T1 The Challenge of Environmental Samples for PCR Detection of Phytopathogenic Bacteria: A Case Study of Citrus Huanglongbing Disease
A1 Morán, Félix
A1 Barbé, Silvia
A1 Bastin, Saskia
A1 Navarro-Herrero, Inmaculada
A1 Bertolini, Edson
A1 López, María M.
A1 Hernández-Suárez, Estrella
A1 Urbaneja, Alberto
A1 Tena, Alejandro
A1 Siverio, Felipe
A1 Marco-Noales, Ester
K1 undesired amplification
K1 16S
K1 real-time PCR
K1 sequence identity
K1 greening
K1 HLB
K1 Trioza erytreae
K1 Tamarixia dryi
K1 H20  Plant diseases
K1 Bacterial flora
K1 Candidatus Liberibacter
K1 HLB
K1 PCR
AB Huanglongbing (HLB) is the most devastating citrus disease and is associated with three bacterial species of the genus ‘Candidatus Liberibacter’ transmitted by insect vectors. The early detection of HLB is based on PCR methods, and it is one of the cornerstones for preventing incursion into disease-free countries. However, the detection of phytopathogenic bacteria with PCR-based methods is problematic in surveys that include a variety of samples of different origins. Here, we first report the proportion of amplifications obtained by two standardized real-time PCR methods for the diagnosis of HLB in various environmental samples that include plants, psyllid vectors, and parasitic wasps of the psyllids. The results of 4915 samples showed that 9.3% of them were amplified by the first rapid screening test and only 0.3% by the more specific tests. Most of the amplifications were associated with parasitic wasps. We designed the primers external to the target regions of both real-time PCR protocols to determine if amplifications belonged to one of three ‘Ca. Liberibacter’ species associated with HLB. The bioinformatic analysis of the sequences obtained with these primers revealed that all these amplifications came from the presence of other prokaryotic organisms in the samples. The primers developed in this study overcome the problem of undesired amplification in environmental samples. Thus, they could be used in future survey protocols to prevent the eradication of negative trees and the generation of unjustified alarms.
PB MDPI
SN 2073-4395
YR 2021
FD 2021
LK http://hdl.handle.net/20.500.11939/6927
UL http://hdl.handle.net/20.500.11939/6927
LA en
NO Morán, F.; Barbé, S.; Bastin, S.; Navarro, I.; Bertolini, E.; López, M. M. et al. (2020). The Challenge of Environmental Samples for PCR Detection of Phytopathogenic Bacteria: A Case Study of Citrus Huanglongbing Disease. Agronomy 2020, 11(1), 10.
DS MINDS@UW
RD Aug 19, 2022