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Morphological and Molecular Characterization of Eggplant

Md. Rezwan Molla
Scientific Officer
Plant Genetic Resources Centre, Bangladesh Agricultural Research Institute(BARI), Joydebpur, Gazipur-1701, Bangladesh

M.A.N. Nazim-ud-Dowla
Scientific Officer
Plant Genetic Resources Centre, Bangladesh Agricultural Research Institute(BARI), Joydebpur, Gazipur-1701, Bangladesh

Md. Shefatur Rahman
Scientific Officer
Plant Genetic Resources Centre, Bangladesh Agricultural Research Institute(BARI), Joydebpur, Gazipur-1701, Bangladesh

Sharmin Rahman
MS student
Bangabandhu Sheikh Mujibur Rahman Agricultural University, Salna, Gazipur

Abstract/ Executive Summary:

Characterization of brinjal (Solanun melogena) accessions on the basis of DNA fingerprinting data in combination with morphological traits has become an efficient tool to link phenotypic and genotypic variation. This work is reporting the utilization of a small set of six previously developed brinjal microsatellite markers for the identification and discrimination of four HYVs and 77 local including four wild brinjal accessions. Using six primers across 85 brinjal accessions a total of 30 alleles were found of which EM155 and EEMS28 showed highest number of alleles (7). The primer EEMS28 motif [(TAA)21] also yielded highest number of PIC value (0.803). Average genetic differentiation (Fst) values and gene flow (Nm) were found 0.651 and 0.134 respectively. Higher level of genetic differentiation and low level of gene flow values in 85 brinjal accessions which are indicative of diversity among the accessions as most of the studied accessions were land races and HYVs. Over all Nei’s genetic distance value (D) from nil to 2.239 among 3570 varieties pair resulting as a means of permutation combination of 85 brinjal accessions. Out of 85 accessions, 83 accessions were identified with at least one and/or combination of 6 primers.

Keywords: Solanun melogena, Morphological traits, DNA fingerprinting, Microsatellite marker, Genetic distance, Gene flow

Location: Plant Genetic Resources Centre, Bangladesh Agricultural Research Institute(BARI), Joydebpur, Gazipur-1701, Bangladesh

Start Date: 01-08-2008

End Date: 01-03-2010

Program Area: Variety and Species

Commodity: Brinjal

Objectives:

1) To reveal variation in morphological traits 2) To identify accessions with unique DNA banding pattern using SSRs marker 3) To estimate genetic diversity 4) To identify duplicate among the collection 5) To establish genetic relationship

Methodology:

Morphological characterization The experiment was conducted using 85 brinjal accessions collected from the PGRC and HRC, BARI. Five seedlings of each accession were transplanted in a single row with 75 cm x 60 cm spacing. Morphological data was recorded according to standard descriptor. Molecular characterization Bulked DNA was isolated from 2-5 fresh leaves of 12 days old seedlings using following the protocol described by Aljanabi and Martnez (1997) and also used by Rahman et al. (2007) with some modifications. The genomic DNA was quantified at 260 nm spectrophotometrically (Spectronic® GENESYS™ 10 Bio). Primer pairs viz. EM134, EM114, EM155, EM133, EEMS28, and EEMS48 with clear and expected amplified product sizes were used for microsatellite analysis. The reaction was performed in a oil free Techne, TC 512 thermal cycler with the following thermal profiles: i) one cycle of 94⁰C for 3 min; 35 cycles of 95⁰C for 0.5 min., 60-61⁰C for 1 min and and 72⁰C for 1 min; and one cycle at 72⁰C for 5 min. ii) a touch down PCR protocol was applied, consisting of a 94⁰C/5 min denaturation, 11 cycles of 94⁰C/0.5 min, 57-58⁰C/1 min decreasing by 0.5⁰C per cycle, 72⁰C/1 min followed by 30 cycles of 94⁰C/0.5 min, 52-53⁰C/1 min and 72⁰C/1 min. PCR-products were electrophoresed on a 6% denaturing polyacrylamide gel containing 19:1 acrylamide: bis-acrylamide and 8M urea. Electrophoresis was done using the SequiGen GT Sequencing Cell (BIO-RAD Laboratories, Hercules, CA, USA). The DNA fragments were visualized following the Promega (Madison, WI) silver-staining protocol. The software DNA FRAG version 3.03 was used to estimate allelic length (Nash, 1991). Expected (He) and observed heterozygosity (Ho) were also calculated as per Nei (1972) formula. Estimation of Nei’s genetic distance values (D) and construction of UPGMA dendrogram was constructed using the software POPGENE (Version 1.31) (Yeh et al., 1999).

Source of Information: Book, MOA

Article Link (Online Journal):

Funding Source:

1.	Budget: Tk. 3,50,000.00
2.	Budget: Tk. 3,50,000.00

Total Budget (Tk.) : Tk. 3,50,000.00

Achievement/Findings:

The set of microsatellite markers used here provides a positive assessment to the ability of SSR marker to produce unique DNA profiles of brinjal accessions. The results of the present study could be applied as baseline information to maintain the appropriate identity and the construction of a database of all brinjal cultivars and their wild relatives grown in Bangladesh and in broad sense, to protect the plant varieties of Bangladesh. The data obtained can also be used for, providing additional genetic information of the agronomic and quality characteristics of brinjal varieties. Inter-mating cultivars from the major distinct gene pools could provide new genetic recombination to exploit in cultivar development programe.

Knowledge Management: