|Table of Contents|

Optimization of Cotyledonary-node Agrobacterium-mediated Soybean Transformation System(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2015年05期
Page:
768-775
Research Field:
Publishing date:

Info

Title:
Optimization of Cotyledonary-node Agrobacterium-mediated Soybean Transformation System
Author(s):
ZHAI Rui; GAO Le; DING Xue-ni; LIAO Wen-lin; ZHENG Ming-jie; LU Zhi-wen; ZHI Hai-jian
Soybean Research Institute of Nanjing Agricultural University/National Center for Soybean Improvement/National Key Laboratory for Crop Genetics and Germplasm Enhancement/Key Laboratory of Biology and Genetic Improvement of Soybean, Ministry of Agriculture, P.R. China, Nanjing 210095, China
Keywords:
Soybean;  Cotyledonary-node;  Agrobacterium tumefaciens;  Optimization.
PACS:
-
DOI:
10.11861/j.issn.1000-9841.2015.05.0768
Abstract:
In the present study, cotyledon nodes obtained from 17 soybean cultivars were infected with Agrobacterium tumefacines EHA105.Subsequently, we optimized the soybean genotype, chlorine sterilization time, explant state, vitality of bacterial strain, concentration of infection liquid, infection time and co-cultivation time. As a result, when the chlorine sterilization time was 14-18 h, the minimal pollution and the highest seed vigor was observed. The vitality of overnight-treated explants in the dark were better than those germinated in the light for 5-7 d. The highest GUS transient rate was achieved when the bacterial concentration at OD600nm was 0.8-1.0 and the infection liquid at OD600nm?was 0.6-0 .8.We also found that the best infection and co-cultivation time were 30 min and 4 d, respectively. Based on the above optimized research, we established a comprehensive transgenic system which resulted in the maximum transformation efficiency of 3.33%. For the comprehensive evaluation about the transient GUS expression under different treatments and the shoot induction rate of 17 soybean varieties, TL-1, HC-3, HC-6 and Williams 82 were the ideal genotypes for transformation.

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Last Update: 2015-11-07