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Huria Marnis

Balai Penelitian Pemuliaan Ikan, Jl. Raya 2 Sukamandi-Pantura Patok Beusi, Sukamandi-Subang

Author-ID : 3672457

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science

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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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