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Loedin, Inez Hortenza Slamet
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Agrobacterium-Mediated Transformation of Javanica Rice Plants With A Cry1b Gene Under The Control of Wound-Inducible Gene Promoter Estiati, Amy; Rachmawati, Syamsidah; Astuti, Dwi; Loedin, Inez Hortenza Slamet
Annales Bogorienses Vol. 11 No. 1 (2007): Annales Bogorienses
Publisher : BRIN

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A cry1B synthetic gene of Bacillus thuringiensis has been used for the transformation of the javanica rice plants cv. Rojolele to confer resistance to an important pest yellow stem borer (Scirpophaga incertulas). Embryogenic callus were co-cultivated with the EHA105 strain of Agrobacterium tumefaciens harbouring binary vector pCAMBIA I301 containing cry1B gene under the control of wound inducible gene promoter (mpi), hygromycin resistance gene (hpt) as a selectable marker and intron-containing b-glucuronidase (gus intron) gene as a reporter gene driven by CaMV35S promoter. Previously. our histochemical assay and PCR analysis had proved the integration of cry1B gene into the genome of rice plants at first generation. However, the existence of the gene should remain stable 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 and showed that integration of cry1B into the genomic DNA of javanica rice plants cv. Rojolele. An effective resistance of transgenic plants against stem borer was verified in bioassays.
Identification and Activity of The Retrotranposon Tos17 in Indonesian Javanica Rice CV. Rojolele and Japonica Rice CV. Gajahmungkur Nugroho, Satya; Loedin, Inez Hortenza Slamet; Ouwerkerk, Pieter B. F.
Annales Bogorienses Vol. 11 No. 1 (2007): Annales Bogorienses
Publisher : BRIN

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Retrotransposons are mobile genetic elements that transpose via an RNA intermediate that is revers transcribed before integration into a new location within the host genome. They are ubiquitous in eukaryotic organisms and constitute a major portion of the nuclear genome (often more than half of the total DNA) in plants. Tos17 is a rice endogenous retrotransposon that has been studied thoroughly. Tos17 has been shown to be an efficient insertional mutagen and saturation mutagenesis tool for gene tagging and functional genomics in Japonica rice cv nipponbare. In Javanica rice, however, the presence and activity of Tos17 has not been described thus far. while in some Indica rice Tos17 has been found to be inactive. Javanica rice, also known as tropical Japonica rice, has many cultivars which may serve as potential genetic resources of great interest for breeding programmes. Here, the presence and activities of retrotransposon Tos17 in Javanica rice cv Rojolele was described and compared to those of Japonica rice and Japonica rice cv Gajahmungkur. We identified five and three copies of Tos17 in Rojolele and Gajahmungkur, respectively, with different activities.
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; Rachmawati, Syamsidah; Loedin, Inez Hortenza Slamet
Annales Bogorienses Vol. 16 No. 1 (2012): Annales Bogorienses
Publisher : BRIN

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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 Dwi Astuti Estiati, Amy Fatimah Zahra Nurhasanah, Ade Nena Ouwerkerk, Pieter B. F. Rachmawati, Syamsidah Satya Nugroho Yuli Sulistyowati