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Fatimah Zahra
Research Center for Biotechnology, Indonesian Institute of Sciences (LIPI)

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Genetic Transformation of Rice cv. Ciherang Using Double T-DNA Vector Harboring cry1Ab Gene Yuli Sulistyowati; Agus Rachmat; Fatimah Zahra; Syamsidah Rachmawati; Satya Nugroho
Annales Bogorienses Vol. 15 No. 1 (2011): Annales Bogorienses
Publisher : National Research and Innovation Agency (BRIN)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ann.bogor.2011.v15.n1.27-32

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.
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 Rachmawati; Inez Hortenza Slamet Loedin
Annales Bogorienses Vol. 16 No. 1 (2012): Annales Bogorienses
Publisher : National Research and Innovation Agency (BRIN)

Show Abstract | Download Original | Original Source | Check in Google Scholar | 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 of 9 (more than 60% damaged tillers ) for non-transgenic plants. However, to confirm the efficacy of cry1B gene to yellow stem borer in natural condition, confined field trial in endemic area of yellow stem borer should be conducted. 
Co-Authors Ade Nena Nurhasanah Agus Rachmat Amy Estiati Inez Hortenza Slamet Loedin Satya Nugroho Syamsidah Rachmawati Yuli Sulistyowati