|Grayburn W. Scott,|
|Vick Brady A,|
Submitted to: Plant Cell Reports
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: September 3, 1994
Publication Date: N/A
Interpretive Summary: This article describes a method to introduce foreign DNA into cultivated sunflower. Fewer manipulations are required than with previously published transformation methods. Following wounding with glass beads, young seedlings are incubated with Agrobacterium tumefaciens, allowing DNA transfer to plant cells. DNA hybridization experiments were used to verify DNA transfer and rule out the possibility of bacterial contamination.
Technical Abstract: A procedure was developed for transformation of Helianthus annuus (sunflower) using Agrobacterium tumefaciens. Cotyledons were removed from young seedlings, and the remaining tissue was uniformly wounded by shaking with glass beads. The wounded tissue was then co-cultivated with a hypervirulent strain of Agrobacterium tumefaciens harboring the binary plasmid pCNL56. Minimal use of defined medium was required, and no callus was observed. The polymerase chain reaction (PCR) and hybridization techniques demonstrated the presence of gusA DNA from pCNL56 in total leaf DNA of six primary transformants and two progeny plants. No Agrobacterium DNA was detected in total DNA from transformed sunflower leaves that was amplified with primers specific to the miaA chromosomal gene of Agrobacterium. Grafting was used to increase the number of seeds present on plants that had undergone tissue culture manipulations.