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All Journal Journal of Tropical Biodiversity and Biotechnology Agrin : Jurnal Penelitian Pertanian Journal of Tropical Crop Science
PUTRANTO, Riza Arief
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science Immunology & microbiology

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Characterization of a Drought-Inducible Dehydrin Promoter from Sugarcane (Saccharum officinarum L.) in Tobacco (Nicotiana tabacum L.) Iskandar, Hayati Minarsih; Suhandono, Sonny; Pambudi, Jembar; Kristianti, Tati; Putranto, Riza Arief; Mose, Windi; Sustiprijatno, Sustiprijatno
Journal of Tropical Crop Science Vol 7 No 01 (2020): Journal of Tropical Crop Science
Publisher : Department of Agronomy and Horticulture, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (910.185 KB) | DOI: 10.29244/jtcs.7.01.28-36

Abstract

Dehydrin (DHN) is known to play an important role in plant response and adaptation to abiotic stresses (drought, high salinity, cold, heat, etc.). Previous research reported the increased expression of DHN in sugarcane stems exposed to drought stress for 15 days which may be controlled by its corresponding stress inducible promoter. The DHN promoter was succesfully isolated from sugarcane variety PSJT 941 (Pr-1DHNSo) and was cloned to pBI121 expression vector fused to a β-glucuronidase (GUS) reporter gene. The aim of this research was the functional testing of the Pr-1DHNSo promoter through transformation into tobacco plant treated with in vitro drought stress. Genetic transformation of Pr-1DHNSo construct was conducted by Agrobacterium tumefaciens. The transformed tobacco was then subjected to drought stress treatment using 40% PEG 6000 for five sequential incubations (0, 12, 24, 48 and 72 hours). The GUS assay reveal that the transformed tobacco treated with drought stress showed a blue color denoting GUS activity in leaf, stem and root tissues and this expression increased along with the length of the drought treatment. The analysis of gusA gene using real time-qPCR normalized to the L25 reference gene also showed that the expression increased in line with the length of time of drought stress. The results presented in this study indicated that the Pr-1DHNSo promoter from sugarcane was expressed and induced by drought stress treatment in tobacco.
Sequence-Structure Comparative and Network-Based Prediction of Drought Gene Candidate Regulator in Elaeis guineensis Permatasari, Galuh Wening; Putranto, Riza Arief; Mardhika, Larasati Dena; Aksa, Annisa Aulia; Setiawati, Yuli; Minarsih, Hayati; Riyadi, Imron; Ernayunita, Ernayunita
Journal of Tropical Biodiversity and Biotechnology Vol 9, No 3 (2024): September
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jtbb.90808

Abstract

Drought poses a significant threat to global food security, particularly impacting crops like oil palm. Selecting genes for genome editing to enhance drought tolerance presents formidable challenges. To ensure that the target gene is chosen correctly and results in the desired character, a pilot study is necessary to determine the target gene for knockout. Two genes drought-related, AtBRL3 and AtOST2, were scrutinized in this context. Aligned with the Elaeis guineensis genome, their neighbouring proteins and gene ontology were analysed to identify potential targets for genome editing. AtBRL3, identified as BRL1 (XP_010913986.1) in E. guineensis, exhibited 58.48% identity and 100% coverage. It interacts with 12 nodes, including BIR1, BRI1, and AT2G20050, crucial for signalling pathways and cellular responses. Molecular function analysis revealed kinase activity. AtOST2 showed high similarity to plasma membrane ATPase/HA1 (XP_010913679.1) in E. guineensis, with 87.46% identity and 100% query cover. It correlated with 14 genes associated with ABA stimulus, stomatal movement, and hormone response. EgBRL1 and EgHA1, resembling AtBRL3 and AtOST2, respectively, emerge as promising targets for developing drought-tolerant oil palm cultivars through gene editing. Nonetheless, further validation through in vitro gRNA target selection and in vivo conversion of OST2/BRL3-containing plasmids in oil palm calluses is indispensable to demonstrate their efficacy in conferring novel drought resistance traits. 
OPTIMASI DAN EFISIENSI TEKNIK ISOLASI RNA DAUN DAN AKAR KELAPA SAWIT (Elaeis guineensis Jacq) Sari, Dini Astika; Martiansyah, Irfan; Mukmin, Restu Prasetya; Hadi, Sapto Nugroho; Syahputra, Indra; Afandi, Dadang; Putranto, Riza Arief
Agrin Vol 23, No 2 (2019): Agrin
Publisher : Jenderal Soedirman University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (191.431 KB) | DOI: 10.20884/1.agrin.2019.23.2.500

Abstract

Tanaman kelapa sawit memiliki kandungan polisakarida dan polifenol yang tinggi. Kontaminasi polisakarida dan polifenol menyebabkan sulitnya proses isolasi RNA dari jaringan tanaman kelapa sawit. Penelitian ini dilakukan untuk mengoptimasi beberapa protokol isolasi RNA tanaman kelapa sawit yang efektif dan efisien. Penelitian dilaksanakan selama tiga bulan, pada bulan Oktober-Desember 2018 di Laboratorium Biokimia dan Biologi Molekuler, Pusat Penelitian Bioteknologi dan Bioindustri Indonesia (PPBBI), Jalan Taman Kencana No. 1 Bogor 16128. Penelitian dilakukan dengan melaksanakan teknik isolasi RNA menggunakan tiga protokol, yaitu modifikasi Cetyl Trimethyl Ammonium Bromide (CTAB), kit isolasi RNA RNeasy Plant Mini Kit (Qiagen), dan kit isolasi NucleoSpin RNAPlant (Macherey-Nagel). Sampel yang digunakan adalah daun dan akar tanaman kelapa sawit berumur kurang dari tiga bulan dengan bobot 0,1 gram dan 2,5 gram yang disesuaikan untuk tiap protokol. Variabel yang diamati adalah konsentrasi (ng/µl), kemurnian (rasio A260/A280 dan A260/A230), dan pita RNA pada elektroforesis gel agarosa. Hasil penelitian menunjukkan, RNA total hasil isolasi protokol NucleoSpin RNAPlant (Macherey-Nagel) memiliki kualitas paling tinggi. Konsentrasi RNA total daun dan akar kelapa sawit yang didapatkan melalui protokol NucleoSpin RNAPlant (Macherey-Nagel) sebesar 338 ng/µl dan 184,4 ng/µl dengan rasio A260/A280 RNA total daun dan akar kelapa sawit sebesar 2,13 dan 2,18 serta rasio A260/A230 sebesar 2,09 dan 2,20. Hasil elektroforesis gel agarosa menunjukkan integritas yang bagus dari RNA total hasil isolasi RNeasy Plant Mini Kit (Qiagen) dan NucleoSpin RNAPlant (Macherey-Nagel), namun terdapat kontaminasi dan smear pada RNA total hasil isolasi CTAB modifikasi 1 dan 2. Kata kunci: kelapa sawit, isolasi RNA, spektrofotometer, elektroforesis gel agarosa
Co-Authors Afandi, Dadang Aksa, Annisa Aulia Ernayunita, Ernayunita Hayati Minarsih Imron Riyadi Indra Syahputra Iskandar, Hayati Minarsih Mardhika, Larasati Dena MARTIANSYAH, Irfan Mose, Windi Mukmin, Restu Prasetya Pambudi, Jembar PERMATASARI, Galuh Wening Sapto Nugroho Hadi Sari, Dini Astika Suhandono, Sonny Sustiprijatno Sustiprijatno, Sustiprijatno TATI KRISTIANTI Yuli Setiawati, Yuli