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Amy Estiati

Research Centre for Biotechnology, Indonesian Institute of Sciences (LIPI)

Author-ID : 3965267

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Immunology & microbiology Medicine & Pharmacology

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Articles

Agrobacterium-mediated transformation of banana Musa acuminata AA cv "“Mas Lampung” with hpt gene using sterile corm as target tissue Ade Nena; Witjaksono Witjaksono; Amy Estiati
ANNALES BOGORIENSES Vol 13, No 1 (2009): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

The protocol for Agrobacterium-mediated transformation of local banana plants cv “Mas Lampung” (AA) has been established. A selectable marker gene (hpt) has been used to study the transformation using in vitro corm slices as target tissues. Banana in vitro corm slices were co-cultivated with the EHA105 strain of Agrobacterium tumefaciens harbouring binary vector pCAMBIA 1301 containing hygromycin resistance gene (hpt) as a selectable marker and intron-containing β-Glucuronidase (gus-intron) gene as a reporter gene driven by CaMV 35S promoter. Polymerase Chain Reaction (PCR) were used to examine the existence of hpt gene in plants resulted from the transformation. Using primer pairs specific for hpt gene, our PCR analysis on leaves showed the presence of the hpt transgene in banana transgenic plants at first generation (T0) of transformation. To prove the existence of hpt gene in the fruits of transgenic banana plants, PCR analysis were also carried out. The data showed that the hpt gene could be amplified from banana fruits of tested samples. These result demonstrates that the Agrobacterium-mediated transformation method used in this experiment has been successful to transfer gene into banana plants. Thus, the transformation method reported here could be used as a standard protocol to transfer another useful genes into local banana plants cv. “Mas Lampung”. Furthermore, the presence of transgene in fruits of banana transgenic plants is important achievement especially for transgene that is expected to be expressed in the fruit including to introduce vaccine genes into banana fruits for edible vaccine.Key words: Agrobacterium, hpt gene, transgenic banana Mas Lampung, Musa acuminate corm slices, PCR

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)

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

Gene flow from genetically modified rice to their weedy and wild relatives and its environmental consequences Amy Estiati; Satya Nugroho
ANNALES BOGORIENSES Vol 13, No 1 (2009): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Rice is one of the most important food crops in the world. Nearly half of the world population consume rice as their staple food (FAO, 2004). With the increasing of world population, the need to provide more food supplies including rice is obvious. Biotechnology is expected to play major roles in the improvement of crop productivity and quality. Unlike transgenic maize, eventhough many research have been done to improve rice performance by genetic engineering, transgenic rice has not been released or commercialized. Among those research to improve rice performance are the atempt to introduce biotic and abiotic stress tolerant traits, herbicide tolerant trait and beta-carotin biosynthetic pathway. Currently, several genetically modified rice cultivars have been and being tested on limited field trials.Keywords : Rice, stress tolerant

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)

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 Ade Nena Nurhasanah Dwi Astuti Fatimah Zahra Inez H S-Loedin Inez H.S Loedin Satya Nugroho Syamsidah Rahmawati Witjaksono Witjaksono Yuli Sulistyowati

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