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Determination of Genetic Variations Among Different Trichoderma Isolates Using Rapd Marker in Bangladesh

M. S. I. Sagar
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

M. B. Meah
Department of Plant Pathology, Bangladesh Agricultural University Mymensingh-2202, Bangladesh. Email: sagarbau@yahoo.com

M. M. Rahman
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

A. K. Ghose
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Abstract/ Executive Summary:

Some Trichoderma isolates were collected from different locations of Bangladesh for evaluating their bio efficiency by determining their genetic variations. PCR-based Random Amplified Polymorphic DNA (RAPD) Marker employing 3 decamer primers produced 29 scorable bands of which all (100%) were polymorphic. The co-efficient of gene differentiation (Gst) was 1.0000 reflecting the existence of high level of genetic diversity among the isolates. The Unweighted Pair Group Method of Arithmetic Means (UPGMA) dendrogram constructed from Nei’s (1972) genetic distance produced 2 main clusters (16 isolates in cluster 1 and 19 isolates in cluster 2). The result indicating their genetic diversity has opened new possibility of using the most efficient and more isolates of Trichoderma in the preparation of effective biopesticide.

Keywords: Genetic diversity, Trichoderma, RAPD

Location: BAU, Mymensingh

Start Date: 00-11-2009

End Date: 00-06-2010

Program Area: Pest Management

Commodity: Insects

Objectives:

To determine genetic variations among some isolates of Trichoderma spp. using RAPD technique for evaluating their efficiency as biopesticides.

Methodology:

An in vitro experiment was conducted at the IPM Laboratory, Department of Plant Pathology and Central Laboratory, Bangladesh Agricultural University, Mymensingh. Part of molecular experiment was conducted at the Biotechnology Laboratory, Bangladesh Institute of Nuclear Agriculture (BINA). Thirty five Trichoderma isolates collected from diverse origin of different hosts and locations in Bangladesh were used in the present experiment. The experiment was designed in a Completely Randomized Design (CRD) with three replications. The activities were performed from November 2009 to June 2010.Molecular characterization of Trichoderma isolates For DNA extraction, mycelial cultures were raised individually in 150 ml conical flasks containing 100 ml potato dextrose broth. Mycelia were collected on filter paper in a Buchner funnel, washed with distilled water, frozen, and lyophilized. After extraction, mycelia were wrapped with aluminium foil for each isolate and kept in refrigerator before genomic DNA isolation. Isolates of Trichoderma spp. were used for the RAPD. The extraction of total genomic DNA of each isolate of Trichoderma was made. As a source of genomic DNA, fresh mycelia samples were collected from the 3 days old culture. Genomic DNA samples of each accession were extracted following phenol: chloroform: isoamyl alcohol extraction and ethanol precipitation method from vigorous and fast actively growing mycelial tissue. DNA was extracted using the mini preparation CTAB method. Genomic DNA isolation Thirty five isolates of Trichoderma spp. were used for the RAPD.Genomic DNA samples of each accession were extracted following phenol: chloroform: isoamyl alcohol extraction and ethanol precipitation method from vigorous and fast actively growing mycelial tissue. Firstly, soft mycelia portion (before spore production in 3 days) of the vigorous and actively growing Trichoderma spp. were cut apart with sterile scissors and forceps. Then ethanol was used to wash scissors and forceps to avoid contamination. The DNA from mycelial tissues was then extracted.

Source of Information: Bangladesh Agril. Univ. 9(1): 9–20, 2011 ISSN 1810-3030

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The result indicating their genetic diversity has opened new possibility of using the most efficient and more isolates of Trichoderma in the preparation of effective biopesticide.

Knowledge Management: Journal