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All Journal HAYATI Journal of Biosciences Vegetalika BIOTROPIA - The Southeast Asian Journal of Tropical Biology Indonesian Journal of Biotechnology Indonesian Journal of Emerging Trends in Community Empowerment
Ence Darmo Jaya Supena
Department Of Biology, Faculty Of Mathematics And Natural Sciences, IPB University, Bogor 16680, Indonesia
Agriculture, Biological Sciences & Forestry Humanities Biochemistry, Genetics & Molecular Biology Chemistry Earth & Planetary Sciences Education Immunology & microbiology Library & Information Science Materials Science & Nanotechnology Veterinary

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Induction of Somatic Embryogenesis in Sengon (Falcataria moluccana) With Thidiazuron and Light Treatments Ari Sunandar; . Dorly; Ence Darmo Jaya Supena
HAYATI Journal of Biosciences Vol. 24 No. 2 (2017): April 2017
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (992.467 KB) | DOI: 10.4308/hjb.24.2.105

Abstract

Falcataria moluccana is important for reforestation and afforestation in Indonesia. However, epidemic of gall rust disease in F. moluccana plantations decreases its productivity. Genetic engineering is an alternative solution to against gall rust disease. Somatic embryogenesis is an efficient in vitro plant regeneration for successful plant improvement through genetic engineering. The objective of this study was to investigate the effect of thidiazuron and light treatments on the induction of somatic embryogenesis of F. moluccana. The effects of thidiazuron concentration (5, 10 or 15 μM) and light (continuous light, 7 days of dark followed by light, or continuous dark) on the induction of somatic embryogenesis in leaf explants were assessed. The highest production of somatic embryos was obtained in 5 μM thidiazuron and dark treatments for 7 days followed by light in Murashige and Skoog medium supplemented with 1.2 g/L proline. Histological analysis in globular and cotyledon stages confirmed that cells had progressed to secondary somatic embryogenesis. This research needs more improvements to become a successful and efficient somatic embryogenesis method and as a potential method for successful plant improvement through genetic engineering in F. moluccana.
Agrobacterium-MEDIATED GENETIC TRANSFORMATION OF SEAWEED Kappaphycus alvarezii USING Gα GENE AND CALLUS CULTURES Erina Sulistiani; Suharsono Suharsono; Ence Darmo Jaya Supena; Miftahudin Miftahudin
BIOTROPIA - The Southeast Asian Journal of Tropical Biology Vol. 26 No. 1 (2019)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (133.176 KB) | DOI: 10.11598/btb.2019.26.1.989

Abstract

Cottonii seaweed (Kappaphycus alvarezii Doty) is one of the most important commercial sources of carrageenans which are widely used in the pharmaceuticals and food industries. A problem in the cultivation of this seaweed is the ice-ice disease, which is caused by extreme changes in environmental conditions such as temperature and seawater salinity. Gene transformation to produce Cottonii seaweed transgenics that are tolerant to environmental stress is a potential solution to this problem. Gα gene encodes for the heterotrimeric G protein α subunit is a gene that plays a role in tolerance to biotic and abiotic environmental stresses. This study aimed to: (a) introduce the Gα gene into the callus cells of K. alvarezii and regenerate transformed callus cells to transgenic plantlets; (b) determine the appropriate concentration of acetosyringone and Agrobacterium tumefaciens strain for gene transfer into the callus of K. alvarezii. The callus cells of K. alvarezii were transformed using Agrobacterium tumefaciens strains LBA4404 and EHA105 carrying the expression vector pGWB502-Gα with a CaMV-35S promoter. The calli and A. tumefaciens were co-cultivated in several concentrations of acetosyringone (20, 40, 60 mg/L). The regeneration of transformed callus cells into transgenic plantlets was successfully performed using the somatic embryogenesis technique. The results showed that the highest percentage of putative transgenic micropropagule formation occurred at the 20-40 mg/L concentration of acetosyringone. Polymerase chain reaction (PCR) analysis on the twenty transgenic plantlets indicated that the Gα gene was successfully introduced into the genomic DNA of all of them. The highest transformation efficiency was in the co-cultivation treatment of 20-40 mg/L acetosyringone (22-28%). The transformation efficiency produced by Agrobacterium tumefaciens EHA105 (23%) was not significantly different from that produced by the LBA4404 (15%).
Detection and quantification of splicing variants of Hd3a gene in oil palm Aqwin Polosoro; Wening Enggarini; Kusumawaty Kusumanegara; Toto Hadiarto; Miftahudin Miftahudin; Ence Darmo Jaya Supena
Indonesian Journal of Biotechnology Vol 29, No 1 (2024)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.88327

Abstract

Alternative splicing is a complex process that contributes to the generation of diverse mRNA and protein isoforms, including in oil palm (Elaeis guineensis). Despite their importance, many functions of alternative splicing genes remain poorly characterized. This study aims to investigate splicing variants of gene encoding Heading date 3a in E. guineensis (EgHd3a) using the GenBank database and ClustalW algorithm. To ensure the data accuracy and reliability of design isoform‐ specific primers, special emphasis is given to primer design techniques and validation using polymerase chain reaction (PCR) and quantitative real‐time (qRT)‐PCR analysis. The designed primers demonstrated high specificity and discrimination between mRNA specimens. Nucleotide variations at the 3’‐end influenced the specificity of primers with the addition of GC composition. Furthermore, qRT‐PCR analysis revealed a strong correlation between Ct values and gene concentration for the isoforms which indicates a reliable amplification of EgHd3a. Although two isoforms, Hd3a‐X2 and Hd3a‐X3, showed slightly higher than acceptable PCR efficiency values, caution is advised to prevent non‐specific amplification. Despite the challenge posed by the limitation of primer positioning due to alternative splicing, the chosen primer proved optimal for analysis. This study highlights the importance of considering alternative splicing in gene quantification experiments and provides insights into the critical steps, methods, and quality control measures necessary for accurately detecting alternative splicing events, contributing to understanding this complex biological process.
Optimasi Ekstraksi RNA dan Teknik Kloning: Studi Kasus Kloning Gen Heading Date 3a pada Kelapa Sawit Aqwin Polosoro; Wening Enggarini; Kusumawaty Kusumanegara; Toto Hadiarto; Miftahudin Miftahudin; Ence Darmo Jaya Supena
Vegetalika Vol 13, No 2 (2024)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/veg.92085

Abstract

Pembungaan memegang peranan penting bagi tumbuhan karena memfasilitasi rekombinasi genetik, sehingga mendukung perkembangan keragaman genetik yang penting. Keluarga protein phosphatidylethanolamine binding proteins (PEBP) memainkan peran penting dalam mengatur waktu pembungaan dan dormansi benih di beragam spesies tanaman. Penelitian ini bertujuan untuk merancang vektor biner dengan membangun pCAMBIA1300 yang menggabungkan rangkaian gen EgHd3a dari kelapa sawit. Proses konstruksi gen meliputi ekstraksi RNA, sintesis cDNA, amplifikasi gen EgHd3a, kloning gen menjadi vektor kloning, subkloning ke dalam vektor biner pCAMBIA1300, dan diakhiri dengan validasi gen melalui analisis sekuens. Pada ekstraksi RNA, metode PCL-Chisam telah terbukti efektif melalui ekstraksi berulang, meningkatkan kualitas dan kuantitas total RNA. Dalam proses kloaning, metode konvensional menghadapi tantangan dalam memilih lokasi pembelahan yang tepat. Untuk mengatasi kendala ini, penggunaan enzim dengan overhang yang kompatibel diusulkan sebagai solusi potensial. Secara khusus, penggantian BamHI dari BglII telah secara efektif mengatasi tantangan ini. Konfirmasi integrasi fragmen gen ke dalam plasmid pCAMBIA1300 dicapai melalui pengurutan. Meskipun perbedaan diidentifikasi dalam rangkaian EgHd3a-2, perubahan ini tidak berdampak pada asam amino yang dikodekan, sehingga menjaga integritas rangkaian protein
Workshop Manajemen dan Substansi Pengelolaan Jurnal Ilmiah Menuju Jurnal Terakreditasi Nasional dan Internasional Muhamad Syazali; Hayatul Khairul Rahmat; I Dewa Ketut Kerta Widana; Ence Darmo Jaya Supena; Rahmat Basuki; Miftahul Huda Fendiyanto; Adhi Kusumadjati; Mo Awwanah
Indonesian Journal of Emerging Trends in Community Empowerment Vol. 1 No. 1 (2023): Juni
Publisher : Perkumpulan Ahli Bimbingan dan Konseling Islam Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Workshop on the management and substance of managing scientific journals towards national and international accredited journals aim to gain an understanding of how the management and substance of management of scientific journals towards national and international accredited journals within the Faculty of Military Mathematics and Natural Sciences, the Republic of Indonesia Defense University. The procedure for implementing activities starts from the preparation, implementation, and closing stages. This activity is very useful for improving the quality of scientific journal management at the Faculty of Military Mathematics and Natural Sciences, the Republic of Indonesia Defense University towards national and international accredited journals. This can also be seen from the response and enthusiasm of the participants who asked when carrying out management workshop activities and the substance of managing scientific journals towards this national and international accredited journal.
Co-Authors Adhi Kusumadjati Aqwin Polosoro Aqwin Polosoro Dorly Dorly Erina Sulistiani Hayatul Khairul Rahmat I Dewa Ketut Kerta Widana Kusumawaty Kusumanegara Kusumawaty Kusumanegara Miftahudin . Miftahudin Miftahudin Miftahudin Miftahudin Miftahul Huda Fendiyanto Mo Awwanah Muhamad Syazali Rahmat Basuki Suharsono Suharsono Sunandar, Ari Toto Hadiarto Toto Hadiarto Wening Enggarini Wening Enggarini