Plant And Animal Genetic Resources Preservation Research Unit Site Logo
ARS Home About Us Helptop nav spacerContact Us En Espanoltop nav spacer
Printable VersionPrintable Version     E-mail this pageE-mail this page
Agricultural Research Service United States Department of Agriculture
Search
  Advanced Search
 
Programs and Projects
Subjects of Investigation
National Animal Germplasm (NAGP)
Preservation and Quality Assessment of Plant Genetic Resources (PGRPP)
 

Research Project: RESEARCH TO DEVELOP STRATEGIES AND TECHNOLOGES FOR PRESERVING PLANT GENETIC DIVERSITY IN EX SITU GENEBANKS

Location: Plant And Animal Genetic Resources Preservation Research Unit

Title: ULTRASTRUCTURAL CHANGES IN MINT MERISTEMS DURING THE CRYOPRESERVATION PROCESS

Authors

Submitted to: Symposium Series
Publication Type: Abstract Only
Publication Acceptance Date: May 5, 2005
Publication Date: June 1, 2005
Citation: Volk, G.M., Caspersen, A.M. Ultrastructural changes in mint meristems during the cryopreservation process. In: Symposium Series Meristems, June 2-5, 2005, Ames, Iowa. p. 21.

Interpretive Summary: Cryopreservation techniques can be used to preserve specific plant genotypes. One millimeter shoot tips are excised and treated with cryoprotectant solutions to remove excess water and introduce solutes that promote glass formation during the cooling process. Shoot tips exposed to liquid nitrogen temperatures are diluted into concentrated sucrose solutions upon thawing. Shoot tips recover and form green shoots in tissue culture within a few weeks. Mint shoot tips exposed to each step of the cryopreservation procedure were embedded in resin, sectioned, and observed using transmission electron microscopy. We demonstrate that despite the significant water loss that occurs during cryoprotective treatments, meristem cells remain intact. These cells are primarily damaged during the warming and dilution treatments of the cryopreservation procedure. We believe that membrane rupture upon warming and cryoprotectant toxicity may be significant sources of damage during cryopreservation. We believe that membrane rupture upon warming and cryoprotectant toxicity may be significant sources of damage during cryopreservation.

Technical Abstract: Cryopreservation techniques can be used to preserve specific plant genotypes. One millimeter shoot tips are excised and treated with cryoprotectant solutions to remove excess water and introduce solutes that promote glass formation during the cooling process. Shoot tips exposed to liquid nitrogen temperatures are diluted into concentrated sucrose solutions upon thawing. Shoot tips recover and form green shoots in tissue culture within a few weeks. Mint shoot tips exposed to each step of the cryopreservation procedure were embedded in resin, sectioned, and observed using transmission electron microscopy. Shoot tip water content during the cryopreservation process was determined using differential scanning calorimetry. Plasmolysis within meristematic cells was minimal, despite the significant loss of water during cryoprotectant treatments. Permeable cryoprotectants such as dimethyl sulfoxide and ethylene glycol replaced most of the free water within the cells. Significant damage was not observed after cryoprotectant treatment and liquid nitrogen exposure; however, shoot tips cooled to liquid nitrogen temperatures, and subsequently warmed and diluted in 1.2 M sucrose were plasmolyzed and appeared to have damaged membranes. Our data suggest that the loss of water during cryoprotectant treatments may not be the primary source of damage during the cryopreservation procedure. We believe that membrane rupture upon warming and cryoprotectant toxicity may be significant sources of damage during cryopreservation.

   

 
Project Team
Walters, Christina
Volk, Gayle
Richards, Christopher
 
Publications
   Publications
 
Related National Programs
  Plant Genetic Resources, Genomics and Genetic Improvement (301)
  Plant Biological and Molecular Processes (302)
 
 
Last Modified: 05/23/2013
ARS Home | USDA.gov | Site Map | Policies and Links 
FOIA | Accessibility Statement | Privacy Policy | Nondiscrimination Statement | Information Quality | USA.gov | White House