title: LONG-TERM PRECONDITIONING OF PLANTLETS: A PRACTICAL METHOD FOR ENHANCING SURVIVAL OF PINEAPPLE (Ananas comosus Merr.) SHOOT TIPS CRYOPRESERVED USING VITRIFICATION.
abstract: BACKGROUND
  The purpose of this study was to develop an efficient cryopreservation protocol for pineapple (Ananas comosus Merr.) shoot tips.
  
  
  OBJECTIVE
  The optimal state of pineapple plantlets was investigated by using sucrose preconditioning to enhance survival after cryostorage.
  
  
  METHODS
  To achieve a suitable state of plantlets before cryopreservation, 0.2 M to 0.4 M sucrose concentrations combined with short- (0-7 days), medium- (15-30 days), and long-term (75-150 days) preconditioning periods were compared.
  
  
  CONCLUSIONS
  The highest survival (100 %) was achieved using the following procedure: intact plantlets underwent long-term preconditioning with 0.2 M sucrose for 135 days, dissected shoot tips were treated with a loading solution containing 2.0 M glycerol + 0.4 M sucrose for 60 min at 25 degree and the shoot tips were dehydrated in PVS2 for 2h at 0 degree C before being plunged in liquid nitrogen. Rewarming was conducted in a water-bath for 30 s at 40 degree C and PVS2 was replaced with a 1.2 M sucrose solution for 30 min at 25 degree C. The shoot tips were transferred on semisolid medium and left in the dark for 1 week, then in dim light for 3 weeks.
authors.affiliation: Department of Biology, National Museum of Natural Science, Taichung, Taiwan.
authors.forename: W H
authors.initials: WH
authors.lastname: Hu
authors.affiliation: Department of Life Sciences, National Chung-Hsing University, Taichung, Taiwan.
authors.forename: S F
authors.initials: SF
authors.lastname: Liu
authors.affiliation: Department of Life Sciences, National Chung-Hsing University, Taichung, Taiwan. siliaw@mail.nchu.edu.tw.
authors.forename: S I
authors.initials: SI
authors.lastname: Liaw
mesh_terms: D031230:Ananas / Q000187:drug effects / Q000502:physiology*; D015925:Cryopreservation / Q000379:methods; D003451:Cryoprotective Agents / Q000494:pharmacology; D005990:Glycerol / Q000494:pharmacology; D018520:Plant Shoots / Q000187:drug effects / Q000502:physiology*; D013395:Sucrose / Q000494:pharmacology; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents; D013395:Sucrose; D005990:Glycerol