ARS | Publication request: POLYMERASE CHAIN REACTION (PCR) PROTOCOLS FOR DETECTING THE BACTERIA THAT CAUSE LEAF SCALD AND RATOON STUNTING DISEASES OF SUGARCANE.

Submitted to: American Society of Sugar Cane Technologists
Publication Type: Abstract Only
Publication Acceptance Date: June 18, 1997
Publication Date: N/A

Technical Abstract: Two pairs of polymerase chain reaction (PCR) primers were developed that primed specific amplification of the 16S and 23S ribosomal intergenic transcribed spacer region from Xanthomonas albilineans (Xa) and Clavibacter xyli subsp. xyli (Cxx), the causal agents of sugarcane leaf scald and ratoon stunting disease, respectively. A PCR protocol using primers PGBL1 and PGBL2 amplified a specific 288 bp DNA product from all Xa strains collected worldwide including representatives of serovars I, II, and III. No amplification was observed from sugarcane bacterial saprophytes, Cxx, or other related Xanthomonas species tested. Results were obtained in less than two hours. Another PCR protocol with primers Cxx1 and Cxx2 amplified a specific 438 bp DNA product from the genomic DNA of 21 Cxx strains collected worldwide. No amplification of DNA from sugarcane bacterial saprophytes, Xa, or other closely related Clavibacter species was observed. DNA products can be amplified directly from cultured Xa and Cxx cells without prior extraction of the genomic DNA. These two pathogen-specific PCR protocols can be used as diagnostic tools for identification and early detection of the two important sugarcane diseases.