EVALUATION OF 16S-23S rDNA SPACER REGION AS A MOLECULAR MARKER TO DIFFERENTIATION OF ACIDOVORAX AVENAE SUBSP. CATTLEYAE AND BURKHOLDERIA GLADIOLI PV. GLADIOLI PATHOGENIC TO ORCHIDS

ABSTRACT

The 16S-23S rDNA spacer region was evaluated as a molecular marker to differentiation of orchids pathogens, Acidovorax avenae subsp. cattleyae and Burkholderia gladioli pv. gladioli. Other species belonging to the two genera such as B. gladioli pv. alliicola, A. avenae subsp. avenae, A. avenae subsp. citrulli, A. anthurii, A. facilis, A. delafieldii and A. konjaci were also analyzed. The PCR amplification of the 16S-23S rDNA spacer region produced a fragment of 1,100 base pairs. The products obtained from all strains were digested with Afa I, Alu I, Hae III and Hpa II and the results showed distinct RFLP patterns for each endonuclease tested. The sequences of 16S-23S rDNA spacer region were aligned and the phylogenetic analysis confirmed the differences between A. avenae subsp. cattleyae e B. gladioli pv. gladioli. The PCR-RFLP technique was very efficient and useful to the identification of these pathogens since they cause very similar symptoms in orchids.

KEY WORDS
PCR-RFLP; phytobacteria diagnosis; Phalaenopsis sp.