Journal of Environmental Biology

Inter and intraspecific diversity of Chironomid larvae

using COI and RAPD markers

 Ramprasad Kuncham¹*, Tha.Thayumanavan² and Gopireddy V. Subba Reddy³

¹Department of Biotechnology, Jawaharlal Nehru Technological University Anantapur, Ananthapuramu-515 002, India

²School of Biotechnology, Dr. G.R. Damodaran College of Science, Coimbatore-641 014, India

³Department of Chemistry, JNTUACE, Pulivendula-516 390, India

*Corresponding Author E-mail: ramprasadbt@gmail.com

Publication Data

Paper received:

13 June 2016

Revised received:

21 July 2016

Re-revised received:

05 August 2016

Accepted:

05 August 2016

Abstract

Cytochrome Oxidase I (COI) and Random Amplified Polymorphic DNA (RAPD) markers have become widely applied tools for quick and accurate identification of species and to resolve intra and inter specific relationship in many insect groups. In the present study, these markers have been used to infer genetic relatedness of intra and inter specific populations collected randomly from different Indian geographic regions. Initially, mitochondrial DNA isolated using modified CTAB method and 480bp fragment of mitochondrial gene cytochrome oxidase subunit I (COI) has been amplified and sequenced. Sanger sequencing data of 19 species of Chironomidae including 20 populations of Chironomous circumdatus obtained and inter and intra specific phylogenetic relationship was investigated using MEGA 6 software. Interspecific divergence was found to be 0.578 as compared to intra specific divergence (0.042). Further, RAPD analysis of 20 population of Chironomous circumdatus was carried out using RAPD markers for reliable information about genetic diversity. Total DNA extracted and PCR amplified using 2 arbitrary decamer primers, which produced 180 reproducible and scorable bands ranged from 300bp to 1300bp. Dendrograms with genetic distance coefficient were performed with the procedure of? Nei and Li (Dice) using UVIsoft analysis software taking low range DNA ruler as molecular weight marker. Relationships inferred from RAPD UPGMA tree were partially similar to the results obtained from sequences of cytochrome oxidase I of mitochondrial DNA (COI).?????????

 Key words

Chironomid larvae, Cytochrome oxidase I, Genetic divergance, RAPD, Mitochondrial DNA

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