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Laboratory Protocols for Phytophthora Sp (Institutional use)
Laboratory Protocols for Phytophthora Sp (Institutional use)
  • A collection of protocols for researching Phytophthora species
  • Use as a baseline for developing and optimizing lab techniques
  • Includes images and procedural descriptions to ensure proper technique
  • Personally subscribe to this entire collection of Phytophthora protocols annually

  • This is an online resource, available on the APS Journals Site, you will receive an email with access instructions within 2 business days.

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    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 new 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 73 individual protocols is divided into sections, including:

    • Phytophthora Detection, Identification, and Isolation
    • Phytophthora Inoculum Production and Inoculation
    • Long-term Storage of Phytophthora spp.
    • Mating Techniques
    • Chemical Sensitivity Studies
    • DNA Techniques
    • 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


    PROTOCOL 01-02.1: Isolation of Phytophthora infestans from tomato leaves

    Kelly Ivors


    PROTOCOL 01-03.1: Isolation of Phytophthora infestans from tomato fruit

    Jean Ristaino and L. Guo


    PROTOCOL 01-04.1: Isolation of Phytophthora infestans from potato leaves

    Kelly Ivors


    PROTOCOL 01-05.1: Isolation of Phytophthora nicotianae from soil

    David Shew and Courtney Gallup


    PROTOCOL 01-06.1: Isolation of Phytophthora nicotianae from tobacco stems and roots

    David Shew


    PROTOCOL 01-07.1: Float incubation technique for Phytophthora and Pythium diagnostics

    Gail Ruhl


    PROTOCOL 01-08.1: Recovering Pythium and Phytophthora spp. from infected plant tissue

    Jason Brock and Glenn Beard


    PROTOCOL 01-09.1: Preparation of hyphal tip Phytophthora cultures

    Sabine Werres


    PROTOCOL 01-10.1: Purification of Phytophthora cultures contaminated with bacteria

    Kelly Ivors


    PROTOCOL 01-11.1: Isolation and detection of Phytophthora spp. using Rhododendron leaf baits

    Kelly Ivors


    PROTOCOL 01-12.1: Rhododendron leaf test to detect Phytophthora spp. in roots, soil, and water

    Sabine Werres and Corina Junker


    PROTOCOL 01-13.1: Pear baiting for isolation of Phytophthora spp. from various substrates

    Daniel Hüberli


    PROTOCOL 01-14.1: Detection of Phytophthora spp. in water supplies

    D. Michael Benson


    PROTOCOL 01-15.1: Qualitative and quantitative recovery of Phytophthora cinnamomi from soil

    Kelly Ivors


    PROTOCOL 01-16.1: Sieving–Centrifugation method using a sucrose solution for extraction of Phytophthora capsici oospores from field soil

    Mohammad Babadoost


    PROTOCOL 01-17.1: Detecting Phytophthora spp. on plant foliage using ELISA

    Arthur Wagner


    PART 2: Phytophthora Inoculum Production and Inoculation


    PROTOCOL 02-01.1: Maintaining Phytophthora infestans in planta

    Kelly Ivors and Dreama Milks


    PROTOCOL 02-02.1: Mass production of Phytophthora nicotianae zoospores using a wet plate method

    Chuan Hong


    PROTOCOL 02-03.1: Production of Phytophthora zoospores

    Kelly Ivors


    PROTOCOL 02-04.1: Induction of sporangia formation and zoospore release for Phytophthora ramorum and other Phytophthora spp.

    Everett Hansen


    PROTOCOL 02-05.1: Obtaining and manipulating zoospores of Phytophthora infestans

    Howard Judelson


    PROTOCOL 02-06.1: Production of Phytophthora sporangia using vinca leaves

    Kelly Ivors and Walter Gutierrez


    PROTOCOL 02-07.1: Rice grain method for Phytophthora inoculum production

    D. Michael Benson and Kala Parker


    PROTOCOL 02-08.1: Vermiculite method for Phytophthora inoculum production

    Kelly Ivors


    PROTOCOL 02-09.1: Modified vermiculite method for Phytophthora inoculum production

    Kelly Ivors


    PROTOCOL 02-10.1: Inoculating detached leaves via a Phytophthora zoospore drop

    Rachel Linzer and Matteo Garbelotto


    PROTOCOL 02-11.1: Detached leaf inoculation for tanoak

    Katherine Hayden and Matteo Garbelotto


    PROTOCOL 02-12.1: Production of Phytophthora inoculum on oat grain

    Kelly Ivors


    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


    PROTOCOL 03-02.1: Long-term storage of Phytophthora cultures in water

    Wendy Sutton, Paul Reeser, and Everett Hansen


    PROTOCOL 03-03.1: Long-term storage of Phytophthora and Pythium isolates using hemp seed

    R. Wick, Nina Shishkoff, and Margery Daughtrey


    PROTOCOL 03-04.1: Long-term storage of Phytophthora isolates using oatmeal agar

    Sabine Werres


    PROTOCOL 03-05.1: Long-term storage of Phytophthora infestans on Rye A agar

    Alison Lees


    PROTOCOL 03-06.1: Long-term preservation of Phytophthora spp. in liquid nitrogen

    Kelly Ivors


    PROTOCOL 03-07.1: Freezing Phytophthora cultures for long-term storage in liquid nitrogen

    Paul Tooley


    PROTOCOL 03-08.1: Storage of Phytophthora infestans using liquid nitrogen

    Kevin Myers and William Fry


    PROTOCOL 03-09.1: Cryopreservation of Phytophthora spp.

    Michael Coffey


    PART 4: Mating Techniques


    PROTOCOL 04-01.1: Mating-type determination of heterothallic Phytophthora species

    David Shew and Courtney Gallup


    PROTOCOL 04-02.1: Phytophthora and Pythium gametangia production using grass blades

    Frank Martin


    PROTOCOL 04-03.1: Determining the viability of Phytophthora oospores

    Kestrel McCorkle, Courtney Gallup, and David Shew


    PROTOCOL 04-04.1: Production of Phytophthora ramorum oogonia in vitro

    Clive Brasier and Susan Kirk


    PART 5: Chemical Sensitivity Studies


    PROTOCOL 05-01.1: In vitro sensitivity of Phytophthora spp. to mefenoxam

    David Shew and Courtney Gallup


    PROTOCOL 05-02.1: Evaluating fungicide sensitivity of Phytophthora capsici in vitro

    Anthony Keinath


    PART 6: DNA Techniques


    PROTOCOL 06-01.1: Preparation of freeze-dried mycelium for DNA extraction from Phytophthora spp.

    Alison Lees


    PROTOCOL 06-02.1: Quick and easy DNA extraction from Phytophthora infestans

    Alison Lees


    PROTOCOL 06-03.1: Modified CTAB extraction of fungal DNA

    Kelly Ivors


    PROTOCOL 06-04.1: DNA extraction from plant material or mycelium using IsoCode Stixs or FTA cards

    Jean Ristaino


    PROTOCOL 06-05.1: Desiccation of plant leaves for molecular detection of Phytophthora spp.

    Pierluigi Bonello


    PROTOCOL 06-06.1: Phytophthora DNA extraction and amplification from dried plant or herbarium samples

    Jean Ristaino


    PROTOCOL 06-07.1: DNA isolation from Phytophthora mycelium using the DNeasy Plant Mini Kit (QIAGEN)

    Kelly Ivors


    PROTOCOL 06-08.1: DNA isolation from Phytophthora mycelium using the DNeasy Plant Mini Kit (QIAGEN)

    Paul Tooley and Marie Carras


    PROTOCOL 06-09.1: DNA extraction from hyphal tips of Phytophthora spp.

    Matteo Garbelotto


    PROTOCOL 06-10.1: DNA extraction for RFLP analysis of Phytophthora infestans

    Steve Goodwin


    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


    PART 7: Media and Solutions for Phytophthora Studies


    PROTOCOL 07-01.1: Nonsterile soil extract solution, 1.5% (NS-SES)

    Steve Jeffers


    PROTOCOL 07-02.1: Water agar (WA)

    Kelly Ivors


    PROTOCOL 07-03.1: PARP(H)-CMA

    Kelly Ivors


    PROTOCOL 07-04.1: PARP(H)-V8A

    Steve Jeffers


    PROTOCOL 07-05.1: 5% clarified V8 PARP(H)

    David Shew


    PROTOCOL 07-06.1: PCH agar

    Kelly Ivors


    PROTOCOL 07-07.1: Selective Phytophthora agar

    Paul Tooley


    PROTOCOL 07-08.1: CARP

    Everett Hansen


    PROTOCOL 07-09.1: CARP-D

    Everett Hansen


    PROTOCOL 07-10.1: CARP+

    Everett Hansen


    PROTOCOL 07-11.1: V8 agar (V8A) or broth

    Steve Jeffers


    PROTOCOL 07-12.1: Super V8 agar (sV8)

    Steve Jeffers


    PROTOCOL 07-13.1: V8 rye agar

    Jean Ristaino


    PROTOCOL 07-14.1: Rye A agar

    Kelly Ivors


    PROTOCOL 07-15.1: Rye B agar

    Jean Ristaino


    PROTOCOL 07-16.1: Oatmeal agar

    Sabine Werres


    PROTOCOL 07-17.1: 5% carrot agar (CA)

    David Shew


    PROTOCOL 07-18.1: Fresh lima bean agar

    Jean Ristaino


    PROTOCOL 07-19.1: Pea sucrose agar (PSA)

    Kelly Ivors


    PROTOCOL 07-20.1: Pea broth

    Peter Bonants


    PROTOCOL 07-21.1: Corn meal PPP agar

    Kelly Ivors


    PROTOCOL 07-22.1: Rye seed medium

    Michael Coffey


    PROTOCOL 07-23.1: Carrot agar (CA) or broth

    Michael Coffey


    PROTOCOL 07-24.1: Pea or lima bean agar or broth

    Michael Coffey


    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


    Publish Date: 2016
    Format: NOTE: Pricing is based on a per-site basis. To purchase subscriptions for two or more sites, please adjust your quantity during checkout.

    Edited by Kelly Ivors

    Laboratory Protocols for Phytophthora Species (Annual Online Subscription for Libraries, Companies, and Institutions)

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