›› 2009, Vol. 21 ›› Issue (6): 0-548.

• 论文 •    

马铃薯3GT基因转化拟南芥引起花色苷大量积累

卢其能1,2,杨 清2,沈春修1
  

  1. 1宜春学院 生命科学与资源环境学院,江西 宜春 336000;2南京农业大学 生命科学学院生化与分子生物学系,江苏 南京 210095
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-11-25 发布日期:2009-11-25

Accumulation of anthocyanins in Arabidopsis thaliana caused by transformation with 3GT gene from wild potato

LU Qi-neng;YANG Qing;SHEN Chun-xiu   

  1. 1 School of Life Sciences,Resources and Environment Sciences,Yichun University,Yichun 336000,China;2 Department of Biochemistry and Molecular Biology,College of Life Sciences,Nanjing Agricultural University,Nanjing 210095,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-11-25 Published:2009-11-25

摘要:

为了验证马铃薯野生种3GT(UDP?鄄glucose:flavonoid 3?鄄0?鄄glucosyltransferase,类黄酮3?鄄O?鄄葡萄糖基转移酶)基因的功能,构建了携带CaMV 35S启动子的表达载体pG3GT并转化农杆菌GV3101。用菌液浸泡花序法对拟南芥进行遗传转化,在含50 mg/L Kan的培养基上对T0代种子进行筛选,先后得到4个阳性幼苗,转化率为0.13%。对移栽成活的3株抗性植株进行PCR检测为阳性,Southern blot分析有2株表现为阳性并为单拷贝整合,其中一株的叶片和茎杆颜色变成紫红色;经花色苷含量的测定表明其含量比野生型植株高出7.01倍。



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关键词: 马铃薯, 遗传转化, 类黄酮3-O-葡萄糖基转移酶基因

Abstract: To verify the function of 3GT(UDP-glucose:flavonoid 3-O-glucosyltransferase) gene isolated from wild potato,the 3GT gene was inserted into the plant expression vector pG3GT with CaMV 35S promoter,and then was introduced into Arabidopsis thaliana by Agrobacterium GV3101 through floral dip method. There were four transformants selected for their ability to grow on medium containing 50 mg/L kanamycin. The frequency of transformants was 0.13%. There were three survival transformants conformed by PCR and two transformants conformed by Southern blotting analysis. Only simple copy of 3GT gene had been transferred into the Arabidopsis thaliana genome conformed by Southern blotting analysis. There was one transformant of them whose stem and leaves turned purple,the content of anthocyanin was 7.01 times of the wild type.

Key words: potato, genetic transformation, UDP-glucose:flavonoid 3-O-lucosyltransferase gene