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Characterization of Lentil (lens Culinaris M.) Germplasm Through Rapd Markers

R. Gupta
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh

L. Hassan
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh

S. N. Begum
Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, Mymensingh

M. M. Islam
Biotechnology Division, Bangladesh Institute of Nuclear Agriculture, Mymensingh

M. S. Alam
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh

Abstract/ Executive Summary:

PCR based random amplified polymorphic DNA (RAPD) markers were used to find out the relationship and diversity among the individuals and to characterize the 22 lentil genotypes. RAPD markers viz. OPC-05, OPB-08 and OPB-10 showed good technical resolution and sufficient variations among the genotypes and produced 24 polymorphic bands. Genetic distance (GD) between lentil genotypes were computed from combined data for the three primers and the values ranged from 0.0000 to 0.4700. Higher genetic distance was observed between LM-24-3 vs. LM-28-2 and LM-75-4 vs. LM-24-3, while the no genetic distance was observed between the genotypes LM-185-2 vs. LM-14-2, LM-185-2 vs. LM-21-6 and ICARDA-23128 vs. ICARDA-23128. The unweighted pair group method of arithmetic mean (UPGMA) dendrogram based on Nei’s genetic distance grouped the 22 genotypes into two main clusters. LM-24-3 was in cluster B and the others in cluster A. All the genotypes produced polymorphic bands with three tested RAPD markers suggesting that the genotypes were different from each other but genetic diversity were narrow among the genotypes. Thus, these RAPD markers could be efficient for comparing the genetic relationships, patterns of variation and measurement of genetic distance among lentil genotypes.

Keywords: Characterization, lentil, germplasm, RAPD markers.

Location: Biotechnology laboratory, Biotechnology Division, BINA, Bangladesh Institute of Nuclear Agriculture, BAU campus, Mymensingh-2202

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Lentil

Objectives:

To find out the relationship and diversity among the individuals and to characterize the 22 lentil genotypes.

Methodology:

The experiment was conducted at the Biotechnology laboratory, Biotechnology Division, BINA, Bangladesh Institute of Nuclear Agriculture (BINA), BAU campus, Mymensingh-2202. 22 lentil genotypes viz. LM-123-7, LM-132-7, LM-28-2, LM-14-2, LM-21-6, LM-24-3, LM-37-8, LM-48-1, LM-99-4, LM-95-3, LM-13-1, LM-156-1, LM-185-2, LM-20-3, LM-67-5 and LM-75-4, ICARDA 2348, ICARDA 38211, ICARDA 23136, ICARDA 23121, ICARDA 23105 and ICARDA 23128 were used for the characterization through RAPD markers. Fresh leaf samples from 28 days old seedlings were used for DNA extraction. Modified SDS mini-prep method was followed to extract DNA from lentil plants. Quantification of the DNA was done through electrophoresis on a 1% agarose gel. Twenty six RAPD primers were tested for their ability to amplify scorable and reproducible DNA fragments. Primers resulting in faint or irreproducible bands were excluded from subsequent analysis. Finally three primers were selected for this study. PCR reactions were performed in a 10 μl reaction mix containing 3.3 μl sterile deionized water, 1 μl Taq polymerase buffer, 1 μl dNTPs (1 mM) mixture, 0.2 μl Taq DNA polymerase, 2 μl (approx. 50 ng) genomic DNA and 2.5 μl (10 mM) primer. Samples were subjected to the following thermal profile for amplification in a thermocycler: 5 min at 95ºC for initial denaturation (pre-heat) followed by 39 cycles of 1 min denaturation at 94ºC, 0.45 min at 34ºC (annealing) and 2 min at 72ºC (extension) and a final extension step at 72ºC for 7 min. Electrophoresis was carried out in 1 X TBE buffer on a 1.5 % agarose gel and amplified fragments were visualized by staining with ethidium bromide. The amplified bands were visually scored as present (1) and absent (0) separately for each individual primer. The scores of bands were pooled to create a single data matrix. This was used to estimate polymorphic loci. Nei’s genetic distance and identity and a dendrogram based on an unpaired group method of arithmetic means (UPGMA), was constructed using POPGENE.

Source of Information: Bangladesh J. Seed Sci. & Tech. 16 (1 &2): 49-59 (2012), ISSN 1029 - 8800

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

All the genotypes produced polymorphic bands with three tested RAPD markers suggesting that the genotypes were different from each other but genetic diversity were narrow among the genotypes. Thus, these RAPD markers could be efficient for comparing the genetic relationships, patterns of variation and measurement of genetic distance among lentil genotypes.

Knowledge Management: Journal