|Table of Contents|

Optimization of Agrobacterium-mediated Genetic Transformation of Soybean with Transient Expression of GUS Gene(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2019年03期
Page:
353-359
Research Field:
Publishing date:

Info

Title:
Optimization of Agrobacterium-mediated Genetic Transformation of Soybean with Transient Expression of GUS Gene
Author(s):
(Key Laboratory of Soybean Biology in Chinese Ministry of Education/Key Laboratory of Biology and Genetics & Breeding for Soybean in Northeast China, Ministry of Agriculture, Northeast Agricultural University, Harbin 150030, China)
Keywords:
Agrobacterium tumefaciens; Soybean genetic transformation; Transformation rate; Transient expression of GUS gene
PACS:
-
DOI:
10.11861/j.issn.1000-9841.2019.03.0353
Abstract:
High transformation efficiency is an important factor for plant molecular breeding. Agrobacterium tumefaciens-mediated transformation is commonly used in plant genetic transformation because its higher heritability and lower copy number. To optimize the soybean genetic transformation system used in our laboratory, we optimized Agrobacterium tumefaciens-mediated transformations of Dongnong 47, Kennong 18 and B12088 soybean varieties of germination time, bean cutting methods, infestation time, co-culture method and co-culture time with the expression of 35S∶GUS of pCAMBIA3301. The results showed that the expression level of GUS was highest when Dongnong 47 was germinated for 1 d, infested for 30 min and Kennong 18 and B12088 germinated for 12 h, infected for 30 min. In other treatments, the results of the three varieties were consistent. The GUS gene expression was highest when we cut the soybean cotyledonary with dipped Agrobacterium and when the cotyledon′s wound is placed face down and co-cultivated for 3-5 d in the dark. Under the optimal conditions, the Kennong 18 showed the highest transformation efficiency. This study provides an improved protocol for Agrobacterium tumefaciens-mediated transformation of soybean cotyledonary nodes that can be utilized for soybean genetic transformation.

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Last Update: 2019-05-30