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CORRELATION OF MICROSATELLITE DNA MARKERS WITH GROWTH TRAITS IN STRIPED CATFISH (Pangasianodon hypophthalmus) Huria Marnis; Evi Tahapari; Jadmiko Darmawan
Indonesian Aquaculture Journal Vol 13, No 2 (2018): (December, 2018)
Publisher : Center for Fisheries Research, Agency for Marine and Fisheries Research and Human Resource

Marker-assisted selection in genetic improvement of striped catfish is useful in the breeding program of the fish. Five microsatellite markers were characterized in the largest (4.03 kg ± 1.67 kg) and smallest (1.41 kg ± 0.22 kg) individuals. Five polymorphic loci were then used to genotype 160 individuals and the associations between their genotypes and growth traits were examined. The result showed that twenty-seven alleles were detected in striped catfish. The number of alleles per locus (NA) ranged from 4 to 7, with an average of 5.4 alleles per locus. The effective number of alleles per locus (NE) ranged from 3.940 to 6.939, with an average of 5.32 alleles per locus. HO and HE ranged from 0.125 to 0.944 (mean value of 0.472) and from 0.564 to 0.775 (mean value of 0.697), respectively. PIC ranged from 0.573 to 0.799 (mean value of 0.706), showing that they were highly polymorphic loci. Only one microsatellites loci (Pg13) that showed significant differences (P<0.01) in the associations between their genotypes and growth traits, while Pg3 and Pg14 were significantly associated with the standard length (P<0.01) and body weight (P<0.05). However, the Pg1 and Pg2 were not significantly associated with the body weight and standard length. Four genotypes of three loci were positively correlated with the growth traits (body weight and standard length) i.e. genotypes 194/194 for Pg3, 227/227 and 229/229 for Pg13, 279/279 for Pg14. These four genotypes can be used to identify growth traits in the molecular marker-based selection of a breeding program.

THE ABILITY OF FAST-GROWING TRANSGENIC AFRICAN CATFISH (Clarias gariepinus) ON PREDATOR AVOIDANCE Huria Marnis; Bambang Iswanto; imron imron; selny Febrida; Raden Roro Sri Pudji Sinarni Dewi
Indonesian Aquaculture Journal Vol 11, No 1 (2016): (June 2016)
Publisher : Center for Fisheries Research, Agency for Marine and Fisheries Research and Human Resource

Research Institute for Fish Breeding has produced transgenic African catfish (Clarias gariepinus) containing stripped catfish growth hormone gene (PccBA-PhGH) with growth 19.86% faster than that of non-transgenic fish. This fish has high potential to be released and utilized for fish farming sector to increase national production. However, there is not yet information about environmental risk of this fish. One of the major fitness traits determining potential environmental risk is predator avoidance. This study aimed to determine the predator avoidance ability of transgenic African catfish in an experimental laboratory condition. In this study, thirty five individuals each of transgenic and non-transgenic with body weight of about 0.1 ± 0.019 g were communally stocked in 60 cm x 40 cm x 40 cm aquarium with limited feeding frequency (ad libitum twice a day). One day after the fish were stocked, the predators were added to each aquarium. The non-transgenic and transgenic with body weight of 1.0 ± 0.024 g were stocked as predators as many as five individual in each aquarium. After approximately two weeks of predation, all remaining fish were collected for transgenic verification by PCR method. Genomic DNA was isolated from fin tissue of individually survivors. The results of this study showed that the transgenic fish had worse predator avoidance and lower cannibal than non-transgenic (P<0.05). There was no significant difference in growth performance between transgenic and non-transgenic African catfish (P>0.05) in limited food. The transgenic fish may have lower fitness than non-transgenic.

GENETIC PERFORMANCE OF THE STRIPED CATFISH (Pangasianodon hypophthalmus Sauvage, 1878) POPULATION DERIVED FROM SELECTIVE BREEDING Huria Marnis; Evi Tahapari; Jadmiko Darmawan
Indonesian Aquaculture Journal Vol 14, No 1 (2019): (June, 2019)
Publisher : Center for Fisheries Research, Agency for Marine and Fisheries Research and Human Resource

Selective breeding to improve growth trait in striped catfish (Pangasianodon hypophthalmus) need genetic performance data. This study was carried out to evaluate genetic performance of different generations of the striped catfish population derived from selective breeding. Fifty fish of each population from four generations selected striped catfish was analyzed using five microsatellite loci (Pg-1, Pg-2, Pg-3, Pg-13, and Pg-14). Microsatellite allele data were analyzed using Microsoft Excel, Arlequin, and Fstat software. A neighborhood joining dendrogram was constructed based on Nei’s distance (Da) matrix with 1,000 bootstrap replications using MEGA7 software. The result showed that totally 31.5 exist, ranged from 4-7. Number of allele was ranged from 5.0 to 5.2, and polymorphic data was from 0.45-0.60. There were some exception, such as the allele of the loci Pg-2—194 bp, Pg-13—227 bp, Pg-13—229 bp, and Pg-14—279 bp; their gene frequencies were increased by generation. Further analyses indicated that most genetic variations arise from individuals within populations (approximately 57.10%). The founder generation closely related to G-0 generation. Likewise, G-1 generation closely related to G-2 generation. This result indicated that selection activity had a very significant impact on the genetic improvement of the selected population.

IDENTIFIKASI ZIGOSITAS IKAN LELE (Clarias gariepinus) TRANSGENIK F-2 YANG MEMBAWA GEN HORMON (PhGH) DENGAN MENGGUNAKAN METODE REALTIME-qPCR Huria Marnis; Bambang Iswanto; Romy Suprapto; Imron Imron; Raden Roro Sri Pudji Sinarni Dewi
Jurnal Riset Akuakultur Vol 11, No 1 (2016): (Maret 2016)
Publisher : Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

Produktivitas ikan budidaya dapat ditingkatkan melalui teknologi transgenesis. Populasi ikan lele transgenik cepat tumbuh telah dihasilkan dan karakter biologisnya telah diketahui. Namun informasi zigositas ikan lele transgenik perlu ditelaah lebih lanjut. Penelitian ini bertujuan untuk mengidentifikasi zigositas ikan lele transgenik F-2. Zigositas ikan lele transgenik diidentifikasi dengan menggunakan metode real-time qPCR (RT-qPCR) dan uji progeni. Identifikasi zigositas melalui uji progeni, dilakukan dengan mendeteksi transgen (PhGH) pada individu-individu F-3 hasil persilangan transgenik F-2 dengan non-transgenik. Hasil penelitian menunjukkan bahwa zigositas pada ikan lele transgenik F-2 dapat diidentifikasi dengan menggunakan metode RT-qPCR. Semua ikan transgenik F-2 adalah heterozigot, dengan nilai 2-Ct yang hampir sama tiap individu F-2, yaitu berkisar 0,80-0,99. Identifikasi zigositas dengan metode RT-qPCR menunjukkan hasil yang sama dengan uji progeni, semua transgenik F-2 tidak menghasilkan 100% anakan F-3 positif transgen. Pada uji progeni, transmisi transgen pada penelitian ini tidak mengikuti hukum segregasi Mendel, dengan kisaran sebesar 5%-40%.Fish farming productivity can be increased by transgenesis technology. On the previous study, transgenic African catfish population fast growing has been produced and its biological characters has been known. However information of transgenic zygosity of catfish should be examined. The aim of this study was to identify the zygosity of F-2 transgenic African catfish. The zygosity of F-2 transgenic was identified by real time-qPCR (RT-qPCR) method and progeny test. Further, identification of zygosity F-2 transgenic African catfish was confirmed by progeny test, while F-2 transgenic African catfish was mated with non-transgenic. Identification of zygosity F-2 transgenic was conducted by detection PhGH gene (transgene) in F-3 transgenic African catfish population. Transgene transmission was evaluated by PCR method. The result showed that the zygosity F-2 transgenic African catfish could be identified by RT-qPCR method. All F-2 transgenic African catfish were heterozygous, where as the 2-Ct value was almost same for all individual, which ranges from 0.80 to 0.99. The result of zygosity identification using RT-qPCR method was as same as that of progeny test. In the progeny test, transgene transmission in this study was non-Mendelian segregation, with ranges of 5%-40%.

TRANSMISI GEN PhGH DAN PERFORMA PERTUMBUHAN IKAN LELE AFRIKA (Clarias gariepinus) TRANSGENIK GENERASI KETIGA Huria Marnis; Bambang Iswanto; Selny Febrida; Imron Imron; Raden Roro Sri Pudji Sinarni Dewi
Jurnal Riset Akuakultur Vol 11, No 3 (2016): (September 2016)
Publisher : Pusat Riset Perikanan, Badan Riset dan Sumber Daya Manusia Kelautan dan Perikanan

Penelitian ini bertujuan untuk mengevaluasi transmisi transgen PhGH dan performa ikan lele Afrika transgenik generasi ketiga (F3) yang meliputi pertumbuhan, rasio efisiensi konversi pakan, konsentrasi hormon pertumbuhan dan hormon IGF-I. Ikan lele transgenik F3 telah diproduksi dengan meyilangkan ikan lele transgenik F2 dengan non-transgenik. Deteksi transgen (PhGH) dilakukan menggunakan metode PCR. Analisis hormon pertumbuhan dan hormon insuline-like growth factor (IGF-I) menggunakan sampel serum darah dan metode enzyme linked immunosorbent assay (ELISA). Hasil penelitian menunjukkan bahwa ikan lele transgenik F3 yang digunakan pada pengujian ini terdeteksi positif membawa transgen dengan ukuran fragmen gen sebesar 1.500 bp. Transmisi transgen dari induk F2 ke F3 berkisar 0-75%. Pertumbuhan bobot populasi ikan lele transgenik F3 lebih tinggi 51,26% dibandingkan dengan ikan lele non-transgenik (P<0,05). Pertumbuhan bobot populasi ikan transgenik mencapai 484±60,3 g, sedangkan pertumbuhan bobot ikan non-transgenik 319,98±65,3 g. Nilai rasio konversi pakan ikan lele transgenik F3 sebesar 0,89 sedangkan non-transgenik 1,30. Hal ini menunjukkan bahwa efisiensi pakan ikan lele transgenik F3 lebih tinggi dibandingkan dengan ikan non-transgenik (P<0,05). Ikan lele transgenik mempunyai konsentrasi hormon pertumbuhan (5,67±2,65 ng/mL) yang lebih tinggi (P<0,05) jika dibandingkan dengan ikan lele non-transgenik (3,00±1,41 ng/mL). Ikan lele transgenik juga memiliki kandungan hormon IGF-I (6,63±0,11 ng/mL) lebih tinggi (P<0,05) dibandingkan dengan ikan lele non-transgenik (5,38±0,63 ng/mL). Tingginya konsentrasi hormon pertumbuhan dan hormon IGF-I dapat mewakili performa pertumbuhan dan efisiensi penggunaan pakan pada ikan lele transgenik.The aim of this study was to determine the transmission of a transgene (PhGH) and to evaluate the performance of F3 transgenic African catfish, such as body weight, feed conversion ratio (FCR), growth hormone and IGF-I hormone profile. F3 transgenic were produced by mating F2 transgenic with non-transgenic fish. Detection of transgene was performed using PCR method. Analysis of the growth hormone and the insulin-like growth factor-I (IGF-I) hormone were conducted by enzyme linked immunosorbent assay (ELISA) method using serum samples. The results showed that the transgenic catfish F3 was positive carrying the transgene 1.500 bp. Transgene transmission from F2 to F3 ranged zero to 75%. The performance of F3 transgenic African catfish was significantly better 51.26% than that the non-transgenic (P<0.05). The body weight of transgenic population (484±255.25 g) was higher than that non-transgenic (319.98±165.27 g). FCR of transgenic fish (0.89) was lower than that non-transgenic (1.30). The growth hormone level of transgenic (5.67±2.65 ng/mL) was higher than that non-transgenic (3.00±1.41 ng/mL), IGF-I hormone level of F3 transgenic (6.63±0.11 ng/mL) was also higher than that non-transgenic (5.38±0.63 ng/mL). High level of growth hormone and IGF-I hormone represented both growth performance and efficiency of feed utilization of transgenic African catfish.

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