Biodiv Sci ›› 2022, Vol. 30 ›› Issue (12): 22266.  DOI: 10.17520/biods.2022266

Special Issue: 土壤生物与土壤健康

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Application of high-throughput sequencing technique in the study of nematode diversity

Yixin Sun1,2, Yingbin Li1, Yuhui Li1,2, Bing Li1,2, Xiaofang Du1,*(), Qi Li1,*()   

  1. 1. Soil Ecology and Agro-ecological Engineering Research Center, Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016
    2. University of Chinese Academy of Sciences, Beijing 100049
  • Received:2022-05-13 Accepted:2022-07-14 Online:2022-12-20 Published:2022-09-09
  • Contact: *E-mail:;


Background & Aims: Soil nematode diversity is one of the hot topic in soil ecology research. However, in-depth studies on soil nematode community composition and diversity are limited by the established taxonomy and methodology. At present, the development of molecular biological techniques has elevated our understanding of the diversity of soil nematodes, but it also included set-backs. This paper summarizes the research progress in molecular biology technology, including denaturing gradient gel electrophoresis (DGGE), terminal restriction fragment length polymorphism (T-RFLP), quantitative real-time PCR (qPCR) and high-throughput sequencing (HTS) technology, and the advantages and disadvantages of high-throughput sequencing in nematode research. We elaborate these techniques based on the extraction of soil nematode DNA, the selection of primers and reference databases, as well as the comparison between high-throughput sequencing and morphological identification analysis.
Prospects: The selections of soil nematode DNA extraction methods, in combination with proper primers and reference databases remain the key point to study nematode diversity. When focusing on nematode classification, we recommend DNA extraction after enrichment of nematodes suspension. Conclusively, researchers can then select the optimal combination to conduct experiments according to their research objectives.

Key words: molecular biological techniques, high-throughput sequencing, soil nematode diversity, nematode DNA extraction, morphological identification