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Gene Transmission, Growth, and Exogeneous Growth Hormone Expression of G2 Transgenic Betta Fish (Betta imbellis)
Corresponding Author(s) : Alimuddin Alimuddin
Jurnal Ilmiah Perikanan dan Kelautan,
Vol. 13 No. 2 (2021): JURNAL ILMIAH PERIKANAN DAN KELAUTAN
Abstract
Highlight Research
- The F2 of GH-transgenic B. imbellis was successfully produced
- The transgene inheritance by the F2 fish was more than 90%
- The growth and body size of transgenic fish was significantly higher than control
- F2 fish reached a larger body size in a shorter period compared to the F1
Abstract
In our previous research, we had successfully produced G0 and G1 Pangasianodon hypophthalmus growth hormone (PhGH) transgenic B. imbellis, native ornamental betta from Indonesia, which its giant-sized variant has valuable price for the breeders. The G0 and G1 transgenic (TG) fish showed higher growth rate and body size compared to the non-transgenic (NT) fish. The study was aimed to produce and evaluate the consistencies of transgene transmission and expression in G2 generation. The growth rate and body size between TG and NT fish was also compared. The G2 generation was produced using crosses between TG and NT G1 fish: ♂TG í— â™€TG, ♂TG í— â™€NT, ♂NT í— â™€TG, and ♂NT í—♀ NT. Fish were reared for 12 weeks, and transgene detection was performed using the polymerase chain reaction method (PCR) on isolated DNA from the caudal fin clips. The endogenous and exogenous GH expression analysis was conducted using the quantitative real-time PCR (qPCR) method. The results showed that the inheritance of the GH transgene by the G2 fish was more than 90% in all transgenic crosses. Endogenous GH was expressed at the same levels in the brain of TG and NT fish, but the exogenous GH expression was highly detected only in the TG fish. The G2 transgenic fish had a higher specific growth rate, up to 31%, compared to the control. The body length of TG crosses were 23−35% higher and had 111−135% higher body weight compared to NT fish. These results showed a promising approached in mass-producing stable lines of giant-sized betta using the GH-transgenic technology.
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- Chen, J., Luo, Q., Bao, H., Liao, L., Li, Y., Zhu, Z., Wang, Y., & Hu, W. (2015). The integration characteristics of the exogenous growth hormone gene in a transgenic common carp (Cyprinus carpio L.) with fast-growth performance. Science Bulletin, 60(19):1654–1660.
- Crouse, C., Davidson, J., May, T., Summerfelt, S., & Good, C. (2021). Production of market-size European strain Atlantic salmon (Salmo salar) in land-based freshwater closed containment aquaculture systems. Aquacultural Engineering, 92:102138.
- Devlin, R. H. (2011). Cellular, molecular, genomics, and biomedical approaches | Growth Hormone Overexpression in Transgenic Fish. In Encyclopedia of Fish Physiology (Vol. 3, pp. 2016–2024). Elsevier Inc.
- Devlin, Robert H., Biagi, C. A., & Yesaki, T. Y. (2004). Growth, viability and genetic characteristics of GH transgenic coho salmon strains. Aquaculture, 236(1–4):607–632.
- Dewi, R. R. S. P. S., Alimuddin, A., Sudrajat, A. O., & Sumantadinata, K. (2012). The effectivity of gene transfer and expression of PhGH in stripped catfish (Pangasianodon hypophthalmus). Jurnal Riset Akuakultur, 7(2):171.
- Dunham, R. A., & Winn, R. N. (2014). Production of Transgenic Fish. In Transgenic Animal Technology: A Laboratory Handbook: Third Edition (pp. 305–334). Elsevier Inc.
- Fu, C., Li, D., Hu, W., Wang, Y., & Zhu, Z. (2007). Growth and energy budget of F2 "all-fish” growth hormone gene transgenic common carp. Journal of Fish Biology, 70(2):347–361.
- Fuentes, E. N., Valdés, J. A., Molina, A., & Björnsson, B. T. (2013). Regulation of skeletal muscle growth in fish by the growth hormone - Insulin-like growth factor system. General and Comparative Endocrinology, 192(June):136–148.
- Gong, Z., Wan, H., Tay, T. L., Wang, H., Chen, M., & Yan, T. (2003). Development of transgenic fish for ornamental and bioreactor by strong expression of fluorescent proteins in the skeletal muscle.
- Biochemical and Biophysical Research Communications, 308(1):58–63.
- Ignatz, E. H., Dumas, A., Benfey, T. J., Hori, T. S., Braden, L. M., Runighan, C. D., Rise, M. L., & Westcott, J. D. (2020). Growth performance and nutrient utilization of growth hormone transgenic female triploid Atlantic salmon (Salmo salar) reared at three temperatures in a land-based freshwater recirculating aquaculture system (RAS). Aquaculture, 519(September 2019):734896.
- Kobayashi, S. ichiro, Alimuddin, Morita, T., Miwa, M., Lu, J., Endo, M., Takeuchi, T., & Yoshizaki, G. (2007). Transgenic Nile tilapia (Oreochromis niloticus) over-expressing growth hormone show reduced ammonia excretion. Aquaculture, 270(1–4):427–435.
- Kurdianto, Alimuddin, Faridah, N., Yoshizaki, G., Nuryati, S., & Setiawati, M. (2016). Growth, Survival, and Body Composition of Transgenic Common Carp Cyprinus carpio 3rd Generation Expressing Tilapia Growth Hormone cDNA. HAYATI Journal of Biosciences, 23(3):150–154.
- Kusrini, E., Alimuddin, A., Zairin Jr, M., & Soelistyowati, D. T. (2018). Foreign growth hormone gene transmission and expression in F1 transgenic betta fish (Betta imbellis). Pakistan Journal of Biotechnology, 15(1):1–9.
- Kusrini, E., Alimuddin, A., Zairin, M. Jr., & Sulistyowati, D. T. (2016). Gene transfer on Betta imbellis through transfection method with different dna concentration. Indonesian Aquaculture Journal, 11(1):1.
- Kusrini, E., Zairin, M.Jr., & Sulistyowati, D.T. (2016). Identification of founder generation of betta fish (Betta imbellis) growth hormone transgenic. Jurnal Riset Akuakultur, 11(3):197–205.
- Ledford, H. (2015). Transgenic salmon leaps the dinner table. Nature, 527:417–418.
- Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 25(4):402–408.
- Nam, Y. K., Noh, J. J., Cho, Y. S., Cho, H. J., Cho, K.-N., Kim, C. G., & Kim, D. S. (2001). Dramatically accelerated growth and extraordinary gigantism of transgenic mud loach Misgurnus mizolepis. Transgenic Research, 10:353–362.
- Nipkow, M., Wirthgen, E., Luft, P., Rebl, A., Hoeflich, A., & Goldammer, T. (2018). Characterization of igf1 and igf2 genes during maraena whitefish (Coregonus maraena) ontogeny and the effect of temperature on embryogenesis and igf expression. Growth Hormone and IGF Research, 40(December 2017):32–43.
- Panijpan, B., Kowasupat, C., Laosinchai, P., Ruenwongsa, P., Phongdara, A., Senapin, S., Wanna, W., Phiwsaiya, K., Kühne, J., & Fasquel, F. (2014). Southeast Asian mouth-brooding Betta fighting fish (Teleostei: Perciformes) species and their phylogenetic relationships based on mitochondrial COI and nuclear ITS1 DNA sequences and analyses. Meta Gene, 2:862–879.
- Panijpan, B., Sriwattanarothai, N., & Laosinchai, P. (2020). Wild Betta fighting fish species in Thailand and other Southeast Asian countries. ScienceAsia, 46(4):382–391.
- Rezaei, M., Basiri, M., Hasani, S.-N., Asgari, B., Kashiri, H., Shabani, A., & Baharvand, H. (2019).
- Establishment of a Transgenic Zebrafish Expressing GFP in the Skeletal Muscle as an Ornamental Fish. Galen Medical Journal, 8(January):1068.
- Triantaphyllopoulos, K. A., Cartas, D., & Miliou, H. (2020). Factors influencing GH and IGF-I gene expression on growth in teleost fish: how can aquaculture industry benefit? Reviews in Aquaculture, 12(3):1637–1662.
References
Chen, J., Luo, Q., Bao, H., Liao, L., Li, Y., Zhu, Z., Wang, Y., & Hu, W. (2015). The integration characteristics of the exogenous growth hormone gene in a transgenic common carp (Cyprinus carpio L.) with fast-growth performance. Science Bulletin, 60(19):1654–1660.
Crouse, C., Davidson, J., May, T., Summerfelt, S., & Good, C. (2021). Production of market-size European strain Atlantic salmon (Salmo salar) in land-based freshwater closed containment aquaculture systems. Aquacultural Engineering, 92:102138.
Devlin, R. H. (2011). Cellular, molecular, genomics, and biomedical approaches | Growth Hormone Overexpression in Transgenic Fish. In Encyclopedia of Fish Physiology (Vol. 3, pp. 2016–2024). Elsevier Inc.
Devlin, Robert H., Biagi, C. A., & Yesaki, T. Y. (2004). Growth, viability and genetic characteristics of GH transgenic coho salmon strains. Aquaculture, 236(1–4):607–632.
Dewi, R. R. S. P. S., Alimuddin, A., Sudrajat, A. O., & Sumantadinata, K. (2012). The effectivity of gene transfer and expression of PhGH in stripped catfish (Pangasianodon hypophthalmus). Jurnal Riset Akuakultur, 7(2):171.
Dunham, R. A., & Winn, R. N. (2014). Production of Transgenic Fish. In Transgenic Animal Technology: A Laboratory Handbook: Third Edition (pp. 305–334). Elsevier Inc.
Fu, C., Li, D., Hu, W., Wang, Y., & Zhu, Z. (2007). Growth and energy budget of F2 "all-fish” growth hormone gene transgenic common carp. Journal of Fish Biology, 70(2):347–361.
Fuentes, E. N., Valdés, J. A., Molina, A., & Björnsson, B. T. (2013). Regulation of skeletal muscle growth in fish by the growth hormone - Insulin-like growth factor system. General and Comparative Endocrinology, 192(June):136–148.
Gong, Z., Wan, H., Tay, T. L., Wang, H., Chen, M., & Yan, T. (2003). Development of transgenic fish for ornamental and bioreactor by strong expression of fluorescent proteins in the skeletal muscle.
Biochemical and Biophysical Research Communications, 308(1):58–63.
Ignatz, E. H., Dumas, A., Benfey, T. J., Hori, T. S., Braden, L. M., Runighan, C. D., Rise, M. L., & Westcott, J. D. (2020). Growth performance and nutrient utilization of growth hormone transgenic female triploid Atlantic salmon (Salmo salar) reared at three temperatures in a land-based freshwater recirculating aquaculture system (RAS). Aquaculture, 519(September 2019):734896.
Kobayashi, S. ichiro, Alimuddin, Morita, T., Miwa, M., Lu, J., Endo, M., Takeuchi, T., & Yoshizaki, G. (2007). Transgenic Nile tilapia (Oreochromis niloticus) over-expressing growth hormone show reduced ammonia excretion. Aquaculture, 270(1–4):427–435.
Kurdianto, Alimuddin, Faridah, N., Yoshizaki, G., Nuryati, S., & Setiawati, M. (2016). Growth, Survival, and Body Composition of Transgenic Common Carp Cyprinus carpio 3rd Generation Expressing Tilapia Growth Hormone cDNA. HAYATI Journal of Biosciences, 23(3):150–154.
Kusrini, E., Alimuddin, A., Zairin Jr, M., & Soelistyowati, D. T. (2018). Foreign growth hormone gene transmission and expression in F1 transgenic betta fish (Betta imbellis). Pakistan Journal of Biotechnology, 15(1):1–9.
Kusrini, E., Alimuddin, A., Zairin, M. Jr., & Sulistyowati, D. T. (2016). Gene transfer on Betta imbellis through transfection method with different dna concentration. Indonesian Aquaculture Journal, 11(1):1.
Kusrini, E., Zairin, M.Jr., & Sulistyowati, D.T. (2016). Identification of founder generation of betta fish (Betta imbellis) growth hormone transgenic. Jurnal Riset Akuakultur, 11(3):197–205.
Ledford, H. (2015). Transgenic salmon leaps the dinner table. Nature, 527:417–418.
Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 25(4):402–408.
Nam, Y. K., Noh, J. J., Cho, Y. S., Cho, H. J., Cho, K.-N., Kim, C. G., & Kim, D. S. (2001). Dramatically accelerated growth and extraordinary gigantism of transgenic mud loach Misgurnus mizolepis. Transgenic Research, 10:353–362.
Nipkow, M., Wirthgen, E., Luft, P., Rebl, A., Hoeflich, A., & Goldammer, T. (2018). Characterization of igf1 and igf2 genes during maraena whitefish (Coregonus maraena) ontogeny and the effect of temperature on embryogenesis and igf expression. Growth Hormone and IGF Research, 40(December 2017):32–43.
Panijpan, B., Kowasupat, C., Laosinchai, P., Ruenwongsa, P., Phongdara, A., Senapin, S., Wanna, W., Phiwsaiya, K., Kühne, J., & Fasquel, F. (2014). Southeast Asian mouth-brooding Betta fighting fish (Teleostei: Perciformes) species and their phylogenetic relationships based on mitochondrial COI and nuclear ITS1 DNA sequences and analyses. Meta Gene, 2:862–879.
Panijpan, B., Sriwattanarothai, N., & Laosinchai, P. (2020). Wild Betta fighting fish species in Thailand and other Southeast Asian countries. ScienceAsia, 46(4):382–391.
Rezaei, M., Basiri, M., Hasani, S.-N., Asgari, B., Kashiri, H., Shabani, A., & Baharvand, H. (2019).
Establishment of a Transgenic Zebrafish Expressing GFP in the Skeletal Muscle as an Ornamental Fish. Galen Medical Journal, 8(January):1068.
Triantaphyllopoulos, K. A., Cartas, D., & Miliou, H. (2020). Factors influencing GH and IGF-I gene expression on growth in teleost fish: how can aquaculture industry benefit? Reviews in Aquaculture, 12(3):1637–1662.