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Dwi Astuti
Research Center for Biotechnology, Indonesian Institute of Sciences (LIPI)

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

Show Abstract | Download Original | Original Source | Check in Google Scholar | 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 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.
Phenotypic Screening of Ds Transposon and Activation-tag Insertional Mutant Rice Population for Drought and Salinity Tolerant Related Traits Satya Nugroho; Anky Zannati; Apriadi Situmorang; Vincentia Esti Windiastri; Dwi Widyajayanti; Carla Frieda Pantouw; Dwi Astuti; Sri Indrayani; Syamsidah Rachmawati; Enung Sri Mulyaningsih; Tri Joko Santoso; Kurniawan Rudi Trijatmiko
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.n1.21-25

Abstract

The studies of rice genes through functional genomics are greatly facilitated by the availability of the complete genome sequences, including the complete physical map of the japonica rice cv. nipponbare. Using the Ac and Ds transposon, that are capable of transposition in various heterologous plants including the monocotyledon rice, combined with the enhancer element for generating activation taged lines, it is possible to discover and isolate functional genes involved in various important agronomical traits; such as those involved in abiotic stress tolerance (drought, high salt) and biotic stresses diseases and pests. We have developed 1,785 first (T0) generation of mutant nipponbare rice by transposons Ac/Ds insertions containing activation-tag, generated by transformation using Agrobacterium-mediated method. Currently, we have generated approximately 1,000 stable lines with transposon Ds and activation-tag insertion ready for screening. Efficient screening methods for mutant Nipponbare rice lines have been established for agronomically important mutant traits. Among the new phenotypes related to important agronomical traits observed were drought and salt tolerant or sensitive, stunted and robust growth, variable root penetration and other interesting traits such as reduced tillering, rolled leaf and thin tiller. 
Co-Authors Amy Estiati Anky Zannati Apriadi Situmorang Carla Frieda Pantouw Dwi Widyajayanti Enung Sri Mulyaningsih Inez Hortenza Slamet Loedin Kurniawan Rudi Trijatmiko Satya Nugroho Sri Indrayani Syamsidah Rachmawati Tri Joko Santoso Vincentia Esti Windiastri