Arabidopsis ovule is the target for Agrobacterium in planta vacuum infiltration transformation
Article first published online: 5 JAN 2002
The Plant Journal
Volume 19, Issue 3, pages 249–257, August 1999
How to Cite
Ye, G.-N., Stone, D., Pang, S.-Z., Creely, W., Gonzalez, K. and Hinchee, M. (1999), Arabidopsis ovule is the target for Agrobacterium in planta vacuum infiltration transformation. The Plant Journal, 19: 249–257. doi: 10.1046/j.1365-313X.1999.00520.x
- Issue published online: 5 JAN 2002
- Article first published online: 5 JAN 2002
- Received 1 March 1999; revised 4 June 1999; accepted 4 June 1999.
The visual marker GUS has been utilized in this study to understand the Arabidopsis thaliana vacuum infiltration transformation process by Agrobacterium tumefaciens. High transformation frequencies of up to 394 transgenic seeds per infiltrated plant were achieved. The results showed that the majority of the transgenic seeds from single infiltrated plants were from independent transformation events based on Southern analysis, progeny segregation, distribution of transgenic seeds throughout the infiltrated plants and the microscopic analysis of GUS expression in ovules of infiltrated plants. GUS expression in mature pollen and anthers was monitored daily from 0 to 12 days post-infiltration. In addition, all ovules from a single infiltrated plant were examined every other day. GUS expression frequencies of up to 1% of pollen were observed 3–5 days post-infiltration, whereas frequencies of up to 6% were detected with ovules of unopened flowers 5–11 days post-infiltration. Most importantly, transgenic seeds were obtained only from genetic crosses using infiltrated plants as the pollen recipient but not the pollen donor, demonstrating Agrobacterium transformation through the ovule pathway.