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1

IRAP and REMAP assessments of genetic similarity in rice

100%
Branco C. J. S., Vieira E. A., Malone G., Kopp M. M., Malone E., Bernardes A., Mistura C. C., Carvalho F. I. F., Oliveira C. A.
Journal of Applied Genetics
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2007
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vol. 48
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issue 2
107-113
EN
Rice is a model genome for cereal research, providing important information about genome structure and evolution. Retrotransposons are common components of grass genomes, showing activity at transcription, translation and integration levels. Their abundance and ability to transpose make them good potential markers. In this study, we used 2 multilocus PCR-based techniques that detect retrotransposon integration events in the genome: IRAP (inter-retrotransposon amplified polymorphism) and REMAP (retrotransposon-microsatellite amplified polymorphism). Markers derived from Tos17, a copia-like endogenous retrotransposon of rice, were used to identify genetic similarity among 51 rice cultivars (Oryza sativa L.). Genetic similarity analysis was performed by means of the Dice coefficient, and dendrograms were developed by using the average linkage distance method. A cophenetic correlation coefficient was also calculated. The clustering techniques revealed a good adjustment between matrices, with correlation coefficients of 0.74 and 0.80, or lower (0.21) but still significant, between IRAP and REMAP-based techniques. Consistent clusters were found for Japanese genotypes, while a subgroup clustered the irrigated Brazilian genotypes.
2

Rice improvement, involving altered flower structure more suitable to cross-pollination, using in vitro culture in combination with mutagenesis

100%
Min S.-K.
Journal of Applied Genetics
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1998
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vol. 39
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issue 2
151-162
EN
Anther and somatic tissue culture in combination with mutagenesis were carried out to evaluate the efficiency of different mutagenic treatments of various in vitro culture materials, and to obtain some promising variants for rice improvement. Results indicated that in japonica rice radiation treatment of dry seeds and young panicles influenced the percentage of green plantlets regeneration from anther culture. Both treatments increased significantly the percentage of regenerated green plantlets in comparison with the control. Irradiation with 30 Gy of rice callus increased also the percentage of regenerated green plantlets. For indica rice, the combination of the suitable dose of gamma rays irradiation on seeds and an improved medium, increased the percentage of callus induction. This approach made it possible to use anther culture in indica rice breeding. Somatic tissue cultures combined with radiation-induced mutagenesis led to the development of a number of promising mutants including some new cytoplasm-nucleus interacting male-sterile lines with almost 100% stigma exsertion. Their development would be of practical significance for increasing the genetic diversity for production of hybrid rice.
3

Generation of marker-free Bt transgenic indica rice and evaluation of its yellow stem borer resistance

75%
Kumar S., Arul L., Talwar D.
Journal of Applied Genetics
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2010
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vol. 51
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issue 3
243-257
EN
We report on generation of marker-free ('lean DNA') transgenic rice (Oryza sativa), carrying minimal gene-expression-cassettes of the genes of interest, and evaluation of its resistance to yellow stem borer Scirpophaga incertulas (Lepidoptera: Pyralidae). The transgenic indica rice harbours a translational fusion of 2 different Bacillus thuringiensis (Bt) genes, namely cry1B-1Aa, driven by the green-tissue-specific phosphoenol pyruvate carboxylase (PEPC) promoter. Mature seed-derived calli of an elite indica rice cultivar Pusa Basmati-1 were co-bombarded with gene-expression-cassettes (clean DNA fragments) of the Bt gene and the marker hpt gene, to generate marker-free transgenic rice plants. The clean DNA fragments for bombardment were obtained by restriction digestion and gel extraction. Through biolistic transformation, 67 independent transformants were generated. Transformation frequency reached 3.3%, and 81% of the transgenic plants were co-transformants. Stable integration of the Bt gene was confirmed, and the insert copy number was determined by Southern analysis. Western analysis and ELISA revealed a high level of Bt protein expression in transgenic plants. Progeny analysis confirmed stable inheritance of the Bt gene according to the Mendelian (3:1) ratio. Insect bioassays revealed complete protection of transgenic plants from yellow stem borer infestation. PCR analysis of T2 progeny plants resulted in the recovery of up to 4% marker-free transgenic rice plants.
4

Transgenic Indica rice to the benefit of Less Developed Countries: Towards pest-resistance, and accumulation of beta-carotene in the endosperm

75%
Potrykus I., Burkhard P., Futterer J., Klotti A., Wunn J.
Biotechnologia
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1996
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issue 1
14-25
EN
We have established gene transfer to IRRI breeding lines to explore the contributions of genetic engineering to sustained and stable production of high quality food.Experiments are in progress on the development of resistance towards Yellow Stem Borer, towards Rice Tungro Virus, towards fungal pests, and towards accumulation of provitamin A in the endosperm. So far we have recovered the first transgenic Indica rice with elevated resistance to sheath blight (Rhizoctonia solani) and with 100% resistance against stem borer (Scirpophaga incertulas).We have also harvested seed from a series of transgenic rice plants harbouring a number of different transgenic DNA sequences representing different anti tungro virus disease strategies.And we have so far, been able to initiated the beta-carotene pathway in the endosperm to the accumulation of phytogene, hoping, that it will be possible to subsequently complete the pathway towards provitamin A.
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