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Investigation of Lepidopteransin Bangladesh by DNA BARCoding of Malaise Trap Collection

Santosh Mazumdar*,
Department of Zoology, University of Chittagong, BANGLADESH.

Paul D. N. Hebert
Centre for Biodiversity Genomics, University of Guelph, 50 Stone Road East, Guelph, Ontario, CANADA

Badrul Amin Bhuiya
Biodiversity Research for Environment & Ecosystem Protection (BREEP), Chattogram-4325, BANGLADESH.

Abstract/ Executive Summary:

Lepidoptera forman essential part and one of the dominant groups of insects of natural terrestrial ecosystem, and they are widely distributed ranging from desert to rainforest, from lowland grasslands to mountain plateaus. The main aim of the present study is to identify the lepidopterans’ diversity, at a site in Bangladesh through DNA barcoding technique (658 bp sequence from the 5′-end of cytochromeoxidase I). Field collections were carried out by a Malaise trap at Chittagong university campus between April 2014 and March 2015. Including first country records of 52 species, 76 genera, 23 subfamilies and 19families, a total of 87 lepidopteran species, 115 genera, 61 subfamilies and 38 families were confirmed. Most new records belong to moth group and they are associated with agricultural fields. All the specimen records, with the Barcode Index Numbers (BINs) (the species proxies), are available on the Barcode of Life Data System (BOLD).

Keywords: Lepidoptera, Malaise trap, DNA barcode, Bangladesh

Location:

Start Date:

End Date:

Program Area: Pest Management

Commodity: Insects

Objectives:

The present work was carried out to reveal lepidopteran diversity by applying DNA barcoding technique of Malaise trap collections as a first report from Bangladesh.

Methodology:

Specimen Collection, Processing, Identification and Specimen Deposition Collections were made by a Townes-style Malaise trap (BioQuip Inc. USA) installed in perceived flight paths at Chittagong University Campus (Lat. 22.46359°N; Long. 91.7808°E) in Bangladesh by following the Standard Operating Protocol for the Global Malaise Trap Program (www.dnabarcoding.ca.). Between March 2014 and February 2015, the samples were harvested weekly in a 500 mL plastic Nalgene bottle that was filled with 375 mL of 95% ethanol and placed in 500 mL of fresh ethanol before storage at -20°C until analysis. Collected insects were analyzed, following standard barcoding protocols (http://ccdb.ca/resources.php), at the Canadian Centre for DNA Barcoding within the Centre for Biodiversity Genomics, University of Guelph, Canada. Collection data, voucher information and taxonomy for each specimen are available in the Barcode of Life Data Systems (http://v3.boldsystems.org index.php/TaxBrowser_Taxonpage 'taxid=125). All the specimens analyzed in this study have been curated at the Centre for Biodiversity Genomics, University of Guelph, Guelph, Ontario, Canada.Molecular Analysis and Data Analysis: A small portion of each specimen (usually 1-3 legs) was removed and used for DNA extraction and from the whole body of smaller taxa, and vouchers were recovered after DNA extraction for imaging and curation. Tissue lysis, DNA extraction, PCR amplification, cycle sequencing and sequence analysis were performed at the Canadian Centre for DNA Barcoding following the standard protocols (CCDB). PCR amplification of COI-5′ was performed with primers C_LepFolF and C_LepFolR (http://www.ccdb.ca/docs/CCDB_PrimerSets). following PCR conditions; 94°C (1 min), 5 cycles at 94°C (40 s), 45°C (40 s), 72°C (1 min); 35 cycles at 94°C (40 s), 51°C (40 s), 72°C (1 min) and a final extension at 72°C (5 min) and amplicons were sequenced using BigDye v3.1 (Applied Biosystems) on an ABI 3730XL. Sequences were assembled, aligned, and edited using CodonCode Aligner (CodonCode Corporation, USA) and submitted to Barcode of Life Data Systems (BOLD) (www.boldsystems.org).

Source of Information: Munis Entomology & Zoology, 16 (1): 296-311

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The current results focus on the DNA barcoding based taxonomy of Malaise trap collection. The study area is still enriched with Bangladeshi lepidopterons. By considering total identified lepidopteron species from entire country, the present study reveals very low portion in lepidopteron diversity. New addition of lepidopteran species from the current study included in total lepidopterans of country records . Hence, it can be proposed to deal with extensive survey of lepidopterans and more DNA barcoding based taxonomic works for accurate confirmation in the future.

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