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Assessment of Genetic Diversity of Brinjal Germplasm by Rapd Markers

Zaharul Islam
Former MS Student
Department of Horticulture, Bangladesh Agricultural University, Mymensingh

M. Monjurul Alam Mondal, PhD
Principal Scientific Officer
Crop Physiology Division, Bangladesh Institute of Nuclear Agriculture, BAU Campus, Mymensingh-2202, Bangladesh

Prof. Dr. M. K. Siddiqua
Professor
Department of Horticulture, Bangladesh Agricultural University, Mymensingh

Md. Mahmudul Hasan
Scientific Officer
Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, BAU Campus, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

Assessment of genetic diversity in a crop species is prerequisite to its improvement. The use of germplasm with distinct DNA profiles help to generate genetically diversified breeding populations. The present study was carried out to investigate the genetic diversity in brinjal using random amplified polymorphic DNA (RAPD). Fifteen brinjal germplasm and three decamer primers were used for random polymorphic DNA assay. A total of 17 fragments were obtained, out of which 12 (70.59%) were polymorphic. Each primer generated 4 to 8 amplified fragments with an average of 5.67 fragments per primer. The highest genetic distance (0.8873) and the lowest genetic identity (0.4118) were observed in Laffa (Elongated) versus Jessore L and Dharola combinations. The lowest genetic distance (0.1525) was observed in several cultivars. The unweighted pair-group method of arithmetic means (UPGMA) dendrogram was constructed from genetic distance and all brinjal cultivars were grouped into five clusters. The genetic diversity of brinjal cultivars reported in this study will be useful when planning future crosses amongst these cultivars.

Keywords: Brinjal, Biodiversity, RAPD marker, Crop improvement

Location: Experimental field of BAU, Mymensingh

Start Date: 22-10-2011

End Date: 28-05-2012

Program Area: Variety and Species

Commodity: Brinjal

Objectives:

To characterize brinjal cultivars collected from different geographical regions of Bangladesh using RAPD markers and to assess the genetic diversity within the germplasm.

Methodology:

Fifteen cultivars were collected from various agro-climatic regions of Bangladesh. Immature and vigorous leaf collected from fourteen days old seedlings were used as the source material of genomic DNA. DNA extraction was done by using the mini preparation modified CTAB method. Altogether, 15 decamer primers of random sequence (Genei, Bangalore, India) were experienced for identifying polymorphisms. After a preliminary screen, three primers, which produced reproducible banding patterns, were selected to amplify the DNA of each brinjal cultivars. A 10 μl PCR reaction mixture was prepared with 1 μlampli Taq polymerase buffer (l0x), 2.5 μlPrimer, 1μ dNTPs,0.2 μlAmpliTaq DNA polymerase, 4 μlGenomic DNA (25 ng/μl) and a suitable amount of sterile deionized water (1.3 μl). DNA amplification was performed as follows: 1 cycle for 3 min at 94°C for initial strand separationfollowed by 45 cycles each of 1 min at 94°C fordenatuartaion, 1 min at 34°C for annealing the DNA double strand and 2 min at 72°C for primer extension. At last 1 cycle for 7 minutes at 72ºC was added to allow complete extension of all amplified fragments followed by drench at 4°C. A 1 kb (100 bp DNA ladder) DNA ladder marker was used as a molecular size marker. PCR reaction products were mixed with 2.5 μl of 10X loading dye were resolved by electrophoresis in 1.4% agarose gel at 80 V for 2 hr followed by staining with ethidium bromide. Gels were photographed in Gel Documentation System (Alphainnotech, USA). All distinct bands or fragments (RAPD markers) were thereby given identification numbers according to their position on gel and scored visually on the basis of their presence (1) or absence (0), separately for each individual and each primer. The scores obtained using all primers in the RAPD analysis were then pooled to create a single data matrix.

Source of Information: Research on Crops, 15 (2): 416-422 (2014)

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

RAPD markers have been proved to be powerful tools for molecular genetic analysis of brinjal cultivars for plant breeding program to assess genetic diversity to allow for the development of new cultivars that are aimed towards the increasing crop productivity and quality. Some cultivars have wide variation with each other and some were similar in respective of some characters. Khatkhatia and Khatkhatia BAU formed same cluster indicating there was no genetic distance between them also representing these two cultivars may not be used for breeding programme. On the other hand, Laffa (Elongated) showed high genetic distance with Jessore L and Dharola cultivars. High genetic variability and significant genetic differentiation between cultivars indicating potential genetic resources of brinjal germplasm in Bangladesh.Therefore, based on RAPD analysis crossing progrmmae among Laffa (Elongated), Jessore L and Dharola might be fruitful for breeding programme. Prior identification of more polymorphic RAPD markers may provide a better characterization of brinjal cultivars in Bangladesh.

Knowledge Management: Journal