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Genetic Diversity Analysis in Brassica Varieties Through Rapd Markers

K. K. Ghosh
Researcher of BAU

M. E. Haque
Scientific Officer, Biotechnology Division
BARI

Abstract/ Executive Summary:

This investigation was aimed at exploring the genetic diversity and relationship among nine Brassica varieties, namely BARI sharisha-12, Agrani, Sampad, BINA sharisha-4, BINA sharisha-5, BARI sharisha-13, Daulot, Rai-5, Alboglabra, using Random Amplified Polymorphic DNA (RAPD) markers. In total, 59 reproducible DNA bands were generated by four arbitrary selected primers, of which 58 (98.03%) bands were proved to be polymorphic. These bands were ranged from 212-30686bp in size. The highest proportion of polymorphic loci and gene diversity values were 37.29% and 0.1373, respectively for BARI sharisha-12 and the lowest proportion of polymorphic loci and gene diversity values were 8.47% and 0.0318, 8.47% and 0.0382 for BINA sharisha-4 and Rai-5, respectively. A dendrogram was constructed using unweighted pair group method of arithmetic mean (UPGMA). The result of cluster analysis indicated that the 9 accessions were capable of being classified into 2 major groups. One group consists of BARI sharisha-12, Agrani, Sampad, Daulot, Rai-5, Alboglabra, where Daulot and Rai-5 showed the lowest genetic distance of 0.049. And another group contain BINA sharisha-4, BINA sharisha-5 and BARI sharisha-13, where BINA sharisha-5 and BARI sharisha-13 showed genetic distance of 0.071.

Keywords: Genetic Diversity, Mustard, RAPD markers

Location: Mymensingh

Start Date: 01-01-2005

End Date: 01-01-2007

Program Area: Variety and Species

Commodity: Mustard

Objectives:

This investigation was aimed to explore genetic diversity in Brassica cultivars.

Methodology:

Nine varieties of Brassica spp. Viz; BARI Sharisha-12, Agrany, Sampad, BINA Sharisha-4, BINA Sharisha-5, BARI Sharisha-13, Daulot, Rai-5, Alboglabra were used in the study. Out of the 9 varieties, 3 were BINA varieties (Agrany, BINA Sharisha-4 and BINA sharisha-5), 4 BARI varieties (BARI Sharisha-12, BARI Sharisha-13, Daulot and Rai-5), 1 was BAU varieties (Sampad) and Alboglabra was exotic line from Germany. Seed germination was performed in the laboratory and the germinated seeds were sown in the pot placed in the green house of the Department of Genetics and Plant Breeding of Bangladesh Agricultural University. Fresh and young leaf samples were collected from 12- day old seedlings. Genomic DNA was extracted from young leaf following Phenol: Chloroform: Isoamyl alcohol (25:24:1; v:v:v) purification and ethanol precipitation method. DNA was precipitated with 800 µl of absolute ethanol where DNA became visible as white strands by flicking the tube several times with fingers. DNA was pelleted by centrifugation and reprecipitation of the DNA solution was done by adding 400 µl of 70% ethanol. The pellets were then air dried and dissolved in 50 µl of TE buffer (10 mM Tris. HCl, 1 mM EDTA, pH=8.0). DNA quality was checked by electrophoresis in 2.5% agarose gel and quantified using a spectrophotometer at 260 nm wave length (Spectronic® GENESISTM). The DNA samples were evaluated both quantitatively and qualitatively using spectrophotometer and agarose gel electrophoresis, respectively. The primers used in RAPD analysis were found to discriminate different varieties and single exotic line (Two randomly chosen individuals from two different varieties to evaluate their suitability for amplification of the DNA sequences) effectively. Among the 14 primers initially tested, 4 primers (OPB-04, OPC-05, OPC-09 and OPD-02) yielded comparatively maximum number of amplification products with high intensity and minimal smearing, good resolution and also clear bands. The number of fragments amplified per primer varied.

Source of Information:

Article Link (Online Journal):

Funding Source:

1.	Budget: 30,000

Total Budget (Tk.) : 30,000

Achievement/Findings:

The highest number and proportion of polymorphic loci was found in the variety of BARI sharisha-12, which was 22 and its percentage was 37.29. By contrast, the lowest values was recorded in varieties: BINA sharisha-4 and Rai-5. The highest and lowest Nei’s (1973) gene diversity values were found in BARI sharisha-12 and BINA sharisha-4.

Knowledge Management: Report/Proceedings