Detection of Podosphaera macularis in Air Samples by Quantitative PCR.

Autor:	Gent DH; Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR 97331.; U.S. Department of Agriculture, Agricultural Research Service, Forage Seed and Cereal Research Unit, Corvallis, OR 97331., Adair NL; U.S. Department of Agriculture, Agricultural Research Service, Forage Seed and Cereal Research Unit, Corvallis, OR 97331., Hatlen RJ; Department of Plant, Soil and Microbial Sciences, Michigan State University, East Lansing, MI 48824., Miles TD; Department of Plant, Soil and Microbial Sciences, Michigan State University, East Lansing, MI 48824., Richardson BJ; Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR 97331., Rivedal HM; U.S. Department of Agriculture, Agricultural Research Service, Forage Seed and Cereal Research Unit, Corvallis, OR 97331., Ross C; Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR 97331., Wiseman MS; Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR 97331.
Jazyk:	angličtina
Zdroj:	Plant disease [Plant Dis] 2024 Sep; Vol. 108 (9), pp. 2820-2829. Date of Electronic Publication: 2024 Sep 09.
DOI:	10.1094/PDIS-04-24-0894-RE
Abstrakt:	Detection and quantification of pathogen propagules in the air or other environmental samples is facilitated by culture-independent assays. We developed a quantitative PCR assay for the hop powdery mildew fungus, Podosphaera macularis , for detection of the organism from air samples. The assay uses primers and a TaqMan probe designed to target species-specific sequences in the 28S large subunit of the nuclear ribosomal DNA. Analytical sensitivity was not affected by the presence of an exogenous internal control or potential PCR inhibitors associated with DNA extracted from soil. The level of quantification of the assay was between 200 and 350 conidia when DNA was extracted from a fixed number of conidia. The assay amplified all isolates of P. macularis tested and had minimal cross-reactivity with other Podosphaera species when assayed with biologically relevant quantities of DNA. Standard curves generated independently in two other laboratories indicated that assay sensitivity was qualitatively similar and reproducible. All laboratories successfully detected eight unknown isolates of P. macularis and correctly discriminated Pseudoperonospora humuli and a water control. The usefulness of the assay for air sampling for late-season inoculum of P. macularis was demonstrated in field studies in 2019 and 2020. In both years, airborne populations of P. macularis in hop yards were detected consistently and increased during bloom and cone development. Competing Interests: The author(s) declare no conflict of interest.
Databáze:	MEDLINE
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