Detection and Quantification of Pythium tracheiphilum in Soil by Multiplex Real-Time qPCR
| Autor: | Hervé Van der Heyden, C. André Lévesque, Odile Carisse, Thérèse Wallon |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
0301 basic medicine Canada Damping off Pythium Lactuca Plant Science Biology Real-Time Polymerase Chain Reaction 01 natural sciences Soil 03 medical and health sciences Multiplex polymerase chain reaction Multiplex DNA Primers Quebec food and beverages Soil classification biology.organism_classification Horticulture 030104 developmental biology Oospore Pythium tracheiphilum Multiplex Polymerase Chain Reaction Agronomy and Crop Science 010606 plant biology & botany |
| Zdroj: | Plant Disease. 103:475-483 |
| ISSN: | 1943-7692 0191-2917 |
| DOI: | 10.1094/pdis-03-18-0419-re |
| Popis: | In Canada, head lettuce (Lactuca sativa capitata) is extensively produced in the muck soils of southwestern Québec. However, yields are increasingly affected by various soilborne pathogens, including Pythium spp., which cause wilt and damping off. In a survey conducted in Québec muck soils in 2010 and 2011, Pythium tracheiphilum Matta was identified as the predominant Pythium sp. in the root of head lettuce showing Pythium stunt symptoms. Therefore, to improve risk assessment and help further understanding of disease epidemiology, a specific and sensitive real-time quantitative polymerase chain reaction (qPCR) assay based on TaqMan-minor groove binder (MGB) technology was developed for P. tracheiphilum. The PCR primers along with a TaqMan-MGB probe were designed from the ribosomal internal transcribed spacer 2 region. A 100-bp product was amplified by PCR from all P. tracheiphilum isolates tested while no PCR product was obtained from 38 other Pythium spp. or from a selection of additional lettuce pathogens tested. In addition to P. tracheiphilum, the assay was multiplexed with an internal control allowing for the individual validation of each PCR. In artificially infested soils, the sensitivity of the qPCR assay was established as 10 oospores/g of dry soil. P. tracheiphilum was not detected in soils in which lettuce has never been grown; however, inoculum ranged from 0 to more than 200,000 oospores/g of dry soil in commercial lettuce fields. Also, disease incidence was positively correlated with inoculum concentration (r = 0.764). The results suggest that inoculum concentration should be considered when making Pythium stunt management decisions. The developed qPCR assay will facilitate reliable detection and quantification of P. tracheiphilum from field soil. |
| Databáze: | OpenAIRE |
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