Establishment of a high efficiency Agrobacterium-mediated transformation system of rice (Oryza sativa L.)
Establishment of a high efficiency Agrobacterium-mediated transformation system of rice (Oryza sativa L.)
复制标题
DOI：
10.1016/j.plantsci.2009.01.013
复制
发表时间：
2009-04-01
期刊：
影响因子：
5.2
通讯作者：
Ozawa, Kenjirou
中科院分区：
文献类型：
--
作者：
Ozawa, Kenjirou
为了培育出具有有用农业性状的转基因水稻植株，功能基因组学对水稻转化技术的要求不断提高。然而，许多水稻品种不能有效地转化农杆菌。我们成功地建立了水稻愈伤组织与农杆菌在3张滤纸上共同培养的高效转化体系，滤纸上用富集的N6或DKN培养基湿润，而不是使用固体培养基。水稻愈伤组织浸泡在农杆菌悬浮液（EHA101，农杆菌浓度OD600 = 0.04）中，在3张直径为9 cm的滤纸上，用5.5 mL N6或DKN共培养液湿润，共培养培养基中添加2,4- d （2 mg/L）、脯氨酸（10 mM）、酪蛋白水解物（300 mg/L）、蔗糖（30 g/L）、葡萄糖（5 g/L）、L-半胱氨酸（100 mg/L）和乙酰丁香酮（15 mg/L），温度25℃，黑暗培养3天。与在固体培养基上共培养愈伤组织的转化效率相比，滤纸法对许多水稻品种的转化效率提高了约5倍，包括那些以前转化率很低的品种。2009爱思唯尔爱尔兰有限公司版权所有。
Technologies for transformation of rice have been developed to meet the requirements of functional genomics in order to enable the production of transgenic rice plants with useful agricultural characters. However, many rice varieties are not efficiently transformed by Agrobacterium. We have succeeded in establishing a highly efficient transformation system in rice by co-cultivating rice calli with Agrobacterium on three filter papers moistened with enriched N6 or DKN media instead of using solid media. Rice calli immersed in Agrobacterium suspension (EHA101, Agrobacterium concentration of OD600 = 0.04) were co-cultured on three pieces of filter paper (9 cm in diameter) moistened with 5.5 mL of N6 or DKN liquid co-cultivation medium supplemented with 2,4-D (2 mg/L), proline (10 mM), casein hydrolysate (300 mg/L), sucrose (30 g/L), glucose (5 g/L), L-cysteine (100 mg/L) and acetosyringone (15 mg/L) at 25 degrees C for 3 days in the dark. Compared with the transformation efficiency of calli co-cultivated on solid media, transformation efficiency was increased by about fivefold by using the filter paper method for many varieties of rice, including those that previously yielded much poor transformation rates. (C) 2009 Elsevier Ireland Ltd. All rights reserved.