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Culture and Exogenous Gene Transformation of Suspension Cells in Soybean(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2015年01期
Page:
36-41
Research Field:
Publishing date:

Info

Title:
Culture and Exogenous Gene Transformation of Suspension Cells in Soybean
Author(s):
FANG Xing;  HU Qian-qian;  WANG Jian-guo;  YUE Na;  MENG Xiang-xun
School of Basic Medicine and Biological Science, Soochow University, Suzhou 215123, Jiangsu)
Keywords:
Soybean;  Suspension cell;  Born acid; Agrobacterium;  Sonication;  Transformation
PACS:
S565.1
DOI:
10.11861/j.issn.1000-9841.2015.01.0036
Abstract:
In the experiment, a soybean suspension cells was initially established with the callus induced from the hypocotyl of sterile seedling of Jilinxiaoli 7 soybean. And then the influence of concentrations of boric acid on the growth of suspension cell cultures (fresh weight) were determinated and the transformation of exogenous GUS gene to the cells were conducted by using Agrobacterium-mediated method. The transformation efficiencies influenced by sonication duration and boric acid concentration were mainly investigated through histochemical method in order to find an appropriate genetic transformation way. The positive transformed cell cultures were further identified by PCR technique and Southern blot hybridization. The results showed that boric acid concentration for normal growth of soybean suspension cell was between 2.0 and 10 mg ? L-1, higher or lower boron medium would inhibited soybean cell growth, especially at a high level of more than 100 mg ? L-1. However, the appropriate increase in boric acid concentration (10-30 mg ? L-1) could improve the transformation efficiency of the cell suspension, which was also helpful to the screening of the transformant in the transformation process. Twenty eight transformants were obtained in 1 mg transformed cells under 30 mg?L-1?boric acid concentration with the assistance of 3 min sonication.

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Last Update: 2015-04-12