novel method for detection of viable zoospores of Pythium in irrigation water

Summary A membrane filtration test has been developed for the detection of viable zoospores of Pythium species. Zoospore suspensions were filtered through 5 (m nitrocellulose membranes and the membranes incubated overnight in 0.07 m glucose, rifamycin (30 mg litre‐1) and pimaricin (100 mg litre‐1)....

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Bibliographic Details
Published in:Annals of applied biology 1997-12, Vol.131 (3), p.427-435
Main Authors: Wakeham, A.J, Pettitt, T.R, White, J.G
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
contaminated nursery irrigation water
contamination
detection
enzyme-linked immunosorbent assay
Fundamental and applied biological sciences. Psychology
Fungal plant pathogens
Generalities. Techniques
immunologic techniques
indirect protein a elisa
irrigation water
membrane filtration test
Phytopathology. Animal pests. Plant and forest protection
Phytophthora cryptogea
plant nurseries
polyclonal antibodies
polyclonal antibody
Pythium
quantitative analysis
rapid methods
serological assay
soil fungi
viability
zoospore
zoospores
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Summary:Summary A membrane filtration test has been developed for the detection of viable zoospores of Pythium species. Zoospore suspensions were filtered through 5 (m nitrocellulose membranes and the membranes incubated overnight in 0.07 m glucose, rifamycin (30 mg litre‐1) and pimaricin (100 mg litre‐1). Zoospore germlings were detected using a polyclonal antiserum, raised to mycelial surface washings of five Pythium spp., and visualised with Sigma fast red. The assay gave positive results for all Pythium spp. tested and also to zoospores of Phytophthora cryptogea. Of 10 fungal species isolated from commercial irrigation water, two were detected by the polyclonal antiserum in ELISA tests but only one produced detectable zoospore germlings. The latter isolate was later identified as a Pythium sp. Irrigation water samples collected from commercial UK nurseries yielded zoospores of both Pythium and Phytophthora spp. which, using the assay, were positively identified. Results indicated greater sensitivity than was seen with conventional plating methods. This is a test which could be adapted for on‐site use in commercial nurseries.
ISSN:0003-4746
1744-7348
DOI:10.1111/j.1744-7348.1997.tb05170.x