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ISHS Acta Horticulturae 521: XXV International Horticultural Congress, Part 11: Application of Biotechnology and Molecular Biology and Breeding - Gene Expression and Molecular Breeding, Genome Analysis.

PCR-BASED CLONING OF CANDIDATE DISEASE RESISTANCE GENES FROM BANANA (MUSA ACUMINATA)

Authors:   L. Wiame, R. Swennen, L. Sági
Keywords:   banana, cloning, disease resistance, polymerase chain reaction
Abstract:
Plant disease resistance genes involved in signal transduction contain domains that are conserved throughout mono- and dicotyledons. Primers have been designed to those domains in the RPS2 gene of Arabidopsis thaliana and the N gene of tobacco. Using these primers for PCR, candidate resistance genes have already been cloned from soybean, potato, rice, barley and Arabidopsis. We apply a similar strategy to clone candidate resistance genes from banana.

PCR performed on banana DNA with the conserved primers resulted in amplification of fragments ranging from 500 to 2000 bp. Several fragments were cloned and sequenced. Based on their nucleotide sequence, the clones could be divided into 10 classes. One class consisted of clones containing a variable number of repeats of one of the conserved primer sequences. Sequence comparison to nucleotide and protein databases revealed homologies in several classes. A short part of class D showed homology to the Arabidopsis RPS2 disease resistance gene. Class E contained a putative intron, while its putative exon revealed high homology to a Cu2+-transporting ATPase.

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