Specific PCR detection and identification of Xylella fastidiosa strains causing citrus variegated chlorosis.
By cloning and sequencing specific randomly amplified polymorphic DNA (RAPD) products, we have developed pairs of PCR primers that can be used to detect Xylella fastidiosa in general, and X. fastidiosa that cause citrus variegated chlorosis (CVC) specifically. We also identified a CVC-specific region of the X. fastidiosa genome that contains a 28-nucleotide insertion, and single base changes that distinguish CVC and grape X. fastidiosa strains. When using RAPD products to develop specific PCR primers, we found it most efficient to screen for size differences among RAPD products rather than presence/absence of a specific RAPD band.
Fruit Laboratory, United States Department of Agriculture, Beltsville Agricultural Research Center, MD 20705, USA.
M R
MR
Pooler
J S
JS
Hartung
D001483:Base Sequence; D002957:Citrus / Q000382:microbiology*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D006090:Gram-Negative Bacteria / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology*; D019105:Random Amplified Polymorphic DNA Technique; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D017931:DNA Primers; D004269:DNA, Bacterial