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Rapd Fingerprinting in Four Different Species of Oyster Mushroom Available at Namdec

Nirod Chandra sarker
National Mushroom Development and Extension Centre, Sobhanbag, Savar, Dhaka

Taufiqur Rahman
National Mushroom Development and Extension Centre, Sobhanbag, Savar, Dhaka

Md. Ruhul Amin
National Mushroom Development and Extension Centre, Sobhanbag, Savar, Dhaka

Md. Bazlul Karim Chaudhury
Ph.D. Student
Jahangirnagar University and OSD, DG Health, Mohakhali, Dhaka, Bangladesh.

M. A. K. Azad
Department of Biotechnology and Genetic Engineering, Islamic University, Kustia.

Md. Amzad Hossain
Biotechnology laboratory, Bangladesh Sugarcane Research Institute, Ishurdi, Pubna.

Abstract/ Executive Summary:

Genetic diversity in four different species of oyster mushrooms was studied based on their random amplification of polymorphic DNA. Four different species of oyster mushroom i.e. Pleurotus djamor (NO-3), Pleurotus florida (NO-4), Pleurotus ostreatus (NO-5, White snow) and Pleurotus salmoneostramineus (NO-6) were taken from National Mushroom Development and Extension Center (NAMDEC), Bangladesh. The dendrogram based on RAPD analysis generated 2 major clusters ‘C₁ ’ and ‘C₂ ’. Cluster ‘C₂ ’ also formed sub cluster SC₁ and SC₂ . Out of six random primers, the maximum polymorphism was observed by primers OPA 09 (18.18%). The two species, Pleurotus djamor (NO-3) and Pleurotus ostreatus (White snow) were observed to be least similar having value 43.0% and constituting a sub cluster SC₂ . The present work revealed that RAPD primers showed different pattern of genetic diversity among the selected different oyster mushrooms.

Keywords: Genetic diversity, Oyster mushroom, RAPD.

Location: National Mushroom Development and Extension Centre, Sobhanbag, Savar, Dhaka.

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Mushroom

Objectives:

To investigate the genetic relationship in four different species of oyster mushrooms.

Methodology:

Four species; Pleurotus djamor (NAMDEC Oyster – NO 3), Pleurotus florida (NAMDEC Oyster – NO 4), Pleurotus ostreatus (NAMDEC Oyster – NO 5) and Pleurotus salmoneostramineus (NAMDEC Oyster – NO 6) were taken from National Mushroom Development and Extension Centre, Sobhanbag, Savar, Dhaka. Filamentous fungi have strong cell walls, which are often difficult to rupture. In the present investigation, Modified method has been used to isolate the total genomic DNA from mushroom. DNA of four species of oyster mushroom were extracted from 0.2-0.3 g fruiting body of each species. It was grinded in extraction buffer (200 mM Tris-HCl, pH-8.5, 250 mM NaCl, 25 mM EDTA, 0.5% SDS) with a morter pestle. The lysates were incubated at 65°C for 40 min in water bath and centrifuge 30 mins at 10000 rpm. DNA was precipitated from the supernatant by adding equal volumes of isopropanol and resultant pallet was washed with 70% ethanol. The DNA pellet was air dried and dissolved in 50 μL TE buffer. DNA quantification was performed and a dilution of 50ng/μl was used in downstream application. Genomic DNA was amplified by the RAPD technique in which six sorts of arbitrary 10-base oligonucleotide primers (Operon technologies Inc.) such as OPA-02 (sequence 5' TGCCGAGCTG 3'); Tube E-1 (sequence 5' GTGACATGCC 3'); OPA-07 (sequence 5' GAAACGGGTG 3'); OPA-09 (sequence 5' GGGTAACGCC 3'); OPA-10 (sequence 5' GTGATCGCAG 3'); OPA-15 (sequence 5' TTCCGAACCC 3'). RAPD-PCR reaction was performed using a thermal cycler with an initial denaturation stage of 5 min at 94°C, followed by 40 cycles of denaturation for 1 min at 94°C, annealing for 1 min at 36°C, extension for 2 min at 72°C and a final extension for 10 min at 72°C. RAPD products were electrophoresed on 1.4% agarose gels in 1X TBE buffer for 1.15 hr at 100v with 1kb DNA ladder as a size marker and stained while agitated in an EtBr solution (0.5% µg/ml). The stained gels were visualized under a UV transilluminator and photographed using Bio-Rad gel documentation system. The amplification products generated by each RAPD primer were scored as ‘1’ or ? ‘0’ for presence or absence of specific allele, respectively. To estimate the similarity and genetics distance among different species, cluster analysis based on Nei’s unweighted pair-group with arithmetic average (UPGMA) was performed using the ‘statistica’ software and a dendrogram was constructed.

Source of Information: Bangladesh J. Mushroom. 5(2): 7-14, 2011 (December)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The mushroom varieties belonging to ecological proximity or different geographical origins can be classified through morphological and molecular markers. The current study demonstrated that, the RAPD analysis is useful for characterization, genetic diversity and identifying relationships among the oyster mushrooms. Study also revealed that, RAPD analysis can be very useful tool for mushroom grower for classification and maintenance of good quality spawns.

Knowledge Management: Journal