DNA Fingerprinting of Mungbean (Vigna radiata L.) genotypes
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Date
2022-08-23
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guntur
Abstract
Thirty elite improved lines of mungbean were characterized morphologically by using PPV&FRA DUS descriptors and studied for genetic variability parameters to know the variability in the studied material. Further, the genotypes were characterized using molecular markers (RAPD, ISSR and SSR) to know diversity and utilizing them in DNA fingerprinting.
The DUS descriptors, plant growth habit and plant habit, plant height, petiole colour, stem colour, stem pubescence, leaf vein colour, flower petal colour, pod colour and seed size showed no variation among the genotypes. The genotypes differed significantly for the descriptors like anthocyanin pigmentation, leaflet lobes, leaf shape, colour and size, time of flowering, premature pod colour, pod pubescence, pod position, curvature of mature pod, pod length, seed colour, lustre and shape among the genotypes indicating their utilization in characterization, registration, protection and purity maintenance.
In the present study, PCV was more than the GCV for all the ten quantitative characters indicating the presence of environmental influence on character expression. Further, the genetic variability was more for the characters viz., no. of branches/ plant, no. of clusters/ plant, no. of pods/ plant and test weight. High heritability and high genetic advance for characters no. of branches/ plant, no. of clusters/ plant, no. of pods/ plant, test weight and seed yield per hectare indicated the additive gene effects role and exploitation of simple selection for these traits improvement.
These genotypes were characterized using Random Amplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeats (ISSR) and Simple Sequence Repeats (SSR) markers for DNA Fingerprinting of these genotypes. Nine RAPD and Seven ISSR and 50 SSR primers were used for the characterization while, Six RAPD, five ISSRs and 44 SSR produced scorable with clear and consistent amplification profiles. six RAPD markers produced 27 amplification products with an average of 4.6 fragments per primer. The size of the band varied from 500 bp to 2600. The PIC values ranged from 0.27 (OPN 9) to 0.50 (OPN 10, OPA 9 and OPA 19) with an
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average of 0.42. The genetic similarity values range was zero to 0.85 indicating the presence of huge genetic diversity at molecular level among the genotypes. The UPGMA dendrogram grouped the thirty genotypes into two clusters. The genotype, LGG 709, formed a separate cluster indicating its divergent nature and utilization in the breeding programmes.
The five ISSR primers produced a total of 23 amplified bands with an average of 4.6 fragments per primer, out of which 14 were found polymorphic with an average of 2.8. The size of the band varied from 500 bp to 3000 bp. PIC values ranged from 0.37 (UBC-879) to 0.50 (UBC-848) with an average of 0.43. The genetic similarity values range was zero to 0.27 indicating the presence of low genetic diversity at molecular level among the studied thirty genotypes. The UPGMA dendrogram grouped the thirty genotypes into two clusters. The genotypes, LGG 698 and LGG 709, formed a separate cluster indicating their utilization in the breeding programmes.
Among the 44 SSR primers amplified, only nine SSR primers produced polymorphism and produced a total of 236 bands. The size of the bands ranged from 160-200bp. PIC values varied from 0.06 to 0.69 with an average of 0.36. The genetic similarity values range was 0.05 to 0.33. The UPGMA dendrogram grouped the thirty genotypes into two clusters. The genotypes, LGG 696, LGG 697 and LGG 698, formed into a separate cluster indicating their divergent nature from the other genotypes.
The simulated DNA Fingerprinting profiles of the genotype, LGG 698, had the unique banding pattern for the primers, CEDG-008 (180bp) and CEDG-015 (180bp), CEDG-056 (180bp), CEDG-092 (180bp), GLLC-108 (180bp), PBALC-13 (180bp), PBALC-217 (180bp) and Vr SSR-014 (200bp) while, bands were absent for the primer CEDG- 068. The genotype, LGG 708, also produced unique profile as two primers, CEDG-056 at 200bp and 160bp and GLLC-108 at 200bp and 180bp, were not amplified indicating the use of these markers for this genotype identification from others. The highly informative primers identified in this study, could be utilized in generating useful molecular descriptors for fingerprinting of mungbean genotypes
Description
DNA Fingerprinting of Mungbean
(Vigna radiata L.) genotypes