Multiplex real‐time PCR assays for the detection and identification of <i>Heterobasidion</i> species attacking conifers in Europe

Ioos, Renaud; Chrétien, Paul L.; Perrault, J.; Jeandel, Claude; Dutech, Cyril; Gonthier, Paolo; Sillo, Fabiano; Hietala, Ari M.; Solheim, Halvor; Hubert, Jacqueline

doi:10.1111/ppa.13071

Cited by 10 publications

(7 citation statements)

References 38 publications

(79 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For example, the primers used for specific detection of E. alphitoides may amplify DNA from powdery mildew species in the same clade, which share highly similar ITS sequences but do not affect oaks (Chen et al, 2015). However, primers targeting single copy genes to differentiate species within the H. annosum complex, designed for diagnostic purposes (Ioos et al, 2019), failed to detect any target in aerobiological samples (results not shown).…”

Section: Discussionmentioning

confidence: 99%

Combining permanent aerobiological networks and molecular analyses for large‐scale surveillance of forest fungal pathogens: A proof‐of‐concept

Aguayo¹,

Husson

Chancerel

et al. 2020

Plant Pathology

Self Cite

View full text Add to dashboard Cite

Forest disease management relies principally on a preventive approach in which epidemiological surveillance plays a crucial role. However, efficient and cost-effective surveillance methods are not currently available for large spatial scales. Nevertheless, aerobiological networks have been set up for several decades in many countries to monitor pollen dispersal and provide real-time assessments of allergenic risk. Here, we suggest that the same approach could be used for the surveillance of forest pathogens. Using molecular methods, we analysed samples from 12 sites of the French aerobiological network, at different dates. Both metabarcoding by high-throughput sequencing (using two markers and two different bioinformatics approaches) and real-time PCR targeting eight important forest pathogens were conducted. To validate the approach, temporal and spatial trends of spore detection were compared with field disease data. The metabarcoding approach demonstrated that many fungal plant pathogens could be found in aerobiological samples. Moreover, five of the eight targeted forest pathogens were detected by real-time PCR, with temporal and spatial trends of spore capture consistent with field data. In particular, Hymenoscyphus fraxineus was detected at high frequency in aerobiological samples in the areas where ash dieback has been present for the longest period of time, and at lower frequency in areas with more recent invasion. Spore detection of seasonal pathogens showed a temporal pattern similar to that of disease reports. Overall, our study provides a proof of concept that permanent aerobiological networks combined with molecular methods may provide a useful tool for large-scale surveillance of forest pathogens.

show abstract

Section: Discussionmentioning

confidence: 99%

Combining permanent aerobiological networks and molecular analyses for large‐scale surveillance of forest fungal pathogens: A proof‐of‐concept

Aguayo¹,

Husson

Chancerel

et al. 2020

Plant Pathology

Self Cite

View full text Add to dashboard Cite

show abstract

“…Up to 30 mg of tissue was processed with DNeasy Plant Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions, to a final elution volume of 50 μl. The real-time PCR assays for detection of Heterobasidion annosum and H. parviporum , the two Heterobasidion species present in Norway, were conducted in singleplex conditions in Takyon Low ROX Probe MasterMix dTTP Blue (Eurogentec, Seraing, Belgium), with primers and probes described by Ioos et al ( 2019 ) and synthesized by Eurogentec.…”

Section: Methodsmentioning

confidence: 99%

“…The H. parviporum qPCR assay included the forward primer 5′-CAATCGTATGGGGTCATTGTAA-3′, the reverse primer 5′-CACATCCGCCATGTCCC-3′ and the probe TAMRA-GATCTGCGAGCCCGACGAACCG-BHQ2. Real-time PCR amplifications were carried out in an Applied Biosystems ViiA 7 system (ThermoFisher, Waltham, MA, USA) with standard, instead of fast, cycling and the same PCR cycling parameters as described by Ioos et al ( 2019 ). Two technical replicates were prepared for each sample.…”

Section: Methodsmentioning

confidence: 99%

Patterns and roles of lignan and terpenoid accumulation in the reaction zone compartmentalizing pathogen-infected heartwood of Norway spruce

Nagy

Norli

Fongen

et al. 2022

Planta

View full text Add to dashboard Cite

Main conclusion Lignan impregnation of the reaction zone wood protects against oxidative degradation by fungi. Traumatic resin canals may play roles in the underlying signal transduction, synthesis, and translocation of defense compounds. Abstract Tree defense against xylem pathogens involves both constitutive and induced phenylpropanoids and terpenoids. The induced defenses include compartmentalization of compromised wood with a reaction zone (RZ) characterized by polyphenol deposition, whereas the role of terpenoids has remained poorly understood. To further elucidate the tree–pathogen interaction, we profiled spatial patterns in lignan (low-molecular-weight polyphenols) and terpenoid content in Norway spruce (Picea abies) trees showing heartwood colonization by the pathogenic white-rot fungus Heterobasidion parviporum. There was pronounced variation in the amount and composition of lignans between different xylem tissue zones of diseased and healthy trees. Intact RZ at basal stem regions, where colonization is the oldest, showed the highest level and diversity of these compounds. The antioxidant properties of lignans obviously hinder oxidative degradation of wood: RZ with lignans removed by extraction showed significantly higher mass loss than unextracted RZ when subjected to Fenton degradation. The reduced diversity and amount of lignans in pathogen-compromised RZ and decaying heartwood in comparison to intact RZ and healthy heartwood suggest that α-conindendrin isomer is an intermediate metabolite in lignan decomposition by H. parviporum. Diterpenes and diterpene alcohols constituted above 90% of the terpenes detected in sapwood of healthy and diseased trees. A significant finding was that traumatic resin canals, predominated by monoterpenes, were commonly associated with RZ. The findings clarify the roles and fate of lignan during wood decay and raise questions about the potential roles of terpenoids in signal transduction, synthesis, and translocation of defense compounds upon wood compartmentalization against decay fungi.

show abstract

“…A new PCR assay has been developed to also distinguish Fusarium circinatum and Caliciopsis pinea that cause comparable symptoms on P. radiata in the initial stages of colonization (Luchi et al 2018). All four European species of Heterobasidion attacking conifers may be simultaneously detected in two reactions (Ioos et al 2019b).…”

Section: Disease Controlmentioning

confidence: 99%

Fast and reliable molecular methods to detect fungal pathogens in woody plants

Luchi

Ioos

Santini

2020

Appl Microbiol Biotechnol

Self Cite

View full text Add to dashboard Cite

Plant diseases caused by pathogenic microorganisms represent a serious threat to plant productivity, food security, and natural ecosystems. An effective framework for early warning and rapid response is a crucial element to mitigate or prevent the impacts of biological invasions of plant pathogens. For these reasons, detection tools play an important role in monitoring plant health, surveillance, and quantitative pathogen risk assessment, thus improving best practices to mitigate and prevent microbial threats. The need to reduce the time of diagnosis has prompted plant pathologists to move towards more sensitive and rapid methods such as molecular techniques. Considering prevention to be the best strategy to protect plants from diseases, this review focuses on fast and reliable molecular methods to detect the presence of woody plant pathogens at early stage of disease development before symptoms occur in the host. A harmonized pool of novel technical, methodological, and conceptual solutions is needed to prevent entry and establishment of new diseases in a country and mitigate the impact of both invasive and indigenous organisms to agricultural and forest ecosystem biodiversity and productivity.

show abstract

Multiplex real‐time PCR assays for the detection and identification of Heterobasidion species attacking conifers in Europe

Cited by 10 publications

References 38 publications

Combining permanent aerobiological networks and molecular analyses for large‐scale surveillance of forest fungal pathogens: A proof‐of‐concept

Combining permanent aerobiological networks and molecular analyses for large‐scale surveillance of forest fungal pathogens: A proof‐of‐concept

Patterns and roles of lignan and terpenoid accumulation in the reaction zone compartmentalizing pathogen-infected heartwood of Norway spruce

Fast and reliable molecular methods to detect fungal pathogens in woody plants

Contact Info

Product

Resources

About