Submitted to: Current Microbiology
Publication Type: Peer reviewed journal
Publication Acceptance Date: 4/5/1999
Publication Date: N/A
Citation: Banks, D., Albibi, R., Chen, J., Lamikanra, O., Jarret, R.L., Smith, B.J. 1999. Specific detection of Xylella fastidiosa Pierce's Disease strains. Current Microbiology. 39:85-88 Interpretive Summary: Pierce's disease of grapevine limits commercial production of bunch grape in the southeastern United States. The bacterial pathogen, Xylella fastidiosa, is difficult to isolate and identify, and strains are pathogenically specialized. Specific identification of Pierce's disease strains is a prerequisite for ecological and epidemiological studies of this important pathogen. A unique random amplified polymorphic DNA was isolated from Pierce's disease infected a grapevine grown in Florida. This fragment was cloned and PCR primers were designed to utile this sequence for Pierce's disease detection. The results of this research will be used by scientists to develop techniques to detect the Pierce's disease bacterium directly in grape tissue. Quick and accurate detection of this pathogen will lead to a better understand of how the bacterium infects grapevines and other hosts. Ultimately farmers will benefit from improved control of this disease.
Technical Abstract: Pierce's disease (PD, Xylella fastidiosa) of grapevine is the primary pathogen limiting vinifera grape production if Florida and other regions of the southeastern United States. Quick and accurate detection of PD strains is essential for PD studies and control. A unique random amplified polymorphic DNA (PD1-1-2) was isolated from a PD strain from Florida. Fragment PD1-1-2 was cloned, sequenced, and found to be 1005 bp in length. PCR primers were designed to utilize these sequence data for PD strain detection. One primer set (XF176f-XF954r) amplified a 779-bp DNA fragment from 34 PD strains including seven pathotypes of X. fastidiosa, but not from strains of Xanthomonas campestris pv. campestris, Xan. vesicatoria or Escherichia coli. A second primer set (XF176f and XF686r) amplified a 511-bp fragment specific to 98 PD strains, but not from strains of citrus variegated chlorosis, mulberry leaf scorch, oak leaf scorch, periwinkle wilt, phony peach, or plum leaf scald. Sequence analysis indicated that RAPD fragment PD1-1-2 contains a Ser-tRNA gene. The PD-specific region includes a TaqI restriction site (TCGA) and is 150 bp downstream of the Ser-tRNA gene.