Advertisement

Journal of Applied Phycology

, Volume 23, Issue 3, pp 409–414 | Cite as

Bioassay to detect Ascophyllum nodosum extract-induced cytokinin-like activity in Arabidopsis thaliana

  • Wajahatullah Khan
  • David Hiltz
  • Alan T. Critchley
  • Balakrishnan PrithivirajEmail author
Article

Abstract

Ascophyllum nodosum, a brown macroalga, is the most widely used seaweed in agriculture. We report a rapid method for the detection of cytokinin-like activity in plants treated with a commercial A. nodosum liquid concentrate (Stimplex®) using a transgenic line of Arabidopsis carrying the ARR5 promoter fused to ß-glucuronidase (GUS) reporter gene. Based on GUS activity assay, an increase in cytokinin-like activity was detected in plants grown in vitro treated with 3 mL L−1 Stimplex®, whereas foliar spray treatments showed similar cytokinin-like activity at a concentration of 5 mL L−1. Histochemical staining showed Stimplex®-induced GUS activity in leaf as well as in the root tissues. Taken together, our results suggest that Stimplex® contains compounds that may elicit endogenous cytokinin-like activity. Furthermore, it is shown that this bioassay can be used for rapid screening of extracts that can stimulate cytokinin-like activities using Arabidopsis AAR5::GUS reporter transgenic plants.

Keywords

Ascophyllum nodosum Cytokinin Elicitor Arabidopsis thaliana Plant growth regulator 

Notes

Acknowledgments

The research reported in this paper was supported by the Atlantic Canada Opportunities Agency (ACOA) and Acadian Seaplants Limited, Dartmouth, Nova Scotia, Canada. WK would like to thank Research Centre, College of Science, King Saud University.

References

  1. Abetz P (1980) Seaweed extracts: have they a place in Australian agriculture or horticulture? J Aust Inst Agr Sci 46:23–29Google Scholar
  2. Aich S, Delbaere LTJ, Chen R (2001) A continuous spectrophotometric assay for β-glucuronidase. Biotechniques 30:846–850PubMedGoogle Scholar
  3. Baardseth E (1970) A square-scanning, two-stage sampling method of estimating seaweed quantities. Rep Norw Inst Seaweed Res 33:1–41Google Scholar
  4. Bokil KK, Mehta VC, Datar DS (1974) Seaweeds as manure II: pot culture manorial experiments on wheat. Phykos 13:1–5Google Scholar
  5. 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 72:248–254PubMedCrossRefGoogle Scholar
  6. Craigie JS (2010) Seaweed extract stimuli in plant science and agriculture. J Appl Phycol. doi: 10.1007/s10811-010-9560-4
  7. Crouch IJ, Van Staden J (1992) Effect of seaweed concentrate on the establishment and yield of greenhouse tomato plants. J Appl Phycol 4:291–296CrossRefGoogle Scholar
  8. Crouch IJ, van Staden J (1993) Evidence for the presence of plant growth regulators in commercial seaweed products. Plant Growth Reg 13:21–29Google Scholar
  9. D’Agostino IB, Deruére J, Kieber JJ (2000) Characterization of the response of the Arabidopsis response regulator gene family to cytokinin. Plant Physiol 124:1706–1717PubMedCrossRefGoogle Scholar
  10. FAO (2006) Yearbook of fishery statistics. Food and Agricultural Organisation of the United Nations, Rome, 98 (1–2)Google Scholar
  11. Featonby-Smith BC, van Staden J (1983) The effect of seaweed concentrate and fertilizer on the growth of Beta vulgaris. Z Pflanzenphysiol 112:155–162Google Scholar
  12. Finnie JF, Van Staden J (1985) Effect of seaweed concentrate and applied hormones on in-vitro cultured tomato roots. J Plant Physiol 120:215–222Google Scholar
  13. Jefferson RA, Wilson KJ (1991) The GUS gene fusion system. In: Gelvin SB, Schilperoort RA (eds) Plant molecular biology manual. Kluwer, Dordrecht, pp 1–33Google Scholar
  14. Khan W, Rayirath UP, Subramanian S, Jithesh MN, Rayorath P, Hodges DM, Critchley AT, Craigie JS, Norrie J, Prithiviraj B (2009) Seaweed extracts as biostimulants of plant growth and development (review). J Plant Growth Regul 28:386–399CrossRefGoogle Scholar
  15. Lobban CS, Harrison PJ (1994) Seaweed ecology and physiology. Cambridge University Press, CambridgeCrossRefGoogle Scholar
  16. Metha VC, Trivedi BS, Bokil KK, Narayana MR (1967) Seaweed as manure, studies on nitrification. Proceedings of Seminar ‘Sea Salt and Plants’ (CSMCR), Bhavnagar, pp 357–365Google Scholar
  17. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  18. Nelson WR, Van Staden J (1984a) The effect of seaweed concentrate on growth of nutrient-stressed greenhouse cucumbers. Hort Sci 19:81–82Google Scholar
  19. Nelson WR, Van Staden J (1984b) The effect of seaweed concentrate on wheat culms. J Plant Physiol 115:433–437Google Scholar
  20. Rayorath P, Narayanan JM, Farid A, Khan W, Palanisamy R, Hankins S, Critchley AT, Prithiviraj B (2008) Rapid bioassays to evaluate the plant growth promoting activity of Ascophyllum nodosum (L.) Le Jol. using a model plant, Arabidopsis thaliana (L.) Heynh. J Appl Phycol 20:423–429CrossRefGoogle Scholar
  21. Romanov GA, Kieber JJ, Schmulling T (2002) A rapid cytokinin response assay in Arabidopsis indicates a role for phospholipase D in cytokinin signaling. FEBS Lett 515:39–43PubMedCrossRefGoogle Scholar
  22. Sanderson KJ, Jameson PE (1986) The cytokinins in a liquid seaweed extract: could they be active ingredients? Acta Hort 179:113–116Google Scholar
  23. Stirk WA, van Staden J (1997) Comparison of cytokinin- and auxin-like activity in some commercially used seaweed extracts. J Appl Phycol 8:503–508CrossRefGoogle Scholar
  24. Stirk WA, Novak O, Strnad M, van Staden J (2003) Cytokinins in macroalgae. Plant Growth Regul 41:13–24CrossRefGoogle Scholar
  25. Stirk WA, Arthur GD, Lourens AF, Novak O, Strnad M, van Staden J (2004) Changes in cytokinin and auxin concentrations in seaweed concentrates when stored at an elevated temperature. J Appl Phycol 16:31–39CrossRefGoogle Scholar
  26. Taylor WR (1957) Marine algae of the northeast coast of North America. University of Michigan Press, Ann ArborGoogle Scholar
  27. Ugarte RA, Sharp G, Moore B (2006) Changes in the brown seaweed Ascophyllum nodosum (L.) Le Jol. plant morphology and biomass produced by cutter rake harvests in southern New Brunswick, Canada. J Appl Phycol 18:351–359CrossRefGoogle Scholar
  28. Ugarte R, Craigie JS, Critchley AT (2010) Fucoid flora of the rocky intertidal of the Canadian Maritimes: Implications for the future with rapid climate change. In: Israel A, Einav R, Seckbach J (eds) Seaweeds and their role in globally changing environments. SpringerGoogle Scholar
  29. Zhang X, Ervin EH (2004) Cytokinin-containing seaweed and humic acid extracts associated with creeping bentgrass leaf cytokinins and drought resistance. Crop Sci 44:1737–1745CrossRefGoogle Scholar
  30. Zodape ST (2001) Seaweed as a biofertilizer. J Sci Ind Res 60:378–382Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Wajahatullah Khan
    • 1
  • David Hiltz
    • 2
  • Alan T. Critchley
    • 2
  • Balakrishnan Prithiviraj
    • 3
    Email author
  1. 1.Department of Biochemistry, College of ScienceKing Saud UniversityRiyadhSaudi Arabia
  2. 2.Acadian Seaplants LimitedDartmouthCanada
  3. 3.Department of Environmental SciencesNova Scotia Agricultural CollegeTruroCanada

Personalised recommendations