[논문]Genetic Analysis of Three River Populations of Catla catla (HAMILTON) Using Randomly Amplified Polymorphic DNA Markers

Islam, M.S.; Ahmed, A.S.I.; Azam, M.S.; Alam, M.S.

doi:10.5713/ajas.2005.453

Genetic Analysis of Three River Populations of Catla catla (HAMILTON) Using Randomly Amplified Polymorphic DNA Markers 원문보기

Asian-Australasian journal of animal sciences, v.18 no.4, 2005년, pp.453 - 457

Islam, M.S. (Department of Biotechnology, Bangladesh Agricultural University) , Ahmed, A.S.I. (Department of Fisheries Biology and Genetics, Bangladesh Agricultural University) , Azam, M.S. (Department of Fisheries Biology and Genetics, Bangladesh Agricultural University) , Alam, M.S. (Department of Fisheries Biology and Genetics, Bangladesh Agricultural University)

Abstract ▼ AI-Helper

The genetic variations in three major river populations viz. the Halda, the Jamuna and the Padma of the Indian major carp, Catla catla were analyzed by Random Amplified Polymorphic DNA (RAPD) markers. Four decamer primers were used for amplifying DNA of 10 individuals from each population. The proportion of polymorphic loci and the gene diversity estimates were 59.4 and 0.20 for the Halda, 37.5 and 0.14 for the Jamuna and 46.9 and 0.16 for the Padma populations respectively indicating the existence of a relatively high level of genetic variation in the Halda river population. The inter-population similarity indices, gene flow and genetic distance values indicated that the Jamuna-Padma population pair of catla was genetically closer than the Halda-Jamuna and the Halda-Padma population pairs in compliance with the geographical distances among them. The coefficient of gene differentiation ($G_{ST}$=0.13) reflects some degree of genetic differentiation among three populations of catla studied. The data suggest that the RAPD technique could be used to discriminate different river populations of catla.

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문제 정의

This study presents a first step towards the investigation of genetic variability among three different riverine stocks of catla by a DNA marker. The Halda stock is genetically more diversified than the Jamuna and the Padma population while the Jamuna and the Padma population are genetically more similar as they are geographically closer.

제안 방법

Ten primers having high intensity of bands and no smearing were further used for amplification of DNA of two individuals from each population. A final subset of four primers (Table 1) exhibiting the highest quality banding patterns and sufficient variability for population analysis were taken for analysis of all the samples.
Experiments were run to optimize DNA, dNTPs and Taq DNA polymerase concentrations and to determine the optimum annealing temperature. Finally, PCR reactions were performed on each DNA sample in a 10 卩l reaction mix containing 1 卩l of 10xTaq polymerase buffer, 2 卩 l of 10 卩M primer, 1 卩 l of 250 卩M dNTPs, 1 unit of Taq DNA polymerase (GENEI, Bengalore, India) and 50 ng of genomic DNA and a suitable amount of sterile deionized water.
Experiments were run to optimize DNA, dNTPs and Taq DNA polymerase concentrations and to determine the optimum annealing temperature. Finally, PCR reactions were performed on each DNA sample in a 10 卩l reaction mix containing 1 卩l of 10xTaq polymerase buffer, 2 卩 l of 10 卩M primer, 1 卩 l of 250 卩M dNTPs, 1 unit of Taq DNA polymerase (GENEI, Bengalore, India) and 50 ng of genomic DNA and a suitable amount of sterile deionized water. DNA amplification was performed in an oil-free thermal cycler (Master Cycler Gradient, Eppendorf).
The Halda stock is genetically more diversified than the Jamuna and the Padma population while the Jamuna and the Padma population are genetically more similar as they are geographically closer. Further studies involving large number of samples and primers need to be conducted to get more precise information about the genetic structure of the three riverine stocks of catla

이론/모형

The scores were then pooled for constructing a single data matrix. This was used for comparing the frequencies of allpolymorphic RAPD markers and estimating Nei’s (1973) gene diversity (h), gene flow (Nm), coefficient of gene differentiation (GST), genetic distance (D) and constricting a UPGMA (unweighted pair group method of arithmetic means) dendrogram among populations with 1000 simulated samples using POPGENE (version 1.31) (Yeh et al., 1999) computer program. Bandsharing based similarity indices between individuals within a population sample (Si) and between population samples (Sij) were calculated for all possible comparisons according to the method of Lynch (1991).
This was used for comparing the frequencies of allpolymorphic RAPD markers and estimating Nei’s (1973) gene diversity (h), gene flow (Nm), coefficient of gene differentiation (GST), genetic distance (D) and constricting a UPGMA (unweighted pair group method of arithmetic means) dendrogram among populations with 1000 simulated samples using POPGENE (version 1.31) (Yeh et al., 1999) computer program.

성능/효과

051. The proportion of polymorphic loci and Nei’s (1973) gene diversity (h) values were found to be higher (59.4% and 0.20 respectively) in the Halda population followed by those of the Padma population (46.9% and 0.16 respectively) whilst the values for the Jamuna river population were lower (37.5% and 0.14 respectively) (Table 4). The coefficient of gene differentiation among three different populations was found to be 0.
On the other hand, the RAPD method uncovered 24 polymorphic loci from four primers selected for population analysis. The result is consistent with the fact that the RAPD technique, being able to screen more easily a larger part of the nuclear genome than allozymes, may assess higher levels of genetic variation.
Among the primers OPB15 gave DNA profiles with more numerous bands than the other three primers (Table 1). The total number of fragments yielded from four primers were 32 of which 24 (75%) were polymorphic (the frequency of the most common allele is 0.95 or lesser) (Table 4). RAPD profiles obtained in this analysis are shown in Figure 2.

참고문헌 (19)

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상세보기
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DOI : 10.5713/ajas.2005.453 [무료]
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