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All Journal ANNALES BOGORIENSES
Syamsidah Rahmawati
Research Centre for Biotechnology, Indonesian Institute of Sciences (LIPI)
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Immunology & microbiology Medicine & Pharmacology

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Segregation Analysis of Transgenic Rice Plants CV Rojolele Harboring Cry1B Gene and Plant Selection for Potential to Yellow Stem Borer Amy Estiati; Yuli Sulistyowati; Fatimah Zahra; Ade Nena Nurhasanah; Syamsidah Rahmawati; Inez H S-Loedin
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: 10.14203/ann.bogor.2012.v16.n1.19-25

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
Agrobacterium-Mediated Transformation of Javanica Rice Plants With A Cry1b Gene Under The Control of Wound-Inducible Gene Promoter Amy Estiati; Syamsidah Rahmawati; Dwi Astuti; Inez H.S Loedin
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 | Full PDF (2157.952 KB) | DOI: 10.14203/ann.bogor.2007.v11.n1.1-5

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.
Co-Authors Ade Nena Nurhasanah Amy Estiati Dwi Astuti Fatimah Zahra Inez H S-Loedin Inez H.S Loedin Yuli Sulistyowati