title: Convenient detection of the citrus greening (huanglongbing) bacterium 'Candidatus Liberibacter asiaticus' by direct PCR from the midrib extract.
abstract: A phloem-limited bacterium, 'Candidatus Liberibacter asiaticus' (Las) is a major pathogen of citrus greening (huanglongbing), one of the most destructive citrus diseases worldwide. The rapid identification and culling of infected trees and budwoods in quarantine are the most important control measures. DNA amplification including conventional polymerase chain reaction (PCR) has commonly been used for rapid detection and identification. However, long and laborious procedures for DNA extraction have greatly reduced the applicability of this method. In this study, we found that the Las bacterial cells in the midribs of infected leaves were extracted rapidly and easily by pulverization and centrifugation with mini homogenization tubes. We also found that the Las bacterial cells in the midrib extract were suitable for highly sensitive direct PCR. The performance of direct PCR using this extraction method was not inferior to that of conventional PCR. Thus, the direct PCR method described herein is characterized by its simplicity, sensitivity, and robustness, and is applicable to quarantine testing.
authors: National Institute of Fruit Tree Science, Tsukuba, Ibaraki, Japan.
authors: Takashi
authors: T
authors: Fujikawa
authors: Shin-Ichi
authors: S
authors: Miyata
authors: Toru
authors: T
authors: Iwanami
mesh_terms: D002957:Citrus / Q000382:microbiology*; D004269:DNA, Bacterial / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence; D010935:Plant Diseases / Q000382:microbiology*; D018515:Plant Leaves / Q000737:chemistry / Q000382:microbiology; D016133:Polymerase Chain Reaction; D012230:Rhizobiaceae / Q000235:genetics*; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial