High throughput cryopreservation of 140 000 Physcomitrella patens mutants

  • Schulte J
  • Reski R
  • 16


    Mendeley users who have this article in their library.
  • 30


    Citations of this article.


A high throughput protocol was established to preserve 140,000 mutants of a moss, Physcomitrella patens, a model plant for functional genomics studies, over liquid nitrogen. Regarding the reliable long-term storage of diverse mutant phenotypes, as well as time and cost effectiveness, each working step was optimized: 1) plant preparation, 2) freezing regime, cryogenic conditions, 3) regrowth after thawing. A prerequisite for maximum regrowth was a 1-week preculture of chopped plant material on a supplemented medium prior to freezing. Cryo vials as preculture vessels resulted in identical regrowth rates, compared to petri dishes. The cryo vial type had a significant influence on regrowth rates. A cooling rate of - 1 degrees C/min down to - 35 degrees C with a 10 min holding time before transferring plants to - 152 degrees C was the most suitable freezing regime. This protocol allows a cryopreservation of 1100 plants during a 5-day working week, practicable by one person. For more than 650 cryopreserved mutants a maximum regrowth rate of 100 % was obtained, independently of mutant phenotypes.

Author-supplied keywords

  • Cryopreservation
  • Functional genomics
  • Germplasm storage
  • Moss
  • Mutant collection
  • Physcomitrella patens (Hedw.) B.S.G.

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Get full text


  • J. Schulte

  • R. Reski

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free