Ahmad Tariq, Ishaq Sidra Erum, Liang Lewen, Xie Ruize, Wang Yinzhao, Wang Fengping
State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240 China.
Key Laboratory of Polar Ecosystem and Climate Change, Ministry of Education, School of Oceanography, Shanghai Jiao Tong University, Shanghai, 200240 China.
Mar Life Sci Technol. 2024 Aug 12;7(3):466-477. doi: 10.1007/s42995-024-00240-2. eCollection 2025 Aug.
Traditional cultivation methods with defined growth media can only isolate and cultivate a small number of microbes. However, much higher microbial diversity has been detected by cultivation-independent tools from a range of natural ecosystems. These represent a large unexplored pool of potentially novel taxa. In this study, a diffusion-based integrative cultivation approach (DICA) was developed to efficiently isolate novel taxonomic candidates from marine sediment. DICA combined a newly designed diffusion-based apparatus called a "microbial aquarium" with modified low-nutrient media. To determine the efficiency of DICA, cultivation results were compared with traditional cultivation approach (TCA). Both cultivation approaches resulted in the isolation of numerous representatives from the phyla , , , and . However, the newly developed DICA also led to the successful cultivation of species from rarely cultivated phyla such as and Based on 16S rRNA analyses, the application of DICA resulted in the successful cultivation of 115 previously uncultured taxa out of a total of 196 isolates. Among these, 39 were identified at the genus level and 4 at the family level, showcasing a novelty ratio of 58%. Conversely, the TCA cultivated 12% (20/165) of novel isolates, with all at species level only. The isolated microbial diversity showed that species recovered by DICA belong to 12 different classes, twice the number produced by TCA. Overall, these results demonstrate that the newly designed DICA produces a high recovery of diverse and previously uncultured bacteria.
The online version contains supplementary material available at 10.1007/s42995-024-00240-2.
使用特定生长培养基的传统培养方法只能分离和培养少量微生物。然而，通过非培养工具在一系列自然生态系统中检测到了更高的微生物多样性。这些代表了大量未被探索的潜在新分类单元库。在本研究中，开发了一种基于扩散的综合培养方法（DICA），以有效地从海洋沉积物中分离新的分类候选物。DICA将一种新设计的基于扩散的装置“微生物水族箱”与改良的低营养培养基相结合。为了确定DICA的效率，将培养结果与传统培养方法（TCA）进行了比较。两种培养方法都从 、 、 和 门中分离出了大量代表菌株。然而，新开发的DICA还成功培养了来自 和 等很少培养的门的物种。基于16S rRNA分析，DICA的应用导致在总共196个分离物中成功培养了115个以前未培养的分类单元。其中，39个在属水平上被鉴定，4个在科水平上被鉴定，新颖率为58%。相反，TCA培养了12%（20/165）的新分离物，且均仅在物种水平。分离出的微生物多样性表明，DICA回收的物种属于12个不同的类，是TCA产生数量的两倍。总体而言，这些结果表明新设计的DICA能高效回收多样且以前未培养的细菌。
在线版本包含可在10.1007/s42995-024-00240-2获取的补充材料。
