title: Recovery of plants from pea and strawberry meristems cryopreserved for 28 years.
abstract: Meristems isolated from in vitro strawberry shoot cultures (Fragaria x ananassa Duch. cv. Redcoat) and in vitro germinated pea seedlings (Pisum sativum L. cv. Century) were treated with cryoprotectants, slowly frozen to -40 degree C (0.6 degree C per min for pea and 0.84 degree C per min for strawberry), and stored in liquid nitrogen in 1979. In 2007, several ampoules containing cryopreserved meristems of each species were thawed, cultured in vitro, and scored for viability. Meristems which differentiated into plants upon thawing and reculture were considered viable. Fifty-nine percent of the 91 strawberry meristems thawed in 2007 were viable after 28 years of cryopreservation, as compared to 56 percent viability after 8 weeks cryopreservation, as reported in the original study. Fourteen percent of the 78 pea meristems thawed in 2007 were viable after 28 years, as compared to 61 percent after 26 weeks of cryopreservation, as reported in the original study. With both strawberry and pea meristems there was variability in viability between ampoules. Viability between thawed ampoules of strawberry meristems ranged from 35 to 80 percent. Four of eight thawed ampoules of pea meristems contained no viable meristems, while the highest viability from a single ampoule was 77 percent. Shoots obtained from surviving meristems were rooted in vitro, transferred to soil, and produced phenotypically normal plants.
authors.affiliation: National Research Council of Canada, Plant Biotechnology Institute, 110 Gymnasium Place, Saskatoon, SK, Canada S7N 0W9. Karen.Caswell@nrc-cnrc.gc.ca
authors.forename: Karen L
authors.initials: KL
authors.lastname: Caswell
authors.forename: Kutty K
authors.initials: KK
authors.lastname: Kartha
mesh_terms: D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D031985:Fragaria / Q000502:physiology*; D018519:Meristem / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D018532:Pisum sativum / Q000502:physiology*; D013997:Time Factors
chemical_list: D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide