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Area of Science:

  • Plant Pathology
  • Mycology
  • Cryobiology

Background:

  • Hop powdery mildew (HPM), caused by *Podosphaera macularis*, is a significant pathogen of hops.
  • HPM is an obligate parasite, necessitating continuous cultivation on living hop plants for strain maintenance.

Purpose of the Study:

  • To establish cryopreservation parameters for HPM spores.
  • To reduce the labor-intensive requirement of maintaining HPM cultures on living plant tissue.

Main Methods:

  • HPM spores (OSU C-100 strain and a mixed strain) were desiccated over silica gel to determine optimal moisture content (MC).
  • Spore viability was assessed by observing hyphal regrowth on glass slides and successful infection of hop leaf discs after cryopreservation in liquid nitrogen.
  • Regrowth was evaluated before and after desiccation and liquid nitrogen exposure.

Main Results:

  • Optimal spore desiccation to 2-3% MC was determined to be crucial for successful cryopreservation.
  • The OSU C-100 strain required 8-10 hours of desiccation, while mixed strains needed 6-8 hours to reach the target MC.
  • Successful hyphal growth and leaf disc infection confirmed spore viability post-cryopreservation.

Conclusions:

  • A protocol for cryopreserving HPM spores was successfully developed.
  • The recommended method involves drying spores to 2-3% MC over silica gel, followed by direct immersion in liquid nitrogen.
  • Rewarming involves sequential 1-minute incubation in 45°C and 20°C water, with viability confirmed on hop leaf discs.