Analisis 16s rDNA-mtDNA sebagai marka genetik variabilitas pertumbuhan ikan kerapu macan (Epinephelus fuscoguttatus) melalui metode PCR-RFLP=Analysis of 16srDNA-mtDNA as variability growth genetic marker ...

Groupers are an important commercial fish species in Indonesia and popular marine food fish of high market value in many parts of the world. The objective of this study was to find the genetic marker which can be used as indicator variability of brown marbled grouper fries. The study was conducted at biotechnology laboratory, Research institute for Mariculture on Bali. The initial study were selecting of fish fry based on the total length and body weight. The average total length of the large, medium, and small larvae were 63.27±2.62 mm, 50.64±1.15 mmand 38.09±1.39 mm respectively. The average of body weight of the large, medium, and small larvae were 4.84±0.81 g, 2.35±0.22 g, and 0.96±0.08 g respectively. The fragment of 16s rDNA were amplified with thermal PCR by using primer Mt-1 : 5\u27-CAT ATT AAA CCC GAA TGA TAT TTT-3\u27 and Mt-2 : 5\u27-ATA ATA GGG TAT CTA ATC CTA GTT T-3\u27. The polymorphism fragment of product amplification of 16s rDNA were digested with four restriction enzyme i.e. EcoRV, HaeIII, Hinf I, and Mbo I. Agarose gel electrophoresis was used to assess the DNA fragment pattern and fragment length polymorphism. GENEPOP program was used to confirm genetic variation and genetic distance values between growth variability of grouper.Result of PCR amplification showed that molecule weight of 16s rDNA fragment amplified was 1,948 bp. Digestion with EcoRV resulted in two fragment, on large, medium, and small, i.e. 1,233 and 715 bp, 1,242 and 706 bp, and 1,241 and 708 by respectively. Similar result also showed on Hinf I, and Mbo I digestion, were two fragment length were obtained at large, medium, and small fry. Digestion with restriction enzyme Hinf I result 1,518 and 430 by (large fry), 1,521 and 427 by (medium fry), 1,521 and 427 by (small fry) fragment. Restriction enzyme Mbo I give the result of 1,324 and 624 by (large fry), 1,320 and 628 by (medium fry), 1,331 and 617 by (small fry). Digestion by using restriction enzyme HaeIII that shown different fragment length pattern between fish fry. The large and medium fry were indicated 4 fragment length (1,020538249and 141 bp), and (991556269and 131 bp) respectively, where as at small fish fry were indicated 3 fragment length i.e. 1,108554and 286 bp. Genetic distance value inter pop with different growth rate were calculated 0.007 between large and medium size, while between large and small were 0.249. The standarized genetic variance (Fst) value about 0.961. The conclusion of this study were confirmed that HaeIII restriction enzyme could digested of 16s rDNA fragment and suggested to use as variability growth genetic marker of brown marbled grouper fry. Keywords : 16s rDNA, variability growth genetic marker, E. fuscoguttatus, PCR-RFL