Evolutionary Ecology and Genetics, University of Kiel.
Evolutionary Ecology and Genetics, University of Kiel; Max-Planck-Institute for Evolutionary Biology;
J Vis Exp. 2022 Aug 17(186). doi: 10.3791/64249.
The nematode Caenorhabditis elegans interacts with a large diversity of microorganisms in nature. In general, C. elegans is commonly found in rotten plant matter, especially rotten fruits like apples or on compost heaps. It is also associated with certain invertebrate hosts such as slugs and woodlice. These habitats are rich in microbes, which serve as food for C. elegans and which can also persistently colonize the nematode gut. To date, the exact diversity and consistency of the native C. elegans microbiota across habitats and geographic locations is not fully understood. Here, we describe a suitable approach for isolating C. elegans from nature and characterizing the microbiota of worms. Nematodes can be easily isolated from compost material, rotting apples, slugs, or attracted by placing apples on compost heaps. The prime time for finding C. elegans in the Northern Hemisphere is from September until November. Worms can be washed out of collected substrate material by immersing the substrate in buffer solution, followed by the collection of nematodes and their transfer onto nematode growth medium or PCR buffer for subsequent analysis. We further illustrate how the samples can be used to isolate and purify the worm-associated microorganisms and to process worms for 16S ribosomal RNA analysis of microbiota community composition. Overall, the described methods may stimulate new research on the characterization of the C. elegans microbiota across habitats and geographic locations, thereby helping to obtain a comprehensive understanding of the diversity and stability of the nematode's microbiota as a basis for future functional research.
秀丽隐杆线虫与自然界中大量的微生物相互作用。一般来说，秀丽隐杆线虫常见于腐烂的植物物质中，特别是腐烂的水果，如苹果，或在堆肥堆中。它也与某些无脊椎动物宿主如鼻涕虫和木虱有关。这些栖息地富含微生物，它们是秀丽隐杆线虫的食物来源，也可以持续定植在线虫肠道中。迄今为止，对于不同栖息地和地理位置的秀丽隐杆线虫原生微生物组的的确切多样性和一致性还不完全清楚。在这里，我们描述了一种从自然界中分离秀丽隐杆线虫并对其肠道微生物群进行特征分析的合适方法。线虫可以很容易地从堆肥材料、腐烂的苹果、鼻涕虫或通过在堆肥堆上放置苹果来吸引它们中分离出来。在北半球，发现秀丽隐杆线虫的最佳时间是从 9 月到 11 月。可以通过将收集的基质材料浸泡在缓冲溶液中，将线虫从收集的基质材料中冲洗出来，然后收集线虫并将其转移到线虫生长培养基或 PCR 缓冲液中进行后续分析。我们进一步说明了如何使用这些样本分离和纯化与线虫相关的微生物，并对处理后的线虫进行 16S 核糖体 RNA 分析，以了解微生物群落组成。总的来说，所描述的方法可能会激发人们对不同栖息地和地理位置的秀丽隐杆线虫微生物组的特征分析的新研究，从而帮助全面了解线虫微生物组的多样性和稳定性，为未来的功能研究提供基础。
