Abdelwahed GHORBEL, Mohsen HANANA, Samia DALDOUL, Laurent DELUC, Celine LEON, Michael HOEFER, François BARRIEU, Romain FOUQUET

Identifcation and characterization of a seed-specifc grapevine dehydrin involved inabiotic stress response within tolerant varieties

To identify and isolate genes related to abiotic stress (salinity and drought) tolerance in grapevine, a candidate gene approachled to the isolation from Cabernet Sauvignon cultivar of a full-length cDNA of dehydrin gene. Te latter, named VvDhn , which is highlyand mainly induced in late embryogenesis in seeds, encodes for a protein of 124 amino acids with a predicted molecular mass of 13.3kDa. Details of the physicochemical parameters and structural properties (molecular mass, secondary structure, conserved domainsand motives, and putative posttranslational modifcation sites) of the encoded protein have also been elucidated. Te expression study ofVvDhn was carried out within plant organs and tissues as well as under drought and salt stresses. VvDhn was not detected in vegetativetissue, whereas it was only expressed during seed development (during late embryogenesis) at extremely high levels and was induced bysalt and drought stresses as well as ABA application. Moreover, salt stress induced VvDhn expression in the tolerant variety (Razegui)but not the sensitive variety (Syrah), which did not display expression variation during stress; VvDhn expression level and salt-stressresponse depend on regulatory mechanisms that are efcient only in the tolerant variety. On the other hand, under drought stressVvDhn was induced in both tolerant and sensitive varieties, with higher levels in the tolerant variety. In addition, stress signal moleculessuch as ABA (applied alone or in combination with saccharose) induced VvDhn expression, even at low levels. Minimal knowledgeabout the role and functionality of this gene is necessary and constitutes a prerequisite for including VvDhn in grapevine abiotic stresstolerance improvement programs.

PDF

___

Allagulova ChR, Gilamov FR, Shakirova FM, Vakhitov VA (2003). Te plant dehydrins: structure and functions. Biochemistry (Moscow) 68: 945951.
Alsheikh MK, Heyen BJ, Randall SK (2003). Ion binding properties of the dehydrin ERD14 are dependent upon phosphorylation. J Biol Chem 278: 4088240889.
Asif MH, Dhawan P, Nath P (2000). A simple procedure for the isolation of high quality RNA from ripening banana fruit. Plant Mol Biol Rep 18: 109115.
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990). Basic local alignment search tool. J Mol Biol 215: 403410.
Bailey TL, Elkan C (1994). Fitting a mixture model by expectation maximization to discover motifs in biopolymers, Proceedings of the Second International Conference on Intelligent Systems for Molecular Biology, pp. 2836, AAAI Press, Menlo Park, California, USA.
Blaskovic S, Blanc M, van der Goot FG (2013). What does S-palmitoylation do to the membrane proteins? FEBS J 280: 27662774.
Campbell SA, Close TJ (1997). Dehydrins: genes, proteins, and associations with phenotypic traits. New Phytol 137: 6174.
Chang S, Puryear J, Cairney J (1993). A simple and efcient method for isolating RNA from pine trees. Plant Mol Biol Rep 11: 113 116.
Chitarra W, Balestrini R, Vitali M, Pagliarani C, Perrone I, Schubert A, Lovisolo C (2014). Gene expression in vessel-associated cells upon xylem embolism repair in Vitis vinifera L. petioles. Planta 239: 887899.
Choi Y-J, Hur YY, Jung S-M, Kim S-H, Noh J-H, Park S-J, Park K-S, Yun H-K (2013). Transcriptional analysis of Dehydrin1 genes responsive to dehydrating stress in grapevines. Hortic Environ Biote 54: 272279.
Church GM, Gilbert W (1984). Genomic sequencing. P Natl Acad Sci USA 81: 19911995.
Close TJ (1996). Dehydrins: emergence of a biochemical role of a family of plant dehydration proteins. Physiol Plantarum 97: 795803.
Close TJ (1997). Dehydrins: a commonality in the response of plants to dehydration and low temperature. Physiol Plantarum 100: 291296.
Cramer G (2010). Abiotic stress and plant responses from the whole vine to the genes. Aust J Grape Wine R 16: 8693.
Cramer GR, Van Sluyter SC, Hopper DW, Pascovici D, Keighley T, Haynes PA (2013). Proteomic analysis indicates massive changes in metabolism prior to the inhibition of growth and photosynthesis of grapevine (Vitis vinifera L.) in response to water defcit. BMC Plant Biol 13: 49.
Daldoul S, Chenenanoui S, Mliki A, Höfer M (2009). Improvement of an RNA purifcation method for grapevine (Vitis vinifera L.) suitable for cDNA library construction. Acta Physiol Plant 31: 871875.
de Lartigue J (2011). Phosphorylation: the master switch of the cell. Oncology live. Published Online: Tursday, December 15, 2011.
Decendit A, Ramawat KG, Wafo P, Defeux G, Badoc A, Mérillon JM (1996). Anthocyanins, catechins, condensed tannins and piceid production in Vitis vinifera cell bioreactor cultures. Biotechnol Lett 18: 659662.
Deinlein U, Stephan AB, Horie T, Luo W, Xu G, Schroeder JI (2014). Plant salt-tolerance mechanisms. Trends Plant Sci 19: 371379.
Deluc L, Barrieu F, Marchive C, Lauvergeat V, Decendit A, Richard T, Carde JP, Mérillon JM, Hamdi S (2006). Characterization of a grapevine R2R2-MYB transcription factor that regulates the phenylpropanoid pathway. Plant Physiol 140: 499511.
Downey M, Harvey J, Robinson S (2003). Analysis of tannins in seeds and skins of Shiraz grapes throughout berry development. Aust J Grape Wine R 9: 1527.
Dure L III (1993). Structural motifs in LEA proteins. In: Close TJ, Bray EA, editors. Plant Responses to Cellular Dehydration During Environmental Stress. Current Topics in Plant Physiology. Vol. 10. Rockville, MD, USA: American Society of Plant Physiologists, pp. 91103.
Gaper NE, Giovanonni JJ, Whatkins CB (2014). Understanding development and ripening of fruit crops in an omics era. Hort Res 1: 14034. doi:10.1038/hortres.2014.34.
Gautier R, Douguet D, Antonny B, Drin G (2008). HELIQUEST: a web server to screen sequences with specifc α-helical properties. Bioinformatics 18: 21012102.
Goday A, Jensen AB, Culianez-Macia FA et al. (1994). Te maize abscisic acid-responsive protein Rab17 is located in the nucleus and interacts with nuclear localization signals. Plant Cell 6: 351360.
Gould N, Doulias PT, Tenopoulou M, Raju K, Ischiropoulos H (2013). Regulation of protein function and signaling by reversible cysteine S-nitrosylation. J Biol Chem 288: 2647326479.
Gupta B, Huang B (2014). Mechanism of salinity tolerance in plants: physiological, biochemical, and molecular characterization. International Journal of Genomics 214: 118.
Hamrouni L (2010). Evaluation de la tolérance au sel chez la vigne en Tunisie. Tèse de Doctorat, Faculté des Sciences de Tunis, pp. 246 (in French).
Hamrouni L, Hanana M, Abdelly C, Ghorbel A (2011). Exclusion du chlorure et inclusion du sodium: deux mécanismes concomittants de tolérance à la salinité chez la vigne sauvage Vitis vinifera subsp. Sylvestris (var. Séjnène). Biotechnol Agron Soc 15: 387400 (in French).
Hanana M, Deluc L, Fouquet R, Daldoul S, Léon C, Barrieu F, Ghorbel A, Mliki A, Hamdi S (2008). Identifcation et caractérisation dun gène de réponse à la déshydratation rd22 chez la vigne (Vitis vinifera L.). CR Biol 331: 569578 (in French).
Hanin M, Brini F, Ebe C, Toda Y, Takeda S, Masmoudi K (2011). Plant dehydrins and stress tolerance: versatile proteins for complex mechanisms. Plant Signaling and Behavior 6: 15031509.
Hara M, Fujinaga M, Kuboi K (2005). Metal binding by citrus dehydrin with histidine-rich Domains. J Exp Bot 56: 2695 2703.
Hennig L (1999). WinGene/WinPep: user-friendly sofware for the analysis of amino acid sequences. Biotechnology (NY) 26: 11701172.
Hewitt EJ (1966). Sand and Water Culture Methods Used in the Study of Plant Nutrition. Technical Communication No. 22. Commonwealth Agricultural Bureaux, Farnham Royal, UK.
Hochberg U, Degu A, Toubiana D, Gendler T, Nikoloski Z, Rachmilevitch S, Fait A (2013). Metabolite profling and network analysis reveal coordinated changes in grapevine water stress response. BMC Plant Biol 13: 184.
Hu L, Wang Z, Du H, Huang B (2010). Diferential accumulation of dehydrins in response to water stress for hybrid and common Bermuda grass genotypes difering in drought tolerance. J Plant Physiol 167: 103109.
Jaillon O, Aury J-M, Noel B, Policriti A, Clepet C, Casagrande A, Choisne N, Aubourg S, Vitulo N, Jubin C et al. (2007). Te grapevine genome sequence suggests ancestral hexaploidization in major angiosperm phyla. Nature 449: 463467.
Kleinwächter M, Radwan A, Hara M, Selmar D (2014). Dehydrin expression in seeds: an issue of maturation drying. Front Plant Sci 5: 402.
Koag M-C, Fenton RD, Wilken S, Close J (2003). Te binding of maize DHN1 to lipid vesicles. Gain of structure and lipid specifcity. Plant Physiol 131: 309316.
Kruger C, Berkowitz O, Stephan UW et al. (2002). A metal binding member of the late embryogenesis abundant protein family transports iron in the phloem of Ricinus communis L. J Biol Chem 277: 2506225069.
Lanigan RM, Sheppard TD (2013). Recent developments in amide synthesis: direct amidation of carboxylic acids and transamination reactions. Eur J Org Chem 2013: 74537465.
Matton DP, Nass N, Clarke AE, Newbigin E (1994). Self- incompatibility: how plants avoid illegitimate ofspring. P Natl Acad Sci USA 91: 1992.
Moriya K, Nagatoshi K, Noriyasu Y, Okamura T, Takamitsu E, Suzuki T, Utsumi T (2013). Protein N-myristoylation plays a critical role in the endoplasmic reticulum morphological change induced by overexpression of protein lunapark, an integral membrane protein of the endoplasmic reticulum. PLoS ONE 8: e78235.
Ollat N, Geny L, Soyer JP (1998). Les boutures fructifères de Vigne: validation dun modèle détude de la physiologie de la Vigne. J Int Sci Vigne Vin 32: 19 (n French).
OIV (2013). International Organization of Vine and Wine: Statistical Report on World Vitiviniculture 2013, pp. 28.
Rorat D (2006). Plant dehydrinstissue location, structure and function. Cell Mol Biol Lett 11: 536556.
Sadder MT, Al-Doss AA (2014). Characterization of dehydrin AhDHN from Mediterranean saltbush (Atriplex halimus). Turk J Biol 38: 469477.
Schwarz F, Aebi M (2011). Mechanisms and principles of N-linked protein glycosylation. Curr Opin Struc Biol 21: 576582.
Seymour GB, Granell A (2014). Fruit development and ripening. J Exp Bot 65: 44894490.
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar K (2005). MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28: 27312739.
Tompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997). Te CLUSTAL X windows interface: fexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25: 48764882.
Vaseva II, Anders I, Feller U (2014a). Identifcation and expression of diferent dehydrin subclasses involved in the drought response of Trifolium repens. J Plant Physiol 171: 213224.
Vaseva II, Anders I, Yuperlieva-Mateeva B, Nenkova R, Kostadinova A, Feller U (2014b). Dehydrin expression as a potential diagnostic tool for cold stress in white clover. Plant Physiol Bioch 78: 4348.
Wang Y, Dasso M (2009). SUMOylation and deSUMOylation at a glance. J Cell Sci 122: 42494252.
Wang Y, Xu H, Zhu H, Tao Y, Zhang G, Zhang L, Zhang C, Zhang Z, Ma Z (2014). Classifcation and expression diversifcation of wheat dehydrin genes. Plant Sci 214: 113120.
Xiao H, Nassuth A (2006). Stress- and development-induced expression of spliced and unspliced transcripts from two highly similar dehydrin 1 genes in V. riparia and V. vinifera. Plant Cell Rep 25: 968977.
Yang Y, He M, Zhu Z, Li S, Xu Y, Zhang C, Singer SD, Wang Y (2012). Identifcation of the dehydrin gene family from grapevine species and analysis of their responsiveness to various forms of abiotic and biotic stress. BMC Plant Biol 12: 140.
Zhang HM, Zhang LS, Liu L, Zhu WN, Yang WB (2013). Changes of dehydrin profles induced by drought in winter wheat at diferent developmental stages. Biol Plantarum 57: 797800.