Detection of RAPD polymorphisms in Musa sp. materials with differential response to the attack of Xanthomonas campestris pv. musacearum
Abstract
To detect RAPD polymorphisms in Musa sp. with differential response to Xanthomonas campestris pv. Musacearum six susceptible genotypes (Pisang Awak, Dwarf Cavendish, Giant Cavendish, FHIA 17, FHIA17 in vitro, FHIA25) and two tolerant (BB and BBB) were evaluated. Conditions for DNA isolation and random amplification of polymorphic DNA (RAPD) were established. DNA was isolated according to Dellaporta et al. (1983), as amended by CIAT (2002), DNA concentration was measured spectrophotometrically. Additionally, A260/A280 and A260/A230 ratios indicated the purity of the molecule. 27 Operon Technology primers (20 sequences in the series OPA, 5 from OPJ and 2 from OPM) were used. The statistical analysis was performed with a hierarchical cluster analysis using Infostat Professional version 2.0, Ward clustering method and the Jaccard distance. Specific band patterns were obtained for each genotype. Six bands were identified as unique for the tolerant genotypes (981 bp with OPA-03, 630 bp with OPA-07, 630 bp with OPA10, 1725 bp with OPJ01, 580 bp with OPM20 y 630 bp with OPJ05). OPA series allowed observation of higher levels of polymorphisms, having PIC values ranging from 0.22 y 0.68. Hierarchical conglomerate analysis separated genotypes in 2 main groups, one including genotypes with the A genome exclusively, and the other with the B genome, except for FHIA 17 in vitro. There were genetic differences between FHIA 17 growing under field conditions and the one regenerated in vitro.
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Copyright (c) 2011 Pi-Hsia Chen, Efraín Salazar, Hilda Fernández, Luis Castro, Antonio Russo, Sundry Vázquez
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