Journal of Plant Growth Regulation

, Volume 38, Issue 2, pp 683–700 | Cite as

Salt Stress Induces Increase in Starch Accumulation in Duckweed (Lemna aequinoctialis, Lemnaceae): Biochemical and Physiological Aspects

  • Marciana Bizerra de MoraisEmail author
  • Adauto Gomes Barbosa-Neto
  • Lilia Willadino
  • Cláudia Ulisses
  • Tercilio Calsa Junior


In this study, antioxidant processes were searched for in macrophyte duckweed to investigate tolerance mechanisms in this species against oxidative damage caused by salinity stress. Biochemical and histological analyses were performed on four Lemna aequinoctialis clones grown in Schenk-Hildebrandt medium, 0.5 × SH, supplemented with 1% sucrose liquid medium containing or not containing NaCl in different NaCl concentrations (0, 25 and 50 mM). For most clones, the salt stress effects caused growth inhibition and antioxidant responses at 50 mM NaCl. Also, starch and reducing sugar accumulations were increased with salt, whereas the photosynthetic pigment content was reduced in clone L. aequinoctialis 5569. The plant growth inhibition reflects the oxidative stress shown by the significant increase in malondialdehyde (MDA) and hydrogen peroxide (H2O2) content. In the L. aequinoctialis 5568 clone, with the highest MDA levels, no antioxidant enzymatic activity was observed. The L. aequinoctialis 5570 clone presented higher ascorbate peroxidase and catalase activities in parallel, indicating that the efficiency of the defence mechanism relies on synchrony between such enzyme activities toward successive elimination of reactive oxygen species and resulting in the assurance of some level of protection of the metabolism from oxidative damage. Considering the moderate salt stress (25 mM), the maintenance of MDA content and small growth inhibition associated with the high starch production suggested the acclimation efficiency of L. aequinoctialis 5570 and 5567 clones, indicating that they may be suitable for cultivation under moderate saline conditions, serving as biofuel feedstock. In addition, this study demonstrates great intraspecific phenotypic plasticity of duckweed, L. aequinoctialis, from closely related clones.


Antioxidant defence Abiotic stress Oxidative stress 



The authors gratefully acknowledge the Universidade Federal de Pernambuco for support and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for a scholarship granted to the first author.

Compliance with Ethical Standards

Conflict of interest

The authors declare no conflicts of interest.


  1. Abraham G, Dhar DW (2010) Induction of salt tolerance in Azolla microphylla Kaulf through modulation of antioxidant enzymes and ion transport. Protoplasma 245:105–111. CrossRefGoogle Scholar
  2. Acosta-Motos JR, Ortuño M, Bernal-Vicente A et al (2017) Plant responses to salt stress: adaptive mechanisms. Agronomy. Google Scholar
  3. Adem GD, Roy SJ, Zhou M, Bowman JP, Shabala S (2014) Evaluating contribution of ionic, osmotic and oxidative stress components towards salinity tolerance in barley. BMC Plant Biol. Google Scholar
  4. Agarwal S, Pandey V (2004) Antioxidant enzyme responses to NaCl stress in Cassia angustifolia. Biol Plant 48:555–560CrossRefGoogle Scholar
  5. 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. Environment 24:1337–1344. Google Scholar
  6. Almeida P, Katsching D, Boer AH (2013) HKT transporters—state of the art. Int J Mol Sci 14:20359–20385. CrossRefGoogle Scholar
  7. Anderson KE, Lowman Z, Stomp A-M, Chang J (2011) Duckweed as a feed ingredient in laying hen diets and its effect on egg production and composition. Int J Poult Sci 10:4–7. CrossRefGoogle Scholar
  8. Appenroth KJ, Krech K, Keresztes A et al (2010) Effects of nickel on the chloroplasts of the duckweeds Spirodela polyrhiza and Lemna minor and their possible use in biomonitoring and phytoremediation. Chemosphere 78:216–223CrossRefGoogle Scholar
  9. Appenroth KJ, Borisjuk N, Lam E (2013) Telling duckweed apart: genotyping technologies for Lemnaceae. Chin J Appl Environ Biol 19:1–10CrossRefGoogle Scholar
  10. 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 the leaves and roots of wild-type and a catalase-deficient mutant of barley. Physiol Plant 104:280–292CrossRefGoogle Scholar
  11. Azevedo R, Carvalho RF, Cia MC, Gratão PL (2011) Sugarcane under pressure: an overview of biochemical and physiological studies of abiotic stress. Trop Plant Biol 4:42–51. CrossRefGoogle Scholar
  12. Azevedo Neto AD, Pereira PPA, Costa DP, Santos ACC (2011) Fluorescência da clorofila como uma ferramenta possível para seleção de tolerância à salinidade em girassol. Rev Cien Agron 42:893–897. CrossRefGoogle Scholar
  13. Balibrea ME, Dell’Amico J, Bolarín MC, Pérez-Alfocea F (2000) Carbon partitioning and sucrose metabolism in tomato plants growing under salinity. Physiol Plant 110:503–511CrossRefGoogle Scholar
  14. Berlyn GP, Miksche JP (1976) Botanical microtechnique and cytochemistry. The Iowa State University Press, AmesCrossRefGoogle Scholar
  15. Bezerra Neto E, Barreto LP (2011) Análises químicas e bioquímicas em plantas. Editora Universitária da UFRPE, RecifeGoogle Scholar
  16. Bonates LC de M (1993) Estudos ecofisiológicos de Orchidaceae da Amazônia II—Anatomia ecologia foliar de espécies com metabolismo CAM de uma campina da Amazônia Central. Acta Amaz 23:315–348CrossRefGoogle Scholar
  17. Borisjuk N, Chu P, Gutierrez R et al (2015) Assessment, validation and deployment strategy of a two-barcode protocol for facile genotyping of duckweed species. Plant Biol 17:42–49. CrossRefGoogle Scholar
  18. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. CrossRefGoogle Scholar
  19. Bukatsch F (1972) Bemerkungen zur doppelfärbung astrablau-safranin. Mikrokosmos 8:255Google Scholar
  20. Chang IH, Cheng KT, Huang PC et al (2012) Oxidative stress in greater duckweed (Spirodela polyrhiza) caused by long-term NaCl exposure. Acta Physiol Plant 34:1165–1176. CrossRefGoogle Scholar
  21. Chaves MM, Flexas J, Pinheiro C (2009) Photosynthesis under drought and salt stress: regulation mechanisms from whole plant to cell. Ann Bot 551–560.
  22. Cheng TS (2011) NaCI-induced responses in giant duckweed (Spirodela polyrhiza). J Aquat Plant Manage 49:62–71Google Scholar
  23. Cheng JJ, Stomp AM (2009) Growing duckweed to recover nutrients from wastewaters and for production of fuel ethanol and animal feed. Clean Soil Air Water 37:17–26. CrossRefGoogle Scholar
  24. Cia MC, Guimarães a CR, Medici LO et al (2012) Antioxidant responses to water deficit by drought-tolerant and -sensitive sugarcane varieties. Ann Appl Biol 161:313–324. CrossRefGoogle Scholar
  25. Cui W, Cheng JJ (2015) Growing duckweed for biofuel production: a review. Plant Biol 17:16–23. CrossRefGoogle Scholar
  26. Cui W, Xu J, Cheng JJ, Stomp AM (2011) Starch accumulation in duckweed for bioethanol production. Biol Eng 3:187–197. CrossRefGoogle Scholar
  27. Fahn A, Cutler DF (1992) Xerophytes. Gebruder Borntraeger, BerlinGoogle Scholar
  28. Fodorpataki L, Bartha L (2008) Differential sensitivity of the photosynthetic apparatus of a freshwater green alga and of duckweed exposed to salinity and heavy metal stress. In: Allen JF, Gantt E, Golbeck J, Osmond B (eds) Photosynthesis. Energy from the sun: 14th international congress on photosynthesis. Springer, New York, pp 1451–1454CrossRefGoogle Scholar
  29. Franceschi VR, Nakata PA (2005) Calcium oxalate in plants: formation and function. Annu Rev Plant Biol 56:41–71. CrossRefGoogle Scholar
  30. Franklin GL (1945) Preparation of thin sections of synthetic resins and wood-resin composites, and a new macerating method for wood. Nature 155:51–51. CrossRefGoogle Scholar
  31. Giannopolitis CN, Ries SK (1977) Superoxide dismutases: I. Occurrence in higher plants. Plant Physiol 59:309–314CrossRefGoogle Scholar
  32. Greenway H, Munns R (1980) Mechanisms of salt tolerance in nonhalophytes. Annu Rev Plant Physiol 31:149–190CrossRefGoogle Scholar
  33. Havir E, McHale N (1987) Biochemical and developmental characterization of multiple forms of catalase in tobacco leaves. Plant Physiol 84:450–455. CrossRefGoogle Scholar
  34. Heath RL, Packer L (1968) Photoperoxidation in isolated Chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198CrossRefGoogle Scholar
  35. Hou W, Chen X, Song G et al (2007) Effects of copper and cadmium on heavy metal polluted waterbody restoration by duckweed (Lemna minor). Plant Physiol Biochem 45:62–69. CrossRefGoogle Scholar
  36. Jensen WA (1962) Botamical histochemistry: principles and practice. W.H. Freeman and Company, San FranciscoGoogle Scholar
  37. Joseph EA, Radhakrishnan VV, Mohanan KV (2015) A study on the accumulation of proline—an osmoprotectant amino acid under salt stress in some native rice cultivars of North Kerala, India. Univ J Agric Res 3:15–22. Google Scholar
  38. Karuppanapandian T, Kim W (2013) Cobalt-induced oxidative stress causes growth inhibition associated with enhanced lipid peroxidation and activates antioxidant responses in Indian mustard (Brassica juncea L.) leaves. Acta Physiol Plant 35:2429–2443. CrossRefGoogle Scholar
  39. Koca H, Bor M, Ozdemir F (2007) The effect of salt stress on lipid peroxidation, antioxidative enzymes and proline content of sesame cultivars urkan. Environ Exp Bot 60:344–351. CrossRefGoogle Scholar
  40. Lam E, Appenroth KJ, Michael T et al (2014) Duckweed in bloom: the 2nd international conference on duckweed research and applications heralds the return of a plant model for plant biology. Plant Mol Biol 84:737–742. CrossRefGoogle Scholar
  41. Landolt E (1986) Biosystematic investigations in the family of duckweeds (Lemnaceae), vol 2. Veroff. Geobot. Inst., RubelGoogle Scholar
  42. Lemoine R, La Camera S, Atanassova R, Dédaldéchamp F et al (2013) Source to sink transport and regulation by environmental factors. Front Plant Sci 4:1–21. CrossRefGoogle Scholar
  43. Lichtenthaler H, Wellburn A (1983) Determinations of total carotenoids and chlorophylls b of leaf extracts in different solvents. Biochem Soc Trans 11:591–592. CrossRefGoogle Scholar
  44. Liu C, Dai Z, Sun H (2017) Potential of duckweed (Lemna minor) for removal of nitrogen and phosphorus from water under salt stress. J Environ Manage 187:497–503. CrossRefGoogle Scholar
  45. Ma YB, Zhu M, Yu CJ et al (2018) Large-scale screening and characterisation of Lemna aequinoctialis and Spirodela polyrhiza strains for starch production. Plant Biol 20:357–364. CrossRefGoogle Scholar
  46. Malavolta E, Vitti GC, Oliveira SA (1989) Avaliação do estado nutricional das plantas—princípios e aplicações. Associação Brasileira para a Pesquisa da Potassa e do Fosfato, PiracicabaGoogle Scholar
  47. Miller GL (1959) Use of dinitrosalicylle acid for determination of reducing sugar. Anal Chem 11:426–428CrossRefGoogle Scholar
  48. Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410CrossRefGoogle Scholar
  49. Moller IM, Jensen PE, Hansson A (2007) Oxidative modifications to cellular components in plants. Annu Rev Plant Biol 58:459–481. CrossRefGoogle Scholar
  50. Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol. Google Scholar
  51. Nakano Y, Asada K (1981) Hydrogen-peroxide is scavenged by ascorbate-specific peroxidase in spinach-chloroplasts. Plant Cell Physiol 22:867–880Google Scholar
  52. Oukarroum A, Bussotti F, Goltsev V, Kalaji HM (2015) Correlation between reactive oxygen species production and photochemistry of photosystems I and II in Lemna gibba L. plants under salt stress. Environ Exp Bot 109:80–88. CrossRefGoogle Scholar
  53. Panda SK, Upadhyay RK (2003) Salt stress injury induces oxidative alterations and antioxidative defence in the roots of Lemna minor. Biol Plant 48:249–253. CrossRefGoogle Scholar
  54. Prasad M, Malec P, Waloszek A et al (2001) Physiological responses of Lemna trisulca L. (duckweed) to cadmium and copper bioaccumulation. Plant Sci 161:881–889CrossRefGoogle Scholar
  55. Prychid CJ, Rudall PJ (1999) Calcium oxalate crystals in monocotyledons: a review of their structure and systematics. Ann Bot 84:725–739. CrossRefGoogle Scholar
  56. Reale L, Ferranti F, Mantilacci S et al (2016) Cyto-histological and morpho-physiological responses of common duckweed (Lemna minor L.) to chromium. Chemosphere 145:98–105. CrossRefGoogle Scholar
  57. Schenk RU, Hildebrandt AC (1972) Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Can J Bot 50:199–204. CrossRefGoogle Scholar
  58. Shabala S, Munns R (2012) Salinity stress: physiological constraints and adaptive mechanisms. CABI, London. Google Scholar
  59. Shepherd G, Urbanetz C (2010) FITOPAC 2.1Google Scholar
  60. Sikorski Ł, Piotrowicz-Cieślak AI, Adomas B (2013) Phytotoxicity of sodium chloride towards common duckweed (Lemna Minor L.) and yellow lupin (Lupinus luteus L.). Arch Environ Prot 39:117–128. CrossRefGoogle Scholar
  61. Sree KS, Appenroth K (2014) Increase of starch accumulation in the duckweed Lemna minor under abiotic stress. Albanian J Agric Sci 11–14Google Scholar
  62. Sree KS, Adelmann K, Garcia C et al (2015) Natural variance in salt tolerance and induction of starch accumulation in duckweeds. Planta 241:1395–1404. CrossRefGoogle Scholar
  63. Szabó-Nagy A, Galiba G, Erdei L (1992) Induction of soluble phosphatases under ionic and non-ionic osmotic stresses in wheat. J Plant Physiol 140:629–633CrossRefGoogle Scholar
  64. Tanou G, Molassiotis A, Diamantidis G (2009) Induction of reactive oxygen species and necrotic death-like destruction in strawberry leaves by salinity. Environ Exp Bot 65:270–281. CrossRefGoogle Scholar
  65. Tkalec M, Prebeg T, Roje V et al (2008) Cadmium-induced responses in duckweed Lemna minor L. Acta Physiol Plant 30:881–890. CrossRefGoogle Scholar
  66. Wang W, Vinocur B, Altman A (2003) Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta 218:1–14. CrossRefGoogle Scholar
  67. Willadino L, Camara TR (2010) Tolerância das plantas à salinidade: aspectos fisiológicos e bioquímicos. Encicl Biosf 6:1–23Google Scholar
  68. Witham FH, Blaydes DF, Devlin RM (1971) Experiments in plant physiology. D.V. Nostrand, New YorkGoogle Scholar
  69. Xu J, Cui W, Cheng JJ, Stomp A-M (2011) Production of high-starch duckweed and its conversion to bioethanol. Biosyst Eng 110:67–72. CrossRefGoogle Scholar
  70. Xue H, Xiao Y, Jin Y et al (2012) Genetic diversity and geographic differentiation analysis of duckweed using inter-simple sequence repeat markers. Mol Biol Rep 39:547–554. CrossRefGoogle Scholar
  71. Yazici I, Türkan I, Sekmen AH, Demiral T (2007) Salinity tolerance of purslane (Portulaca oleracea L.) is achieved by enhanced antioxidative system, lower level of lipid peroxidation and proline accumulation. Environ Exp Bot 61:49–57. CrossRefGoogle Scholar
  72. Zhao X, Moatesa GK, Wellnera N et al (2014) Chemical characterisation and analysis of the cell wall polysaccharides of duckweed (Lemna minor). Carbohydr Polym 111:410–418. CrossRefGoogle Scholar
  73. Ziegler P, Adelmann K, Zimmer S et al (2015) Relative in vitro growth rates of duckweeds (Lemnaceae)—the most rapidly growing higher plants. Plant Biol 17:33–41. CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Programa de Pós-graduação em Ciências Naturais (PPGCN)Universidade do Estado do Rio Grande do Norte (UERN)MossoróBrazil
  2. 2.Departamento de GenéticaUniversidade Federal de PernambucoRecifeBrazil
  3. 3.Departamento de BiologiaUniversidade Federal Rural de PernambucoRecifeBrazil

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