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Combined selective gamma irradiation and pulverized soil inoculation for ecologically relevant soil microfauna studies
Applied Soil Ecology ( IF 4.8 ) Pub Date : 2021-09-15 , DOI: 10.1016/j.apsoil.2021.104223
Junwei Hu 1 , Mesfin Tsegaye Gebremikael 1 , Bjorn Tytgat 2 , Kenneth Dumack 3 , Ummehani Hassi 1 , Pezhman Salehi Hosseini 1 , Steven Sleutel 1 , Elie Verleyen 2 , Stefaan De Neve 1
Affiliation  

The separate and collective role of soil nematodes and protists at community level in soil biogeochemical processes remains poorly quantified due to the lack of appropriate methodologies to study them independently from other soil biota under conditions emulating natural soil. Here we present a methodological exploration for the selective removal of nematodes and protists using gamma irradiation and the subsequent reinoculation with metazoan-free soil powder inoculum that allows reconstructing a complex microbial food web comprising the native microflora and soil protist communities, without other fauna. Nematodes, and culturable flagellates and ciliates were removed from 6 kGy onwards, while a small fraction of the amoebae survived (8% of the entire protist population) at 16 kGy but were totally removed at 27 kGy. Reinoculation with pulverized soil powder successfully established a protist community of similar size and composition as the control soil as assessed by both amplicon sequencing-based detection and microscopic observation after cultivation. A viable native microflora community remained from 6 to 16 kGy doses, but microbial activity was completely inhibited at 27 kGy. We suggest that, in ecological relevant experiments where selective removal of nematodes or other fauna is the purpose and protist community is meant to be kept intact, gamma irradiation needs to be followed routinely by soil powder inoculation in order to restore the suppressed protist community. This experiment also opens up new perspectives for studying the ecological roles of the entire protist community or specific groups thereof (flagellates and ciliates) in real soil and in the presence of a viable native microflora community.



中文翻译:

用于生态相关土壤微型动物研究的选择性伽马辐射和粉状土壤接种相结合

由于缺乏适当的方法来在模拟天然土壤的条件下独立于其他土壤生物群对它们进行研究,因此土壤线虫和原生生物在土壤生物地球化学过程中在社区水平上的单独和集体作用仍然很难量化。在这里,我们提出了一种方法学探索,用于使用伽马辐射选择性去除线虫和原生生物,然后用无后生动物的土壤粉末接种物重新接种,从而重建复杂的微生物食物网,包括本地微生物群落和土壤原生生物群落,没有其他动物群。线虫、可培养鞭毛虫和纤毛虫从 6 kGy 开始被去除,而一小部分变形虫在 16 kGy 下存活(占整个原生生物种群的 8%),但在 27 kGy 下被完全去除。通过基于扩增子测序的检测和培养后的显微观察评估,用粉碎的土壤粉末重新接种成功地建立了与对照土壤大小和组成相似的原生生物群落。在 6 到 16 kGy 剂量下,一个可行的本地微生物群落保持不变,但微生物活动在 27 kGy 时被完全抑制。我们建议,在以选择性去除线虫或其他动物群为目的且原生生物群落保持完整的生态相关实验中,伽马辐射需要定期进行土壤粉末接种,以恢复被抑制的原生生物群落。

更新日期:2021-09-15
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