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All Journal Indonesian Journal of Agricultural Science JURNAL BIOLOGI INDONESIA JURNAL HAMA DAN PENYAKIT TUMBUHAN TROPIKA ANNALES BOGORIENSES Biosaintifika: Journal of Biology & Biology Education Jurnal DISPROTEK Catha Saintifica Annales Bogorienses
Yuli Sulistyowati
Research Center for Biotechnology-LIPI Jalan Raya Bogor Km.46 Cibinong 16911, Phone (021)8754587 Fax. (021)8754588,
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Civil Engineering, Building, Construction & Architecture Computer Science & IT Education Electrical & Electronics Engineering Energy Immunology & microbiology Industrial & Manufacturing Engineering Medicine & Pharmacology Other

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Journal : ANNALES BOGORIENSES

 1 
Genetic transformation of rice cv. Ciherang using double T-DNA vector harboring cry1Ab gene Sulistyowati, Yuli; Rachmat, Agus; Zahra, Fatimah; Rahmawati, Syamsidah; Nugroho, Satya
ANNALES BOGORIENSES Vol 15, No 1 (2011): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (383.64 KB) | DOI: 10.1234/37

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.Keyword: double T-DNA, transgenic rice, Cry1Ab, transformation
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; Rahmawati, Syamsidah; S-Loedin, Inez H
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: 10.1234/42

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
Co-Authors Agus Rachmat Albaz Ulumudin, Muhammad ALEX HARTANA Ambar Yuswi Perdani, Ambar Yuswi Cahyani, Yani Didy Sopandie Dody Priadi Enung Sri Mulyaningsih Eryanto, Aji Estiati, Amy Estiati, Amy Faqih, Nasyiin Fatchur Roehman Fatimah Zahra Fiqolbi Nuro Pohan, Fiqolbi Nuro Herlambang, Sigit Hery Widijanto Inez H. Slamet -Loedin Loedin, Inez Hortenza Slamet Mushthofa Nurhasanah, Ade Nena Nurhasanah, Ade Nena Paradisa, Yashanti Berlinda Rachmawati, Syamsidah Rizky Ramadhon, Ahmad S-Loedin, Inez H Satoto Satoto Satya Nugroho Sintho Wahyuning Ardie Solly Aryza Sri Indrayani, Sri Syamsidah Rahmawati, Syamsidah TRI KOESOEMANINGTYAS