DNA markers for identification of Pseudomonas syringae pv. actinidiae.
The specific DNA fragment was screened by RAPD analysis of Pseudomonas syringae pv. actinidiae, as well as similar strains that were isolated from kiwifruits. The primer C24 detected a fragment that is specific in P. syringae pv. actinidiae. This fragment was cloned. The pathovar-specific fragment was detected from a Southern blot analysis of the genomic DNAs of P. syringae pv. actinidiae using the cloned fragment as a probe. The sequence size of the cloned fragment was determined as 675 bp. A DNA Database search suggested that the fragment was a novel one. Approximately 9 kb of a single fragment was detected only in the P. syringae pv. actinidiae by a Southern blot analysis of the genomic DNAs of P. syringae pv. actinidiae. Similar strains were also detected with the use of the cloned fragment as a probe. Since the genomic DNAs were digested with HindIII without a cleavage site, the result reveals that the cloned fragment exists on the genome of P. syringae pv. actinidiae as a single copy. A pair of primers that produced a 492 bp single fragment (only in the strains of P. syringae pv. actinidiae) were synthesized, based on the pathovar-specific sequences of the cloned fragment of P. syringae pv. actinidiae. The development of the primers and probe made it possible to diagnose the bacterial canker infection from leaves or trunks of kiwifruit trees before any symptom appeared on the tree.
Koh
Young Jin
YJ
Major of Applied Biology, School of Environmental and Agricultural Science, Sunchon National University, Korea.
Nou
Ill Sup
IS
D029042:Actinidia / Q000382:microbiology*; D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D005638:Fruit / Q000382:microbiology; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology*; D012150:Polymorphism, Restriction Fragment Length; D011549:Pseudomonas / Q000145:classification* / Q000235:genetics*; D019105:Random Amplified Polymorphic DNA Technique
D004269:DNA, Bacterial