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Genetic Diversity Analysis of Ginger Genotypes Using Rapd Markers

Md. Mahmudul Hasan
Scientific Officer
Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, BAU Campus, Mymensingh-2202, Bangladesh

M. Monjurul Alam Mondal, PhD
Principal Scientific Officer
Crop Physiology Division, Bangladesh Institute of Nuclear Agriculture, BAU Campus, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

The present investigation was undertaken to assess the genetic diversity among eight ginger genotypes using RAPD markers. A total of 16 distinct DNA fragments ranging from 100–1000 bp were amplified by using three selected primers of which 10 (62.50%) were polymorphic. BARI ada-1 was more homogenous than others and Syedpuri was found less homogenous showing the low intra-variety similarity value (75.56). The genotype ‘Syedpuri’ was found as more diversified from the viewpoint of lowest intra-variety similarity index value, highest gene diversity, proportion of polymorphic loci and highest level of genetic variation. The cluster analysis indicated that the eight genotypes were grouped into two major clusters. ‘Indian’ alone formed the first major cluster while the second major cluster had seven genotypes and was divided into two minor clusters. China and Sherpuri genotype pair was very close to each other with the lowest genetic distance (0.03). On the other hand, Indian and Syedpuri pair was more distant to each other with the highest genetic distance (0.55). RAPD analysis revealed a considerable level of polymorphism among the studied genotypes. The genetic variation thus detected has significance for ginger improvement programs.

Keywords: Genetic diversity, Ginger, RAPD marker, Crop improvement

Location: Experimental field of BAU, Mymensingh

Start Date: 10-03-2010

End Date: 30-05-2010

Program Area: Variety and Species

Commodity: Ginger

Objectives:

To assess the genetic diversity among eight ginger genotypes using RAPD markers.

Methodology:

Eight ginger genotypes collected from different regions of Bangladesh were used in this study. Genomic DNA was extracted from fresh leaves by using the cetyltrimethyl ammonium bromide (CTAB) method. The DNA samples were evaluated both quantitatively and qualitatively by 1% agarose gel electrophoresis and genomic DNA concentration was estimated by spectrophotometer at a wavelength of 260 nm using the Spectronic® GenesisTM. Twelve primers, corresponding to kits A, B, C, D and E were initially tested and finally three primers (OPA-05, OPC-01 and OPE-02) exhibiting higher quality bands with minimal smearing and good resolution was selected for DNA amplification. Finally, three primers were selected for the analysis of the whole sample of the eight genotypes. PCR reactions were performed for each DNA sample with single primer in a 10 µl reaction mix containing 1 µl Taq DNA polymerase buffer (10X), 2.5 µl primer (10 µM), 1 µl dNTPs (250 µM each), 1 unit Taq DNA polymerase (0.3 µl) and 100 ng (4 µl) of genomic DNA and a suitable amount of sterile deionized water. DNA amplification was performed in an oil-free thermal cycler. Amplicons were analyzed electrophoretically on 1.5% agarose gel containing ethidium bromide in 1X TBE buffer at 120 V for 1 hr. Distinct RAPD bands were given identification numbers according to their position on gel and scored visually on the basis of their presence (1) or absence (0) separately for each individual and each primer. The scores were then pooled to construct a single data matrix. This was used for estimating polymorphic loci and the UPGMA dendrogram of the populations using the POPGENE software package. The similarity index values between the RAPD profiles of any two individuals on the same gel were calculated from RAPD markers according to the following formula: Similarity index (SI) = 2 N_xy/(N_x + N_y).

Source of Information: Life Science Journal, 11 (8): 90-94 (2014)

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

This investigation presented a sufficient amount of variation among the studied genotypes indicated that the RAPD method could be used as an effective tool for molecular genetic analysis of various genotypes of ginger. However, large number of samples from all the AEZ of Bangladesh and higher number of primers would be useful to draw a more definite conclusion. Through this study, we have assessed, for the first time the genetic relationship among some ginger genotypes of Bangladesh. The result of the present study can be used as a guideline for future diversity assessment and genetic analysis of ginger genotypes.

Knowledge Management: Journal