Phytoparasitica (2002) 30, 473-485

P.H. Wang, L.M. Boo, Y.S. Lin and Y. Yeh (2002)
Specific detection of Pythium aphanidermatum from hydroponic nutrient solution by booster PCR with DNA primers developed from mitochondrial DNA
Phytoparasitica 30 (5), 473-485
Abstract: Pythium aphanidermatum causes damping-off and root rot of vegetable crops in hydroponic systems. A DNA probe was isolated and modified from a library of HindIII-digested mito-chondrial DNA of P. aphanidermatum that strongly hybridized to DNA of P. aphanidermatum and weakly hybridized to DNA of Pythium deliense. Cross-hybridizing sequences were absent from DNA of plants and other related fungi. The probe detected as little as 5 ng of P. aphanidermatum DNA and 250 ng of P. deliense DNA in slot-blot assays. P. aphanidermatum was detected by a hybridization assay of total DNA extracted directly from infected roots. A pair of oligonucleotide primers P1 and RP2, which allowed amplification of a specific 0.65 kb DNA fragment of P. aphanidermatum using polymerase chain reaction (PCR), was designed from a specific DNA probe. Specific amplification of this fragment from P. aphanidermatum was highly sensitive, detecting template DNA as low as 0.1 pg total DNA by booster PCR. Specific booster PCR amplification using P1 and RP2 was successful in detecting P. aphanidermatum in naturally infected nutrient solution and roots of vegetables in a field hydroponic system.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
Link to article at publishers website
Database assignments for author(s): Pi-Han Wang

Research topic(s) for pests/diseases/weeds:
identification/taxonomy
surveys/sampling/distribution

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Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Pythium aphanidermatum
	Pythium deliense