Quantification of Phytophthora capsici Oospores in Soil by Sieving-Centrifugation and Real-Time Polymerase Chain Reaction

Plant Disease
C PavonK N Lambert

Abstract

A procedure was developed to quantify Phytophthora capsici oospores in soil by combining a sieving-centrifugation method and a real-time quantitative polymerase chain reaction (QPCR) assay. Five soil samples representing three different soil textures were infested with oospores of P. capsici to produce 101, 102, 103, 104, or 105 spores per 10 g of air-dried soil. Each 10-g sample of infested soil was suspended in 400 ml of water and then passed through 106-, 63-, and 38-μm metal sieves. The filtrate was then passed through a 20-μm mesh filter. Materials caught on the filter were washed with water into two 50-ml centrifuge tubes and spun for 4 min (900 × g). The pellet was suspended in 30 ml of 1.6 M sucrose solution and centrifuged for 45 s (190 × g). The supernatant was passed through the 20-μm mesh filter. The sucrose extraction process of oospores was repeated five times to maximize oospore extraction. Materials caught on the 20-μm mesh filter were washed with water into a 50-ml tube and spun for 4 min (900 × g). The pellet was suspended in 1 ml of water, and the number of oospores was determined with a haemocytometer. The relationship between number of oospores recovered from the soil and number of oospores incorporated int...Continue Reading

References

Oct 1, 1996·Genome Research·U E GibsonP M Williams
Nov 5, 1997·Applied and Environmental Microbiology·I G Wilson
Mar 21, 1998·Applied and Environmental Microbiology·J B RistainoG Parra
Jun 17, 2000·Journal of Virological Methods·A J EunS Wong
Nov 10, 2001·Annual Review of Phytopathology·Robert R MartinC André Lévesque
Feb 16, 2002·Methods : a Companion to Methods in Enzymology·K J Livak, T D Schmittgen
Mar 1, 2003·Journal of Microbiological Methods·Martin FilionSuha H Jabaji-Hare
Jan 1, 1997·Phytopathology·S A MillerA F Schmitthenner
Dec 1, 2004·Plant Disease·Mary K Hausbeck, Kurt H Lamour

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Citations

Mar 1, 2013·Plant Disease·Paul Vincelli, Bernadette Amsden
Apr 8, 2014·Phytopathology·Amara R DunnChristine D Smart
Aug 1, 2009·Plant Disease·Mohammad Babadoost, Thomas A Zitter
Nov 1, 2013·Plant Disease·M Babadoost, C Pavon
Aug 28, 2020·Frontiers in Plant Science·Melissa Si AmmourVittorio Rossi

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