Author(s):

R Larinfeli, GT Behere, DM Firake, Bhagawati Sharma, Amrita Banerjee and T Rajesh

Abstract: Significant morphological similarities within or in between species of insects has made reliable taxonomic identification difficult. DNA barcoding has appeared to be a useful tool in resolving the issues related to the identification of taxonomically difficult insect species. The North eastern region of India is one of the mega biodiversity hotspots in the world and the climatic conditions of the region are highly conducive for reproduction and multiplication of insects. The efforts were undertaken to study the biodiversity of insects and to develop molecular database by developing DNA barcodes of natural enemies of cole crops insect pests in mid hills of Meghalaya. Limited information is available on natural enemies in cole crops ecosystem in mid –hills of Meghalaya. A total of 17 natural enemies belonging to three insect orders viz., Coleoptera (4) Predator, Hymenoptera (5) (1 nymphal/pupal; 3 Larval parasitoid; 1 Pupal parasitoid), Diptera (8) (5 Predator, 3 pupal parasitoid) Hemiptera (2) were documented during the year 2014-2015. Natural enemies especially Predators of aphids viz., Coccinellid beetles, Syrphid flies were recorded, Parasitoids observed were; nymphal/adult Parasitoid of aphids, Diaeretiella rapae, larval parasitoid of cabbage butterfly, Hyposoter sp and Cotesia glomerata and Pupal parasitoid including Brachymeria femorata and Tachinidae sp. The collected species were identified by established taxonomic keys, by taxonomists and/or molecular basis. DNA was successfully extracted from multiple specimens of 17 insect species and molecular assays were also undertaken for presence of Wolbachia infection. The Wolbachia infected specimens were discarded and not used for further analysis. The DNA barcodes were successfully developed for 17 species by sequencing partial Cytochrome oxidase I (COI) gene of mitochondrial DNA. The molecular identity of the insect species was established through BLAST-n at NCBI. The total nucleotide length of barcodes varied from species to species (501bp to 682bp). All the analysed sequences were submitted to National Centre for Biotechnology Information (NCBI) and Accession numbers were obtained (KT175578 to KT 175606). The comprehensive taxonomical and molecular database developed in this study for a total of 17 species observed in cole crop ecosystem could be used as diagnostic guide at both morphological and molecular level.

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