Cryopreservation of immature seeds of Bletilla striata by vitrification.
An efficient protocol was established for the cryopreservation of immature seeds of a terrestrial orchid, Bletilla striata. Immature seeds collected 2-4 months after pollination (MAP) were treated using three different cryogenic procedures: (1) direct plunging into liquid nitrogen, (2) vitrification, and (3) vitrification with preculture. When immature seeds collected 3 MAP and 4 MAP were precultured for 3 days on New Dogashima medium supplemented with 0.3 M sucrose and cryopreserved by vitrification, the survival rate after preservation, as assessed by staining with 2,3,5-triphenyltetrazolium chloride, was 92% and 81%, respectively. Immature seeds thus treated showed no decrease in germination rate relative to untreated immature seeds, and they developed into normal plantlets in vitro.
Hirano
T
T
Laboratory of Plant Cell Technology, Faculty of Horticulture, Chiba University, 648 Matsudo, Matsudo-City, Chiba, 271-8510, Japan.
Godo
T
T
Mii
M
M
Ishikawa
K
K
D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D018744:DNA, Plant / Q000032:analysis; D018525:Germination; D029595:Orchidaceae / Q000166:cytology* / Q000254:growth & development; D012639:Seeds / Q000737:chemistry / Q000166:cytology*; D014867:Water / Q000032:analysis
D018744:DNA, Plant; D014867:Water