农杆菌介导GUS基因对多年生黑麦草转化的研究  

Study on Agrobacterium-mediated GUS transformation for Lolium perenne L.

作　　者：张振霞[1] 刘萍[1] 杜雪玲[2] 苏乔[3] 杨中艺[1] 

机构地区：[1]中山大学生命科学学院 [2]安徽淮北煤炭师范学院生物系,安徽淮北235000 [3]大连理工大学环境与生命学院,辽宁大连116024

出　　处：《广西植物》2007年第1期121-126,共6页Guihaia

基　　金：国家转基因植物研究与产业化专项(J2002-B-006)~~

摘　　要：通过检测愈伤组织中GUS基因的瞬间表达，研究农杆菌LBA4404／pCAMBIA1301介导多年生黑麦草的转化体系。通过对多年生黑麦草瞬间表达率的比较，确立了其遗传转化的最佳优化条件。研究发现，多年生黑麦草不同品种的转化率在25％～45％之间变化。多年生黑麦草遗传转化最佳优化条件是预培养10d的胚性愈伤组织、浓度为0．5～0．8OD的农杆菌菌液以及2d共培养时间。在共培养基中添加100μmol／L乙酰丁香酮能有效地提高植物瞬间表达率。两种侵染处理方法比较结果为滤纸滴加法比浸泡法更优。转化后对愈伤组织的干燥处理能抑制农杆菌过度繁殖，能改善愈伤状态，有利于提高转化率。Agrobacterium tumefaciens mediated transformation system of Lolium perenne was studied by using GUS gene transient expression method. Mature seed derived callus from L. perenne was transformed by GUS gene containing A. tumefaciens LBA4404/pCAMBIA1301. Frequency of the transformation was detected, and the optimal conditions for L. perenne genetic transformation were determined. It was found that the transformation frequency varied from 25 % ～45 % according to different genotypes （variety）. The optimal conditions to get higher transformation efficiency were 2 days co-cultivation with 0. 5～0. 8 OD A. turnefaciens solution in dark after 10 days pre-culturing. The transformation frequency was significantly improved by adding 100 μmol/L acetosyingone into co-cultivation medium. Adding A. turnefaciens solution on surface of filter paper was found to be a better infecting method of A. tumefaciens rather than suspending in A. tumefaciens solution. The partial desiccation treatment after transformation was beneficial to prevent over-growth of A. turnefaciens and increase the transformation efficiency.

关 键 词：多年生黑麦草 农杆菌转化 GUS基因 

分 类 号：Q943[生物学—植物学]