Nurhasanah, Ade Nena
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Journal : Annales Bogorienses

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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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. 
Insect Bioassay in Biosafety Containment to Select Transgenic Rice (Oryza sativa L.) Harboring Cry1B Gene Resistant to Yellow Stem Borer (Scrirpophaga incertulas Walk.) Estiati, Amy; Nurhasanah, Ade Nena; Nugroho, Satya
Annales Bogorienses Vol. 17 No. 2 (2013): Annales Bogorienses
Publisher : BRIN

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Development of rice varieties resistant to yellow stem borer (YSB) is very crucial. Agrobacterium-mediated transformation of cry1B gene under wound inducible gene promoter mpi (maize proteinase inhibitor) into a local rice variety Rojolele had been conducted. PCR analysis proved that cry1B gene had been integrated into plant genome of 3R25 and 3R5 rice lines. Segregation analysis using PCR for cry1B gene of the two putative transgenic rice lines at third (T2), fourth (T3), fifth (T4), and sixth (T5) generations of 3R25 and 3R5 lines proved that 3R25.7.27, 3R25.7.13.8.2, 3R25.7.13.8.6, 3R25.7.13.8.8, 3R5.26.2, and 3R5.26.5 are homozygous lines for cry1B gene. Insect bioassay on three randomly picked homozygous transgenic rice lines to study the efficacy of cry1B gene toward YSB was conducted in biosafety containment by infestingYSB larvae at first instar into 3R25.7.27, 3R25.7.13.8.6, and 3R5.26.5 transgenic rice lines, using non--transgenic Rojolele, IR64 and IR74 as susceptible controls. The results showed that the percentages of deadhearts symptoms of 3R25.7.27, 3R5.26.2, and 3R25.7.13.8.6 rice lines were lower than those of the susceptible control lines with scores of 0.1 and 0, respectively. While the scores of all three susceptible control plants were 9. The results proved that lines 3R25.7.27, 3R25.7.13.8.6, and 3R5.26.2 were categorized as resistant lines while the non-transgenic Rojolele, IR64, and IR74 were categorized as susceptible lines. The results also showed that the cry1B gene was expressed and produced insecticidal protein CRY1B which were active against YSB to protect rice plant toward YSB infestation.