ORIGINAL ARTICLE Protein changes in the shoot-tips of vanilla (Vanilla planifolia) in response to osmoprotective treatments Marı ´a Teresa Gonza ´ lez-Arnao 1 • Armando Guerrero-Rangel 2 • Octavio Martı ´nez 3 • Silvia Valde ´ s-Rodrı ´guez 2 Received: 16 August 2016 / Accepted: 23 January 2018 Ó Society for Plant Biochemistry and Biotechnology 2018 Abstract Cryogenic storage of vanilla shoot-tips represents the safest biotechnological strategy for the long-term conservation of the vanilla germplasm, but successful cryopreservation depends on its tolerance to both dehydration stress imposed by cry- oprotective treatments and thermal stress produced by immersion in liquid nitrogen. In this work, we evaluated the impact of various osmoprotective treatments on protein expression patterns in vanilla (Vanilla planifolia) shoot-tips subjected to successive dehydration steps prior to cryopreservation. Two-dimensional electrophoretic protein profiles of shoot-tips dis- sected from in vitro grown plants and preconditioned on semisolid media with 0.3 M sucrose for one day, and shoot-tips preconditioned, loaded with a solution of 0.4 M sucrose and 2 M glycerol, and subsequently exposed to plant vitrification solution 3 (50% (w/v) sucrose and 50% (w/v) glycerol), were compared with non-treated dissected shoot-tips. We observed an increase in the expression level of six protein spots (fold change exceeding 1.5) and a decrease (fold change not exceeding 0.6) of ten protein spots after preconditionig treatment, whereas the profiles after preconditioning, loading and exposure to vitrification solution showed an increase in the expression level of 21 protein spots and a decrease in the expression level of 13. Most proteins identified were down-regulated and belonged to groups of biosynthesis, folding, and protein degradation. Many others were related to energetic metabolism, defense, and cell structure. These preliminary results contribute to knowledge of the proteome of this species and partially clarify its sensitivity to osmotic dehydration treatments. Keywords Dehydration stress Á Osmoprotection Á Protein profiles Á Proteomics Á Two-dimensional electrophoresis Á Vanilla Abbreviations 2-DE Bidimensional electrophoresis DTT Dithiothreitol EDTA Ethylenediaminetetraacetic acid IPG Immobilized pH gradients IEF Isoelectric focusing LC Liquid chromatography LTQ Linear ion trap MS/MS Mass spectrometry in tandem PVS3 Plant vitrification solution 3 Q-TOF Quadrupole time-of-flight Introduction Vanilla is a very emblematic orchid that grows in tropical regions around the world (Divakaran et al. 2006). The vanilla genus contains about 110 species, including V. planifolia Jack that is considered the most relevant because it produces vanillin. This traditional flavoring is widely used in numerous foods and beverage products. In addition, it is in high demand in the cosmetics, pharmaceutical, and agrochemical industries (Sinha et al. 2008). Vanilla plan- ifolia originated in Me ´xico (Soto-Arenas 2003); however, Electronic supplementary material The online version of this article (https://doi.org/10.1007/s13562-018-0442-1) contains supplementary material, which is available to authorized users. & Silvia Valde ´s-Rodrı ´guez svaldes@ira.cinvestav.mx 1 Facultad de Ciencias Quı ´micas, Universidad Veracruzana, Prolongacio ´n Oriente 6, No. 1009, CP 94340 Orizaba, Veracruz, Mexico 2 Cinvestav-Unidad Irapuato, Km 9.6 Libramiento Norte Carretera Irapuato Leo ´n, CP 36824 Irapuato, Guanajuato, Mexico 3 Laboratorio Nacional de Geno ´mica para la Biodiversidad (Langebio), Cinvestav, Apartado Postal 629, C.P. 36500 Irapuato, Guanajuato, Mexico 123 Journal of Plant Biochemistry and Biotechnology https://doi.org/10.1007/s13562-018-0442-1