Influence of bacterial density during preculture on Agrobacterium-mediated transformation of tomato |
| |
Authors: | Nan Gao Weishou Shen Yu Cao Yanhua Su Weiming Shi |
| |
Affiliation: | (1) State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, 210008 Nanjing, China;(2) Graduate School of the Chinese Academy of Sciences, 100039 Beijing, China;(3) College of Chemistry and Environmental Science, Nanjing Normal University, 210097 Nanjing, China |
| |
Abstract: | An improved bacterial preculture protocol for Agrobacterium-mediated genetic transformation was developed for an economic tomato cultivar (Solanum lycopersicum L. cv. Zhongshu No. 4). Frequencies of transient gene expression and stable transformation were influenced by the density of Agrobacterium preculture and not the density of Agrobacterium used for infection. The improved protocol presented in this study depends on the use of an overnight-grown Agrobacterium preculture density of OD600 nm = 1.0, diluted 1/10th with Luria-Bertani (LB) liquid medium, and grown for an additional 4 h. Cultures are collected and resuspended in a liquid cocultivation medium-I, adjusted to OD600 nm = 0.1. Using this modified Agrobacterium preparation, transient β-glucuronidase expression was higher than 90%, and transformation efficiency reached 44.7%. This improved transformation is simple, repeatable, does not require a feeder layer, and most notably, the transformation frequency is stable and highly efficient. |
| |
Keywords: | Density of bacterial cells nptII Solanum lycopersicum L. Transient β -glucuronidase (GUS) expression |
本文献已被 SpringerLink 等数据库收录! |
|