Journal of Environmental Biology

Identification of selected earthworm species of Northeast India using DNA barcoding

S.D. Hijam¹, R.S. Thounaojam², M.C. Kalita³ and N.C. Talukdar^1,4*

¹Institute of Bioresources and Sustainable Development, Takyelpat Institutional Area, Imphal-795 001, India

²Department of Life Sciences (Zoology), Manipur University, Imphal-795 003, India

³Department of Biotechnology, Gauhati University, Guwahati-781 014, India

⁴Faculty of Science, Assam Down Town University, Panikhaiti-781 026, India

*Corresponding Author Email : talukdar_narayan@rediffmail.com

Received: 12.05.2021                                                                                                   Revised: 17.01.2022                                                                                      Accepted: 22.02.2022

Abstract

Aim: This study aimed at the species identification of selected indigenous earthworms of Manipur and Assam, Northeast India along with an exotic species using morpho-anatomical study and DNA barcoding.

Methodology: Indigenous species of earthworms were collected from Imphal and Jorhat, North-eastern part of India. The exotic species of earthworm were collected from Indian Council of Agricultural Research Complex, Manipur. The samples were collected by digging and hand sorting method. Identification of samples was done by both conventional and molecular methods. Molecular characterization was accomplished through PCR amplification of the mitochondrial cytochrome oxidase I (COI) genes. Automatic sequencing reactions were performed for the amplified PCR products on ABI3100 Genetic Analyser (Applied Biosystems).

Results: Out of five specimens (EM1, EM2, EM4, EG5 and EM6) examined through morpho-anatomical studies, three were identified to species level while the other two were identified to their genus level only. Out of EM1 and EM2 specimens in the genus Perionyx as per the morpho-anatomical studies, DNA barcoding could deduce the EM2 specimen up to the species level as P. excavatus. The exotic EM6 specimen morphologically identified as Eisenia fetida showed 99% COI gene sequence similarity with both E. fetida and E. andrei but its sequence divergence with E. andrei was less than 1%, so, it belonged to E. andrei.

Interpretation: This study shows the reliability of clubbing DNA barcoding experiments with classical taxonomy in supplementing and strengthening the traditional taxonomy for accurate identification of earthworms.

Key words: Earthworms, Mitochondrial cytochrome oxidase I gene, Morpho-anatomical studies, Species identification

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