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Molecular Characterization of Five Selected Brinjal (solanum Melongena L) Genotypes Using Ssr Markers

A. K. M. Khorsheduzzaman
Entomology Section, Horticulture Research Centre (HRC), Bangladesh Agricultural Research Institute (BARI), Joydebpur, Gazipur 1701

M. Z. Alam
Department of Entomology, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur 1706, Bangladesh

M. M. Rahman
Department of Entomology, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur 1706, Bangladesh

M. A. K. Mian
Depatrment of Genetics and Plant Breeding, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur 1706, Bangladesh

M. I. H. Mian
3Department of Plant Pathology, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur 1706, Bangladesh

M. M. Hossain
4Department of Horticulture, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur 1706, Bangladesh

Abstract/ Executive Summary:

Five brinjal (Solanum melongena L.) genotypes were selected for characterization using Simple Sequence Repeats (SSR) markers. All the genotypes showed considerable variation in respect of morphological, anatomical and biochemical aspects. For study of relatedness, plant genomic DNA was extracted by CTAB based method using 11 randomly selected primers produced from Calgene Inc. USA. The primers developed 22 bands through PCR amplification out of which 15 from 3 primers and were polymorphic. Genetic similarities of SSR profiles were estimated based on Jaccard’s coefficient value. The dendrogram generated two clusters and they were clearly distinct and separated from each other. Cluster-I consisted of genotypes TURBO and BL009; and cluster-II comprised of genotypes EG058, EG075 and ISD006. Genotype TURBO and BL009 were identified as the diverse genotype and showed a maximum of 17% dissimilarity from EG058, EG075 and ISD006. The similarity value ranged from 0.83 to 1.00 which indicated the presence of narrow range of genetic diversity at molecular level but have still a possibility of crossing among the genotypes of two clusters. The banding pattern of different genotypes could be utilized as reference for further comparisons.

Keywords: Brinjal (Solanum melongena L.), Molecular characterization, SSR markers

Location: Center for Plant Molecular Biology (CPMB)

Start Date: 00-06-2005

End Date: 00-12-2005

Program Area: Variety and Species

Commodity: Brinjal

Objectives:

The present study was undertaken to know the relatedness among the selected brinjal genotypes in molecular level.

Methodology:

The study on molecular characterization of selected brinjal genotypes using SSR marker technique was carried out at the Center for Plant Molecular Biology (CPMB), Tamil Nadu Agricultural University (TNAU), Coimbatore, India during June to December 2005. Plant material used Among the five brinjal (Solanum spp.) genotypes, BL009 and ISD006 were collected from Bangladesh Agricultural Research Institute (BARI) and EG058, TURBO and a susceptible check (EG075) were from Asian Vegetable Research and Development Centre (AVRDC), Taiwan. All genotypes were grown in the transgenic greenhouse of the CPMB and 20 days old seedlings were used for the study. DNA extraction Plant genomic DNA was extracted using standard methods given by Nagarajan and Kumar (2002) with appropriate modifications. Fresh 20-day-old leaves were taken, weighed and cut into small pieces and were ground in liquid nitrogen with pre-chilled pestle and mortar and the powder was transferred into centrifuge tubes carrying 25ml pre-heated (65 0C) 2% Cityl-triammonium-bromide (CTAB) extraction buffer. The tubes were incubated at 65 0C for an hour followed by addition of 15ml chloroform:isoamyl alcohol (24:1). The samples were centrifuged at 10,000 rpm for 10 minutes at room temperature. Upper aqueous phase was precipitated with 2-volumes of ice cold ethanol and 0.1 volumes of 3M sodium acetate (pH=5.2) and centrifuged at 15,000 rpm for 15 minutes. The DNA was recovered as pellet which was washed in 70% ethanol. The concentration of DNA was determined spectrophotometrically by taking UV absorbance 260 nm. The DNA concentration was rechecked by running samples in 0.80% agarose gel at 100 volt for an hour in Tris-Base Extraction (TBE) buffer along with lambda DNA standard followed by polymerase chain reaction (PCR). Agarose gel electrophorasis Amplification products obtained through PCR were loaded into individual channels of 1.2% agarose horizontal gel in TBE buffer (pH=8.0). Electrophorasis was carried out at 100 volts for an hour. Gels were stained with ethidium bromide (2μg/l) and were observed in a transilluminator under UV light. The λ DNA double digested with Hind and EcoR I (Sigma D 9281) were used as DNA markers for comparing the molecular weights. Data collection and analysis Data were taken using standard scoring sheet supplied by Bangalore genee. The number of DNA bands formed by each sample (genotype) using different primers were recorded and analyzed using NTSYS-pc version 2.02 software. Principal Coordinate Analysis/Principal Component Analysis (PCA) was done as per method described by Rohlf (2001). Attribute of the SSR primers with repeat motif, primer sequence, expected length and annealing temperature used in the present study.

Source of Information: Bangladesh J. Genet Pl. Breed., 21(1): 01-06, 2008

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

It is revealed from the results that, the morphological variability is one of the primary factors, which contribute to the level of genetic or molecular diversity available in the genotypes. In the present investigation, five brinjal genotypes were analyzed through different randomly selected primers and represented low genetic variability. Therefore, a higher level of crossing possibility is prevailing between among genotypes of two clusters.

Knowledge Management: Journal