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All Journal Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Jurnal AgroBiogen BERITA BIOLOGI
Atmitri Sisharmini
Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian, Jl. Tentara Pelajar No. 3A, Bogor 16111 Telp. (0251) 8337975
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Economics, Econometrics & Finance

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Delivering of Over-Expression Construct OsWRKY76 Candidate Gene in Rice cv. Nipponbare through Agrobacterium tumefaciens Apriana, Aniversari; Sisharmini, Atmitri; Enggarini, Wening; Sudarsono, Sudarsono; Khumaida, Nurul; Trijatmiko, Kurniawan R.
Jurnal AgroBiogen Vol 7, No 1 (2011): Jurnal AgroBiogen
Publisher : Jurnal AgroBiogen

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Abstract

Plant genetic improvement can be done through classical breeding or genetic engineering. WRKY is a transcription factor involved in regulating plant defense responses. OsWRKY76 gene is located in a narrow segment of chromosome 9 which is identified previously to be related to wide spectrum resistance in rice. A sequence of OsWRKY76 (+1.200 bp) has available in the gene bank and it makes possible to isolate, clone, and construct the gene into over-expression vector. The aim of this research was to assemble an over-expression construct of OsWRKY76 candidate gene and introduce it into rice through Agrobacterium-mediated transformation. A construct of pCAMBIA-1301::35S::OsWRKY76 has been successfully assembled and transformed into embryogenic calli of rice cv. Nipponbare using A. tumefaciens strain Agl-1 and EHA 105. A number of 126 independent lines has been produced, in which Agl-1 showed 3.8 times more efficient than EHA 105. PCR analysis of randomly selected 25 independent lines showed that all of them positively contained hptII gene, a selectable marker used in the over-expression construct of the OsWRKY76 candidate gene. Based on the result, it could be concluded that the over-expression construct of OsWRKY76 candidate gene have been successfully introduced into the tissue of Nipponbare.
RESPON PADI TRANSGENIK CV. NIPPONBARE GENERASI T1 YANG MENGANDUNG GEN Oryza sativa DEHYDRATION-RESPONSE ELEMENT BINDING 1A (OsDREB1A) TERHADAP CEKAMAN SALINITAS [Response of T1 Generation Transgenic Rice cv. Nipponbare Containing an Oryza sativa Dehydration-response Element Binding 1A (OsDREB1A) Gene to Salinity Stress] Santoso, Tri Joko; Apriana, Aniversari; Sisharmini, Atmitri; Trijatmiko, Kurniawan Rudi
BERITA BIOLOGI Vol 11, No 2 (2012)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (495.332 KB) | DOI: 10.14203/beritabiologi.v11i2.494

Abstract

Salinity is one of the abiotic constraints in the cultivation of rice crop. One of the reasons agricultural land becomes saline is due to the intrusion of seawater into the mainland as a result of global climate change. Dehydration-responsive element binding (DREB) gene is a plant -specific transcription factor gene that have important role in regulating plant responses to abiotic stresses, including high salinity. Transgenic rice plants cv. Nipponbare carrying OsDREB1A gene have been generated. However, study of the response of putative transgenic plants to salinity has not been done. The research objective is to study the response of T1 generation Nipponbare-OsDREB1A transgenic rice plants to salinity stress. The result showed that the response of putative transgenic rice Nipponbare-OsDREB1A to salinity stress 25 mM and 150 mM NaCl indicated a level of tolerance varies from highly sensitive to highly tolerance. These variations were possibly occurred because of the segregation state of the T1 generation transgenic rice. Based on damage symptom scoring and PCR analysis provided information that transgenic rice plant cv. Nipponbare-OsDREB1A which showed positive PCR had a very high tolerance to salinity stress 150 mM compared with non-transgenic rice cv. Nipponbare.
Optimasi Konsentrasi Asetosiringon dan Higromisin dalam Transformasi Genetik Padi Fatmawati dengan Perantaraan Agrobacterium tumefaciens Sisharmini, Atmitri; Purwoko, Bambang Sapta; Khumaida, Nurul; Trijatmiko, dan Kurniawan Rudi
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol. 46 No. 3 (2018): Jurnal Agronomi Indonesia
Publisher : Indonesia Society of Agronomy (PERAGI) and Department of Agronomy and Horticulture, Faculty of Agriculture, IPB University, Bogor, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (413.29 KB) | DOI: 10.24831/jai.v46i3.19445

Abstract

Protocols for genetic transformation of rice have been widely developed, however the protocols are not universal and inapplicable for all types of rice plants directly. Transformation protocol on rice cv. Fatmawati needs to be developed to generate transgenic lines. The present research was carried out to optimize genetic transformation protocol in rice cv. Fatmawati mediated by Agrobacterium tumefaciens harboring pCambia1301 construct using immature embryo as an explant. The experiment was arranged in a completely randomized design. Factors influencing efficiency of transformation, i.e., sensitivity of callus to hygromycin antibiotic, acetosyringone concentration used in cultivation medium, hygromycin concentration for transformant selection were optimized. The results showed that genetic transformation of rice cv. Fatmawati mediated by A. tumefaciens using immature embryos have been successfully carried out with several parameters. Addition of 100 µM acetosyringone in co-cultivation medium and 30 mg L-1 hygromycin for transformant callus selection were optimal for genetic transformation of rice cv. Fatmawati mediated by A. tumefaciens. Transformation efficiency was found to be 7.84% based on the lines carrying the hpt gene. This result would be a valuable reference in genetic transformation of rice cv. Fatmawati using target genes.Keywords: immature embryo, Oryza sativa, pCambia1301, transformation efficiency
Co-Authors Aniversari Apriana Bambang Sapta Purwoko Kurniawan R. Trijatmiko Kurniawan Rudi Trijatmiko, Kurniawan Rudi Nurul Khumaida Sudarsono TRI JOKO SANTOSO Trijatmiko, dan Kurniawan Rudi Wening Enggarini