Pierce's disease (PD, Xylella fastidiosa) of grapevine is the primary pathogen limiting vinifera grape production in 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.
Pierce's disease (PD) strains of Xylella fastidiosa were identified by random amplified polymorphic DNA (RAPD) fingerprinting. Two random primers including OPA-03 (agtcagccac) and OPA-11 (caatcgccgt) were found to be efficient for differentiating PD strains isolated from a vineyard in North Florida in 1996 (129 strains) and 1997 (29 strains) from non-PD strains of X. fastidiosa (citrus variegated chlorosis, mulberry leaf scorch, periwinkle wilt, plum leaf scald, and phony peach) and strains from Xanthomonas campestris pv. vesicatoria and Escherichia coli. This study shows that RAPD fingerprinting is a useful tool to supplement the conventional symptoms-colony morphology-slow growth identification procedure routinely used to identify the PD pathogen. z
Pierce's disease (PD) strains of Xylella fastidiosa were identified by random amplified polymorphic DNA (RAPD) fingerprinting. Two random primers including OPA‐03 (agtcagccac) and OPA‐11 (caatcgccgt) were found to be efficient for differentiating PD strains isolated from a vineyard in North Florida in 1996 (129 strains) and 1997 (29 strains) from non‐PD strains of X. fastidiosa (citrus variegated chlorosis, mulberry leaf scorch, periwinkle wilt, plum leaf scald, and phony peach) and strains from Xanthomonas campestris pv. vesicatoria and Escherichia coli. This study shows that RAPD fingerprinting is a useful tool to supplement the conventional symptoms‐colony morphology‐slow growth identification procedure routinely used to identify the PD pathogen.
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