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Lipophilic components of the brown seaweed, Ascophyllum nodosum, enhance freezing tolerance in Arabidopsis thaliana

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Abstract

Extracts of the brown seaweed Ascophyllum nodosum enhance plant tolerance against environmental stresses such as drought, salinity, and frost. However, the molecular mechanisms underlying this improved stress tolerance and the nature of the bioactive compounds present in the seaweed extracts that elicits stress tolerance remain largely unknown. We investigated the effect of A. nodosum extracts and its organic sub-fractions on freezing tolerance of Arabidopsis thaliana. Ascophyllum nodosum extracts and its lipophilic fraction significantly increased tolerance to freezing temperatures in in vitro and in vivo assays. Untreated plants exhibited severe chlorosis, tissue damage, and failed to recover from freezing treatments while the extract-treated plants recovered from freezing temperature of −7.5°C in in vitro and −5.5°C in in vivo assays. Electrolyte leakage measurements revealed that the LT50 value was lowered by 3°C while cell viability staining demonstrated a 30–40% reduction in area of damaged tissue in extract treated plants as compared to water controls. Moreover, histological observations of leaf sections revealed that extracts have a significant effect on maintaining membrane integrity during freezing stress. Treated plants exhibited 70% less chlorophyll damage during freezing recovery as compared to the controls, and this correlated with reduced expression of the chlorphyllase genes AtCHL1 and AtCHL2. Further, the A. nodosum extract treatment modulated the expression of the cold response genes, COR15A, RD29A, and CBF3, resulting in enhanced tolerance to freezing temperatures. More than 2.6-fold increase in expression of RD29A, 1.8-fold increase of CBF3 and two-fold increase in the transcript level of COR15A was observed in plants treated with lipophilic fraction of A. nodosum at −2°C. Taken together, the results suggest that chemical components in A. nodosum extracts protect membrane integrity and affect the expression of stress response genes leading to freezing stress tolerance in A. thaliana.

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Abbreviations

NMR:

Nuclear magnetic resonance

FYM:

Farmyard manure

COR:

Cold responsive

ANE:

Ascophyllum nodosum extract

TMS:

Tetramethylsilane

NIH:

National Institutes of Health

PME:

Pectin methyl esterase

CBF:

CRT/DRE binding factor

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Acknowledgments

BP’s lab is supported by the grants from Atlantic Canada Opportunities Agency (ACOA), Natural Sciences and Engineering Research Council of Canada (NSERC), Nova Scotia Department of Agriculture & Marketing (NSDAF) and Acadian Seaplants Limited. PR is grateful to Kalyani Prithiviraj, Nova Scotia Agricultural College for her help with real time PCR experiments.

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Authors and Affiliations

  1. Department of Plant and Animal Sciences, Nova Scotia Agricultural College, 58 River Road, P.O. Box 550, Truro, NS, B2N 5E3, Canada

    Prasanth Rayirath, Bernhard Benkel & Balakrishnan Prithiviraj

  2. Atlantic Food and Horticulture Research Centre, Agriculture and Agri-Food Canada, Kentville, NS, B4N 1J5, Canada

    D. Mark Hodges & Paula Allan-Wojtas

  3. Institute for Marine Biosciences, National Research Council of Canada, Halifax, NS, B3H 3Z1, Canada

    Shawna MacKinnon

  4. Acadian Seaplants Limited, 30 Brown Ave., Dartmouth, NS, B3B 1X8, Canada

    Alan T. Critchley

Authors
  1. Prasanth Rayirath
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  2. Bernhard Benkel
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  3. D. Mark Hodges
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  4. Paula Allan-Wojtas
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  5. Shawna MacKinnon
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  6. Alan T. Critchley
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  7. Balakrishnan Prithiviraj
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Correspondence to Balakrishnan Prithiviraj.

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425_2009_920_MOESM1_ESM.tif

Supplementary Figure 1. Pectin methyl esterase activity in plants treated with ANE (1.0 g L-1) orethyl acetate sub-fractions (1.0 g L-1) and untreated controls. (TIFF 6357 kb)

Supplementary material 2 (DOC 37 kb)

Supplementary material 3 (DOC 47 kb)

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Rayirath, P., Benkel, B., Mark Hodges, D. et al. Lipophilic components of the brown seaweed, Ascophyllum nodosum, enhance freezing tolerance in Arabidopsis thaliana . Planta 230, 135–147 (2009). https://doi.org/10.1007/s00425-009-0920-8

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