Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Functional characterization of an acidic SK3 dehydrin isolated from an Opuntia streptacantha cDNA library

Abstract

Cactus pears are succulent plants of the Cactaceae family adapted to extremely arid, hot and cold environments, making them excellent models for the study of molecular mechanisms underlying abiotic stress tolerance. Herein, we report a directional cDNA library from 12-month-old cladodes of Opuntia streptacantha plants subjected to abiotic stresses. A total of 442 clones were sequenced, representing 329 cactus pear unigenes, classified into eleven functional categories. The most abundant EST (unigen 33) was characterized under abiotic stress. This cDNA of 905 bp encodes a SK3-type acidic dehydrin of 248 amino acids. The OpsDHN1 gene contains an intron inserted within the sequence encoding the S-motif. qRT-PCR analysis shows that the OpsDHN1 transcript is specifically accumulated in response to cold stress, and induced by abscisic acid. Over-expression of the OpsDHN1 gene in Arabidopsis thaliana leads to enhanced tolerance to freezing treatment, suggesting that OpsDHN1 participates in freezing stress responsiveness. Generation of the first EST collection for the characterization of cactus pear genes constitutes a useful platform for the understanding of molecular mechanisms of stress tolerance in Opuntia and other CAM plants.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Abbreviations

ABA:

Abscisic acid

CAM:

Crassulacean acid metabolism

ORF:

Open reading frame

OpsDHN1 :

Opuntia streptacantha dehydrin 1

qRT-PCR:

Quantitative reverse transcriptase-polymerase chain reaction

References

  1. Allagulova ChR, Gimalov FR, Shakirova FM, Vakhitov VA (2003) The plant dehydrins: structure and putative functions. Biochemistry (Mosc) 68:945–951

  2. Alsheikh MK, Heyen BJ, Randall SK (2003) Ion binding properties of the dehydrin ERD14 are dependent upon phosphorylation. J Biol Chem 278:40882–40889

  3. Bae EK, Lee H, Lee JS, Noh EW (2009) Differential expression of a poplar SK2-type dehydrin gene in response to various stresses. BMB Rep 42:439–443

  4. Bassett CL, Wisniewski ME, Artlip TS, Richart G, Norelli JL, Farrell RE Jr (2009) Comparative expression and transcript initiation of three peach dehydrin genes. Planta 230:107–118

  5. Battaglia M, Olvera-Carrillo Y, Garciarrubio A, Campos F, Covarrubias AA (2008) The enigmatic LEA proteins and other hydrophilins. Plant Physiol 148:6–24

  6. Bies-Etheve N, Gaubier-Comella P, Debures A, Lasserre E, Jobet E, Raynal M, Cooke R, Delseny M (2008) Inventory, evolution and expression profiling diversity of the LEA (late embryogenesis abundant) protein gene family in Arabidopsis thaliana. Plant Mol Biol 67:107–124

  7. Black C, Osmond C (2003) Crassulacean acid metabolism photosynthesis: ‘working the night shift’. Photosynth Res 76:329–341

  8. Bokor M, Csizmok V, Kovacs D, Banki P, Friedrich P, Tompa P, Tompa K (2005) NMR relaxation studies on the hydrate layer of intrinsically unstructured proteins. Biophys J 88:2030–2037

  9. Borovskii GB, Stupnikova IV, Antipina AI, Vladimirova SV, Voinikov VK (2002) Accumulation of dehydrin-like proteins in the mitochondria of cereals in response to cold, freezing, drought and ABA treatment. BMC Plant Biol 2:5

  10. Brini F, Hanin M, Lumbreras V, Amara I, Khoudi H, Hassairi A, Pages M, Masmoudi K (2007) Overexpression of wheat dehydrin DHN-5 enhances tolerance to salt and osmotic stress in Arabidopsis thaliana. Plant Cell Rep 26:2017–2026

  11. Busk PK, Jensen AB, Pages M (1997) Regulatory elements in vivo in the promoter of the abscisic acid responsive gene rab17 from maize. Plant J 11:1285–1295

  12. Campbell SA, Close TJ (1997) Dehydrins: genes, proteins, and associations with phenotypic traits. New Phytol 137:61–74

  13. Chung E, Kim SY, Yi SY, Choi D (2003) Capsicum annuum dehydrin, an osmotic-stress gene in hot pepper plants. Mol Cells 15:327–332

  14. Close TJ (1996) Dehydrins: emergence of a biochemical role of a family of plant dehydration proteins. Physiol Plant 97:795–803

  15. Close TJ (1997) Dehydrins: a commonalty in the response of plants to dehydration and low temperature. Physiol Plant 100:291–296

  16. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

  17. Cushman JC (2001) Crassulacean acid metabolism. A plastic photosynthetic adaptation to arid environments. Plant Physiol 127:1439–1448

  18. Dure L 3rd (1993) A repeating 11-mer amino acid motif and plant desiccation. Plant J 3:363–369

  19. Fan Z, Wang X (2006) Isolation and characterization of a novel dehydrin gene from Capsella bursa-pastoris. Mol Biol (Mosk) 40:52–60

  20. Felsenstein J (1989) PHYLIP—Phylogeny Inference Package (Version 3.2). Cladistics 5:164–166

  21. Garay-Arroyo A, Colmenero-Flores JM, Garciarrubio A, Covarrubias AA (2000) Highly hydrophilic proteins in prokaryotes and eukaryotes are common during conditions of water deficit. J Biol Chem 275:5668–5674

  22. Griffith MP (2004) The origins of an important cactus crop, Opuntia ficus-indica (Cactaceae): new molecular evidence. Am J Bot 91:1915–1921

  23. Hara M, Terashima S, Kuboi T (2001) Characterization and cryoprotective activity of cold-responsive dehydrin from Citrus unshiu. J Plant Physiol 158:1333–1339

  24. Hara M, Terashima S, Fukaya T, Kuboi T (2003) Enhancement of cold tolerance and inhibition of lipid peroxidation by citrus dehydrin in transgenic tobacco. Planta 217:290–298

  25. Hara M, Fujinaga M, Kuboi T (2005) Metal binding by citrus dehydrin with histidine-rich domains. J Exp Bot 56:2695–2703

  26. Hara M, Shinoda Y, Tanaka Y, Kuboi T (2009) DNA binding of citrus dehydrin promoted by zinc ion. Plant Cell Environ 32:532–541

  27. Heyen BJ, Alsheikh MK, Smith EA, Torvik CF, Seals DF, Randall SK (2002) The calcium-binding activity of a vacuole-associated, dehydrin-like protein is regulated by phosphorylation. Plant Physiol 130:675–687

  28. Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil. California Agricultural Experiment Station Circular 347, Berkeley

  29. Hundertmark M, Hincha DK (2008) LEA (late embryogenesis abundant) proteins and their encoding genes in Arabidopsis thaliana. BMC Genomics 9:118

  30. Ingram J, Bartels D (1996) The Molecular basis of dehydration tolerance in plants. Annu Rev Plant Physiol Plant Mol Biol 47:377–403

  31. Koag MC, Fenton RD, Wilkens S, Close TJ (2003) The binding of maize DHN1 to lipid vesicles Gain of structure and lipid specificity. Plant Physiol 131:309–316

  32. Kosová K, Prášil I, Vítámvás P (2010) Role of dehydrins in plant stress response. In: Pessarakli M (ed) Handbook of plant and crop stress, 3rd edn. Books in soils, plants, and the environment. CRC Press, New York, pp 239–285

  33. Kovacs D, Kalmar E, Torok Z, Tompa P (2008) Chaperone activity of ERD10 and ERD14, two disordered stress-related plant proteins. Plant Physiol 147:381–390

  34. Lee SC, Lee MY, Kim SJ, Jun SH, An G, Kim SR (2005) Characterization of an abiotic stress-inducible dehydrin gene, OsDhn1 in rice (Oryza sativa L.). Mol Cells 19:212–218

  35. Lüttge U (2004) Ecophysiology of crassulacean acid metabolism (CAM). Ann Bot 93:629–652

  36. Mouillon JM, Gustafsson P, Harryson P (2006) Structural investigation of disordered stress proteins: comparison of full-length dehydrins with isolated peptides of their conserved segments. Plant Physiol 141:638–650

  37. Mueller JK, Heckathorn SA, Fernando D (2003) Identification of a chloroplast dehydrin in leaves of mature plants. Int J Plant Sci 164:S35–S42

  38. Nobel PS (1997) Recent ecophysiological findings for Opuntia ficus-indica. JPACD 2:89–96

  39. Nobel PS, Bobich EG (2002) Enviromental Biology. In: Nobel PS (ed) Cacti: biology and uses. University of California Press, California, pp 57–74

  40. Nylander M, Svensson J, Palva ET, Welin BV (2001) Stress-induced accumulation and tissue-specific localization of dehydrins in Arabidopsis thaliana. Plant Mol Biol 45:263–279

  41. Ohlrogge J, Benning C (2000) Unraveling plant metabolism by EST analysis. Curr Opin Plant Biol 3:224–228

  42. Peng Y, Reyes JL, Wei H, Yang Y, Karlson D, Covarrubias AA, Krebs SL, Fessehaie A, Arora R (2008) RcDhn5, a cold acclimation-responsive dehydrin from Rhododendron catawbiense rescues enzyme activity from dehydration effects in vitro and enhances freezing tolerance in RcDhn5-overexpressing Arabidopsis plants. Physiol Plant 134:583–597

  43. Pimienta-Barrios E (1994) Prickly pear (Opuntia spp.): a valuable fruit crop for the semi-arid lands of Mexico. J Arid Environ 28:1–11

  44. Puhakainen T, Hess MW, Makela P, Svensson J, Heino P, Palva ET (2004) Overexpression of multiple dehydrin genes enhances tolerance to freezing stress in Arabidopsis. Plant Mol Biol 54:743–753

  45. Rabas AR, Martin CE (2003) Movement of water from old to young leaves in three species of succulents. Ann Bot 92:529–536

  46. Reyes JL, Campos F, Wei H, Arora R, Yang Y, Karlson D, Covarrubias AA (2008) Functional dissection of hydrophilins during in vitro freeze protection. Plant Cell Environ 12:1781–1790

  47. Rodriguez-Kessler M, Ruiz OA, Maiale S, Ruiz-Herrera J, Jimenez-Bremont JF (2008) Polyamine metabolism in maize tumors induced by Ustilago maydis. Plant Physiol Biochem 46:805–814

  48. Rorat T (2006) Plant dehydrins—tissue location, structure and function. Cell Mol Biol Lett 11:536–556

  49. Rorat T, Szabala BM, Grygorowicz WJ, Wojtowicz B, Yin Z, Rey P (2006) Expression of SK3-type dehydrin in transporting organs is associated with cold acclimation in Solanum species. Planta 224:205–221

  50. Saavedra L, Svensson J, Carballo V, Izmendi D, Welin B, Vidal S (2006) A dehydrin gene in Physcomitrella patens is required for salt and osmotic stress tolerance. Plant J 45:237–249

  51. Sahu BB, Shaw BP (2009) Isolation, identification and expression analysis of salt-induced genes in Suaeda maritima, a natural halophyte, using PCR-based suppression subtractive hybridization. BMC Plant Biol 9:69

  52. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, vol 2, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

  53. Shakirova F, Allagulova C, Bezrukova M, Aval’baev A, Gimalov F (2009) The role of endogenous ABA in cold-induced expression of the TADHN dehydrin gene in wheat seedlings. Russ J Plant Physiol 56:720–723

  54. Shen Y, Tang M-J, Hu Y-L, Lin Z-P (2004) Isolation and characterization of a dehydrin-like gene from drought-tolerant Boea crassifolia. Plant Sci 166:1167–1175

  55. Silva-Ortega CO, Ochoa-Alfaro AE, Reyes-Aguero JA, Aguado-Santacruz GA, Jimenez-Bremont JF (2008) Salt stress increases the expression of p5cs gene and induces proline accumulation in cactus pear. Plant Physiol Biochem 46:82–92

  56. Sun X, Lin HH (2010) Role of plant dehydrins in antioxidation mechanisms. Biologia 5:755–759

  57. Thomashow MF (1999) Plant cold acclimation: freezing tolerance genes and regulatory mechanisms. Annu Rev Plant Physiol Plant Mol Biol 50:571–599

  58. Tompa P, Kovacs D (2010) Intrinsically disordered chaperones in plants and animals. Biochem Cell Biol 88:167–174

  59. Tunnacliffe A, Wise MJ (2007) The continuing conundrum of the LEA proteins. Naturwissenschaften 94:791–812

  60. Wang WX, Vinocur B, Shoseyov O, Altman A (2001) Biotechnology of plant osmotic stress tolerance: physiological and molecular considerations. Acta Hort 560:285–292

  61. Weiss J, Egea-Cortines M (2009) Transcriptomic analysis of cold response in tomato fruits identifies dehydrin as a marker of cold stress. J Appl Genet 50:311–319

  62. Xu J, Zhang Y, Guan Z, Wei W, Han L, Chai T (2008a) Expression and function of two dehydrins under environmental stresses in Brassica juncea L. Mol Breed 21:431–438

  63. Xu J, Zhang YX, Wei W, Han L, Guan ZQ, Wang Z, Chai TY (2008b) BjDHNs confer heavy-metal tolerance in plants. Mol Biotechnol 38:91–98

  64. Yin Z, Rorat T, Szabala BM, Ziólkowska A, Malepszy S (2006) Expression of a Solanum sogarandinum SK3-type dehydrin enhances cold tolerance in transgenic cucumber seedlings. Plant Sci 170:1164–1172

  65. Zhang Y, Mian MAR, Chekhovskiy K, So S, Kupfer D, Lai H, Roe BA (2005) Differential gene expression in Festuca under heat stress conditions. J Exp Bot 56:897–907

  66. Zhang Y, Li J, Yu F, Cong L, Wang L, Burkard G, Chai T (2006) Cloning and expression analysis of SKn-type dehydrin gene from bean in response to heavy metals. Mol Biotechnol 32:205–218

Download references

Acknowledgments

This work was supported by SAGARPA (2004-C01-216) and the CONACYT (Investigación Ciencia Básica 2008-103106) fundings. We are grateful to Dr. Paul Riley for a grammatical review.

Author information

Correspondence to J. F. Jiménez-Bremont.

Electronic supplementary material

Below is the link to the electronic supplementary material.

http://bioinf.cs.ucl.ac.uk/disopred/
http://www.cbs.dtu.dk/services/NetPGene/

Suppl. Fig. S3 Twelve-month-old O. streptacantha plants were grown either in semi-hydroponic conditions or in a commercial soil mixture, and subjected to different abiotic stresses during 17 d: control, salinity (N), heat-salinity (HN) (semi-hydroponic); control, cold (C), heat-cold (HC), heat-drought (HD) and drought-heat-cold (DHC) (soil mixture)

Suppl. Fig. S4 a Phenotype of the 35S::OpsDHN1-1 transgenic line and the wild type (Col-0) after 21 d of recovery from salt stress treatment (150 mM NaCl). b Survival rate of the 35S::OpsDHN1-1 transgenic line and Col-0 plants after 21 d of NaCl stress recovery. Data are mean ± SE (n = 10) from 5 replicates. The experiments were conducted twice obtaining similar results

Supplementary material 1 (DOC 615 kb)

Supplementary material 2 (DOC 51 kb)

Supplementary material 3 (DOC 64 kb)

Supplementary material 4 (DOC 34 kb)

Supplementary material 5 (DOC 370 kb)

Supplementary material 6 (DOC 215 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Ochoa-Alfaro, A.E., Rodríguez-Kessler, M., Pérez-Morales, M.B. et al. Functional characterization of an acidic SK3 dehydrin isolated from an Opuntia streptacantha cDNA library. Planta 235, 565–578 (2012). https://doi.org/10.1007/s00425-011-1531-8

Download citation

Keywords

  • Abiotic stress
  • Cactus pear
  • cDNA library
  • Cold stress
  • Dehydrin