构建矩阵式高通量扩散室和培养方法分离培养深海沉积物中小单孢菌群

The microbe species known to be cultured occupied merely 0.01-0.1% of marine microbes, and about 1% of terrestrial microbes. However, these uncultured microorganisms displayed tremendous biodiversity. The abundant natural microbes are valuable resource of drug discovery, functional microbe mining, and solving sustainable development problems. Thus, it is an emergency to conduct innovation researches on culture technology..In recent years, a large number of uncultured novel microbial species were available in lab culture conditions, due to the application of some innovative isolation & cultivation methods, such as extinction culturing, diffusion growth chamber & trap culturing, and co-cultivation method and so forth. In brief, the development of effective methods, technologies, and devices plays a crucial role for culturing the uncultured microorganisms..Recently, dozens of novel microbe species displaying multiple bioactivities from the deep sea sediments have been isolated in our group. Meanwhile, our metagenomic research confirmed there is a wealth of microbes were still unisolated, thereinto, especially the abundant Micromonospora. The Micromonospora can produce various uniquely bioactive substances of great development value. Therefore, basing on the in situ cultivation and diffusion growth chamber culturing methods, this project plans to develop a high-throughput method with diffusion chamber array for isolation, cultivation and characterization of the uncultured microorganisms from marine sediments, and then study their physiological mechanism. Moreover, the isolation of Micromonospora will be particularly paid attention to.

自然环境中的微生物，是药物发现、功能微生物挖掘和解决重大可持续发展问题的资源宝库。已知可培养的微生物仅是海洋微生物的0.01-0.1%，陆地微生物的1%左右，本研究组近年来从深海沉积物中分离到了大量放线菌资源，其中获得了数十株具多种生物活性的新种，但研究发现，这仅是极少部分，其中仍有大量微生物未能获得分离和培养，特别是其中丰富的小单孢菌群。小单孢菌能产生多种独特的生物活性物质，具极大的开发价值。因而开展培养技术的创新研究成为当务之急。本项目希望利用原位培养法和扩散室技术对这些来自深海样品再次分离，并设计一种矩阵式扩散室，建立一种高通量原位培养新方法和新装置，以提高分离效率，获得更加多样性的未培养的微生物新种和新化合物，为药物发现、难培养微生物分离、合成生物学元件筛选等研究提供一种技术手段，并研究这些难培养微生物与环境、生物、化学分子之间的生态生理机制。

自然环境中的微生物，是药物发现、功能微生物挖掘和解决重大可持续发展问题的资源宝库。已知可培养的微生物仅是海洋微生物的0.01-0.1%，陆地微生物的1%左右，因此，研究微生物分离培养新技术新方法成为开发自然界中微生物资源的先决条件。本项目通过建立一套分离培养微生物的新装置和研究配套的分离培养新方法，形成一套分离环境中微生物的培养新技术。最终研制出一套矩阵式高通量扩散室装置和模拟自然环境条件的分离培养微生物方法。这套新技术具有高通量、微型化特点，很容易与市场上成熟的专门处理微孔板的自动化设备兼容。基于这套装置和方法，分离庐山土样和黄海海泥时，细胞回收率比常规方法提高了数百倍，并观察到一些特别的规律和现象，如：（1）培养分离出的微生物菌落微小，即形成微菌落；（2）微菌落形态有丰富的多样性，许多形态是常规分离方法中看不到的。因此，这种微生物分离培养新方法新装置显示了分离难培养微生物的潜在价值。