Browsing by Author "Godagama, G.R.M.N."

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    Determination of Morphological and Genetic Diversity of Wild Guppy (Poecilia reticulata) in Sri Lanka across the MHC Complex with Special Reference to Class IIB Region
    (Uva Wellassa University of Sri Lanka, 2013) Godagama, G.R.M.N.; Fernando, T.S.R.; Bulumulla, P.B.A.I.K.; Jayamanne, S.C.
    Wild guppies have potential in developing various strains with attractive colour patterns, tail types and tolerance to wide range of environment conditions, resistance to disease conditions due to high immunity. Application of molecular genetic markers, are important to identify diversity among wild guppies which are economically beneficial to ornamental industry and to implement conservation of these valuable genetic resources. Major histocompatibility complex (MHC) genes are highly polymorphic gene family and exon 2 of class II B gene is functionally important in immunity and disease resistance. Hence, in the present study attempts are made to assess the genetic and morphological diversity of wild guppy of Sri Lanka with special reference to immune related MCH class II B gene. A total of 238 wild guppies were collected from 10 regions to represent different agro-ecological zones of the country. The standard length in between 13-24 mm was selected (179 fishes) to collect morphological data and genomic DNA was extracted from muscle tissue using Chelex 100 DNA extraction kit. A PCR based method was used to amplify exone 2 region of candidate gene with forward (5’GTG GAT TTC AGA GAA TAT GCA 3’) and reverse (5’ TGA TTT ATC CAG AGC GGT TTG 3’) primers. Touch down PCR was followed to amplification in the temperature range of 47 to 45 . Selected fish sample consisted of 43.6% of male fishes and 56.4% female fishes. Significant association existed in tail types and colour patterns versus region. Highest variation of tail pattern types was recorded from Buttala region and 40.8% of guppies consisted round tail type. High variation of colour pattern is observed from Badulla region. 58.7% fishes had brownish gray colour pattern and 43% had golden upper body colour pattern. Variation of upper body colour in all regions was similar. PCR protocol was optimized. There was a morphological diversity between wild guppy fishes in different regions of Sri Lanka. Exon II in MHC class IIB region was amplified and optimized PCR protocol for further studies. Sequence variation based on Single Nucleotide Polymorphism (SNPs) and differences of immune response of wild guppy population is yet to be analyzed.
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    Determination of Morphological and Genetic Diversity of Wild Guppy (Poecilia Reticulata) in Sri Lanka Across the Major Histocompatibility Complex with Special Reference to Class IIB Region
    (Uva Wellassa University of Sri Lanka, 2013) Godagama, G.R.M.N.
    Wild guppies have potential in developed various strains with attractive colour patterns, tail type and tolerated in a wide range of environment conditions, resistance to disease conditions due to high immunity. Application of molecular genetic markers, are important to conservation of genetic resources and finding morphological diversity of wild guppies economically beneficial to ornamental industry. Major histocompatibility complex (MHC) genes are highly polymorphic gene family and exon 2 gene is functionally important in immune and disease resistance. Conservation by incorporating immune genes is important to understand wild life immune genetic. Wild guppies were collected from 10 regions according to Agro climatological regions, 238 fishes were collected as total sample of the study and stranded length in between 13-24mm were selected (179 fishes) to gathered of morphological information, Genomic DNA was extracted from muscle tissue using Chelex 100 DNA extraction kit, targeted region was amplified using forward (5'GTG GAT TTC AGA GAA TAT GCA 3') and reverse (5' TGA TTT ATC CAG AGC GGT TTG 3'). Touch down PCR was followed to amplification in the temperature ranging 45°C to 47°C. Selected fish sample was consisted 43.6% of male fishes and 56.4% female fishes. Significant association was exists tail type and colour pattern versus region. Highest variation of tail pattern types was recorded from Buttala region and from whole sample of guppies 40.8% were consisted round tail type. Badulla region fishes were consisted high variation of colour patterns. 58.7% fishes had brownish gray colour pattern. 43% golden upper body colour pattern type fish was consisted from whole sample, variation of upper body colour in all regions were similar. PCR protocol was optimized. There was a morphological diversity between wild guppy fishes in different regions of Sri Lanka. Exon II in MHC class IIB region was amplified and optimized PCR protocol for further studies. Preliminary and sequence variation based on Single Nucleotide Polymorphism (SNPs) of wild guppy population is yet to be done by Single Stranded Conformation Polymorphism. Key words: Major Histocompatibility complex, Wild guppy, Morphological diversity, Gentic diversity, Immunity