GENETIC VARIATION OF THREE POPULATIONS KISSING GOURAMI (Helostoma temminckii) FROM WEST JAVA, SOUTH KALIMANTAN AND JAMBI EVALUATED USING RAPD MARKERS

Otong Zenal Arifin*, Wahyulia Cahyanti and Anang Hari Kristanto
 
Institute of Research and Development for Freshwater Aquaculture
Ministry of Marine Affairs and Fisheries
 Jl. Sempur No. 1, Bogor 16154, Indonesia
zenalarifin@gmail.com

Tambakan or kissing gouramy (Helostoma temminckii) is well known as freshwater species from Southeast Asia. In Indonesia tambakan is an economic important comodity. The status of aquaculture total production (ponds, cage, and rice field) of tambakan tend to decrease nowadays, to increase the production of taambakan, breeding programs are strongly required. The objective of this study is to identify the genetic variation of three tambakan population from West Java, Jambi, and South Kalimantan province using RAPD.

Genomic DNA was isolated from 150 mg of pectoral fin. Primer was obtained from the Oligonucleotide Synthesis Laboratory of the Operon Technologies Inc. (Kit C, OPC-02), the primer pair contains 8 bp of DNA seq (TATCCCCAT). Initial denaturation was 2 min at 94 °C, followed by 40 cycles of 60 s at 94 °C, 60 s at 36 °C, 120 s at 72 °C. An additional cycle of 7 min at 72 °C was used for primer extension; the polymerase chain reaction (PCR) was performed in a thermal cycler (Biometra). The number of polymorphic loci, percentage of polymorphic loci, expected heterozygosity (He) and degree of population differentiation (Fst) were calculated to estimate genetic diversity on these three populations Helostoma temminckii. Genetic distances dendrogram was constructed using the TFPGA / Total For Population Genetic Analysis.

Percentages of polymorphic bands (PPB) and heterozygosity for three populations are shown in Table 1 . South Kalimantan population represented the greatest level of variability (PPB = 81,25) and heterozygosity value (He = 0,3544), whereas population Jambi represented the lowest level of variability (PPB = 50,00) and heterozygosity value (He = 0.1898) with primers OPC-02. To assess the overall distribution of diversity within and between populations, an amova was performed from the distance matrix showed low significant (P< 0.05).  According to the results, genetic differences are lower within populations, not genetic differences between South Kalimantan, Jambi and West Java collections (Table 1).

In order to represent the relationships among populations, a cluster analysis (upgma) was used to generate a dendrogram based on pairwise distances between populations. The three populations (West Java, South Kalimantan and Jambi) are clearly into one main cluster.