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Assessment of Genetic Variability of Selective Mango (magnifera Indica) Varieties by Rapd Analysis

F. Alam
Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208. Bangladesh

M.A. Mannan
Agrotechnology Discipline, Khulna University, Khulna-9208. Bangladesh.

S.M.M. Rahman
Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208. Bangladesh

Abstract/ Executive Summary:

Genetic variation of selective mango varieties was analyzed by RAPD. Seven selective mango varieties, namely, Himsagar, Gobindo Bhog, Local (from seed), Bombay Lota, Gopal Bhog, Rani Pasand and Aswina were used to estimate genetic variability and relatedness among these varieties. DNA was extracted from young leaves using modified CTAB method and extracted DNA was quantified by agarose gel electrophoresis. All of ten RAPD primers showed distinct polymorphic bands which were enough to clarify the genetic variability among these mango varieties. The observed polymorphism among loci were above 90%. The highest pair wise genetic variability (1.25) was observed between Himsagar and Aswina and the minimum genetic variability (0.24) was observed between Gobindo Bhog and Rani Pasand as well as between Rani Pasand and Aswina. The highest gene frequency was 1.0000 at allele in locus S7e. The UPGMA dendogram showed the segregation and the difference of evolutionary changes of these varieties. The dendrogram showed two main clusters: Himsagar, Gobindo Bhog, Local (From seed), Bombay Lota in cluster I; Gopal Bhog, Rani Pasand and Aswina in cluster II. This is the preliminary research regarding genetic varieties, more varieties and inclusion of more primers will give the genetic variation of mango varieties in broad aspect.

Keywords: RAPD, Genetic diversity, Polymorphic loci, Mango.

Location: Germplasm Center of Khulna University maintained by Agrotechnology Discipline, Khulna University.

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Variability, Mango

Objectives:

Genetic diversity in crop species is a fundamental tool in hybrid variety production programs. The breeders need prior knowledge about the genetic background of the breeding materials and varieties. The aims of this study were to analyze the genetic variability of the most commonly cultivated mango varieties in Bangladesh and to measure the pair wise genetic distance among the selected mango varieties.

Methodology:

Extraction of DNA from mango leaves: Seven high yielding mango varieties, namely Himsagar, Gobindo Bhog, Local (From seed), Bombay Lota, Gopal Bhog, Rani Pasand and Aswina were collected from Germplasm Center of Khulna University maintained by Agrotechnology Discipline, Khulna University. Genomic DNA was extracted from very young fresh leaf samples and DNA was extracted by modified CTAB method. The DNA was quantified visually comparing with standard A. DNA by agarose mini-gel method. Ten primers of random sequence (Table 1) were screened to evaluate their suitability for amplifying DNA sequences, which could be accurately scored.

Amplification of genomic DNA: The amplification conditions were based on Williams et al. (1990) with some modifications. PCR reactions were performed on each DNA sample in a 10 Al reaction. Each mix containing 3 ng genomic DNA, 2.5 µl PCR supermix, 0.5 AI of Taq DNA polymerase, 2.0 111 ( 400 picomol) of primer, 0.5 IA of 50 mM MgC12 and 2.5 µl of distilled water. DNA amplification was performed by thermal cycler (Eppendorf Mastercycler Gradient, German). The PCR samples were overlaid with one drop of mineral oil. The amplification program included an initial denaturation step of 94°C for 2 minutes followed by 40 cycles of 1 minute denaturation at 94°C, 1 minute annealing at 36°C and 2 minutes extention at 72°C. The last cycle was followed by 5 minutes extention at 72°C and finally held at 4°C. PCR products from each sample were confirmed by running 2% agarose gel in 1X TAE buffer at 40V for 80 minutes. Loading dye was added to the PCR products and loaded in the wells. Molecular weight markers (DNA ladder) were also loaded on corner lane of the gel. After electrophoresis, the gel was taken out carefully from the electrophoresis chamber and washed with 7µl (10 mg/ml) solution of ethidium bromide mixed with distilled water (500 ml) and placed on high performance ultraviolet light box (UV transilluminator) for checking the DNA bands. The DNA was observed as band and photographed using a digital camera. All distinct bands or fragments (RAPD markers) were thereby 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 obtained using all primers in the RAPD analysis were then pooled to create a single data matrix. This was, then, used to estimate polymorphic loci (Nei, 1973), gene diversity, population differentiation (Gst), gene flow (Nm), genetic distance (D) and to construct a Unweighted Pair Group Method of Arithmetic Means (UPGMA) dendrogram among populations using a computer program, POPGENE (Version 1.31).

Source of Information: South Asian J. Agric. 2009. 4 (1&2): 63-66 ISSN 1991-0037

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The highest pair-wise genetic variability (1.25) was observed between Himsagar and Aswina and the minimum genetic variability (0.24) was observed between Gobindo Bhog and Rani Pasand as well as between Rani Pasand and Aswina. The highest gene frequency was 1.0000 at allelel in locus S7e. The UPGMA dendogram showed the segregation and the difference of evolutionary changes of these varieties. The dendogram showed two main clusters: Himsagar, Gobindo Bhog, Local (From seed), Bombay Lota in cluster I; Gopal Bhog, Rani Pasand and Aswina in cluster II. This is the preliminary research regarding genetic varieties, more varieties and inclusion of more primers will give the genetic variation of mango varieties in broad aspect.

Knowledge Management: Journal