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Saptono Nugroho, Saptono
Program Studi Sarjana Destinasi Pariwisata, Fakultas Pariwisata, Universitas Udayana
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Journal : JURNAL BIOLOGI INDONESIA

 1 
Perbandingan Tiga Metode Transformasi Agrobacterium Untuk Pencarian Gen-gen Terkait Toleransi Kekeringan Menggunakan Transposon Ac/Ds pada padi cv. Batutegi Mulyaningsih, E. S.; Aswidinnoor, H.; Sopandie, D.; Ouwerkerk, P.B. F.; Nugroho, S.; Loedin, I.H. Salmet
JURNAL BIOLOGI INDONESIA Vol 6, No 3 (2010): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v6i3.3144

Abstract

ABSTRACTTransformation Strategy for Indonesian Indica Rice in Attempt to Discover Drought-TolerantRelated Genes Using of Transposon Ac/Ds. Attempt to identify, isolate the gene, and study forgene function for several agronomical traits have been done including some drought toleranttraits. Japonica rice cultivars have been used due to its higher efficiencies compared withindica cultivars. Two plasmids namely pNU400 and pUR224 were used to generate mutants ofthese cultivars (Batutegi dan Kasalath cultivars). Those plasmids contain an element calledActivator (Ac) and Dissociator (Ds) respectively. The pNU400 contains GFP (green flourescensprotein) as a selectable marker, whereas the pUR224 contains hygromycine resistant gene andgusA as a reporter gene. Each plasmid was transformed into rice genome of Batutegi andKasalath cultivars by Agrobacterium mediated transformation using three methods oftransformation (A, B and C). The transformation method A was not suitable for both cultivars,where none of plantlets were produced from pNU400 and pUR224 plasmids. The transformationmethod B produced some plantlets from the Kasalath cultivar only using pUR224 plasmid.The transformation method C was the best method to produce transgenic plants from bothcultivars (Batutegi and Kasalath), using both plasmids (pNU400 and pUR224). The PCR analysisshowed that 19 and 9 plants of Batutegi and Kasalath contained both gusA and hpt genesrespectively. None of those plants contained of gusA gene. Southern blot analysis revealed 3independent lines from Batutegi dan 7 independent lines from Kasalath. The integration of Actransposon was analyzed based on expression gfp gene when observed under UV dark reader.This research has proved that indica rice cultivars, especially the Batutegi cultivar of Indonesianorigin, could be transformed. The cultivar could be used as plant model for the indicatransformation.Key words: transformation, drought tolerant, indica rice, Ac/Ds transposons, Agrobacterium.
PERBANDINGAN TIGA METODE TRANSFORMASI AGROBACTERIUM UNTUK PENCARIAN GEN-GEN TERKAIT TOLERANSI KEKERINGAN MENGGUNAKAN TRANSPOSON AC/DS PADA PADI CV. BATUTEGI Mulyaningsih, E. S.; Aswidinnoor, H.; Sopandie, D.; Ouwerkerk, P.B. F.; Nugroho, S.; Loedin, I.H. Salmet
JURNAL BIOLOGI INDONESIA Vol 6, No 3 (2010): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v6i3.3144

Abstract

ABSTRACTTransformation Strategy for Indonesian Indica Rice in Attempt to Discover Drought-TolerantRelated Genes Using of Transposon Ac/Ds. Attempt to identify, isolate the gene, and study forgene function for several agronomical traits have been done including some drought toleranttraits. Japonica rice cultivars have been used due to its higher efficiencies compared withindica cultivars. Two plasmids namely pNU400 and pUR224 were used to generate mutants ofthese cultivars (Batutegi dan Kasalath cultivars). Those plasmids contain an element calledActivator (Ac) and Dissociator (Ds) respectively. The pNU400 contains GFP (green flourescensprotein) as a selectable marker, whereas the pUR224 contains hygromycine resistant gene andgusA as a reporter gene. Each plasmid was transformed into rice genome of Batutegi andKasalath cultivars by Agrobacterium mediated transformation using three methods oftransformation (A, B and C). The transformation method A was not suitable for both cultivars,where none of plantlets were produced from pNU400 and pUR224 plasmids. The transformationmethod B produced some plantlets from the Kasalath cultivar only using pUR224 plasmid.The transformation method C was the best method to produce transgenic plants from bothcultivars (Batutegi and Kasalath), using both plasmids (pNU400 and pUR224). The PCR analysisshowed that 19 and 9 plants of Batutegi and Kasalath contained both gusA and hpt genesrespectively. None of those plants contained of gusA gene. Southern blot analysis revealed 3independent lines from Batutegi dan 7 independent lines from Kasalath. The integration of Actransposon was analyzed based on expression gfp gene when observed under UV dark reader.This research has proved that indica rice cultivars, especially the Batutegi cultivar of Indonesianorigin, could be transformed. The cultivar could be used as plant model for the indicatransformation.Key words: transformation, drought tolerant, indica rice, Ac/Ds transposons, Agrobacterium.
Co-Authors A. Supardi Alinda Thalia Anak Agung Gde Rama Purwani Yoga Anak agung Tirta Putra Wiranata Arichtia Praevianti Taolin Arida, Nyoman Sukma Asri Kristiansi Laga Hae Bhaskara, Gde Indra Chrissanty, Nadia D. Hudiyanti D. Sopandie Dewi Sasmita Dio Pratama Dyah Estu Larasati Ester Venni Olita Saragih Fahrul Fahrul Ferry Aryanto Padabain Filomena Harsiana Fransisco Situmorang Gede Mahendra Saputra Gilang, Wayan Gede H. Aswidinnoor I Agus Yudi Pardiana I Gede Gian Saputra, I Gede I Gusti Agung Oka Mahagangga I Gusti Ayu Arya Ariningsih I Komang Agus Moni Artadi I Made Adikampana, I Made I Made Sendra I Nyoman Gunarsa I Nyoman Jamin Ariana I Nyoman Sukma Arida I Nyoman Sunarta I P. Sudana I Putu Anom I Wayan Gede Wahyu Pradnyana Ida Ayu Suryasih Ida Ayu Suryasih, Ida Ayu Ida Bagus Suryawan Joanne, Marie Josephine Kesya Marcella Tjampan Lena Haryanti Lidia Uli Christiani Karmelia Hutagalung Loedin, I.H. Salmet Loedin, I.H. Salmet Luh Putu Aritiana Kumala Pratiwi Luh Putu Kerti Pujani Luh Putu Vira Cintya Dewi MADE KUSUMA NEGARA Mahagangga, I.G.A.O. Mon, Maria Yosefina Rosari Mulyaningsih, E. S. Mulyaningsih, E. S. Nasution, Annio Indah Lestari Ni Komang Otami Astuti Widiandari Ni Luh Karmini Ni Wayan Eka Suryaningsih Ni Wayan Purnami Rusadi Nur Kumala Sari Ouwerkerk, P.B. F. Ouwerkerk, P.B. F. Pande I Putu Angga Saputra Philipus Wangku Putra, Putu Guntur Pramana Putu Widya Darmayanti Ricky Leonardo Pehulisya Semara, Sang Made Dwipa Shafa Raya Cahyana Shandy Shandy, Shandy Sihombing, Lisbet Situmorang, Fransisco Theresia Loviannauli P Ursulla Mariska Maduma Silaban