Development of an efficient agrobacterium-mediated gene targeting system for rice and analysis of rice knockouts lacking granule-bound starch synthase (Waxy) and β1,2-xylosyltransferase

Plant & Cell Physiology
Kenjirou OzawaFumio Takaiwa

Abstract

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 with 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 (β1,2-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.

References

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Citations

Oct 24, 2012·Applied Biochemistry and Biotechnology·Shweta Mehrotra, Vinod Goyal
Aug 21, 2013·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Taiji KawakatsuFumio Takaiwa
Apr 13, 2012·Plant & Cell Physiology·Liliana M Costa
Dec 12, 2012·Plant & Cell Physiology·Liliana M Costa
Aug 14, 2013·Journal of Biosciences·Yayuan JiangChangxiang Zhu
Jun 24, 2015·Plant Biotechnology Journal·Fumio TakaiwaTakachika Hiroi
Dec 12, 2012·Scientific Reports·Yuhya WakasaFumio Takaiwa
May 31, 2015·Plant Science : an International Journal of Experimental Plant Biology·Sho TakanoKiyoaki Kato
Jan 21, 2015·Frontiers in Plant Science·Zenpei ShimataniRie Terada
Aug 16, 2016·Plant & Cell Physiology·Kana MiyataTomomi Nakagawa
Mar 31, 2016·Plant Cell Reports·Teodoro Cardi, C Neal Stewart

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