Exploring the physiological basis of cryopreservation success and failure in clonally propagated in vitro crop plant germplasm

Harding, Keith; Johnston, Jason W.; Benson, Erica E.

doi:10.2137/145960609789267524

Cited by 42 publications

(25 citation statements)

References 27 publications

(38 reference statements)

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“…DNA methylation is required for regular development of plants and may have an impact on vigour and morphogenesis (Chan et al 2005). Some studies have discussed putative effects of changes in DNA methylation on genomic DNA stability and rearrangements in micropropagated and cryopreserved stocks (Harding 2004;Harding et al 2009;Peredo et al 2008). Modification of epigenetic features due to the stress during in vitro culture could possibly result in altered phenotypes in the recovered plants.…”

Section: Epigenetic Factorsmentioning

confidence: 99%

Potato Shoot Tip Cryopreservation. A Review

2010

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Potato is one of the most important crops worldwide. Genetic resources of potato (Solanum tuberosum L. ssp. tuberosum) and related cultivated species are conserved through storage of tubers, in vitro plants and in cryopreservation. Cryopreservation, storage in or above liquid nitrogen, is the best option to maintain vegetatively propagated plants in the long term. The present review gives comprehensive information about various cryopreservation techniques for potato published from 1977 until the present. It discusses factors that affect the process and success of cryopreservation, such as donor culture conditions, preculture, cooling, warming and post-culture treatments. Studies are presented that analyse the histological and ultrastructural changes after different cryopreservation steps and the morphological pathways during regeneration of plants after rewarming. The maintenance of genetic stability in potato after cryopreservation has also been demonstrated by various phenotypic and molecular methods. The first thermal analyses on potato shoot tips are presented using differential scanning calorimetry to analyse the state of water during cooling and warming. Biochemical analyses of different compounds, such as soluble sugars and proteins, have been performed to understand and improve existing cryogenic methods. Potato is an example where successful virus elimination has been obtained via cryopreservation of shoot tips (cryotherapy). There are already cryopreserved collections of potato shoot tips in

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Section: Epigenetic Factorsmentioning

confidence: 99%

Potato Shoot Tip Cryopreservation. A Review

2010

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“…Preconditioning of plants has also been shown to increase antioxidant levels in plants prior to cryopreservation (Baek & Skinner, 2003;Dai et al, 2009;Harding et al, 2009;Zhao & Blumwald, 1998). Baek and Skinner (2003) analysed the expression of antioxidant genes in wheat species after cold acclimation and found increased expression of antioxidant enzymes, such as SOD and catalase.…”

Section: Preconditioning and Cold Acclimationmentioning

confidence: 99%

Current Issues in Plant Cryopreservation

Kaczmarczyk¹,

Funnekotter²,

Menon³

et al. 2012

Current Frontiers in Cryobiology

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“…The actual optimum depends on a variety of additional parameters such as the geographical and ecological origin of the plant material (Berjak et al 2011), the genotype of the cell material (Schäfer-Menuhr and Schumacher 1997), and its age and physiological status (Harding et al 2009), and thus can be much more complex than illustrated above (Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

“…The strength of the pretreatment is a combination of the concentration of the applied substance(s) and their residence time. The physiological changes during pretreatment were identified as a conglomerate of competences in cold adaptation, including the maintenance of membrane stability and the scavenging of free radicals after thawing (Volk 2010;Harding et al 2009), severely affecting cryopreservability.…”

Section: Introductionmentioning

confidence: 99%

“…In projects such as CRYMCEPT (EU 5th framework programme, Project Reference: QLK5-CT-2002-01279), the physico-and biochemical state of the considered plant species and tissues needed for successful cryopreservation was elaborated in some detail (see Panis and Lambardi 2006 for an overview). With more information available from fundamental research on the causal relationships determining the cryopreservability of plant materials (Swennen et al 2011;Carpentier et al 2007), awareness increased that physiological factors strongly determine the success of survival (Harding 2004;Harding et al 2009). …”

Section: Introductionmentioning

confidence: 99%

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Impact of pr-10a overexpression on the cryopreservation success of Solanum tuberosum suspension cultures

et al. 2012

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Although many genes are supposed to be a part of plant cell tolerance mechanisms against osmotic or salt stress, their influence on tolerance towards stress during cryopreservation procedures has rarely been investigated. For instance, the overexpression of the pathogenesis-related gene 10a (pr-10a) leads to improved osmotic tolerance in a transgenic cell culture of Solanum tuberosum cv. Désirée. In this study, a cryopreservation method, consisting of osmotic pretreatment, cryoprotection with DMSO and controlled-rate freezing, was used to characterize the relation between cryopreservation success and pr-10a expression in suspension cultures of S. tuberosum wild-type cells and cells overexpressing pathogenesis-related protein 10a (Pr-10a). By varying the sorbitol concentration, thus modifying the strength of the osmotic stress during the pretreatment phase, it can be shown that the wild type can successfully be cryopreserved only in a relatively narrow range of sorbitol concentrations, while the pr-10a overexpression leads to an enhanced cryopreservation success over the whole range of applied sorbitol concentrations. Together with transcription data we show that the pr-10a overexpression causes an enhanced osmotic tolerance, which in turn leads to enhanced cryopreservability, but also indicates a role of pr-10a in signal transduction. An increased cryopreservability of the transgenic cell line occurs for pretreatments longer than 24 h. Since both genotypes, characterized by distinct baseline levels of expression, exhibited similar patterns of expression induction, the induction of pr-10a appears to be a key step in the stress signal transduction of plant cells under osmotic stress.

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Exploring the physiological basis of cryopreservation success and failure in clonally propagated in vitro crop plant germplasm

Cited by 42 publications

References 27 publications

Potato Shoot Tip Cryopreservation. A Review

Potato Shoot Tip Cryopreservation. A Review

Current Issues in Plant Cryopreservation

Impact of pr-10a overexpression on the cryopreservation success of Solanum tuberosum suspension cultures

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