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Molecular characterization of tropical strawberry genotypes

M.A. Sweety
Department of Horticulture, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur-1706, Bangladesh

M.M. Hossain
Department of Horticulture, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur-1706, Bangladesh

M.A. Hoque
Department of Horticulture, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur-1706, Bangladesh

M. Hasan
Department of Genetics and Plant Breeding, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur-1706, Bangladesh

N. A. Ivy
Department of Genetics and Plant Breeding, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur-1706, Bangladesh

M.N. Islam
Department of Botany, University of Dhaka, Dhaka-1000, Bangladesh

R.H. Sarker
Department of Botany, University of Dhaka, Dhaka-1000, Bangladesh

S. Mitra
Department of Botany, Barisal University, Barisal

Abstract/ Executive Summary:

Genetic variability of tropical strawberry genotypes was determined through Ramdom Amplified Polymorphic DNA (RAPD) primers. Out of 14 RAPD primers six showed reproducible and polymorphic bands. The results revealed that the maximum polymorphic bands were produced by the primer OPB 12. Ten genotypes were differentiated into two clusters on the basis of Unweighted Pair Group Method With Arithmetic Averages (UPGMA). Genotypic variation based on molecular characterization indicated that genotypes belonging to two different clusters depend on their genetic component. So, selection of parents from different clusters will provide the maximum heterosis in yield.

Keywords: Dendrogram, Electrophoresis, Polymorphic marker

Location: Department of Horticulture, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur, Bangladesh

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Strawberry

Objectives:

To find out the variations at molecular level of the newly introduced strawberry genotypes in Bangladesh.

Methodology:

The following ten genotypes were used for polymorphism study: V₁- FA 006, V₂- BARI strawberry 1, V₃- FA 015, V₄-FA 024, V₅- FA 016, V₆- FA 005, V₇- FA 007, V₈- FA 008, V₉- FA 023 and V₁₀- FA 017. DNA extraction and RAPD amplification: DNA was isolated using the modified CTAB method and DNA samples were evaluated both quantitatively and qualitatively using spectrophotometer (SPECORD 50, Analytikjena, Germany) at 260 nm and agarose gel electrophoresis, respectively. Then, RAPD amplification reactions were performed in a total volume of 25 μl mixing with 2.5 μl Taq Buffer A10X (Tris with 15 mM MgCl₂), 0.5 μ l dNTPs 2.5 mM, 0.2 μ l Taq DNA polymerase 5U/μl, 1.0 μl primer, 19.8 μl sterile deionized distilled water and 1.0 μl DNA template. The results obtained with this primer were later confirmed by six other standard primers, OPA-02, OPA-03, OPB?05, OPB-06, OPB-11 and OPB 12 (Operon Technologies Inc., Alameda, CA, USA), which yielded satisfying results in the same conditions. PCR amplification for RAPD: PCR mplifications were carried out in a PerkinElmer thermal cycler 480 (Perkin' Elmer, Milan, Italy). In each thermal cycling a negative control (water instead of template) was included to rule out amplification products due to external contamination. All amplifications were repeated twice for each sample on 1% agarose gel. Electrophoresis of the amplified products and documentation: The amplified products were separated electrophoretically on 1% agarose gel. The gel was prepared using 1.0 g agarose powder containing ethidium bromide and 100 ml 1 × TAE buffer. Agarose gel electrophoresis was conducted in 1 × TAE buffer at 50 Volts and 100 mA for 1.5 hrs. One molecular weight marker 1 kb DNA ladder was electrophoresed alongside the RAPD reactions. DNA bands were observed on UV transilluminator and photographed by a gel documentation system. RAPD data analysis: Following electrophoresis, the size of amplification products were estimated by comparing the migration of each amplified fragment with that of a known size fragments of 1 kb molecular weight marker. All distinct bands or fragments (RAPD marker) were there by given identification numbers according to their position on the gel and scored visually on the basis of their presence (1) or absence (0), separately for each individual varieties and each primer. The scores obtained using all primers in the RAPD analysis were then combined to create a single data matrix. Jaccards similarity and dissimilarity coefficients were then calculated between pairs of tracks and strains were grouped by using the Unweighted Pair Group Method With Arithmetic Averages (UPGMA), with a dendrogram construction utility software.

Source of Information: Plant Tissue Cult. & Biotech. 27(1): 33?39 (June)

Article Link (Online Journal):

Funding Source:

1.

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Genotypic variations based on molecular characterization indicated that genotypes belonging to different clusters depend on their genetic components itself, but not at geographical origin at all. Therefore, it could be concluded that for further research program, especially for hybridization, genotypes selected from different clusters would provide maximum heterosis. Ten variants of strawberry have been differentiated into two main clusters after molecular characterization. It could be concluded that for further research program, especially for hybridization, genotypes selected from different clusters would provide maximum heterosis regarding the yield.

Knowledge Management: Journal