Development of a TaqMan Real-Time PCR Assay for Quantification of Airborne Conidia of <i>Botrytis squamosa</i> and Management of Botrytis Leaf Blight of Onion

Carisse, Odile; Tremblay, D. M.; Lévesque, C. André; Gindro, Katia; Ward, Pierre; Houde, Alain

doi:10.1094/phyto-99-11-1273

Cited by 73 publications

(53 citation statements)

References 37 publications

(56 reference statements)

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“…However, qPCR has been also used to quantify the airborne inoculum of fungal pathogens, such as Sclerotinia sclerotiorum, Leptosphaeria spp., Puccinia striiformis, and Botrytis squamosa (38,39,40,41), which cause serious disease in arable crops, and also for the forest tree pathogen Fusarium circinatum (20,42). Many fungal diseases are initiated by airborne inoculum that lands on susceptible hosts under favorable environmental conditions.…”

Section: Discussionmentioning

confidence: 99%

Rapid Detection of Ceratocystis platani Inoculum by Quantitative Real-Time PCR Assay

Luchi¹,

Ghelardini²,

Belbahri

et al. 2013

Appl Environ Microbiol

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Ceratocystis platani is the causal agent of canker stain of plane trees, a lethal disease able to kill mature trees in one or two successive growing seasons. The pathogen is a quarantine organism and has a negative impact on anthropogenic and natural populations of plane trees. Contaminated sawdust produced during pruning and sanitation fellings can contribute to disease spread. The goal of this study was to design a rapid, real-time quantitative PCR assay to detect a C. platani airborne inoculum. Airborne inoculum traps (AITs) were placed in an urban setting in the city of Florence, Italy, where the disease was present. Primers and TaqMan minor groove binder (MGB) probes were designed to target cerato-platanin (CP) and internal transcribed spacer 2 (ITS2) genes. The detection limits of the assay were 0.05 pg/l and 2 fg/l of fungal DNA for CP and ITS, respectively. Pathogen detection directly from AITs demonstrated specificity and high sensitivity for C. platani, detecting DNA concentrations as low as 1.2 ؋ 10 ؊2 to 1.4 ؋ 10 ؊2 pg/l, corresponding to ϳ10 conidia per ml. Airborne inoculum traps were able to detect the C. platani inoculum within 200 m of the closest symptomatic infected plane tree. The combination of airborne trapping and real-time quantitative PCR assay provides a rapid and sensitive method for the specific detection of a C. platani inoculum. This technique may be used to identify the period of highest risk of pathogen spread in a site, thus helping disease management.

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Section: Discussionmentioning

confidence: 99%

Rapid Detection of Ceratocystis platani Inoculum by Quantitative Real-Time PCR Assay

Luchi¹,

Ghelardini²,

Belbahri

et al. 2013

Appl Environ Microbiol

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“…At the damage thresholds of five or ten lesions/leaf, conidia quantification with qPCR assay was more reliable at predicting disease risk than by microscopical counting of conidia. The results indicated that the qPCR protocol was reliable for quantifying B. squamosa airborne inoculum in commercial onion fields, providing a dependable basis for disease risk assessment (Carrise et al 2009). …”

Section: Detection Of Fungal Pathogens In Airmentioning

confidence: 96%

Detection of Fungal Pathogens in the Environment

Narayanasamy¹

2010

Microbial Plant Pathogens-Detection and Disease Diagnosis:

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The presence of fungal pathogens in the environment has to be detected and also quantified rapidly and precisely, because the inoculum for infection of crop plants comes from the pathogen propagules present in the soil, water, air and alternative host plant species. Soilborne pathogens may have different degrees of saprophytic ability, utilizing the organic matter present in the soil for their survival in the absence of crop plants. They produce different structures such as chlamydospores and sexual spores that are long-lived and are capable of surviving in the soil for several years. Fungal structures may be carried by irrigation water or rain water from one part of the field to other parts or to different fields. It has been possible to detect and identify the fungal pathogens in the irrigation water, recycled water used for growing hydroponic plants and also in wash water in the storage facilities for fruits and vegetables. The pathogens infecting aerial plant parts/organs are generally disseminated by wind to different locations. Traditionally spore traps have been used for assessing the spore load of air. Various biological, immunological and nucleic acid-based techniques have been employed for the detection, identification and quantification of pathogen propagules in the environment. Significant improvements have been made in the sensitivity and specificity of detection of fungal pathogens by applying immunoassys and nucleic acid-based methods that are capable of providing reproducible results rapidly and reliably. The relative usefulness and limitations of the detection techniques applied for the detection of fungal pathogens in the environment are discussed.Studies on the ecology of plant hosts provide information on the influence of environment on the growth and development of plants. Likewise, the influence of the environment on the microbial plant pathogens is studied to understand the extent of population build up resulting in the incidence of diseases in different agroecological conditions. Crop husbandry techniques aim to increase the crop yield to the maximum levels. But some of these techniques like monoculture and excessive application of nutrients may provide favorable conditions for pathogen development. Epidemiology deals with effects of biotic and abiotic environments on disease development in plant populations. Microbial pathogens multiply at different rates in a given set of environmental conditions and exhibit wide variations in their pathogenic potential (aggressiveness). The crop plants also have varying growth

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“…Spore sampling protocols were implemented for P. viticola sporangia (Kast, 1997) and E. necator conidia (Carisse et al, 2009a). Spore sampling was greatly improved by the use of molecular tools in order to speed up the counting process, as proposed by Carisse et al (2009b) to develop real-time quantification of the airborne conidia of B. cinerea.…”

Section: Scoutingmentioning

confidence: 99%

A critical review of plant protection tools for reducing pesticide use on grapevine and new perspectives for the implementation of IPM in viticulture

Pertot¹,

et al. 2017

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a b s t r a c tSeveral pests and diseases have grapevine as their favourite host and the vineyard as preferred environment, so an intensive pesticide schedule is usually required to meet qualitative and quantitative production standards. The need to prevent the negative impact of synthetic chemical pesticides on human health and the environment and the consumer expectations in term of chemical residues in food stimulated the research of innovative tools and methods for sustainable pest management. The research project PURE (www.pure-ipm.eu) was a Europe-wide framework, which demonstrated that several solutions are now available for the growers and evaluated several new alternatives that are under development or almost ready for being applied in practice. Although the use of resistant/tolerant varieties is not yet feasible in several traditional grape growing areas, at least part of the synthetic chemical pesticides can be substituted with biocontrol agents to control pests and pathogens and/or pheromone mating disruption, or the number of treatments can be reduced by the use of decision support systems, which identify the optimal timing for the applications. This review presents the state of the art and the perspectives in the field of grapevine protection tools and strategies.

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Development of a TaqMan Real-Time PCR Assay for Quantification of Airborne Conidia of Botrytis squamosa and Management of Botrytis Leaf Blight of Onion

Cited by 73 publications

References 37 publications

Rapid Detection of Ceratocystis platani Inoculum by Quantitative Real-Time PCR Assay

Rapid Detection of Ceratocystis platani Inoculum by Quantitative Real-Time PCR Assay

Detection of Fungal Pathogens in the Environment

A critical review of plant protection tools for reducing pesticide use on grapevine and new perspectives for the implementation of IPM in viticulture

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