H. Aswidinnoor
Departemen Agronomi dan Hortikultura, Fakultas Pertanian Institut Pertanian Bogor, Kampus Darmaga, Bogor 16680

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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.
Perbanyakan Padi Fl Interspesifik untuk Bahan Silang Balik (Back Cross) Syukur, M.; Aswidinnoor, H.; Suharsono, ,
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol. 31 No. 2 (2003): Buletin Agronomi
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 (1112.995 KB) | DOI: 10.24831/jai.v31i2.1466

Abstract

A lot of plant materials are needed for successful back cross program of interspecific FI rice to their recurrent parents. Multiplication of the steril FI plants through tiller propagation is not adequate. The research aims to develop technique of vegetatif multiplication of interspesific hybrid rice utilizing nodal segments. In the vegetatif multiplication experiment, several factors were examined as treatments: nutrition culture (MS, Yoshida, and water), stum position (first, second and third) and growth media (vermiculite, husk charcoal, and sand). The plant materials are interspecific FI rice i.e. Ranah Sanra (genom AA) x O. officinalis 100870 (genom CC), Hawara Bunar (genom AA) x O. Punctata 9101411 (genom BB), Grogol (genom AA) x O. punctata 9101411 (genom BB), CT 6510-24-1-3 (genom AA) x O. malamphuzaensis 100957 (genom BBCC). Results of the study indicated that MS and Yoshida nutrition, stum without sheath, base of stum and sand media gave better growth more than other treatments. Key words: Vegetatif multiplication, FI interspecific