Rapid diagnostic tools for Phytophthora on Horticultural Crops

Description

Manual includes the agenda for the Phytophthora diagnostics workshop and the lab protocols for Phytophthora diagnostic tools.

List of lab protocols:

  1. Field collection of Phytophthora
  2. Isolation and morphological observations of unknown
    species
  3. Use Lucid Key to identify unknown species
  4. Use US and NL Phytophthora database and NCBI
    sequence to identify unknown species
  5. DNA extraction – Direct PCR from fresh/dried tissue
    (PINF/ITS)
  6. DNA extraction methods: simple to complex and kits
  7. PCR-RFLP of ITS amplicons
  8. Digest PCR-RFLP of ITS amplicons; Run gel for the ITS
    PCR products from Lab5
  9. Phytophthora identification with TaqMan real time PCR
  10. Run gels for PCR-RFLP of ITS amplicons from Lab8
  11. Pack Lock Probe (PLP): Ligation & Exonuclease
  12. PLP: SYBRGreen PCR of ligated samples; PCR-RFLP
    analysis for Lab10
  13. PLP: Hybridizations
  14. Novel detection methods
  15. PLP: Stain microarray and Interpretation; “Unknown
    species revealed” to students

Value Chain

Pest management

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