Agrobacterium-mediated transformation of apricot (Prunus armeniaca L.) leaf explants.
A protocol for Agrobacterium-mediated stable transformation for scored, whole leaf explants of the apricot (Prunus armeniaca) cultivar Helena was developed. Regenerated shoots were selected using a two-step increased concentrations of paromomycin sulphate. Different factors affecting survival of transformed buds, including possible toxicity of green fluorescent protein (GFP) and time of exposure to high cytokine concentration in the regeneration medium, were examined. Transformation efficiency, based on PCR analysis of individual putative transformed shoots from independent lines was 5.6%, when optimal conditions for bud survival were provided. Southern blot analysis on four randomly chosen PCR-positive shoots confirmed the presence of the nptII transgene. This is the first time that stable transformation of an apricot cultivar is reported and constitutes also one of the few reports on the transformation of Prunus cultivars.
Petri
César
C
Mejora de Plantas, CEBAS-CSIC, Aptd. de Correos 164, 30100, Murcia, Spain.
Wang
Hong
H
Alburquerque
Nuria
N
Faize
Mohamed
M
Burgos
Lorenzo
L
D015139:Blotting, Southern; D003470:Culture Media / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D010303:Paromomycin / Q000494:pharmacology; D018515:Plant Leaves / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D016133:Polymerase Chain Reaction; D027861:Prunus / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D012038:Regeneration / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D012231:Rhizobium / Q000235:genetics*; D014170:Transformation, Genetic
D003470:Culture Media; D049452:Green Fluorescent Proteins; D010303:Paromomycin