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All Journal Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati Indonesian Journal of Biotechnology ANNALES BOGORIENSES
Syamsidah Rahmawati, Syamsidah
Pusat Penelitian Bioteknologi-LIPI, Jl. Raya Bogor KM 46, Cibinong 16911
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology

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Comparative Analysis of Rice Transformation Using Agrobacterium tumefaciens and Rhyzobium leguminosarum Rahmawati, Syamsidah; Jefferson, Osmat Azzam; Sopandie, Didy; ., Suharsono; Slamet-Loedin, Inez Hortense
Indonesian Journal of Biotechnology Vol 15, No 1 (2010)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (287.394 KB)

Abstract

This study was aimed to study the effectiveness of Rhizobium transformation system compared to the most widely used Agrobacterium mediated transformation system on three rice cultivars, Ciherang (Indica), Nipponbare (Japonica), and Rojolele (Javanica). Six day old calli induced from immature embryos were inoculated with Rhizobium leguminosarum bv trifolii ANU845 and Agrobacterium tumefaciens LBA288 that harbored with vector pCAMBIA 5106. This plasmid contained a minimum set of transfer machinery genes and had a gusplus and an hptII gene driven by 35S CaMV promoter in the T-DNA. The results showed that the transformation frequencies (number of PCR positive plants per number of calli inoculated) ranging from 0 to 12.05 % depend on the genotype and transfer agent used. The highest transformation frequency (12.05%) was obtained in Ciherang transformed with R. leguminosarum. Most of the transgenic rice obtainedby Rhizobium transformation were normal in morphology and fertile similar to those obtained by Agrobacterium transformation. Integration, expression and inheritance of transgenes were demonstrated by molecular and genetic analysis in T0 and T1 generations.Key words : Rhizobium leguminosarum, immature embryos, Agrobacterium tumefaciens
Guide for Authors AB Vol 21 No 2 (2017) Rahmawati, Syamsidah
ANNALES BOGORIENSES Vol 21, No 2 (2017): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (204.083 KB) | DOI: 10.14203/ab.v21i2.320

Abstract

Genetic transformation of rice cv. Ciherang using double T-DNA vector harboring cry1Ab gene Sulistyowati, Yuli; Rachmat, Agus; Zahra, Fatimah; Rahmawati, Syamsidah; Nugroho, Satya
ANNALES BOGORIENSES Vol 15, No 1 (2011): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (383.64 KB) | DOI: 10.1234/37

Abstract

Rice stem borer (Scirpophaga sp.) is considered as one of the primary pests causing yield decrease in Indonesia. Genetic engineering is considered as one of the most effective way to improve the resistance of rice plants to yellow stem borer since no resistance gene has been found in rice and its wild relatives. A double T-DNA binary vector p2TDNAcryIAb wich carry two independent T-DNA was used to transform Indonesian elite rice cv Ciherang using Agrobacterium-mediated transformation to generate transgenic rice that free from selectable marker genes. One T-DNA containing a selectable marker hygromycin phosphotransferase (hpt) gene and the other carrying the cry1Ab gene. As many as 37 putative transgenic plants from 14 embryos were generated. PCR analysis demonstrated that 22 out of 37 putative transgenic rice contain cry1Ab gene. All Ciherang rice carrying the cry1Ab gene expressing Cry1Ab protein as shown by the immunostrip assay. Further work will be carried out to determine the transgenes copy number, to select marker free transgenic rice, and to evaluate the resistance of transgenic rice against neonates larvae of yellow stem borer.Keyword: double T-DNA, transgenic rice, Cry1Ab, transformation
Guide for Authors AB Vol 22 No 1 (2018) Rahmawati, Syamsidah
ANNALES BOGORIENSES Vol 22, No 1 (2018): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (246.986 KB) | DOI: 10.14203/ab.v22i1.353

Abstract

Segregation Analysis of Transgenic Rice Plants CV Rojolele Harboring Cry1B Gene and Plant Selection for Potential to Yellow Stem Borer Estiati, Amy; Sulistyowati, Yuli; Zahra, Fatimah; Nurhasanah, Ade Nena; Rahmawati, Syamsidah; S-Loedin, Inez H
ANNALES BOGORIENSES Vol 16, No 1 (2012): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (698.51 KB) | DOI: 10.1234/42

Abstract

Transgenic rice plants harboring resistant gene to yellow stem borer, cry1B gene had been obtained. However, the cry1B gene in second generation of transgenic rice lines still segregating following Mendelian ratio 3:1. For further use in the breeding programs, it is important to ensure that the gene is dominant gene and the plants are homozygous for  cry1B gene. Selection  of homozygous transgenic rice lines containing  cry1B  gene at third, fourth and fifth generation had been conducted by PCR.  The presence of  cry1B  gene was determined by showing the PCR product of 1.9 kb. Segregation analysis proved that six transgenic rice  lines i.e. 3R7.8.15.1, 3R7.8.15.9, 3R7.8.15.10, 3R7.8.15.15, 3R7.8.15.21, 3R7.8.15.29 are homozygous lines for  cry1B  gene.   Moreover, bioassay studies at vegetative stage on six homozygous transgenic rice lines showed that  these transgenic rice lines are potential resistant to yellow stem borer comparing with non-transgenic plants, with the score of 0 (indicated no symptom) for six transgenic lines and score of9 (more than 60% damaged tillers ) for non-transgenic plants. However, to confirm the efficacy of  cry 1B  gene to yellow stem borer in naturalcondition, confined field trial in endemic area of yellow stem borer should be conducted. Keywords: bioassay, cry1B gene, transgenic rice, yellow stem borer
Back Cover AB Vol 21 No 2 (2017) Rahmawati, Syamsidah
ANNALES BOGORIENSES Vol 21, No 2 (2017): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (202.587 KB) | DOI: 10.14203/ab.v21i2.321

Abstract

Back Cover AB Vol 22 No 1 (2018) Rahmawati, Syamsidah
ANNALES BOGORIENSES Vol 22, No 1 (2018): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (154.375 KB) | DOI: 10.14203/ab.v22i1.354

Abstract

Front Cover AB Vol 21 No 2 (2017) Rahmawati, Syamsidah
ANNALES BOGORIENSES Vol 21, No 2 (2017): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ab.v21i2.317

Abstract

Agrobacterium-Mediated Transformation of Javanica Rice Plants With A Cry1b Gene Under The Control of Wound-Inducible Gene Promoter Estiati, Amy; Rahmawati, Syamsidah; Astuti, Dwi; Loedin, Inez H.S
ANNALES BOGORIENSES Vol 11, No 1 (2007): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/18

Abstract

A  cry1B synthetic gene of Bacillus thuringiensis has been used  for  the transformation of the javanica rice plants  cv.  Rojolele  confer resistance  to  an  important  pest,  yellow  stem  borer  (Scirpophaga  incertulas). Embryogenic callus were co-cultivated with  the EHA 105 strain of Agrobacterium tumefaciens harbouring binary vector CAMB  IA 130 I containing cry  / B gene under  the control of wound inducible gene promoter  (mpi), hygromycin resistance gene (hpt)  as a selectable marker and intron-containing β-glucuronidase (gus - intron) gene al) a reporter gene  driven  by  CaMV35S  promoter.  Previously.  our  histochemical  assay  and  P  R  analyses.  had  proved  th integration of  cry1B gene  into  the genome of rice plants at first generation .  However,  the existence of the gene should remain  table  throughout generation.  In  this study,  the presence of the cry1B transgene in  rice  transgenic plants at  second generation was confirmed by Polymerase Chain Reaction (PCR) .  Insertion of  the cry1B gene in the genome of PCR positive plants was  verified by Southern blot analysis is and showed that integration of cry1  B  into the genomic DNA of  javanica rice plants cv. Rojolele.  An effective resistance of transgenic  plants against  stem borer was verified in  bioassays.
Front Cover AB Vol 22 No 1 (2018) Rahmawati, Syamsidah
ANNALES BOGORIENSES Vol 22, No 1 (2018): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ab.v22i1.350

Abstract

Co-Authors Agus Rachmat Anky Zannati Apriadi Situmorang Chairunnisa Chairunnisa Didy Sopandie Dwi Astuti Dwi Widyajayantie Enung Sri Mulyaningsih Erdayani, Eva Estiati, Amy Estiati, Amy Fatimah Zahra INEZ HORTENSE SLAMET-LOEDIN Kurniawan Rudi Trijatmiko, Kurniawan Rudi Loedin, Inez H.S Nurhasanah, Ade Nena Osmat Azzam Jefferson, Osmat Azzam Pantouw, Carla F. S-Loedin, Inez H Satya Nugroho SLAMET-LOEDIN, INEZ H. Sri Indrayani, Sri Suharsono . TRI JOKO SANTOSO Vincentia Esti Windiastri, Vincentia Esti Yuli Sulistyowati