Rosmarinic acid ameliorates the negative effects of salinity in in vitro-regenerated potato explants (Solanum tuberosum L.)

  • Hoda Eskandari
  • Naemah Al-Mansour
  • Ali Akbar Ehsanpour
Original Article


Salinity causes massive loss of economic crops due to the build-up of toxic salts in shoot. Rosmarinic acid (RA) has been known to prevent the inevitable damage to vital biological molecules under stress. There is an increasing interest, therefore, in the use of polyphenols in crop stress physiology studies. An in vitro regeneration protocol was established using explants of the potato cultivar White Desiree to assess tolerance to salinity at a morphological, cellular, and biochemical level. Additionally, we examined the potential ameliorative effects of RA on growth in salt-stressed potato explants. Based on our observations, we propose a model for the ameliorative effects of a caffeic acid conjugate of α-hydroxyhydrocaffeic acid. Explants of White Desiree showed differential responses to salt and RA supplementation. Salt-stressed and control explants grown in RA-supplemented media showed a pronounced expansion of leaf area, suggesting direct effects of RA on cell elongation in potato explants. Root elongation was less affected by salinity in comparison to shoot elongation. Moderate levels of RA resulted in stimulatory effects on growth. The microscopic examination of leaves and stems indicated the presence of unique trichomes at the early stages of explant growth under RA supplementation, which varied considerably in size, shape, and distribution in response to stress. Our data indicated that RA significantly decreased osmolyte content under stress. Salinity resulted in the biosynthesis of high levels of free proline, carbohydrates, and malondialdehyde (MDA). However, the lower contents of MDA in salt-stressed explants treated with RA indicate that RA possesses anti-lipid peroxidation properties. Our observations provide evidence for the stimulatory effects of RA on cellular growth and the protection of membranes from ion toxicity.


Malondialdehyde Oxidative stress Potato explant Rosmarinic acid Salinity 



Authors of this study would like to thank University of Isfahan for providing facilities for in vitro potato generation. We express our sincere thanks to Miss Ahlam Al-Kadi from the NanoScope Center at Kuwait University for her help in sample preparation, and Mr. Mohamed Rajab for SEM microscopy.


  1. Abeles FB, Morgan PW, Saltveit ME Jr (2012) Ethylene in plant biology. Academic Press, LondonGoogle Scholar
  2. Adedeji O, Ajuwon O, Babawale O (2007) Foliar epidermal studies, organographic distribution and taxonomic importance of trichomes in the family Solanaceae. Int J Botany 3:276–282CrossRefGoogle Scholar
  3. Ahmad HH, Hamza AH, Hassan AZ, Sayed AH (2013) Promising therapeutic role of Rosmarinus officinalis successive methanolic fraction against colorectal cancer. Int J Pharm Pharm Sci 5:164–170Google Scholar
  4. Anjum SA, Farooq M, Xie X, Liu X, Ijaz MF (2012) Antioxidant defense system and proline accumulation enables hot pepper to perform better under drought. Sci Hort 140:66–73CrossRefGoogle Scholar
  5. Ashraf M (2002) Salt tolerance of cotton: some new advances. CRC Crit Rev Plant Sci 21:1–30CrossRefGoogle Scholar
  6. Bai J-P, Gao H-J, Yang H-Y, Lou Y, Zhang J-L, Wang D, Zhang J-L (2016) Comparison of ultrastructural and physiological changes of potato (Solanum tuberosum L.) plantlets subjected to salt and modeling drought stresses. Acta Physiol Plant 38:1–9CrossRefGoogle Scholar
  7. Bates L, Waldren R, Teare I (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39:205–207CrossRefGoogle Scholar
  8. Bayuelo-Jimenez JS, Craig R, Lynch JP (2002) Salinity tolerance of species during germination and early seedling growth. Crop Sci 42:1584–1594CrossRefGoogle Scholar
  9. Brown C (2005) Antioxidants in potato. Am J Potato Res 82:163–172CrossRefGoogle Scholar
  10. Cai-Hong P, Su-Jun Z, Zhi-Zhong G, Bao-Shan W (2005) NaCl treatment markedly enhances H2O2-scavenging system in leaves of halophyte Suaeda salsa. Physiol Plant 125:490–499CrossRefGoogle Scholar
  11. Costa P, Gonçalves S, Andrade PB, Valentão P, Romano A (2011) Inhibitory effect of Lavandula viridis on Fe2+-induced lipid peroxidation, antioxidant and anti-cholinesterase properties. Food Chem 126:1779–1786CrossRefPubMedGoogle Scholar
  12. Ehsanpour A, Jones M (2000) Evaluation of direct shoot regeneration from stem explants of potato (Solanum tuberosum L.) cv. Delaware by thidiazuron (TDZ). J Sci Technol Agric Natural Resour 4:47–54Google Scholar
  13. Elaleem KGA, Modawi RS, Khalafalla MM (2009) Effect of plant growth regulators on callus induction and plant regeneration in tuber segment culture of potato (Solanum tuberosum L.) cultivar Diamant. Afr J Biotechnol 8:2529–2534Google Scholar
  14. FAO (2015) World food and agriculture 2015. FAO, RomeGoogle Scholar
  15. FAO (2016) The state of food and agriculture. Climate change, agriculture and food security. Food and Agriculture Organization of the United Nations, Rome (ISBN 978-92-5-109374-0)Google Scholar
  16. Fernández-Segura E, Canizares FJ, Cubero MA, Warley A, Campos A (1999) Changes in elemental content during apoptotic cell death studied by electron probe X-ray microanalysis. Exp Cell Res 253:454–462CrossRefPubMedGoogle Scholar
  17. Fry W (2008) Phytophthora infestans: the plant (and R gene) destroyer. Mol Plant Pathol 9:385–402CrossRefPubMedGoogle Scholar
  18. Fryer MJ, Oxborough K, Mullineaux PM, Baker NR (2002) Imaging of photo-oxidative stress responses in leaves. J Exp Bot 53:1249–1254PubMedGoogle Scholar
  19. Fu Q, Yang R, Wang H, Zhao B, Zhou C, Ren S, Guo Y-D (2013) Leaf morphological and ultrastructural performance of eggplant (Solanum melongena L.) in response to water stress. Photosynthetica 51:109–114CrossRefGoogle Scholar
  20. Furtado RA, Oliveira BR, Silva LR, Cleto SS, Munari CC, Cunha WR, Tavares DC (2015) Chemopreventive effects of rosmarinic acid on rat colon carcinogenesis. Eur J Cancer Prev 24:106–112CrossRefPubMedGoogle Scholar
  21. Gelmesa D, Dechassa N, Mohammed W, Gebre E, Monneveux P, Bündig C, Winkelmann T (2017) In vitro screening of potato genotypes for osmotic stress tolerance. Open Agric 2:308–316Google Scholar
  22. González-Vallinas M, Reglero G, Ramirez de Molina A (2015) Rosemary (Rosmarinus officinalis L.) extract as a potential complementary agent in anticancer therapy. Nutr Cancer 67:1223–1231CrossRefGoogle Scholar
  23. Gustavsson J, Cederberg C, Sonesson U, Emanuelsson A (2013) The methodology of the FAO study: “Global Food Losses and Food Waste-extent, causes and prevention”-FAO, 2011. Gӧteborg, SwedenGoogle Scholar
  24. Guyer A, De Vrieze M, Bӧnisch D, Gloor R, Musa T, Bodenhausen N, Bailly A, Weisskopf L (2015) The anti-Phytophthora effect of selected potato-associated Pseudomonas strains: from the laboratory to the field. Front Microbiol 6(1309):1–13Google Scholar
  25. Hakkim F, Kalyani S, Essa M, Girija S, Song H (2011) Production of rosmarinic acid in Ocimum sanctum (L.) cell suspension cultures by the influence of growth regulators. Int J Biol Med Res 2:1158–1161Google Scholar
  26. Hansen J, Nielsen B, Nielsen SV (1999) In vitro shoot regeneration of Solanum tuberosum cultivars: interactions of medium composition and leaf, leaflet, and explant position. Potato Res 42:141–151CrossRefGoogle Scholar
  27. Hao W, Guo H, Zhang J, Hu G, Yao Y, Dong J (2014) Hydrogen peroxide is involved in salicylic acid-elicited rosmarinic acid production in Salvia miltiorrhiza cell cultures. Sci World J 2014:1–7CrossRefGoogle Scholar
  28. Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts: I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198CrossRefPubMedGoogle Scholar
  29. Hernández JA, Ferrer MA, Jiménez A, Barceló AR, Sevilla F (2001) Antioxidant System and O2/H2O2 production in the apoplast of pea leaves. Its relation with salt-induced necrotic lesions in minor veins. Plant Physiol 127:817–831CrossRefPubMedPubMedCentralGoogle Scholar
  30. Horton D, Sawyer RL (1985) The potato as a world food crop, with special reference to developing areas. In: Li PH (ed) Potato physiology. Academic Press, London, pp 1–34Google Scholar
  31. Huttunen P, Kärkkäinen K, Løe G, Rautio P, Ågren J (2010) Leaf trichome production and responses to defoliation and drought in Arabidopsis lyrata (Brassicaceae). Ann Bot Fenn 47:199–207CrossRefGoogle Scholar
  32. James J, Alder N, Mühling K, Läuchli A, Shackel K, Donovan L, Richards J (2006) High apoplastic solute concentrations in leaves alter water relations of the halophytic shrub, Sarcobatus vermiculatus. J Exp Bot 57:139–147CrossRefPubMedGoogle Scholar
  33. Jamil M, Lee CC, Rehman SU, Lee DB, Ashraf M, Rha ES (2005) Salinity (NaCl) tolerance of Brassica species at germination and early seedling growth. Electron J Environ Agric Food Chem 4:970–976Google Scholar
  34. Jin B-R, Chung K-S, Cheon S-Y, Lee M, Hwang S, Hwang SN, Rhee K-J, An H-J (2017) Rosmarinic acid suppresses colonic inflammation in dextran sulphate sodium (DSS)-induced mice via dual inhibition of NF-\kappa B and STAT3 activation. Sci Rep 7:1–11CrossRefGoogle Scholar
  35. Khatun N, Bari M, Islam R, Huda S, Siddque N, Rahman M, Mullah M (2003) Callus induction and regeneration from nodal segment of potato cultivar Diamant. J Biol Sci 3:1101–1106CrossRefGoogle Scholar
  36. Kim H-J, Seo E-Y, Kim J-H, Cheong H-J, Kang B-C, Choi D-I (2012) Morphological classification of trichomes associated with possible biotic stress resistance in the genus Capsicum. Plant Pathol J 28:107–113CrossRefGoogle Scholar
  37. Kissoudis C, van de Wiel C, Visser RG, van der Linden G (2014) Enhancing crop resilience to combined abiotic and biotic stress through the dissection of physiological and molecular crosstalk. Front Plant Sci 5:1–20CrossRefGoogle Scholar
  38. Koca H, Ozdemir F, Turkan I (2006) Effect of salt stress on lipid peroxidation and superoxide dismutase and peroxidase activities of Lycopersicon esculentum and L. pennellii. Biol Plant 50:745–748CrossRefGoogle Scholar
  39. Latijnhouwers M, Ligterink W, Vleeshouwers VG, van West P, Govers F (2004) A Gɑ subunit controls zoospore motility and virulence in the potato late blight pathogen Phytophthora infestans. Mol Microbiol 51:925–936CrossRefPubMedGoogle Scholar
  40. Levy D, Veilleux RE (2007) Adaptation of potato to high temperatures and salinity—a review. Am J Potato Res 84:487–506CrossRefGoogle Scholar
  41. Lianes A, Bertazza G, Palacio G, Luna V (2013) Different sodium salts cause different solute accumulation in the halophyte Prosopis strombulifera. Plant Biol 15:118–125CrossRefGoogle Scholar
  42. Lichtenthaler HK (1987) [34] Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol 148:350–382CrossRefGoogle Scholar
  43. Lichtenthaler HK, Wellburn AR (1983) Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochem Soc Trans 11:591–592CrossRefGoogle Scholar
  44. Margineanu A-M, Molnár I, Rakosy-Tican E (2014) Trichomes types analysis and their density in parental species Solanum tuberosum and S. chacoense and their derived somatic hybrids. Analele Stiintifice ale Universitatii “Al I Cuza” din Iasi 60:33Google Scholar
  45. Martinez CA, Maestri M, Lani EG (1996) In vitro salt tolerance and proline accumulation in Andean potato (Solanum spp.) differing in frost resistance. Plant Sci 116:177–184CrossRefGoogle Scholar
  46. Meloni DA, Oliva MA, Martinez CA, Cambraia J (2003) Photosynthesis and activity of superoxide dismutase, peroxidase and glutathione reductase in cotton under salt stress. Environ Exper Bot 49:69–76CrossRefGoogle Scholar
  47. Mohamed AA, Matter MA, Saker MM (2010) Effect of salt stress on some defense mechanisms of transgenic and wild potato clones (Solanum tuberosum L.) grown in vitro. Nature 12:8Google Scholar
  48. Moore J, Yousef M, Tsiani E (2016) Anticancer effects of rosemary (Rosmarinus officinalis L.) extract and rosemary extract polyphenols. Nutrients 8:731CrossRefPubMedCentralGoogle Scholar
  49. Munns R (2002) Comparative physiology of salt and water stress. Plant Cell Environ 25:239–250CrossRefPubMedGoogle Scholar
  50. Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol 59:651–681CrossRefPubMedGoogle Scholar
  51. Murashige T, Skoog F (1962) Manipulation of organ initiation in plant tissue cultures. Bot Bull Acad Sin 18:1–24Google Scholar
  52. Murshed R, Najla S, Albiski F, Kassem I, Jbour M, Al-Said H (2015) Using growth parameters for in vitro screening of potato varieties tolerant to salt stress. J Agric Sci Technol 17:483–494Google Scholar
  53. Neumann PM (1995) Inhabitation of root growth by salinity stress: toxicity or an adaptive biophysical response. In: Baluska F, Ciamporova M, Gasparikova O, Barlow PW (eds) Structure and function of roots. Kluwer Academic, The Netherlands, pp 299–304CrossRefGoogle Scholar
  54. Ozturk H, Ozturk H, Terzi EH, Ozgen U, Duran A, Uygun I (2014) Protective effects of rosmarinic acid against renal ischaemia/reperfusion injury in rats. J Pak Med Assoc 64:260–265PubMedGoogle Scholar
  55. Park WT, Arasu MV, Al-Dhabi NA, Yeo SK, Jeon J, Park JS, Lee SY, Park SU (2016) Yeast extract and silver nitrate induce the expression of phenylpropanoid biosynthetic genes and induce the accumulation of rosmarinic acid in Agastache rugosa cell culture. Molecules 21:426CrossRefPubMedGoogle Scholar
  56. Pearson D (1976) The chemical analysis of foods, 7th edn. Longman Group Ltd, Harlow, p xiiGoogle Scholar
  57. Rahman M, Islam R, Hossain M, Haider S (2008) Differential response of potato under sodium chloride stress conditions in vitro. J Biosci 16:79–83Google Scholar
  58. Reginato MA, Castagna A, Furlán A, Castro S, Ranieri A, Luna V (2014) Physiological responses of a halophytic shrub to salt stress by Na2SO4 and NaCl: oxidative damage and the role of polyphenols in antioxidant protection. AoB Plants 6:plu042CrossRefPubMedPubMedCentralGoogle Scholar
  59. Roomans GM (1988) Introduction to X-ray microanalysis in biology. J Electron Microsc Tech 9:3–17CrossRefPubMedGoogle Scholar
  60. Saeedipour S (2013) Relationship of grain yield, ABA and proline accumulation in tolerant and sensitive wheat cultivars as affected by water stress. Proc Natl Acad Sci India Sect B 83:311–315CrossRefGoogle Scholar
  61. Sahraroo A, Babalar M, Mirjalili MH, Moghaddam MRF, Ebrahimi SN (2014) In-vitro callus induction and rosmarinic acid quantification in callus culture of Satureja khuzistanica Jamzad (Lamiaceae). Iran J Pharm Res 13:1447PubMedPubMedCentralGoogle Scholar
  62. Sharmila R, Manoharan S (2012) Anti-tumor activity of rosmarinic acid in 7, 12-dimethylbenz (a) anthracene (DMBA) induced skin carcinogenesis in Swiss albino mice. Indian J Exp Biol 50:187PubMedGoogle Scholar
  63. Shirin F, Hossain M, Kabir M, Roy M, Sarker S (2007) Callus induction and plant regeneration from internodal and leaf explants of four potato (Solanum tuberosum L.) cultivars. World J Agric Sci 3:01–06Google Scholar
  64. Skepper JN, Karydis I, Garnett MR, Hegyi L, Hardwick SJ, Warley A, Mitchinson MJ, Cary NR (1999) Changes in elemental concentrations are associated with early stages of apoptosis in human monocyte-macrophages exposed to oxidized low-density lipoprotein: an X-ray microanalytical study. J Pathol 188:100–106CrossRefPubMedGoogle Scholar
  65. Tache A, Radu G-L, Litescu S-C (2012) Assessment of role of rosmarinic acid in preventing oxidative process of low density lipoproteins. Chem Pap 66:1166–1170CrossRefGoogle Scholar
  66. Thordal-Christensen H, Zhang Z, Wei Y, Collinge DB (1997) Subcellular localization of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barley—powdery mildew interaction. Plant J 11:1187–1194CrossRefGoogle Scholar
  67. Toth J, Mrlianova M, Tekelova D, Koremova M (2003) Rosmarinic acid an important phenolic active compound of Lemon Balm (Melissa officinalis). Acta Fac Pharm Univ Comen 50:139–146Google Scholar
  68. Tsedaley B (2014) Late blight of potato (Phytophthora infestans) biology, economic importance and its management approaches. J Biol Agric Healthc 25:215–226Google Scholar
  69. Vinocur B, Altman A (2005) Recent advances in engineering plant tolerance to abiotic stress: achievements and limitations. Curr Opin Biotechnol 16:123–132CrossRefPubMedGoogle Scholar
  70. Wang PJ, Hu CY (1985) Potato tissue culture and its application in agriculture. In: Li PH (ed) Potato physiology. Academic, London, pp 503–577Google Scholar
  71. Warley A (1997) X-ray microanalysis for biologists. Ashgate Publishing, FarnhamGoogle Scholar
  72. Werner JE, Finkelstein RR (1995) Arabidopsis mutants with reduced response to NaCl and osmotic stress. Physiol Plant 93:659–666CrossRefGoogle Scholar
  73. Xu W, Yang F, Zhang Y, Shen X (2016) Protective effects of rosmarinic acid against radiation-induced damage to the hematopoietic system in mice. J Rad Res 57:356–362CrossRefGoogle Scholar
  74. Yemm E, Willis A (1954) The estimation of carbohydrates in plant extracts by anthrone. Biochem J 57:508CrossRefPubMedPubMedCentralGoogle Scholar
  75. Zaman MS, Ali GM, Muhammad A, Farooq K, Hussain I et al (2015) In vitro screening of salt tolerance in potato (Solanum tuberosum L.) varieties. Sarhad J Agric 31:106–113CrossRefGoogle Scholar
  76. Zhang J, Kirkham M (1996) Lipid peroxidation in sorghum and sunflower seedlings as affected by ascorbic acid, benzoic acid, and propyl gallate. J Plant Physiol 149:489–493CrossRefGoogle Scholar
  77. Zhang W, Roomans GM (1998) Volume-induced chloride transport in HT29 cells studied by X-ray microanalysis. Microsc Res Tech 40:72–78CrossRefPubMedGoogle Scholar
  78. Zhu J-K (2001) Plant salt tolerance. Trends Plant Sci 6:66–71CrossRefPubMedGoogle Scholar

Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2018

Authors and Affiliations

  1. 1.Department of BiologyUniversity of IsfahanIsfahanIran
  2. 2.Department of Biological Sciences, Faculty of ScienceUniversity of KuwaitKuwait CityKuwait

Personalised recommendations