EVALUATION OF DIFFERENT METHODS OF DNA EXTRACTION FOR DETECTION OF BACTERIUM Xanthomonas campestris pv. Campestris IN CABBAGE LEAVES

Abstract

The bacterium Xanthomonas campestris pathovar campestris (Xcc) as the causal agent of black rot of cruciferous plants causes considerable losses in agricultural yield all over the world. The control of black rot is difficult as well as the determination of Xcc on the basis of morphological parameters or by pathogenicity testing. Ten different possibilities for extraction bacterial DNA followed by PCR detection method were tested to optimize PCR protocol. On the basis of ISTA validated method, three sets of primers UBP 1052F-BACR, DLH 120-125 and ZUP 2309-2310 were used. The results of measured concentration and quality of DNA and efficacy for PCR amplification were compared. Finally, three approaches for DNA extraction within Xanthomonas campestris pv. campestris detection protocol were recommended – commercial kits used for isolation from tissues by Macherey-Nagel and MO BIO and kit intended for microbial cultures by MO BIO.

Keywords

• black rot
• Brassicaceae
• specific PCR