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Characterization of the potato upreg1gene, encoding a mutated ADP-glucose pyrophosphorylase large subunit, in transformed rice

Lee, Si-Myung, Lee, Yoen-Hee, Kim, Hyun-uk, Seo, Suk-chul, Kwon, Sun-jong, Cho, Hyun-suk, Kim, Su-Il, Okita, Thomas, Kim, Donghern
Plant cell, tissue, and organ culture 2010 v.102 no.2 pp. 171-179
Oryza sativa, rice, grain crops, genetic transformation, transgenes, nucleotidyltransferases, protein subunits, transfer DNA, genetic vectors, promoter regions, Agrobacterium tumefaciens, culture media, genome, Southern blotting, seed set, inflorescences, dry matter partitioning, aerial parts, photosynthesis, glufosinate
The potato upreg1, which encodes a mutated ADP-glucose pyrophosphorylase (AGPase) large subunit, was introduced into rice to evaluate its potential to enhance sink-driven yield productivity in this crop. We also wished to elucidate the activities of the up-regulated allosteric variants of potato AGPase large subunit gene in rice. A T-DNA vector containing the upreg1 gene under the control of the rice glutelin promoter was constructed with a MAR sequence and transformed into rice using Agrobacterium-mediated transformation. Transgenic plants were selected on medium supplemented with phosphinothricin and confirmed by the application of herbicide. A total of 38 transgenic plants were subsequently obtained in which the integration upreg1 into the rice genome was confirmed by Southern blotting. The exogenous AGPase in transgenic rice plants showed a high affinity for 3-phosphoglycerate activator and a low affinity for the orthophosphate inhibitor, as observed in lettuce. The transgenic rice also showed increases in the number of grains per particle, the number of panicles per plant, and also in the fresh weight of the above-ground mass of plant which was about 15% higher than non-transgenic ‘Nak-dong'. The number of seeds per tiller was also found to be about 10% higher in the transgenic plants. However, the net photosynthesis rate showed very little difference in the transgenic rice, and we could not therefore confirm any linkage with the deregulation of allosteric effects. Based on these results, upreg1 mutant genes can be used for the genetic improvement of plant AGPases other than potato and to effectively increase crop yield productivity.