, Volume 26, Issue 5, pp 613–624 | Cite as

Evaluation of photosynthetic performance and carbon isotope discrimination in perennial ryegrass (Lolium perenne L.) under allelochemicals stress

  • M. Iftikhar Hussain
  • Manuel J. Reigosa


Ferulic (FA) and p-hydroxybenzoic acid (pHBA) are commonly found as phenolic compounds (PHC) in many forage and cereal crops. Although the effects of these PHC on seedling growth are relatively explored, not many information is available regarding the phytotoxicity on ecophysiological processes of perennial ryegrass adult plants. The experiment was conducted with the aim to evaluate the phytotoxic potential of PHC on the seedling growth, leaf water relation, chlorophyll fluorescence attributes and carbon isotope discrimination adult plants of perennial ryegrass (Lolium perenne L.). The results clearly indicated that PHC behaved as potent inhibitors of chlorophyll fluorescence yield (Fv/Fm) in leaves of L. perenne and plants showed poor tolerance against allelochemicals stress. Quantum yield (ΦPSII), chlorophyll fluorescence quenching (qP) and non-photochemical quenching (NPQ) were decreased following exposure to FA and pHBA. The portion of absorbed photon energy that was thermally dissipated (D) in L. perenne was decreased. Exposure of the L. perenne seedlings to FA and pHBA stress led to a decrease in fresh/dry weight, relative water content and leaf osmotic potential. Carbon isotope composition ratio (δ13C) was significantly less negative than the control following treatment with FA or pHBA. The results suggested that PHC uptake was a key step for the effectiveness of these secondary metabolites and their phytotoxicity on L. perenne adult plants was mainly due to the alteration of leaf water status accompanied by photosystem II damage. Acquisition of such knowledge may ultimately provide a better understanding about the mode of action of the tested compounds.


Phenolic compounds Physiological growth Ryegrass Natural herbicide Shoot growth Phytotoxicity 



Ratio of intercellular CO2 concentration from leaf to air


Portion of absorbed photon energy thermally dissipated


Photon energy absorbed by PSII antennae and trapped by “closed” PSII reaction centers


Maximal fluorescence


Minimal fluorescence


Variable fluorescence level from light-adapted state


Maximal fluorescence level from dark-adapted leaves


Initial fluorescence level from dark-adapted leaves


Variable fluorescence level from dark-adapted leaves


Leaf osmotic potential


Maximum quantum yield of PSII


Non-photochemical fluorescence quenching


Efficiency of photosystem II photochemistry in the dark-adapted state


Fraction of photon energy absorbed by PS II antennae trapped by “open” PS II reaction centers


Quinone type electron acceptor A


Photochemical quenching


Relative water content


Composition of carbon isotope ratios


Carbon isotope discrimination


Intrinsic water use efficiency



We thank Dr. Aldo Barreiro, Carlos Bolano and Paula Lorenzo with field and laboratory assistance. We are grateful to Jesús Estévez Sío for their technical assistance with isotope ratio mass spectroscopy. We thank Dr. Nuria Pedrol for valuable help in leaf osmotic potential measurement.

Author Contributions

All authors contributed to write this work. M.I: acquisition of data, efficiency of photosystem PSII and leaf fluorescence analysis and interpretation of data, drafting of manuscript, M.J.R: isotopic ratio mass spectroscopy measurement, analysis and interpretation of data, M.J.R: study conception and design and critical revision. The manuscript was approved by all of the authors.

Conflict of interest

The authors declare that they have no competing interests.


  1. Abenavoli MR, Sorgon’ AA, Muscolo A (2001) Morphophysiological changes in tissue culture of Petunia hybrida in response to the allelochemical coumarin. Allelop J 8:171–178Google Scholar
  2. Akhkha A, Boutraa T, Kalaji H, Ahmad P, Dąbrowski P (2013) Chlorophyll fluorescence: a potential selection criterion for drought tolerance in selected durum wheat (Triticum durum Desf.) cultivars. NanoPhotoBioScences 1:147–156Google Scholar
  3. Al- Hamdi B, Inderjit, Olofsdotter M, Streibig JC (2001) Laboratory bioassay for phytotoxicity: An example from wheat straw. Agron J 93:43–48Google Scholar
  4. Allakhverdiev SI, Klimov VV, Nagata T, Nixon P, Shen J-R (2008) Special issue: recent perspectives of photosystem II: structure, function and dynamics. Photosyn Res 98:1–700CrossRefGoogle Scholar
  5. Baker NR (2008) Chlorophyll fluorescence: a probe of photosynthesis in vivo. Annu Rev Plant Biol 59:89–113CrossRefGoogle Scholar
  6. Baker NR, Rosenqvist E (2004) Applications of chlorophyll fluorescence can improve crop production strategies: an examination of future possibilities. J Expt Bot 55:1607–1621CrossRefGoogle Scholar
  7. Barkosky RR, Einhellig FA (2003) Allelopathic interference of plant-water relationships by para-hydroxybenzoic acid. Bot Bull Acad Sin 44:53–58Google Scholar
  8. Barkosky RR, Einhellig FA, Butler J (2000) Caffeic acid-induced changes in plant-water relationships and photosynthesis in leafy spurge (Euphorbia esula). J Chem Ecol 26:2095–2109CrossRefGoogle Scholar
  9. Bernat G, Schreiber U, Sendtko E, Stadnichuk IN, Rexroth S, Rogner M, Koenig F (2012) Unique properties vs. common themes: the atypical cyanobacterium Gloeobacter violaceus PCC 7421 is capable of state transitions and blue-light-induced fluorescence quenching. Plant Cell Physiol 53:528–542CrossRefGoogle Scholar
  10. Bharati A, Kar M, Sabat SC (2016) Artemisinin inhibits chloroplast electron transport activity: Mode of action. Plos One 7:e38942CrossRefGoogle Scholar
  11. Bilger W, Schreiber U, Bock M (1995) Determination of the quantum efficiency of photosystem II and of non-photochemical quenching of chlorophyll fluorescence in the field. Oecologia 102:425–432CrossRefGoogle Scholar
  12. Blum U, Gerig TM (2006) Interrelationships between p-coumaric acid, evapotranspiration, soil water content, and leaf expansion. J Chem Ecol 32:1817–1834CrossRefGoogle Scholar
  13. Borawska-Jarmułowicz B, Mastalerczuk G, Pietkiewicz S, Kalaji MH (2014) Low temperature and hardening effects on photosynthetic apparatus efficiency and survival of forage grass varieties. Plant Soil Environ 60:177–183Google Scholar
  14. Dąbrowski P, Baczewska AH, Pawluśkiewicz B, Paunovc M, Alexantrov V, Goltsev V, Kalaji MH (2016) Prompt chlorophyll a fluorescence as a rapid tool for diagnostic changes in PSII structure inhibited by salt stress in Perennial ryegrass. J Photochem Photobiol B: Biol 157:22–31CrossRefGoogle Scholar
  15. Dayan FE, Cantrell CL, Duke SO (2009) Natural products in crop protection. Bioorg Med Chem 17:4022–4034CrossRefGoogle Scholar
  16. Dayan FE, Romagni JG, Duke SO (2000) Investigating the mode of action of natural phytotoxins. J Chem Ecol 26:2079–2094CrossRefGoogle Scholar
  17. De Albuquerque MB, Santos RC, Lima LM, Melo Filho PDA, Nogueira RJMC, Câmara CAG, Ramos A (2011) Allelopathy, an alternative tool to improve cropping systems. A review. Agron Sustain Dev 31:379–395CrossRefGoogle Scholar
  18. Demmig-Adams B, Adams III WW, Barker DH, Logan BA, Bowling DR, Verhoeven AS (1996) Using chlorophyll fluorescence to assess the fraction of absorbed light allocated to thermal dissipation of excess excitation. Physiol Plant 98:253–264CrossRefGoogle Scholar
  19. Devi SR, Prasad MNV (1996) Influence of ferulic acid on photosynthesis of maize: analysis of CO2 assimilation, electron transport activities, fluorescence emission and photophosphorylation. Photosynthetica 32:117–127Google Scholar
  20. Doblinski PMF et al. (2003) Peroxidase and lipid peroxidation of soybean roots in response to p-coumaric and p-hydroxybenzoic acids. Braz Arch Biol Tech 46:193–198CrossRefGoogle Scholar
  21. Doblinski PMF, Ferrarese MDLL, Huber DA, Scapim CA, Braccini ADL, Ferrarese-Filho O (2003) Peroxidase and lipid peroxidation of soybean roots in response to p-coumaric and p-hydroxybenzoic acids. Braz Arch Biol Technol 46:193–198CrossRefGoogle Scholar
  22. Duke SO, Dayan FE, Rimando AM, Schrader KK, Aliotta G, Oliva A, Romagni JG (2002) Chemicals from nature for weed management. Weed Sci 50:138–151CrossRefGoogle Scholar
  23. Eichenberg D, Ristok C, Kroeber W, Bruelheide H (2014) Plant polyphenols - implications of different sampling, storage and sample processing in biodiversity-ecosystem functioning experiments. J Chem Ecol 30:676–692CrossRefGoogle Scholar
  24. Farquhar GD, Ehleringer JR, Hubick KT (1989) Carbon isotope discrimination and photosynthesis. Ann Rev Plant Physiol Plant Mol Biol 40:503–537CrossRefGoogle Scholar
  25. Farquhar GD, Richards RA (1984) Isotopic composition of plant carbon correlates with water use efficiency of wheat genotypes. Aust J Plant Physiol 2:539–552CrossRefGoogle Scholar
  26. Fracheboud Y, Leipner J (2003) The application of chlorophyll fluorescence to study light, temperature and drought stress. In: De Ell JR, Tiovonen PMA (eds) Practical applications of chlorophyll fluorescence in plant biology. Kluwer Academic, Boston, MA, p 125–150CrossRefGoogle Scholar
  27. Fricke W, Peters WS (2002) The biophysics of leaf growth in salt-stressed barley. A study at the cell level. Plant Physiol 129:374–388CrossRefGoogle Scholar
  28. Genty B, Briantais JM, Baker NR (1989) The relationship between quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence. Biochim Biophys Acta 990:87–92CrossRefGoogle Scholar
  29. Gronle A, Heb J, Böhm H (2015) Weed suppressive ability in sole and intercrops of pea and oat and it is interaction with ploughing and crop interference in organic farming. Org Agric 5:39–51CrossRefGoogle Scholar
  30. Guy RD, Reid DM, Krouse HR (1986) Factors affecting 13C/12C ratios of inland halophytes. II. Ecophysiological interpretations of patterns in the field. Can J Bot 64:2700–2707CrossRefGoogle Scholar
  31. Heap I (2012) The international survey of herbicide resistant weeds. Accessed 29 Jan 2012.
  32. Hussain MI, González L, Reigosa MJ (2008) Germination and growth response of four plant species towards different allelochemicals and herbicides. Allelop J 22:101–110Google Scholar
  33. Hussain MI, González L, Reigosa MJ (2010) Phytotoxic effect of allelochemicals and herbicides on photosynthesis, growth and carbon isotope discrimination in Lactuca sativa. Allelop J 26:157–174Google Scholar
  34. Hussain MI, Reigosa MJ (2011a) Allelochemical stress inhibits growth, leaf water relations, PSII photochemistry, non-photochemical fluorescence quenching and heat energy dissipation in three C3 perennial species. J Exp Bot 62:4533–4545CrossRefGoogle Scholar
  35. Hussain MI, Reigosa MJ (2011b) A chlorophyll fluorescence analysis of photosynthetic efficiency, quantum yield and fractions of photon energy in PSII antennae of Lactuca sativa exposed to cinnamic acid. Plant Physiol Biochem 49:1290–1298CrossRefGoogle Scholar
  36. Hussain MI, Reigosa MJ (2012) Seedling growth, leaf water status and signature of stable carbon isotopes in C3 perennials exposed to natural phytochemicals. Aust J Bot 60:676–684Google Scholar
  37. Inderjit Dakshini KMM (1995) Allelopathic potential of an annual weed, Polypogon monspeliensis, in crops in India. Plant Soil 173:251–257CrossRefGoogle Scholar
  38. Jahns P, Holzwarth AR (2012) The role of the xanthophyll cycle and of lutein in photoprotection of photosystem II. Biochim Biophys Acta 1817:182–193CrossRefGoogle Scholar
  39. Kalaji HM, Jajoo A, Oukarroum A, Brestic M, Zivcak M, Samborska A, Cetner MD, Łukasik I, Goltsev V, Ladle RJ (2016b) Chlorophyll a fluorescence as a tool to monitor physiological status of plants under abiotic stress conditions. Acta Physiol Plant 38:102. doi: 10.1007/s11738-016-2113-y CrossRefGoogle Scholar
  40. Kalaji HM, Łoboda T (2007) Photosystem II of barley seedlings growing under cadmium and lead stress. Plant Soil Environ 53:511–516Google Scholar
  41. Kalaji HM, Oukarroum A, Alexandrov V, Kouzmanova M, Brestic M, Zivcak M, Samborska IA, Cetner MD, Allakhverdiev SI, Goltsev V (2014) Identification of nutrient deficiency in maize and tomato plants by in vivo chlorophyll a fluorescence measurements. Plant Physiol Bioch 81:16–25Google Scholar
  42. Kalaji HM, Schansker G, Brestic M et al. (2016a) Frequently asked questions about chlorophyll fluorescence, the sequel. Photosyn Res. doi: 10.1007/s11120-016-0318-y
  43. Kramer DM, Johnson G, Kiirats O, Edwards G (2004) New fluorescence parameters for the determination of QA redox state and excitation energy. Photosyn Res 79:209–218CrossRefGoogle Scholar
  44. Macías FA, Oliveros-Bastidas A, Marín D, Chinchilla N, Castellano D, Molinillo JM (2014) Evidence for an allelopathic interaction between rye and wild oats. J Agric Food Chem 62:9450–9457CrossRefGoogle Scholar
  45. Maxwell K, Johnson GN (2000) Chlorophyll fluorescence: a practical guide. J Exp Bot 51:659–668Google Scholar
  46. McCarroll D, Loader NJ (2004) Stable isotopes in tree rings. Quat Sci Rev 23:771–801CrossRefGoogle Scholar
  47. Mersie W, Singh M (1993) Phenolic acids affect photosynthesis and protein synthesis by isolated leaf cells of velvet-leaf. J Chem Ecol 19:1293–1301CrossRefGoogle Scholar
  48. Omasa K, Takayama K (2003) Simultaneous measurement of stomatal conductance, non-photochemical quenching, and photochemical yield of photosystem II in intact leaves by thermal and chlorophyll fluorescence imaging. Plant Cell Physiol 44:1290–1300CrossRefGoogle Scholar
  49. Pereira WE, de Siqueira DL, Martínez CA, Puiatti M (2000) Gas exchange and chlorophyll fluorescence in four citrus rootstocks under aluminium stress. J Plant Physiol 157:513–520CrossRefGoogle Scholar
  50. Reigosa MJ, Pazos-Malvido E (2007) Phytotoxic effects of 21 plant secondary metabolites on Arabidopsis thaliana germination and root growth. J Chem Ecol 33:1456–1466CrossRefGoogle Scholar
  51. Robertson A, Overpeck L, Rind D, Mosley-Thompson E, Zielinski O, Lean J, Koch D, Penner J, Tegen L, Healy R (2001) Hypothesized c1imate forcing time series for the last 500 years. J Geophys Res 106:14783–14803CrossRefGoogle Scholar
  52. Vaughan D, Ord BG (2006) Influence of phenolic acids on morphological changes in roots of Pisum sativum. J Sci Food Agri 52:289–299CrossRefGoogle Scholar
  53. Wallstedt A, Dubé SL, Nilsson M-C (2002) Photosynthetic functions of leaves affected by the bibenzyl batatasin-III. In: Reigosa MJ, Pedrol N (eds) Allelopathy: From molecules to ecosystems. Science Publishers Inc, Enfield, p 45–58Google Scholar
  54. Ye SF, Yu JQ, Peng YH, Zheng JH, Zou LY (2004) Incidence of Fusarium wilt in Cucumis sativus L. is promoted by cinnamic acid, an autotoxin in root exudates. Plant Soil 263:143–150Google Scholar
  55. Yu JQ, Matsui Y (1994) Phytotoxic substances in root exudates of cucumber (Cucumis sativus L.). J Chem Ecol 20:21–31CrossRefGoogle Scholar
  56. Zhou YH, Yu JQ (2006) Allelochemicals and photosynthesis. In: Reigosa MJ, Pedrol N, González L (eds) Allelopathy: A physiological process with ecological implications. Springer, Dordrecht, p 127–139CrossRefGoogle Scholar
  57. Živčák M, Olšovská K, Slamka P, Galambošová J, Rataj V, Shao HB, Brestič M (2014) Application of chlorophyll fluorescence performance indices to assess the wheat photosynthetic functions influenced by nitrogen deficiency. Plant Soil Environ 60:204–209Google Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Plant Biology and Soil ScienceUniversidad de VigoVigoSpain

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