, Volume 250, Issue 3, pp 691–700 | Cite as

β-Pinene inhibited germination and early growth involves membrane peroxidation

  • Nadia Chowhan
  • Harminder Pal Singh
  • Daizy R. Batish
  • Shalinder Kaur
  • Nitina Ahuja
  • Ravinder K. Kohli
Original Article


β-Pinene, an oxygenated monoterpene, is abundantly found in the environment and widely occurring in plants as a constituent of essential oils. We investigated the phytotoxicity of β-pinene against two grassy (Phalaris minor, Echinochloa crus-galli) and one broad-leaved (Cassia occidentalis) weeds in terms of germination and root and shoot growth. β-Pinene (0.02–0.80 mg/ml) inhibited the germination, root length, and shoot length of test weeds in a dose–response manner. The inhibitory effect of β-pinene was greater in grassy weeds and on root growth than on shoot growth. β-Pinene (0.04–0.80 mg/ml) reduced the root length in P. minor, E. crus-galli, and C. occidentalis over that in the control by 58–60, 44–92, and 26–85 %, respectively. In contrast, shoot length was reduced over the control by 45–97 % in P. minor, 48–78 % in E. crus-galli, and 11–75 % in C. occidentalis at similar concentrations. Further, we examined the impact of β-pinene on membrane integrity in P. minor as one of the possible mechanisms of action. Membrane integrity was evaluated in terms of lipid peroxidation, conjugated diene content, electrolyte leakage, and the activity of lipoxygenases (LOX). β-Pinene (≥0.04 mg/ml) enhanced electrolyte leakage by 23–80 %, malondialdehyde content by 15–67 %, hydrogen peroxide content by 9–39 %, and lipoxygenases activity by 38–383 % over that in the control. It indicated membrane peroxidation and loss of membrane integrity that could be the primary target of β-pinene. Even the enhanced (9–62 %) activity of protecting enzymes, peroxidases (POX), was not able to protect the membranes from β-pinene (0.04-0.20 mg/ml)-induced toxicity. In conclusion, our results show that β-pinene inhibits root growth of the tested weed species through disruption of membrane integrity as indicated by enhanced peroxidation, electrolyte leakage, and LOX activity despite the upregulation of POX activity.


β-Pinene Lipid peroxidation Electrolyte leakage Membrane disruption Lipoxygenases Peroxidases 



Nadia Chowhan and Nitina Ahuja are thankful to University Grants Commission (UGC, New Delhi, India) for financial assistance in the form of BSR Fellowship.

Conflict of interest

The authors declare no conflict of interest.


  1. Amaral JA, Knowles R (1998) Inhibition of methane consumption in forest soils by monoterpenes. J Chem Ecol 24:723–734CrossRefGoogle Scholar
  2. Amora Y, Chevionb M, Levinea A (2000) Anoxia pretreatment protects soybean cells against H2O2-induced cell death: possible involvement of peroxidases and of alternative oxidase. FEBS Lett 477:175–180PubMedCrossRefGoogle Scholar
  3. Andrews J, Adam SR, Burton KS, Evered CE (2002) Subcellular localization of peroxidise in tomato fruit skin and the possible implications for the regulation of fruit growth. J Exp Bot 53:2185–2191PubMedCrossRefGoogle Scholar
  4. Axelrod B, Cheesbrough TM, Laakso S (1981) Lipoxygenase from soybeans. Meth Enzymol 71:441–451CrossRefGoogle Scholar
  5. Badger MR (1985) Photosynthetic oxygen exchange. Annu Rev Plant Physiol 36:27–53CrossRefGoogle Scholar
  6. Bainard LD, Isman MB, Upadhyaya MK (2006) Phytotoxicity of clove oil and its primary constituent eugenol and the role of leaf epicuticular wax in the susceptibility to these essential oils. Weed Sci 54:833–837CrossRefGoogle Scholar
  7. Bajji M, Kinet J-M, Lutts S (2002) The use of the electrolyte leakage method for assessing cell membrane stability as a water stress tolerance test in durum wheat. Plant GrowthRegul 36:61–70CrossRefGoogle Scholar
  8. Batish DR, Singh HP, Kohli RK, Kaur S (2008) Eucalyptus essential oil as natural pesticide. For Ecol Manage 256:2166–2174CrossRefGoogle Scholar
  9. Bertin N, Staudt M, Hansen U, Seufert G, Ciccioli P, Foster P, Fugit JL, Torres L (1997) Diurnal and seasonal course of monoterpene emission from Quercus ilex (L.) under natural conditions—application of light and temperature algorithms. Atmos Environ 31:135–144CrossRefGoogle Scholar
  10. Burgos NR, Talbert RE (2000) Differential activity of allelochemicals from Secale cereale in seedling bioassays. Weed Sci 47:481–485Google Scholar
  11. Chowhan N, Singh HP, Batish DR, Kohli RK (2011) Phytotoxic effects of β-pinene on early growth and associated biochemical changes in rice. Acta Physiol Plant 33:2369–2376CrossRefGoogle Scholar
  12. Dayan FE, Romagni JG, Duke SO (2000) Investigating the mode of action of natural phytotoxins. J Chem Ecol 26:2079–2093CrossRefGoogle Scholar
  13. Dayan FE, Cantrell CL, Duke SO (2009) Natural products in crop protection. Bioorg Med Chem 17:4022–4034PubMedCrossRefGoogle Scholar
  14. Dudareva N, Negre F, Nagegowda AD, Orlova I (2006) Plant volatiles: recent advances and future perspectives. Crit Rev Plant Sci 25:417–440CrossRefGoogle Scholar
  15. Duke SO, Kenyon WH (1993) Peroxidizing activity determined by cellular leakage. In: Boger P, Sandann G (eds) Target assays for modern herbicides and related phytotoxic compounds. Lewis, Boca Raton, pp 61–66Google Scholar
  16. Duke SO, Dayan FE, Rimando AM, Schrader KK, Alliota G, Oliva A, Romagni JG (2002) Chemicals from nature for weed management. Weed Sci 50:1338–1351CrossRefGoogle Scholar
  17. Elstner EH (1982) Oxygen activation and oxygen toxicity. Annu Rev Plant Physiol 33:73–96CrossRefGoogle Scholar
  18. Foyer CH, Descourvières P, Kunert KJ (1994) Protection against oxygen radicals: an important defence mechanism studied in transgenic plants. Plant Cell Environ 17:507–523CrossRefGoogle Scholar
  19. Geron C, Ramussen R, Arnts RR, Guenther A (2000) A review and synthesis of monoterpene speciation from forests in the United States. Atmos Environ 34:1761–1781CrossRefGoogle Scholar
  20. Geron C, Guenther A, Greenberg J, Loescher HW, Clark D, Baker B (2002) Biogenic volatile organic compound emissions from a low land tropical wet forest in Costa Rica. Atmos Environ 36:3793–3802CrossRefGoogle Scholar
  21. Guenther A, Hewitt CN, Erickson D, Fall R, Geron C (1995) A global model of natural volatile organic compound emissions. J Geophys Res 100:8873–8892CrossRefGoogle Scholar
  22. Halliwell B, Gutteridge JM (1989) Free radicals in biology and medicine. Clarendon, OxfordGoogle Scholar
  23. Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplast. 1. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198PubMedCrossRefGoogle Scholar
  24. Kaur S, Singh HP, Mittal S, Batish DR, Kohli RK (2010) Phytotoxic effects of volatile oil from Artemisia scoparia against weeds and its possible use as a bioherbicide. Ind Crops Prod 32:54–61CrossRefGoogle Scholar
  25. Kaur G, Singh HP, Batish DR, Kohli RK (2012) A time course assessment of changes in reactive oxygen species generation and antioxidant defense in hydroponically grown wheat in response to lead ions (Pb2+). Protoplasma. doi: 10.1007/s00709-011-0353-7
  26. Kegge W, Pierik R (2010) Biogenic volatile compounds and plant competition. Trends Plant Sci 15:126–132PubMedCrossRefGoogle Scholar
  27. Klingler H, Frosch S, Wagner E (1991) In vitro effects of monoterpenes on chloroplast membranes. Photosynth Res 28:109–118CrossRefGoogle Scholar
  28. Kordali S, Cakir A, Sutay S (2007) Inhibitory effects of monoterpenes on seed germination and seedling growth. Z Naturforsch 62c:207–214Google Scholar
  29. Maalekuu K, Elkind Y, Leikin-Frenkel A, Lurie S, Fallik E (2006) The relationship between water loss lipid content membrane integrity and LOX activity in ripe pepper fruit after storage. Postharvest Biol Technol 42:248–255CrossRefGoogle Scholar
  30. Maness P-C, Smolinski S, Blake DM, Huang Z, Wolfrum EJ, Jacoby WA (1999) Bactericidal activity of photocatalytic TiO2 reaction: toward an understanding of its killing mechanism. Appl Environ Microbiol 65:4094–4098PubMedGoogle Scholar
  31. Mucciarelli M, Camusso W, Bertea CM, Bossi S, Maffei M (2001) Effect of (+)-pulegone and other oil components of Mentha×piperita on cucumber respiration. Phytochemistry 57:91–98PubMedCrossRefGoogle Scholar
  32. Muller CH (1965) Inhibitory terpenes volatilized from Salvia shrubs. Bull Torr Bot Club 92:38–45CrossRefGoogle Scholar
  33. Mutlu S, Atici O, Esim N, Mete E (2011) Essential oils of catmint (Nepeta meyeri Benth.) induce oxidative stress in early seedlings of various weed species. Acta Physiol Plant 33:943–951CrossRefGoogle Scholar
  34. Nigam S, Schewe T (2000) Phospholipase A(2)s and lipid peroxidation. Biochim Biophys Acta 1488:167–181PubMedCrossRefGoogle Scholar
  35. Nishida N, Tamotsu S, Nagata N, Saito C, Sakai A (2005) Allelopathic effects of volatile monoterpenoids produced by Salvia leucophylla: inhibition of cell proliferation and DNA synthesis in the root apical meristem of Brassica campestris seedlings. J Chem Ecol 31:1187–1203PubMedCrossRefGoogle Scholar
  36. Owen SM, Peñuelas J (2005) Opportunistic emissions of volatile isoprenoids. Trends Plant Sci 10:420–426PubMedCrossRefGoogle Scholar
  37. Paavolainen L, Kitunen V, Smolander A (1998) Inhibition of nitrification in forest soils by monoterpenes. Plant Soil 205:147–154CrossRefGoogle Scholar
  38. Peñuelas J, Llusia J, Estiarte M (1995) Terpenoids: a plant language. Trends Ecol Evol 10:289PubMedCrossRefGoogle Scholar
  39. Peñuelas J, Staudt M (2010) BVOCs and global change. Trends Plant Sci 15:133–144PubMedCrossRefGoogle Scholar
  40. Romagni JG, Allen SN, Dayan FE (2000) Allelopathic effects of volatile cineoles on two weedy plant species. J Chem Ecol 26:303–313CrossRefGoogle Scholar
  41. Santos WD, Ferrarese MLL, Finger A, Teixeira CAN, Ferrarese FO (2004) Lignification and related enzymes in Glycine max root growth inhibition by ferulic acid. J Chem Ecol 30:1203–1212PubMedCrossRefGoogle Scholar
  42. Siedow JN (1991) Plant lipoxygenase: structure and function. Annu Rev Plant Physiol Plant Mol Biol 42:145–188CrossRefGoogle Scholar
  43. Singh HP, Batish DR, Kohli RK (2002a) Allelopathic effects of two volatile monoterpenes against bill-goat weed (Ageratum conyzoides L.). Crop Prot 21:347–350CrossRefGoogle Scholar
  44. Singh HP, Batish DR, Kaur S, Ramezani H, Kohli RK (2002b) Comparative phytotoxicity of four monoterpenes against Cassia occidentalis. Ann Appl Biol 14:1111–1116Google Scholar
  45. Singh HP, Batish DR, Kohli RK (2003) Allelopathic interactions and allelochemicals: new possibilities for sustainable weed management. Crit Rev Plant Sci 22:239–311CrossRefGoogle Scholar
  46. Singh HP, Batish DR, Kaur S, Arora K, Kohli RK (2006) α-Pinene inhibits growth and induces oxidative stress in roots. Ann Bot 98:1261–1269PubMedCrossRefGoogle Scholar
  47. Singh HP, Batish DR, Kohli RK, Arora K (2007) Arsenic induced root growth inhibition in mung bean (Phaseolus aureus Roxb.) is due to oxidative stress resulting from enhanced lipid peroxidation. Plant Growth Regul 53:65–73CrossRefGoogle Scholar
  48. Singh HP, Kaur S, Mittal S, Batish DR, Kohli RK (2009) Essential oil of Artemisia scoparia inhibits plant growth by generating reactive oxygen species and causing oxidative damage. J Chem Ecol 35:154–162PubMedCrossRefGoogle Scholar
  49. Song FM, Ge XC, Zheng Z (2001) The roles of active oxygen species and lipid peroxidation in the resistance of cotton seedlings to Fusarium wilt. Acta Phytopathol Sin 31:110–116Google Scholar
  50. Stone JR, Yang S (2006) Hydrogen peroxide: a signalling messenger. Antioxidant Redox Signalling 8:243–270CrossRefGoogle Scholar
  51. Tworkoski T (2002) Herbicide effects of essential oils. Weed Sci 50:425–431CrossRefGoogle Scholar
  52. White CS (1991) The role of monoterpenes in soil nitrogen cycling processes in ponderosa pine: results from laboratory bioassays and field studies. Biogeochemistry 12:43–68CrossRefGoogle Scholar
  53. White CS (1994) Monoterpenes: their effects on ecosystem nutrient cycling. J Chem Ecol 20:1381–1406CrossRefGoogle Scholar
  54. Williams RD, Hoagland RE (1982) The effects of naturally occurring phenolic compounds on seed germination. Weed Sci 30:206–212Google Scholar
  55. Yoshimura H, Sawai Y, Tamotsu S, Sakai A (2011) 18-Cineole inhibits both proliferation and elongation of BY-2 cultured tobacco cells. J Chem Ecol 37:320–328PubMedCrossRefGoogle Scholar
  56. Zunino MP, Zygadlo JA (2004) Effect of monoterpenes on lipid oxidation in maize. Planta 219:303–309PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Nadia Chowhan
    • 1
  • Harminder Pal Singh
    • 2
  • Daizy R. Batish
    • 1
  • Shalinder Kaur
    • 2
  • Nitina Ahuja
    • 1
  • Ravinder K. Kohli
    • 1
  1. 1.Department of BotanyPanjab UniversityChandigarhIndia
  2. 2.Department of Environment StudiesPanjab UniversityChandigarhIndia

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