-National Environmental Engineering Research Institute, Nagpur, Maharashtra, India.
Microbiol Spectr. 2022 Jun 29;10(3):e0000722. doi: 10.1128/spectrum.00007-22. Epub 2022 Apr 25.
Culture-independent sequence data from various environmental samples have revealed an immense microbial diversity of environmental, clinical, and industrial importance that has not yet been cultured. Cultivation is imperative to validate findings emerging from cultivation-independent molecular data and exploit the isolated organisms for biotechnological purposes. Efforts have been made to boost the cultivability of microbes from environmental samples by use of a range of techniques and instrumentation. The manuscript presents a novel yet simple and innovative approach to improving the cultivability of natural microorganisms without sophisticated instrumentation. By employing gradient centrifugation combined with serial dilution ("two-dimensional cell separation"), significantly higher numbers of genera (>2-fold higher) and species (>3-fold higher) were isolated from environmental samples, including soil, anaerobic sludge, and landfill leachate, than from using serial dilution alone. This simple and robust protocol can be modified for any environment and culture medium and provides access to untapped microbial diversity. In the manuscript, we have developed a novel yet simple and innovative approach to improving the cultivability of natural microorganisms without sophisticated instrumentation. The method used gradient centrifugation combined with serial dilution (two-dimensional cell separation) to improve taxum recovery from samples. This simple and robust protocol can be modified for any environment and culture medium and provides access to untapped microbial diversity. This approach can be incorporated with less labor and complexity in laboratories with minimal instrumentation. As cultivation is a workflow that is well suited to lower-resource microbiology labs, we believe improvements in cultivability can increase opportunities for scientific collaborations between low-resource labs and groups focused on high-resource cultivation-independent methodologies.
从各种环境样本中获得的非培养序列数据揭示了大量具有环境、临床和工业重要性的微生物多样性，这些微生物尚未被培养。培养对于验证非培养分子数据得出的发现以及利用分离出的生物体进行生物技术应用是至关重要的。已经采取了各种技术和仪器来提高环境样本中微生物的可培养性。本文提出了一种新颖而简单的方法，无需使用复杂的仪器来提高天然微生物的可培养性。通过采用梯度离心与连续稀释（“二维细胞分离”）相结合的方法，从环境样本（包括土壤、厌氧污泥和垃圾渗滤液）中分离出的属数量（增加了 2 倍以上）和种数量（增加了 3 倍以上）明显高于单独使用连续稀释的方法。这种简单而强大的方案可以根据任何环境和培养基进行修改，从而可以利用未开发的微生物多样性。在本文中，我们开发了一种新颖而简单的方法，无需使用复杂的仪器即可提高天然微生物的可培养性。该方法使用梯度离心与连续稀释（二维细胞分离）相结合，从样品中提高了分类群的回收。这种简单而强大的方案可以根据任何环境和培养基进行修改，从而可以利用未开发的微生物多样性。这种方法可以在仪器设备较少的实验室中以较少的劳动力和复杂性进行整合。由于培养是一种非常适合低资源微生物学实验室的工作流程，我们相信提高可培养性可以为低资源实验室和专注于高资源非培养方法的小组之间的科学合作提供更多机会。
