Journal
PLANT BIOLOGY
Volume 6, Issue 2, Pages 119-127Publisher
WILEY
DOI: 10.1055/s-2004-817796
Keywords
cryopreservation; functional genomics; germplasm storage; moss; mutant collection; Physcomitrella patens (Hedw.); BSG
Categories
Ask authors/readers for more resources
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 degreesC/min down to -35degreesC with a 10 min holding time before transferring plants to -152 degreesC 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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available