Ws. Grayburn et Ba. Vick, TRANSFORMATION OF SUNFLOWER (HELIANTHUS-ANNUUS L) FOLLOWING WOUNDING WITH GLASS-BEADS, Plant cell reports, 14(5), 1995, pp. 285-289
A procedure was developed for transformation of Helianthus annuus (sun
flower) using Agrobacterium tumefaciens. Cotyledons were removed from
young seedlings, and the remaining tissue was uniformly wounded by sha
king with glass beads. The wounded tissue was then co-cultivated with
hypervirulent strain of Agrobacterium tumefaciens harboring the binary
plasmid pCNL56. Minimal use of defined medium was required, and no ca
llus was observed. The polymerase chain reaction (PCR) followed by DNA
hybridization demonstrated the presence of gusA DNA from pCNL56 in to
tal leaf DNA of 6 primary transformants and 2 progeny plants. No Agrob
acterium DNA was detected in total DNA from transformed sunflower leav
es that was amplified with primers specific to the miaA chromosomal ge
ne of Agrobacterium. Foreign DNA was also detected in the next generat
ion. beta-Glucuronidase (GUS) activity was demonstrated for 5 of the T
-2 transgenic plants. Grafting was used to increase the number of seed
s present on plants that had undergone tissue culture manipulations.