Transformation of coffee (Coffea Arabica L. cv. Catimor) with the cry1ac gene by biolistic, without the use of markers.
The transformation of coffee plantlets with the cry1ac gene of Bacillus thuringiensis was achieved by biolistic using either the whole pUBC plasmid or only the ubi-cry1ac-nos genetic cassette. The cry1ac gene was inserted into coffee plants in order to confer resistance to the leaf miner Leucoptera coffeella, an insect responsible for considerable losses in coffee crops. Bearing in mind that the genetic cassettes used for this study lack reporter genes and/or selection marker genes, the parameters for the transformation procedure by biolistic were previously standardised with a plasmid carrying the gus reporter gene. The presence of the cry1ac gene in young plantlet tissues was determined by PCR, Southern blot and reverse transcription-PCR. Our results show that the obtainment of viable coffee plantlets, transformed by bombardment with the cry1ac gene and without selection markers nor reporter genes, is feasible.
De Guglielmo-Cróquer
Z
Z
Instituto de Biología Experimental, Universidad Central de Venezuela, Caracas, Venezuela.
Altosaar
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I
Zaidi
M
M
Menéndez-Yuffá
A
A
D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics*; D019470:Biolistics / Q000379:methods; D015153:Blotting, Western; D040503:Coffea / Q000235:genetics*; D004731:Endotoxins / Q000235:genetics*; D006460:Hemolysin Proteins / Q000235:genetics*; D007915:Lepidoptera; D010935:Plant Diseases / Q000469:parasitology / Q000517:prevention & control; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction; D014170:Transformation, Genetic / Q000235:genetics*
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis