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United States Department of Agriculture

Agricultural Research Service

Research Project: MANAGEMENT OF TEMPERATE FRUIT NUT AND SPECIALTY CROP GENETIC RESOURCES

Location: National Clonal Germplasm Repository (Corvallis, Oregon)

Title: Choosing and applying cryopreservation protocols to new species or tissues

Author
item Reed, Barbara

Submitted to: Acta Horticulturae
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: April 15, 2010
Publication Date: July 15, 2011
Citation: Reed, B.M. 2011. Choosing and applying cryopreservation protocols to new species or tissues. Acta Horticulturae. 908:363-372.

Interpretive Summary: Cryopreservation, long-term storage in liquid nitrogen, is now a viable option for storage of plant cells, tissues, seeds and embryos. Facilities throughout the world are beginning to safeguard a wide range of plant propagules in long-term storage. Careful planning of the details of storage will make the cryopreserved collections both useful and more valuable. The general principles apply to all types of storage, but the specifics for each stage of planning will vary with the institution. Standard protocols are used as the basis for most plant cryopreservation. Each of these methods has some basic steps that can be modified to make them effective for many types of plants. The choice of technique used for storage depends on the needs of the facility involved. Personnel, equipment, expertise, plant type, and available facilities may influence which technique is most appropriate. Finding a protocol for use with a new plant species may be as simple as testing available protocols for similar plants. Adjustments at critical points allow relatively quick adaptation of standard protocols to new groups of plants. These parameters may also provide clues for future protocol improvements.

Technical Abstract: Some have proposed that it is necessary to develop new cryopreservation protocols for each new plant or tissue. We have found that standard protocols can be applied to many plants with few if any changes. Screening of groups of plants in a genus show that many protocols are easily applied to large groups of plants. The protocol to use can be chosen from those developed for similar plants or several standard protocols can be tested. We often compare controlled rate cooling, PVS2 vitrification, and encapsulation-dehydration techniques. Comparison of these techniques on diverse germplasm of pear, grass, blueberry and mint provided clear choices of the best protocol to use for storing large collections. Once a protocol is chosen, some critical points can be adjusted to improve the plant response as needed. Each of these methods has some basic steps that can be modified to make them effective for many types of plants. Finding a protocol for use with a new plant species may be as simple as testing available protocols for similar plants. Adjustments at critical points allow relatively quick adaptation of standard protocols to new groups of plants. Preliminary knowledge of the plant species can also provide tools for choosing a technique and certain parameters may help predict success or failure for a particular group of plants. These parameters may also provide clues for future protocol improvements.

Last Modified: 8/21/2014
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