摘要
利用根癌农杆菌将含多基因(hyg选择标记基因、PTA基因、GFP和GUS报告基因)的pCAMBIA1304载体导入水稻品种(501R、中花9号和日本晴)的幼胚愈伤组织,分别在含35、45和65mg/L潮霉素浓度的筛选培养基上筛选获得抗性愈伤。后经PCR检测,从415株T0代再生植株中选出92株(其中501R、中花9号和日本晴分别为4株、43株和45株)。对这些转基因后代植株进行Southernblotting分析表明:PTA基因已经整合进植物基因组中,潮霉素抗感实验结果表明大多数转基因植株能够将外源基因稳定遗传给后代并能表达。
The pCAMBIA1304 vector that carried multi-gene(hygromycin gene,pta gene,mGFP and GUS genes) was transferred into calli,which were derived from immature embryos of three rice varieties(Japonica rice No.9 of Zhonghua and Ribenqing and Indica rice 501) by Agrobacterium tumefaciens.These calli were cultured on selective media,which contained 65,45 and 35 mg/?L hygromycin,respectively,and the resistant calli were obtained.Ninety-two positive transgenic plants(forty-three No.9 of Zhonghua,forty-five Ribenqing and four 501) were identified by PCR from 415 T_0 regeneration plants.Southern blotting assay for these transgenic progenies indicated that the pta gene was successfully introduced and integrated into the genome of the receptors.The result of hygromycin resistance showed that the outer-gene can be stably inherited and expressed in transgenic plants.
出处
《西南农业学报》
CSCD
2005年第3期229-233,共5页
Southwest China Journal of Agricultural Sciences
基金
国家863计划项目(2003AA212030)
关键词
多基因
幼胚愈伤组织
转基因植株
multi-gene
immature embryo calli
transgenic plants
