Plant Growth Regulation

, Volume 78, Issue 2, pp 195–204 | Cite as

Response of Ricinus communis L. to in vitro water stress induced by polyethylene glycol

  • Marina Medeiros de Araújo Silva
  • Lilia Willadino
  • Déborah Yara A. Cursino dos Santos
  • Antonio Fernando M. Oliveira
  • Terezinha Rangel Camara
Original paper


Two castor bean cultivars were subjected to water stress induced by polyethylene glycol 6000 (PEG) under in vitro conditions. After 15 days, a reduced percentage of surviving plants, leaf abscission and greater leaf cuticular wax content were observed, accompanied by reduced water loss through the leaves. PEG also caused a decrease in soluble protein content and increases in the levels of hydrogen peroxide (H2O2) and malondialdehyde, as well as increases in the specific activities of superoxide dismutase, ascorbate peroxidase and catalase enzymes. These findings suggest that the mechanisms of tolerance of these genotypes are associated with their ability to activate key enzymes of the antioxidant defense system. However, the adaptive responses induced in response to stress were not sufficient to improve plant performance during the acclimatization period, and their survival rate dropped nearly 50 % relative to control treatment.


Acclimatization Antioxidant enzymes Castor bean Cuticular wax Oxidative stress 



The authors wish to thank the Embrapa Algodão for providing the castor seeds; the Núcleo de Plataforma Tecnológica—NPT/CPqAM/FIOCRUZ and Laboratório de Micros-copia—LIKA/UFPE, for SEM analysis. As well, the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for doctoral fellowship granted to the first author.

Conflict of interest

The authors declare no conflicts of interest.


  1. Ahmad MSA, Javed F, Ashraf M (2007) Iso-osmotic effect of NaCl and PEG on growth, cations and free proline accumulation in callus tissue of two indica rice (Oryza sativa L.) genotypes. Plant Growth Regul 53:53–63CrossRefGoogle Scholar
  2. Alexieva V, Sergiev I, Mapelli S, Karanov E (2001) The effect of drought and ultraviolet radiation on growth and stress markers in pea and wheat. Plant Cell Environ 24:1337–1344CrossRefGoogle Scholar
  3. Ali-Ahmad M, Hughes HG, Safadi F (1998) Studies on stomatal function, epicuticular wax and stem-root transition region of polyethylene glycol-treated and nontreated in vitro grape plantlets. In vitro Cell Dev Biol Plant 34:1–7CrossRefGoogle Scholar
  4. Andrade FP, Freire EC, Lima EF, Silva GA, Silva LC, Dourado RMF (2010) BRS Nordestina. Embrapa Algodão, Campina Grande, pp 1–6Google Scholar
  5. Azevedo RA, Alas RM, Smith RJ, Lea PJ (1998) Response of antioxidant enzymes to transfer from elevated carbon dioxide to air and ozone fumigation, in leaves and roots of wild-type and catalase-deficient mutant of barley. Physiol Plant 104:280–292CrossRefGoogle Scholar
  6. Baťková P, Pospíšilová J, Synková H (2008) Production of reactive oxygen species and development of antioxidative systems during in vitro growth and ex vitro transfer. Biol Plant 52:413–422CrossRefGoogle Scholar
  7. Bergmann JC, Tupinambá DD, Costa OYA, Almeida JRM, Barreto CC, Quirino BF (2013) Biodiesel production in Brazil and alternative biomass feedstocks. Renew Sust Energ Rev 21:411–420CrossRefGoogle Scholar
  8. Berrs LSJ, Sizer IW (1952) A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 195:133–140Google Scholar
  9. Bidabadi SS, Mahmood M, Baninasab B, Ghobadi C (2012) Influence of salicylic acid on morphological and physiological responses of banana (Musa acuminata cv. ‘Berangan’, AAA) shoot tips to in vitro water stress induced by polyethylene glycol. Plant Omics J 5:33–39Google Scholar
  10. Bradford MM (1976) A rapid and sensitive method for the quantization of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  11. Bruce TJA, Matthes MC, Napier JA, Pickett JA (2007) Stressful “memories” of plants: evidence and possible mechanisms. Plant Sci 173:603–608CrossRefGoogle Scholar
  12. Chandra S, Bandopadhyay R, Kumar V, Chandra R (2010) Acclimatization of tissue cultured plantlets: from laboratory to land. Biotechnol Lett 32:1199–1205CrossRefPubMedGoogle Scholar
  13. Dami I, Hughes H (1995) Leaf anatomy and water loss of in vitro PEG-treated ‘Valiant’ grape. Plant Cell Tiss Org Cult 42:179–184CrossRefGoogle Scholar
  14. Dami I, Hughes HG (1997) Effects of PEG-induced water stress on in vitro hardening ‘Valiant’grape. Plant Cell Tiss Org Cult 47:101CrossRefGoogle Scholar
  15. Demidchik V (2015) Machanisms of oxaditive stress in plants: from classical chemistry to cell biology. Environ Exp Bot 109:2012–2228CrossRefGoogle Scholar
  16. Farmer EE, Mueller JM (2013) ROS-mediated lipid peroxidation and RES-activated signaling. Annu Rev Plant Biol 64:429–450CrossRefPubMedGoogle Scholar
  17. George S, Jatoi SA, Siddiqui SU (2013) Genotypic differences against PEG simulated drought stress in tomato. Pak J Bot 45:1551–1556Google Scholar
  18. Giannopolitis CN, Ries SK (1977) Superoxide dismutases: I. Occurrence in higher plants. Plant Physiol 59:309–314PubMedCentralCrossRefPubMedGoogle Scholar
  19. Grzesiak M, Filek M, Barbasz A, Kreczmer B, Hartikainen H (2013) Relationships between polyamines, ethylene, osmoprotectants and antioxidant enzymes activities in wheat seedlings after short-term PEG- and NaCl-induced stress. Plant Growth Regul 69:177–189CrossRefGoogle Scholar
  20. Hassankhah A, Vahdati K, Lotfi M, Mirmasoumi M, Preece J, Assareh M-H (2014) Effects of ventilation and sucrose concentrations on the growth and plantlet anatomy of micropropagated persian walnut plants. Int J Hortic Sci Technol 1:111–120Google Scholar
  21. Hazarika BN (2006) Morpho-physiological disorders in in vitro culture of plants. Sci Hortic 108:105–120CrossRefGoogle Scholar
  22. Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplast. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198CrossRefPubMedGoogle Scholar
  23. Jaleel CA, Riadh K, Gopi R, Manivannan P, Inès J, Al-Juburi HJ, Chang-Xing Z, Hong-Bo S, Panneerselvam R (2009) Antioxidant defense responses: physiological plasticity in plants under abiotic constraints. Acta Physiol Plant 31:427–436CrossRefGoogle Scholar
  24. Kabiri R, Farahbakhsh H, Nasibi F (2012) Salicylic acid ameliorates the effects of oxidative stress induced by water deficit in hydroponic culture of Nigella sativa. J Stress Physiol Biochem 8:13–22Google Scholar
  25. Karimi S, Abbaspour H, Sinaki JM, Makarian H (2012) Effects of water deficit and chitosan spraying on osmotic adjustment and soluble protein of cultivars castor bean (Ricinus communis L.). J Stress Physiol Biochem 8:160–169Google Scholar
  26. Kumar K, Rao IU (2012) Morphophysiologicals problems in acclimatization of micropropagated plants in ex vitro conditions—a review. J Ornam Hort Plants 2:271–283Google Scholar
  27. Macková J, Vašková M, Macek P, Hronková M, Schreiber L, Šantrůček J (2013) Plant response to drought stress simulated by ABA application: changes in chemical composition of cuticular waxes. Environ Exp Bot 86:70–75CrossRefGoogle Scholar
  28. Magdalita PM, Damasco OP, Adkins SW (2010) Effects of medium replenishment and acclimatization techniques on growth and survival of embryo cultured coconut seedlings. Philipp Sci Lett 3:1–9Google Scholar
  29. Majada JP, Sierra MI, Sánchez-Tamés R (2001) Air exchange rate affects the in vitro developed leaf cuticle of carnation. Sci Hortic 87:121–130CrossRefGoogle Scholar
  30. Melo MM, Barbosa MR, Dias ALF, Willadino L, Camara TR (2014) Pré-condicionamento in vitro de plantas de cana-de-açúcar (Saccharum spp.) para tolerância ao estresse salino. Rev Bras Eng Agríc Ambient 18:27–33Google Scholar
  31. Milani M, Nóbrega MBM, Gondim TMS, Andrade FPA, Suassuna ND, Coutinho WM (2007) BRS Energia. Embrapa Algodão, Campina Grande, pp 1–6Google Scholar
  32. Møller IM, Jensen PE, Hansson A (2007) Oxidative modifications to cellular components in plants. Annu Rev Plant Biol 58:459–481CrossRefPubMedGoogle Scholar
  33. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497CrossRefGoogle Scholar
  34. Nakano Y, Assada K (1981) Hydrogen peroxide is scavenged by ascorbate-especific peroxidase en spinach chloroplasts. Plant Cell Physiol 22:867–880Google Scholar
  35. Osório ML, Gonçalves S, Coelho N, Osorio J, Romano A (2013) Morphological, physiological and oxidative stress markers during acclimatization and field transfers of micropropagated Tuberaria major plants. Plant Cell Tiss Org Cult 115:85–97CrossRefGoogle Scholar
  36. Pospíšilová J, Synková H, Haisel D, Semorádová Š (2007) Acclimation of plantlets to ex vitro conditions: effects of air humidity, irradiance, CO2 concentration and abscisic acid (a review). Acta Hortic 748:29–38CrossRefGoogle Scholar
  37. Rai MK, Kalia RK, Singh R, Gangola M, Dhawan AK (2011) Developing stress tolerant plants through in vitro selection—an overview of the recent progress. Environ Exp Bot 71:89–98CrossRefGoogle Scholar
  38. Rodrigues RFO, Oliveira F, Fonseca AM (2002) As folhas de Palma Christi—Ricinus communis L. Euphorbiaceae Jussieu. Lecta 20:183–194Google Scholar
  39. Rodziewicz P, Swarcewicz B, Chmielewska K, Wojakowska A, Stobiecki M (2014) Influence of abiotic stresses on plant proteome and metabolome changes. Acta Physiol Plant 36:1–19CrossRefGoogle Scholar
  40. Saéz PL, Bravo LA, Latsague MI, Sánchez ME, Ríos DG (2012) Increased light intensity during in vitro culture improves water loss control and photosynthetic performance of Castanea sativa grown in ventilated vessels. Sci Hortic 138:7–16CrossRefGoogle Scholar
  41. Sen A, Alikamanoglu S (2013) Antioxidant enzyme activities, malondialdehyde, and total phenolic content of PEG-induced hyperhydric leaves in sugar beet tissue culture. In Vitro Cell Dev Biol Plant 49:396–404CrossRefGoogle Scholar
  42. Shaheen AM (2002) Morphological variation within Ricinus communis L. in Egypt: fruit, leaf, seed and pollen. Pak J Biol Sci 5:1202–1206CrossRefGoogle Scholar
  43. Sharma P, Dubey RS (2005) Drought induces oxidative stress and enhances the activities of antioxidant enzymes in growing rice seedlings. Plant Growth Regul 46:209–221CrossRefGoogle Scholar
  44. Shepherd T, Griffiths W (2006) The effects of stress on plant cuticular waxes. New Phytol 171:469–499CrossRefPubMedGoogle Scholar
  45. Soni P, Rizwan M, Bhatt KV, Mohapatra T, Singh G (2011) In-vitro response of Vigna aconitifolia to drought stress induced by PEG-6000. J Stress Physiol Biochem 7:108–121Google Scholar
  46. Tsago Y, Andargie M, Takele A (2013) In vitro screening for drought tolerance in different sorghum (Sorghum bicolor (L.) Moench) varieties. J Stress Physiol Biochem 9:72–83Google Scholar
  47. Vermeer CP, Nastold P, Jetter R (2003) Homologous very-long-chain 1,3-alkanediols and 3-hydroxyaldehydes in leaf cuticular waxes of Ricinus communis L. Phytochemistry 62:433–438CrossRefPubMedGoogle Scholar
  48. Yang SH, Yeh DM (2008) In vitro leaf anatomy, ex vitro photosynthetic behaviors and growth of Calathea orbifolia (Linden) Kennedy plants obtained from semi-solid medium and temporary immersion system. Plant Cell Tiss Organ Cult 93:201–207CrossRefGoogle Scholar
  49. Yeats TH, Rose JKC (2013) The formation and function of plant cuticles. Plant Physiol 163:5–20PubMedCentralCrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Marina Medeiros de Araújo Silva
    • 1
  • Lilia Willadino
    • 2
  • Déborah Yara A. Cursino dos Santos
    • 3
  • Antonio Fernando M. Oliveira
    • 4
  • Terezinha Rangel Camara
    • 5
  1. 1.National Institute for the Semi-AridBrazilian Ministry of Science, Technology and InnovationCampina GrandeBrazil
  2. 2.Department of BiologyFederal Rural University of PernambucoRecifeBrazil
  3. 3.Department of BotanyUniversity of São PauloSão PauloBrazil
  4. 4.Department of BotanyFederal University of PernambucoRecifeBrazil
  5. 5.Department of ChemistryFederal Rural University of PernambucoRecifeBrazil

Personalised recommendations