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"Telah dilakukan penelitian yang bertujuan untuk melakukan transformasi gen Osdep1-Tc ke kalus padi cv. Taipei 309 menggunakan Agrobacterium tumefaciens. Transformasi gen Osdep1-Tc dilakukan menggunakan A. tumefaciens strain LBA4404 yang membawa plasmid rekombinan pCAMBIA1301-Osdep1-Tc, mengandung gen reporter (gus), gen nptII dan hptI. Gen Osdep1-Tc yang telah dikloning ke vektor pengklonaan pGEM-T Easy pada penelitian sebelumnya digunakan sebagai sampel untuk kemudian isubkloning ke pCAMBIA 1301 dan ditransformasikan ke dalam Escherichia coli DH5α sehingga dihasilkan vektor rekombinan pCAMBIA-Osdep1-Tc. Vektor rekombinan kemudian dielektroporasi ke A. tumefaciens dan ditransformasi ke kalus embriogenik padi. Aktivitas GUS pada kalus berhasil dideteksi 3 hari setelah infeksi dengan A. tumefaciens. Analisis PCR kalus transforman menunjukkan bahwa gen hptI berhasil terintegrasi dengan stabil pada kelima kalus uji.

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Research about transformation of Osdep1-Tc gene into rice calli cv. Taipei 309 using Agrobacterium tumefaciens had been done. Transformation of Osdep1-Tc was carried out using A. tumefaciens strain LBA4404, harbored recombinant plasmid pCAMBIA1301-Osdep1-Tc, which contained a reporter gene (gus), hptI and nptII gene. Osdep1-Tc gene had been cloned previously into the pGEM-T Easy cloning vector. The gene was being subcloned into pCAMBIA 1301 and transformed into Escherichia coli DH5α in order to obtain recombinant vectors pCAMBIA-Osdep1-Tc. Furthermore, the recombinant vectors was electroporated into A. tumefaciens and transformed into rice embryogenic calli. GUS activity in rice calli was detected 3 days after infection with A. tumefaciens. PCR analysis of the transformant calli revealed that all five calli tested showed a succeeded stable integration of hptI gene."

"Salinitas merupakan salah satu cekaman abiotik yang mengancam produksi padi di Indonesia. Dalam rangka mendukung program ketahanan pangan, BB-Biogen telah melakukan pengembangan varietas padi Ciherang toleran salinitas hingga generasi BC4F2 dan BC5F1. Tanaman generasi BC4F2 dan BC5F1 Ciherang-OsDREB1A selanjutnya memerlukan serangkaian pengujian. Pertama, penapisan toleransi terhadap salinitas tinggi untuk menyeleksi tanaman yang menunjukkan sifat toleran terhadap salinitas tinggi. Kedua, analisis molekuler yang mencakup analisis integrasi, ekspresi OsDREB1A, dan Southern blot. Hasil penapisan salinitas terhadap BC4F2 dan BC5F1 Ciherang-OsDREB1A selama 26 hari dengan EC akhir berkisar 18 mS/cm telah berhasil menyeleksi 134 individu putatif transgenik dari total 543 tanaman uji. Hasil analisis integrasi menggunakan primer hptII menujukkan 73 dari 134 tanaman putatif transgenik memiliki hasil positif hptII. Seluruh tanaman yang positif PCR hptII juga terdeteksi memiliki hasil positif PCR menggunakan primer kombinasi 35S-496-F/OsDREB1A-R, mengindikasikan kestabilan integrasi transgen tetap terjaga selama persilangan. Hasil analisis ekspresi OsDREB1A menunjukkan terdapat variasi level ekspresi OsDREB1A antar individu Ciherang transgenik. Hasil analisis Southern blot menunjukkan jumlah salinan T-DNA berjumlah sekitar 6--8 kopi. Galur terbaik berdasarkan hasil analisis molekuler adalah BC5F1-K14-23-3 dan BC4F2-K13-11-3.

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Salinity is one of the abiotic stresses that threaten rice production in Indonesia. In order to support the food security programs, BB-Biogen has started doing development of salinity tolerant rice varieties Ciherang up to BC4F2 and BC5F1 generations. BC4F2 and BC5F1 generations of Ciherang-OsDREB1A transgenic lines require a series of tests for verifying the salinity tolerance and stability of transgene integration. First, screening for selecting the Ciherang-OsDREB1A transgenic lines that revealed tolerance to high salinity. Second, molecular analysis that includes analysis of integration, OsDREB1A expression, and Southern blot. The screening result of Ciherang-OsDREB1A transgenic lines BC4F2 and BC5F1 for 26 days with final EC approximately 18 mS/cm, had been successfully selected 134 putative transgenic plants of a total 543 tested plants.

PCR analysis results showed that 73 of 134 putative transgenic plants had PCR positive using hptII-F/hptII-R primer. Plants were detected positive in PCR analysis using hptII-F/hptII-R primer were also positive in PCR analysis using specific primer 35S-496-F/OsDREB1A-R, indicating that the stability of the transgene integration is maintained during the crossing. The results of OsDREB1A expression analysis showed that there were variations in expression levels among individuals Ciherang-OsDREB1A transgenic lines. The results of Southern blot analysis showed that the T-DNA copy number around 6--8 copies. Best lines based on the results of molecular analysis is BC5F1-K14-23-3 and BC4F2-K13-11-3."

"Telah dilakukan penelitian yang bertujuan untuk melakukan transformasi gen OsERA1 ke kalus padi cv. Taipei 309 menggunakan Agrobacterium tumefaciens. Gen OsERA1 adalah gen yang berperan dalam meningkatkan sensitifitas dari sel penjaga pada stomata terhadap asam absisat. Transformasi gen OsERA1 dilakukan menggunakan Agrobacterium tumefaciens strain LBA4404 yang membawa plasmid rekombinan pCAMBIA1301-OsERA1. Gen OsERA1 telah dikloning sebelumnya pada vektor pengklonaan pGEM-T Easy. Gen dipotong dengan enzim restriksi BamHI dan SalI. Gen diligasikan dengan pCAMBIA 1301 dan ditransformasikan ke dalam Escherichia coli DH5α dihasilkan vektor rekombinan pCAMBIA-ERA1. Plasmid vektor rekombinan pCAMBIA-ERA1 belum berhasil ditransformasi ke dalam Agrobacterium tumefaciens dengan elektroporasi tetapi berhasil dilakukan dengan particle bombardment.

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Research about transformation of OsERA1 gene into rice calli cv. Taipei 309 using Agrobacterium tumefaciens had been done. OsERA1 gene is a gene that has response to enhance sensitivity of guard cell to absicid acid. Transfer of OsERA1gene into calli was carried out using Agrobacterium tumefaciens strain LBA4404 harboring recombinant plasmid pCAMBIA1301-OsERA1. OsERA1 gene had been cloned previously in the pGEM-T Easy cloning vector and for inserting into pCAMBIA1301, the gene was digested from cloning vector using restriction enzymes BamHI and SalI. Furthermore, the gene was ligated into vector pCAMBIA 1301 and transformed into Escherichia coli DH5α in order to obtain recombinant plasmid pCAMBIA-OsERA1. The recombinant plasmid pCAMBIAOsERA1has not sucessfully transformed with electroporation into Agrobacterium tumefaciens yet, but it can be transformed by particle bombardment method."

"Tanaman padi merupakan penghasil beras yang merupakan makanan pokok mayoritas masyarakat dunia. Produksi tanaman padi di Indonesia masih belum optimal. Metode rekayasa genetika dengan mengintroduksi gen CONSTANS (CO) dari tanaman Arabidopsis thaliana ke dalam tanaman padi kultivar Nipponbare digunakan untuk usaha peningkatan produktivitas padi. Gen CO diketahui mampu menginduksi terjadinya pembungaan pada tanaman. Penelitian bertujuan untuk mengetahui integrasi gen AtCO, pengaruh gen terhadap karakter agronomi, dan ekspresi dari gen AtCO pada tanaman padi Nipponbare transgenik generasi T2 dengan analisis morfologi dan molekular. Hasil pengamatan morfologi menunjukkan bahwa tanaman padi transgenik memiliki karakter agronomi yang lebih baik dibandingkan dengan tanaman padi kontrol, namun tidak menunjukkan adanya perbedaan umur berbunga yang signifikan. Hasil pengamatan dengan teknik Polymerase Chain Reaction (PCR) menunjukkan 169 dari 227 tanaman padi transgenik memiliki integrasi gen higromisin dan CO. Ekspresi gen CO terdeteksi rendah pada 3 dari 4 sampel padi transgenik dengan teknik Reverse Transcription PCR (RT-PCR). Hasil teknik southern blotting yang dilakukan pada 16 sampel padi transgenik menunjukkan masing-masing 6 salinan T-DNA dari 13 sampel padi transgenik.

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Rice is a staple food which is consume by the majority of world’s population. Rice production in Indonesia has not been optimal yet. A method of genetic engineering by introducing CONSTANS (CO) gene of Arabidopsis thaliana into the rice Nipponbare cultivar was used to improve rice productivity. CO gene known to induce early flowering time in plant. The aims of the experiment were to study the AtCO gene integration, the influence of AtCO gene on agronomic traits, and the expression of AtCO gene in transgenic rice Nipponbare AtCO T2 generation by morphological and molecular analysis.

Morphological observations showed that transgenic plant’s agronomic traits were better than controls, but there is no significant difference in flowering time between transgenic rice Nipponbare plants and controls. Observations with Polymerase Chain Reaction (PCR) technique showed that 169 of 227 transgenic rice Nipponbare plants have hygromycin and CO gene integration. CO gene expression detected in 3 of 4 transgenic rice plants samples using Reverse Transcription PCR (RT-PCR) technique. The southern blotting technique in 16 samples of transgenic plants showed that 13 samples have multi-copies transgen integration (6 copies)."