caa a22 4500 467929823 CHVBK 20180406152940.0 cr unu---uuuuu 170328e20060601xx s 000 0 eng 10.1007/s10658-006-9009-4 doi (NATIONALLICENCE)springer-10.1007/s10658-006-9009-4 Genome-based PCR Primers for Specific and Sensitive Detection and Quantificationof Xylella fastidiosa [Elektronische Daten] [Marta Francis, Hong Lin, Juan Rosa, Harshavardhan Doddapaneni, Edwin Civerolo] Xylella fastidiosa is an important pathogen of many commercial crops. Detection of X. fastidiosa is difficult due to low concentrations of the bacteria in insects and asymptomatic plant tissue, and non-uniform distribution in infected plants. A dual purpose conventional PCR and quantitative PCR (TaqMan™) system was developed for the generic detection of X. fastidiosa strains. Primers HL5 and HL6, designed to amplify a unique region common to the sequenced genomes of four Xylella strains, amplified a 221bp fragment from strains associated with Pierce's disease of grapes, almond leaf scorch, and oleander leaf scorch disease and from DNA from an Xf strain associated with citrus variegated chlorosis. Standard curves were obtained using concentrations of Xylella ranging from 5 to 105 cells per reaction in water and grape extracts and 10-105 cells in insect DNA. Regression curves were similar, with correlation coefficients of r 2 > 0.97. In quantitative PCR, Ct values ranged between 20 and 36 cycles for 5-105 bacterial cells per reaction. No amplicons were obtained with several non-Xf bacterial strains tested including related plant pathogenic, grape endophytic bacteria and endosymbiotic bacteria isolated from glassy-winged sharpshooters. The method was evaluated for clinical diagnosis of Xf in grapes, almonds and insect vectors. The procedure described is reliable for detection of the pathogen with a high degree of sensitivity and specificity. Springer, 2006 almond leaf scorch nationallicence citrus variegated chlorosis nationallicence conventional PCR nationallicence oleander leaf scorch nationallicence Pierce's disease of grapes nationallicence quantitative PCR nationallicence ALSD : almond leaf scorch disease nationallicence CTAB : hexadecyltrimethylammonium bromide nationallicence GWSS : glassy-winged sharpshooter nationallicence ITS : internal transcribed spacer nationallicence OLSD : oleander leaf scorch disease nationallicence PCR : polymerase chain reaction nationallicence PD : Pierce's disease nationallicence Q-PCR : quantitative PCR nationallicence Xf : Xylella fastidiosa nationallicence Francis Marta Department of Plant Pathology, University of California Davis, 95616, Davis, CA, USA aut Lin Hong USDA-ARS, San Joaquin Valley Agricultural Sciences Center, 9611 So. Riverbend Ave., 93648, Parlier, CA, USA aut Rosa Juan Department of Entomology, University of California, 92521, Riverside, CA, USA aut Doddapaneni Harshavardhan Department of Viticulture and Enology, University of California Davis, 95616, Davis, CA, USA aut Civerolo Edwin USDA-ARS, San Joaquin Valley Agricultural Sciences Center, 9611 So. Riverbend Ave., 93648, Parlier, CA, USA aut European Journal of Plant Pathology Kluwer Academic Publishers 115/2(2006-06-01), 203-213 0929-1873 115:2<203 2006 115 10658 https://doi.org/10.1007/s10658-006-9009-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10658-006-9009-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Francis Marta Department of Plant Pathology, University of California Davis, 95616, Davis, CA, USA aut NATIONALLICENCE 700 1- Lin Hong USDA-ARS, San Joaquin Valley Agricultural Sciences Center, 9611 So. Riverbend Ave., 93648, Parlier, CA, USA aut NATIONALLICENCE 700 1- Rosa Juan Department of Entomology, University of California, 92521, Riverside, CA, USA aut NATIONALLICENCE 700 1- Doddapaneni Harshavardhan Department of Viticulture and Enology, University of California Davis, 95616, Davis, CA, USA aut NATIONALLICENCE 700 1- Civerolo Edwin USDA-ARS, San Joaquin Valley Agricultural Sciences Center, 9611 So. Riverbend Ave., 93648, Parlier, CA, USA aut NATIONALLICENCE 773 0- European Journal of Plant Pathology Kluwer Academic Publishers 115/2(2006-06-01), 203-213 0929-1873 115:2<203 2006 115 10658 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer