title: Dehydrin expression as a potential diagnostic tool for cold stress in white clover.
abstract: Cold acclimation is important for crop survival in environments undergoing seasonal low temperatures. It involves the induction of defensive mechanisms including the accumulation of different cryoprotective molecules among which are dehydrins (DHN). Recently several sequences coding for dehydrins were identified in white clover (Trifolium repens). This work aimed to select the most responsive to cold stress DHN analogues in search for cold stress diagnostic markers. The assessment of dehydrin transcript accumulation via RT-PCR and immunodetection performed with three antibodies against the conserved K-, Y-, and S-segment allowed to outline different dehydrin types presented in the tested samples. Both analyses confirmed that YnKn dehydrins were underrepresented in the controls but exposure to low temperature specifically induced their accumulation. Strong immunosignals corresponding to 37-40 kDa with antibodies against Y- and K-segment were revealed in cold-stressed leaves. Another 'cold-specific' band at position 52-55 kDa was documented on membranes probed with antibodies against K-segment. Real time RT-qPCR confirmed that low temperatures induced the accumulation of SKn and YnSKn transcripts in leaves and reduced their expression in roots. Results suggest that a YnKn dehydrin transcript with GenBank ID: KC247805 and the immunosignal at 37-40 kDa, obtained with antibodies against Y- and K-segment are reliable markers for cold stress in white clover. The assessment of SKn (GenBank ID: EU846208) and YnSKn (GenBank ID: KC247804) transcript levels in leaves could serve as additional diagnostic tools.
authors.affiliation: Plant Stress Molecular Biology Department, Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 21, 1113 Sofia, Bulgaria; Institute of Plant Sciences and Oeschger Centre for Climate Change Research (OCCR), University of Bern, Altenbergrain 21, CH-3013 Bern, Switzerland. Electronic address: irina.vaseva@abv.bg.
authors.forename: Irina Ivanova
authors.initials: II
authors.lastname: Vaseva
authors.affiliation: Institute of Plant Sciences and Oeschger Centre for Climate Change Research (OCCR), University of Bern, Altenbergrain 21, CH-3013 Bern, Switzerland.
authors.forename: Iwona
authors.initials: I
authors.lastname: Anders
authors.affiliation: Plant Stress Molecular Biology Department, Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 21, 1113 Sofia, Bulgaria.
authors.forename: Bistra
authors.initials: B
authors.lastname: Yuperlieva-Mateeva
authors.affiliation: Plant Stress Molecular Biology Department, Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 21, 1113 Sofia, Bulgaria.
authors.forename: Rosa
authors.initials: R
authors.lastname: Nenkova
authors.affiliation: Plant Stress Molecular Biology Department, Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 21, 1113 Sofia, Bulgaria.
authors.forename: Anelia
authors.initials: A
authors.lastname: Kostadinova
authors.affiliation: Institute of Plant Sciences and Oeschger Centre for Climate Change Research (OCCR), University of Bern, Altenbergrain 21, CH-3013 Bern, Switzerland.
authors.forename: Urs
authors.initials: U
authors.lastname: Feller
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D003080:Cold Temperature; D018506:Gene Expression Regulation, Plant; D015151:Immunoblotting; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013312:Stress, Physiological; D029921:Trifolium / Q000235:genetics* / Q000378:metabolism
chemical_list: D015415:Biomarkers; D010940:Plant Proteins; C070556:dehydrin proteins, plant
keywords: Cold stress; Dehydrins; Immunoblot; RT-PCR; White clover