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induction of heat shock response and acquisition of thermotolerance in callus cultures of Gerbera jamesonii
Authors:Nelson Marmiroli  Francesco M Restivo  Chris J Smith  Gabriella Di Cola  Elena Maestri  Francesca Tassi
Institution:(1) Department of Environmental Sciences, University of Parma, Viale delle Scienze, 1-43100 Parma, Italy;(2) Institute of Genetics, University of Parma, Viale delle Scienze, 1-43100 Parma, Italy;(3) Faculty of Science, Health and Medical Studies, North East Wales Institute, CH5 4BR Deeside, Clwyd, United Kingdom;(4) HI-TECH High Technology in Biology and Medicine srl, Borgo del Parmigianino 5, 1-43100 Parma, Italy
Abstract:Summary The heat shock (HS) response in callus cultures of the ornamental plant Gerbera jamesonii H. Bolus var. hybrida was analyzed. A HS at 35° C or 40° C for 4 h induced (a) the synthesis of several heat shock proteins (HSPs), especially in the small molecular weight range and some spots corresponding to HSP70 components, and (b) an increase in the steady state levels of some specific mRNAs. At the nonstressing temperature (26° C), a sustainable level of translation for HSP70 was indeed carried out, as confirmed by immunological analysis with a monoclonal antibody against cotton HSP70. The steady state levels of mRNAs measured before and after a HS by Northern hybridization showed an increase with the heterologous probes HSP17.4, HSP17.6, and HSP21, whereas the probes HSC70 and HSP70 did not show any difference between the levels of control and HS-mRNAs. A pretreatment at 35° C, which induced a set of HSPs in the callus cultures, decreased the cell damage upon exposure to a temperature of 45° C as determined either with a regrowth test or by the tetrazolium reduction assay. Typically, as with the whole plants, callus of Gerbera jamesonii possessed the ability to respond to HS both by inducing HSPs and by developing an acquired thermotolerance.
Keywords:HSPs  HS-mRNAs  heterologous probes  anti HSP70 antibody  thermotolerance  regrowth test  TTC assay
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