We have developed a high-frequency method for Agrobacterium-mediated gene targeting by combining an efficient transformation system using rice suspension-cultured calli and a positive/negative selection system. Compared to the conventional transformation system using calli on solid medium, transformation using suspension-cultured calli resulted in a 5- to 10- fold increase in the number of resistant calli per weight of starting material after positive/negative selection. Homologous recombination occurred in about 1.5 % of the positive/negative selected calli. To evaluate the efficacy of our method, we show in this report that knockout rice plants containing either a disrupted Waxy (granule-bound starch synthase) or a disrupted Xyl (beta1,2-xylosyltransferase) gene can be easily obtained by homologous recombination. Study of gene function using homologous recombination in higher plants can now be considered routine work as a direct result of this technical advance.
Development of an Efficient Agrobacterium-mediated Gene Targeting System for Rice and Analysis of Rice Knockouts Lacking Granule-bound Starch Synthase (Waxy) and beta1,2-Xylosyltransferase.: "Publication Date: 2012 Feb 10 PMID: 22327484
Authors: Ozawa, K. - Wakasa, Y. - Ogo, Y. - Matsuo, K. - Kawahigashi, H. - Takaiwa, F.
Journal: Plant Cell Physiol
(Via Plant and Cell Physiology.)