The world’s interest in Phytophthora species has been growing in recent years—to the point where more research projects on this important pathogen are expected.
This compilation of working protocols for Phytophthora research was created to meet this demand and serve the growing need for a coherent collection of methods to study members of this genus.
Phytophthora Protocols is a fully online and continually updated resource that includes step-by-step instructions and images to ensure the accuracy and consistency of your Phytophthora research techniques.
Phytophthora Protocols can be used as a baseline for optimizing lab research, developing custom research techniques, and other research applications. This collection of 79 individual protocols is divided into sections, including:
- Phytophthora Detection, Identification, and Isolation
- Phytophthora Inoculum Production and Inoculation
- Long-term Storage of Phytophthora spp.
- Chemical Sensitivity Studies
- Media and Solutions for Phytophthora Studies
This continually growing, customized collection of protocols is ideal for anyone researching Phytophthora or other oomycetes, as well as plant disease diagnosticians and state/federal plant health regulators. Users can access them in several ways:
Laboratory Protocols for Phytophthora Species
(Annual Online Subscription for Libraries, Companies, and Institutions)
Front Matter
Kelly Ivors
PART 1: Phytophthora Detection, Identification, and Isolation
PROTOCOL 01-01.1: Identifying species of Phytophthora based on morphology
Steven Jeffers
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PROTOCOL 01-02.1: Isolation of Phytophthora infestans from tomato leaves
Kelly Ivors
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PROTOCOL 01-03.1: Isolation of Phytophthora infestans from tomato fruit
Jean Ristaino and L. Guo
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PROTOCOL 01-04.1: Isolation of Phytophthora infestans from potato leaves
Kelly Ivors
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PROTOCOL 01-05.1: Isolation of Phytophthora nicotianae from soil
David Shew and Courtney Gallup
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PROTOCOL 01-06.1: Isolation of Phytophthora nicotianae from tobacco stems and roots
David Shew
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PROTOCOL 01-07.1: Float incubation technique for Phytophthora and Pythium diagnostics
Gail Ruhl
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PROTOCOL 01-08.1: Recovering Pythium and Phytophthora spp. from infected plant tissue
Jason Brock and Glenn Beard
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PROTOCOL 01-09.1: Preparation of hyphal tip Phytophthora cultures
Sabine Werres
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PROTOCOL 01-10.1: Purification of Phytophthora cultures contaminated with bacteria
Kelly Ivors
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PROTOCOL 01-11.1: Isolation and detection of Phytophthora spp. using Rhododendron leaf baits
Kelly Ivors
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PROTOCOL 01-12.1: Rhododendron leaf test to detect Phytophthora spp. in roots, soil, and water
Sabine Werres and Corina Junker
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PROTOCOL 01-13.1: Pear baiting for isolation of Phytophthora spp. from various substrates
Daniel Hüberli
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PROTOCOL 01-14.1: Detection of Phytophthora spp. in water supplies
D. Michael Benson
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PROTOCOL 01-15.1: Qualitative and quantitative recovery of Phytophthora cinnamomi from soil
Kelly Ivors
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PROTOCOL 01-16.1: Sieving–Centrifugation method using a sucrose solution for extraction of Phytophthora capsici oospores from field soil
Mohammad Babadoost
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PROTOCOL 01-17.1: Detecting Phytophthora spp. on plant foliage using ELISA
Arthur Wagner
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PART 2: Phytophthora Inoculum Production and Inoculation
PROTOCOL 02-01.1: Maintaining Phytophthora infestans in planta
Kelly Ivors and Dreama Milks
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PROTOCOL 02-02.1: Mass production of Phytophthora nicotianae zoospores using a wet plate method
Chuan Hong
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PROTOCOL 02-03.1: Production of Phytophthora zoospores
Kelly Ivors
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PROTOCOL 02-04.1: Induction of sporangia formation and zoospore release for Phytophthora ramorum and other Phytophthora spp.
Everett Hansen
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PROTOCOL 02-05.1: Obtaining and manipulating zoospores of Phytophthora infestans
Howard Judelson
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PROTOCOL 02-06.1: Production of Phytophthora sporangia using vinca leaves
Kelly Ivors and Walter Gutierrez
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PROTOCOL 02-07.1: Rice grain method for Phytophthora inoculum production
D. Michael Benson and Kala Parker
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PROTOCOL 02-08.1: Vermiculite method for Phytophthora inoculum production
Kelly Ivors
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PROTOCOL 02-09.1: Modified vermiculite method for Phytophthora inoculum production
Kelly Ivors
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PROTOCOL 02-10.1: Inoculating detached leaves via a Phytophthora zoospore drop
Rachel Linzer and Matteo Garbelotto
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PROTOCOL 02-11.1: Detached leaf inoculation for tanoak
Katherine Hayden and Matteo Garbelotto
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PROTOCOL 02-12.1: Production of Phytophthora inoculum on oat grain
Kelly Ivors
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PART 3: Long-Term Storage of Phytophthora spp.
PROTOCOL 03-01.1: Long-term storage of Phytophthora nicotianae isolates in water
David Shew and Courtney Gallup
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PROTOCOL 03-02.1: Long-term storage of Phytophthora cultures in water
Wendy Sutton, Paul Reeser, and Everett Hansen
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PROTOCOL 03-03.1: Long-term storage of Phytophthora and Pythium isolates using hemp seed
R. Wick, Nina Shishkoff, and Margery Daughtrey
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PROTOCOL 03-04.1: Long-term storage of Phytophthora isolates using oatmeal agar
Sabine Werres
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PROTOCOL 03-05.1: Long-term storage of Phytophthora infestans on Rye A agar
Alison Lees
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PROTOCOL 03-06.1: Long-term preservation of Phytophthora spp. in liquid nitrogen
Kelly Ivors
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PROTOCOL 03-07.1: Freezing Phytophthora cultures for long-term storage in liquid nitrogen
Paul Tooley
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PROTOCOL 03-08.1: Storage of Phytophthora infestans using liquid nitrogen
Kevin Myers and William Fry
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PROTOCOL 03-09.1: Cryopreservation of Phytophthora spp.
Michael Coffey
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PART 4: Mating Techniques
PROTOCOL 04-01.1: Mating-type determination of heterothallic Phytophthora species
David Shew and Courtney Gallup
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PROTOCOL 04-02.1: Phytophthora and Pythium gametangia production using grass blades
Frank Martin
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PROTOCOL 04-03.1: Determining the viability of Phytophthora oospores
Kestrel McCorkle, Courtney Gallup, and David Shew
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PROTOCOL 04-04.1: Production of Phytophthora ramorum oogonia in vitro
Clive Brasier and Susan Kirk
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PART 5: Chemical Sensitivity Studies
PROTOCOL 05-01.1: In vitro sensitivity of Phytophthora spp. to mefenoxam
David Shew and Courtney Gallup
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PROTOCOL 05-02.1: Evaluating fungicide sensitivity of Phytophthora capsici in vitro
Anthony Keinath
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PART 6: DNA Techniques
PROTOCOL 06-01.1: Preparation of freeze-dried mycelium for DNA extraction from Phytophthora spp.
Alison Lees
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PROTOCOL 06-02.1: Quick and easy DNA extraction from Phytophthora infestans
Alison Lees
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PROTOCOL 06-03.1: Modified CTAB extraction of fungal DNA
Kelly Ivors
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PROTOCOL 06-04.1: DNA extraction from plant material or mycelium using IsoCode Stixs or FTA cards
Jean Ristaino
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PROTOCOL 06-05.1: Desiccation of plant leaves for molecular detection of Phytophthora spp.
Pierluigi Bonello
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PROTOCOL 06-06.1: Phytophthora DNA extraction and amplification from dried plant or herbarium samples
Jean Ristaino
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PROTOCOL 06-07.1: DNA isolation from Phytophthora mycelium using the DNeasy Plant Mini Kit (QIAGEN)
Kelly Ivors
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PROTOCOL 06-08.1: DNA isolation from Phytophthora mycelium using the DNeasy Plant Mini Kit (QIAGEN)
Paul Tooley and Marie Carras
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PROTOCOL 06-09.1: DNA extraction from hyphal tips of Phytophthora spp.
Matteo Garbelotto
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PROTOCOL 06-10.1: DNA extraction for RFLP analysis of Phytophthora infestans
Steve Goodwin
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PROTOCOL 06-11.2: Molecular identification of Phytophthora isolates using a DNA sequence-based approach and the Phytophthora Database
Seogchan Kang, Michele Mansfield, Bongsoo Park, and Frank Martin
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PART 7: Media and Solutions for Phytophthora Studies
PROTOCOL 07-01.1: Nonsterile soil extract solution, 1.5% (NS-SES)
Steve Jeffers
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PROTOCOL 07-02.1: Water agar (WA)
Kelly Ivors
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PROTOCOL 07-03.1: PARP(H)-CMA
Kelly Ivors
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PROTOCOL 07-04.1: PARP(H)-V8A
Steve Jeffers
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PROTOCOL 07-05.1: 5% clarified V8 PARP(H)
David Shew
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PROTOCOL 07-06.1: PCH agar
Kelly Ivors
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PROTOCOL 07-07.1: Selective Phytophthora agar
Paul Tooley
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PROTOCOL 07-08.1: CARP
Everett Hansen
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PROTOCOL 07-09.1: CARP-D
Everett Hansen
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PROTOCOL 07-10.1: CARP+
Everett Hansen
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PROTOCOL 07-11.1: V8 agar (V8A) or broth
Steve Jeffers
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PROTOCOL 07-12.1: Super V8 agar (sV8)
Steve Jeffers
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PROTOCOL 07-13.1: V8 rye agar
Jean Ristaino
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PROTOCOL 07-14.1: Rye A agar
Kelly Ivors
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PROTOCOL 07-15.1: Rye B agar
Jean Ristaino
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PROTOCOL 07-16.1: Oatmeal agar
Sabine Werres
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PROTOCOL 07-17.1: 5% carrot agar (CA)
David Shew
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PROTOCOL 07-18.1: Fresh lima bean agar
Jean Ristaino
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PROTOCOL 07-19.1: Pea sucrose agar (PSA)
Kelly Ivors
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PROTOCOL 07-20.1: Pea broth
Peter Bonants
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PROTOCOL 07-21.1: Corn meal PPP agar
Kelly Ivors
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PROTOCOL 07-22.1: Rye seed medium
Michael Coffey
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PROTOCOL 07-23.1: Carrot agar (CA) or broth
Michael Coffey
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PROTOCOL 07-24.1: Pea or lima bean agar or broth
Michael Coffey
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Back Matter
References
Kelly Ivors
Archived Protocols
PROTOCOL 06-11.1: Molecular identification of Phytophthora isolates using a DNA sequence-based approach and the Phytophthora Database
Seogchan Kang and Michele Mansfield
Buy Single ProtocolPublish Date: 2016
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Edited by Kelly Ivors
Laboratory Protocols for Phytophthora Species
(Annual Online Subscription for Libraries, Companies, and Institutions)