Efficient genetic transformation of red raspberry, Rubus ideaus L.

Abstract

We have developed an efficient transformation system for red raspberry (Rubus ideaus L.) using Agrobacterium mediated gene transfer. Using this system we have successfully introduced a gene that encodes an enzyme, S-adenosylmethionine hydrolase (SAMase), in raspberry cultivars Meeker (MK), Chilliwack (CH) and Canby (CY). Leaf and petiole expiants were inoculated with disarmed Agrobacterium tumefaciens strain EHA 105 carrying either of two binary vectors, pAG1452 or pAG1552, encoding gene sequences for SAMase under the control of the wound and fruit specific tomato E4 promoter. Primary shoot regenerants on selection medium were chimeral containing both transformed and non-transformed cells. Non-chimeral transgenic clones were developed by iterative culture of petiole, node and leaf explants, on selection medium, from successive generations of shoots derived from the primary regenerants. Percent recovery of transformants was higher with the selection marker gene hygromycin phosphotransferase (hpt), than with neomycin phosphotransferase (nptII). Transformation frequencies of 49.6%, 0.9% and 8.1% were obtained in cultivars Meeker, Chilliwack and Canby respectively from petiole expiants using hygromycin selection. Genomic integration of transgenes was confirmed by Southern hybridization. Transgenic plants from a total of 218 independent transformation events (161 MK, 4 CH, 53 CY) have been successfully established in soil.

Transgenic rose lines harboring an antimicrobial protein gene, Ace-AMP1 , demonstrate enhanced resistance to powdery mildew ( Sphaerotheca pannosa )

“…Means are ratings of disease development on five whole leaves (five leaflets per leaf) per branch, and three branches per plant somatic embryogenesis), this approach avoids the recovery of chimeric transgenic plants. This finding is especially important for plant species where the genotypic identity can be maintained by vegetative propagation (Mathews et al 1995). Although considerable variability in the level of transcription was observed among individual transformants (Fig.…”

The influence of FeEDDHA in red raspberry cultures during shoot multiplication and adventitious regeneration from leaf explants

“…However, some transgenic plants of raspberry have been obtained (Mathews et al, 1995;McNicol and Graham, 1999;Mezzetti et al, 2004). Significant differences between genotypes have been reported in micropropagation (Reed, 1990) as well as in adventitious regeneration efficiency (Turk et al, 1994;Graham et al, 1997).…”

Effect of gelling agents and antibiotics on adventitious bud regeneration from In vitro leaves of pear

“…Gellan gum, a gelling agent composed of glucuronic 1To whom correspondence should be addressed at INRA, Station d'Am~].ioration des Esp~ces Fruiti~res et OrnementaIes, BP57, 49071 Beaucouz6, France. acid, rhamnose, and glucose, has been used with success in transformation experiments on several Rosaceous crops: apple (Maheswaran et al, 1992;De Bondt et al, 1996), peach (Ye et al, 1994), and red raspberry (Mathews et al, 1995). We present here the compared effects of gellan gum and agar on adventitious bud regeneration from leaves of eight pear genotypes, and the histological analysis of the early events of bud regeneration in both cases.…”