草业学报2011，Vol.20Issue(1)：102-110,9.
甘露醇及AgNO3对根癌农杆菌介导的多年生黑麦草遗传转化的影响
The effects of mannitol and AgNO3 for perennial ryegrass transformation mediated by Agrobacterium tume faciens
摘要
Abstract
Perennial ryegrass (Lolium perenne), a graminaceous crop, is one of the most widely distributed grass in the temperate regions for forage and turf. However, its transformation mediated by Agrobacterium tumefaciens is largely recalcitrant and lagged. In this paper, an improved method for perennial ryegrass mediated by A. tumefaciens was developed. And the transgenic plants were successfully obtained. Embryogenic calli induced from mature embryos of a perennial ryegrass variety Tove were transformed by A. tumefaciens strain EHA105 containing the plasmid pCAMBIA2301. In this protocol, 0.1 mol/L mannitol or 5 mg/L silver nitrate (AgNO3) were added in preculture and coculture media. After selected with paromomycin and detected with PCR and GUS expression, we got 38 transgenic plants in all. The transgenic frequency was increased when 0.1 mol/L mannitol or 5 mg/L AgNO3 or 5 mg/L AgNO3 ＋0.1 mol/L mannitol was added in the pre-cultivation and co-cultivation media, which was 1.96, 1.59 and 2.95 times compared to control respectively. Moreover,the transgenic frequency was highest when the two materials were jointly used. The results indicated that mannitol and AgNO3 could improve the A. tumefaciens mediated transformation in perennial ryegrass evidently.关键词
多年生黑麦草/根癌农杆菌/遗传转化/愈伤组织/硝酸银/甘露醇Key words
perennial ryegrass/ Agrobacterium tumefaciens/ genetic transformation/ callus induction/ silver nitrate / mannitol分类
农业科技引用本文复制引用
乔定君,毛萍,马欣荣,杨宏..甘露醇及AgNO3对根癌农杆菌介导的多年生黑麦草遗传转化的影响[J].草业学报,2011,20(1):102-110,9.基金项目
国家863计划项目(2008AA10Z409,2009AA10Z108)和中国科学院知识创薪工程重要方向项目(KSCX2-YW-N-046)资助. （2008AA10Z409,2009AA10Z108）
