Evaluation of transgenic 'Chardonnay' (Vitis vinifera) containing magainin genes for resistance to crown gall and powdery mildew.
Magainins, short peptides with broad-spectrum antimicrobial activity in vitro, were assayed for their ability to confer resistance to pathogens in transgenic grapevines. Embryogenic cell suspensions of 'Chardonnay' (Vitis vinifera L.) were co-transformed by microprojectile bombardment with a plasmid carrying the npt-II gene and a second plasmid harboring either a natural magainin-2 (mag2) or a synthetic derivative (MSI99) gene. Magainin genes and the marker gene were driven by Arabidopsis ubiquitin-3 and ubiquitin-11 promoters, respectively. A total of 10 mag2 and 9 MSI99 regenerated lines were studied by Southern blot hybridization, which showed 1-6 transgene integration events into the plant genome. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) revealed a variable range in transcription levels among mag2 and MSI99 lines. A positive correlation between number of integration events and transcription level was observed (p<0.05). Plants were acclimated and challenged in the greenhouse with either Agrobacterium vitis strains (bacterial crown gall pathogen) at 10(8) cfu/ml or Uncinula necator (fungal powdery mildew pathogen) at 10(5) conidia/ml for evaluation of disease resistance. A total of 6 mag2 and 5 MSI99 lines expressing the antimicrobial genes exhibited significant reductions of crown gall symptoms as compared to non-transformed controls. However, only two mag2 lines showed measurable symptom reductions in response to U. necator, but not strong resistance. Our results suggest that the expression of magainin-type genes in grapevines may be more effective against bacteria than fungi. Additional strategies to enhance transgene expression and the spectrum of resistance to grape diseases are suggested.
Department of Horticultural Sciences, New York State Agricultural Experiment Station, Cornell University, Geneva, NY 14456, USA.
José R
JR
Vidal
Julie R
JR
Kikkert
Mickael A
MA
Malnoy
Patricia G
PG
Wallace
John
J
Barnard
Bruce I
BI
Reisch
D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides / Q000235:genetics* / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics; D015139:Blotting, Southern; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics* / Q000502:physiology; D010941:Plant Tumors / Q000235:genetics* / Q000382:microbiology*; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012231:Rhizobium / Q000472:pathogenicity; D027843:Vitis / Q000235:genetics* / Q000378:metabolism / Q000382:microbiology
D023181:Antimicrobial Cationic Peptides; C446524:MSI-99 peptide; D010455:Peptides; D011994:Recombinant Proteins