In planta Agrobacterium-Mediated Transformation of Rice

作     者：Kumrop RATANASUT Weerawan ROD-IN Kawee SUJIPULI 

作者机构：Department of Agricultural Science Faculty of Agriculture Natural Resources and Environment Naresuan University Phitsanulok 65000 Thailand 

出 版 物：《Rice science》 (水稻科学（英文版）)

年 卷 期：2017年第24卷第3期

页      面：181-186页

核心收录：

学科分类：0710[理学-生物学] 0831[工学-生物医学工程（可授工学、理学、医学学位）] 09[农学] 0901[农学-作物学] 0836[工学-生物工程] 0902[农学-园艺学] 

基　　金：supported by the research grant (Grant No.R2556B036) from Naresuan University  Thailand 

主　　题：Agrobacterium-mediated transformation floral-dip floral-drop transformation efficiency in planta rice 

摘      要：The floral-dip transformation, the simplest technique, is no requirement of tissue culture procedure, and can directly transfer the interest gene into plant reproductive cells. It has been successfully applied to various plant species. In this study, the optimal conditions of a floral-dip method for production of transgenic rice variety RD41 were explored. The simple and effective inoculation medium was composed of Murashige and Skoog(MS) medium, 5% sucrose, 44 nmol/L benzylaminopurine, and 0.075% surfactant Tween-20 with pH 5.7. The transformation efficiencies of Agrobacterium tumefaciens strains AGL1 and EHA105 were compared with the Agrobacterium density at OD_(600) = 0.8–1.0 and the co-cultivation at 25 ℃ for 48 h. A. tumefaciens strain EHA105 gave slightly higher transformation efficiency than AGL1, with statistically non-significant difference. The floral-drop transformation using the optimal floral-dip conditions showed higher transformation efficiency than the floral-dip method, but the dropped flowers turned brown and died within 2 d. Production of transgenic rice variety RD41 by the floral-dip method was achieved using A. tumefaciens strain EHA105 with the optimal conditions. Screening for the gus A gene by PCR using the gus A specific primers in the T_0 lines, there were 4 transgenic lines from 286 T_0 lines(1.4% transformation efficiency). However, histochemical glucuronidase(GUS) assay demonstrated that only three of four transgenic lines exhibited gus A expression. These results indicated that floral-dip transformation is a potential tool for production of the transgenic rice, which can be used for molecular breeding via genetic engineering in the future.