Detection of Pseudomonas syringae pv. actinidiae using polymerase chain reaction (PCR) primers based on the 16S–23S rDNA intertranscribed spacer region and comparison with PCR primers based on other gene regions

摘要: Several published polymerase chain reaction (PCR) primers to identify Pseudomonas syringae pv. actinidiae, the causal organism of bacterial canker kiwifruit, were found not be specific. Two new sets PCR primers, PsaF1/R2 and PsaF3/R4, designed complementary a portion 16S–23S rDNA intertranscribed spacer (ITS) regions. These amplified DNA fragment from strains P. but 56 bacteria six genera 17 species, except for strain tea pathogen, theae. When tested against extracted further 20 Japan, Korea, Italy USA deposited in culture collections as all cultures produced expected product 280 bp with 175 bp PsaF3/R4. Results multilocus sequence analysis using five housekeeping genes (gyrB, acnB, rpoD, pgi cts) showed that none these was phylogenetically similar actinidiae. In contrast actinidiae type strain, positive determinative tests ice nucleation syringomycin production. It is suggested incorrectly identified possible distinguish theae ITS, gyrB, or cts gene regions design primers. Because unlikely on PsaF3/R4 are recommended screening isolated kiwifruit tissue.