Submitted to: American Phytopathological Society Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: August 9, 1997
Publication Date: N/A
Technical Abstract: Our current PCR protocol for Xanthomonas albilineans (Xa) amplifies a unique 366 bp DNA fragment from all Xa strains tested. However, products of 280, 420, and 460 bp, respectively, are also amplified from three bacterial saprophytes occasionally found in sugarcane tissues. To design new primers that are highly specific to Xa, these PCR products were cloned into a plasmid vector and sequenced. The NCBI BLASTN program was used to search for non-redundant GenBANK+EMBL+DDBJ+PDB database sequences that were highly homologous to the Xa sequence. Using computer programs for multiple sequence alignment and primer design, we developed a pair of new Xa-specific PCR primers. When these new primers were substituted into the current PCR protocol, an expected product of 288 bp was amplified from Xa only. This specificity was confirmed on all Xa strains collected worldwide including representatives of serovars I, II, and III. No amplification was sobserved from the sugarcane saprophytes or any other bacterial species tested.