Skip to main content
ARS Home » Pacific West Area » Corvallis, Oregon » National Clonal Germplasm Repository » Research » Publications at this Location » Publication #165601
Title: EVALUATION OF CRITICAL POINTS IN TECHNOLOGY TRANSFER OF CRYOPRESERVATION PROTOCOLS TO INTERNATIONAL PLANT CONSERVATION LABORATORIES

Author
item Reed, Barbara
item KOVALCHUK, IRINA - INSTITUTE OF PLANT PHYS.
item KUSHNARENKO, SVETLANA - INSTITUTE OF PLANT PHYS
item MEIER-DINKEL, ANDREAS - LOWER SAXONY INSTITUTE
item SCHOENWEISS, KATJA - LOWER SAXONY INSTITUTE
item PLUTA, STANISLAW - RESEARCH INSTITUTE OF POM
item STRACZNSKA, KRYSTYNA - RESEARCH INSTITUTE OF POM
item BENSON, ERICA - UNIVERSITY OF ABERTAY

Submitted to: CryoLetters
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/1/2004
Publication Date: 8/30/2004
Citation: Reed, B.M., Kovalchuk, I., Kushnarenko, S., Meier-Dinkel, A., Schoenweiss, K., Pluta, S., Stracznska, K., Benson, E.E. 2004. Evaluation of critical points in technology transfer of cryopreservation protocols to international plant conservation laboratories. CryoLetters. July/August edition. 25:341-352.

Interpretive Summary: Long-term storage of plant tissues in liquid nitrogen is now used for long-term conservation of vegetatively-propagated crops. Development of standard techniques for cryopreservation is important for successful transsfer of storage protocols to diverse laboratories. Evaluation of the critical points of each technique will greatly assist in developing internationally-used protocols. The goals of this project were to assess critical points of two major cryopreservation techniques during the transfer of two common protocols to international laboratories; analyze viability for each technique and location; and develop recommendations based on the assessments and data from the participating laboratories. Investigators from Germany, Kazakhstan, Poland and UK participated in a 2-week methods training workshop after which the techniques were tested in the home laboratories of the participants. After one year, site visits by the technology trainers identified critical points in the protocols. The most critical factors in all laboratories were culture health, operator skills and experience, and clarification of the technical details of the procedures. The laboratories repeated the experiments using corrected procedures. Final results showed that correction of critical factors improved the post-storage recovery in all the laboratories. The study developed recommendations based on critical-point evaluations and assessments of recovery in three experimental data sets.

Technical Abstract: Cryopreservation of plant tissues in liquid nitrogen is now used for long-term conservation of vegetatively-propagated crops. Development of standard techniques for cryopreservation is important to the international plant-conservation community for successful implementation of storage protocols in diverse and internationally dispersed laboratories. Evaluation of the critical points of each preservation technique will greatly assist in developing and validating internationally-used cryopreservation protocols. The goals of this project were to assess critical points of PVS2-vitrification and encapsulation-dehydration protocols during transfer to international laboratories; analyze post-storage viability for each technique and location; and develop recommendations based on the assessments and data from the participating laboratories. Investigators from Germany, Kazakhstan, Poland and UK participated in a 2-week training workshop in cryopreservation methods after which the techniques were tested in the home laboratories of the participants. After one year, site visits by the technology trainers identified critical points in the protocols. The most critical factors in all laboratories were culture health, operator skills and experience, and clarification of the technical details of the procedures. The laboratories repeated the experiments using corrected procedures. Final results showed that correction of critical factors improved the post-storage recovery in all the laboratories. The study developed recommendations based on critical-point evaluations and assessments of recovery in three experimental data sets.