title: Impact of pr-10a overexpression on the cryopreservation success of Solanum tuberosum suspension cultures.
abstract: 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.
authors.affiliation: Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Inhoffenstr. 7b, 38124 Braunschweig, Germany. Lea.Vaas@dsmz.de
authors.forename: Lea A I
authors.initials: LA
authors.lastname: Vaas
authors.forename: Maja
authors.initials: M
authors.lastname: Marheine
authors.forename: Stephanie
authors.initials: S
authors.lastname: Seufert
authors.forename: Heinz Martin
authors.initials: HM
authors.lastname: Schumacher
authors.forename: Heiko
authors.initials: H
authors.lastname: Kiesecker
authors.forename: Elke
authors.initials: E
authors.lastname: Heine-Dobbernack
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D015925:Cryopreservation / Q000379:methods*; D005615:Freezing; D015870:Gene Expression / Q000187:drug effects; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011198:Solanum tuberosum / Q000166:cytology* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D013012:Sorbitol / Q000494:pharmacology; D013535:Suspensions
chemical_list: D010940:Plant Proteins; D013535:Suspensions; D013012:Sorbitol