Journal
BRAZILIAN JOURNAL OF BIOLOGY
Volume 70, Issue 2, Pages 387-393Publisher
INT INST ECOLOGY
DOI: 10.1590/S1519-69842010000200022
Keywords
coffee; biolistic; genetic cassette; cry1ac gene; transformation without markers
Categories
Funding
- FONACIT
- CDCH
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The transformation of coffee plantlets with the cry1ac gene of Bacillus thuringiensis was achieved by Holistic using either the whole pUBC plasmid or only the ubi-ery/ac-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 Holistic 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.
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