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
References
Arnon DI (1949) Copper enzymes in isolated chloroplasts; polyphenoloxidse in Beta vulgaris. Plant Physiol 24:1–15
Ashrafi K, Chang FY, Watts JL, Fraser AG, Kamath RS, Ahringer J, Ruvkun G (2003) Genome-wide RNAi analysis of Caenorhabditis elegans fat regulatory genes. Nature 421:268–272
Blunden G (1991) Agricultural uses of seaweeds and seaweed extracts. In: Guiry MD, Blunden G (eds) Seaweed resources in Europe: uses and potential. Wiley, Chichester, pp 65–81
Bray EA, Bailey-Serres J, Weretilnyk E (2000) Responses to abiotic stresses. In: Gruissem W, Buchannan B, Jones R (eds) Biochemistry and molecular biology of plants. American Society of Plant Biologists, Rockville, pp 1158–1249
Burchett S, Fuller MP, Jellings AJ (1998) Application of seaweed extract improves winter hardiness of winter barley cv Igri. Abstracts—plant cell topics. York Meeting
Crouch I, Van Staden J (1993) Evidence for the presence of plant growth regulators in commercial seaweed products. Plant Growth Regul 13:21–29
Downie B, Dirk LMA, Hadfield KA, Wilkins TA, Bennett AB, Bradford KJ (1998) A gel diffusion assay for quantification of pectin methylesterase activity. Anal Biochem 264:149–157
Featonby-Smith BC, van-Staden J (1983) The effect of seaweed concentrate on the growth of tomatoes in nematode infested soil. Scient Hort 20:137–146
Gilmour SJ, Hajela RK, Thomashow MF (1988) Cold acclimation in Arabidopsis thaliana. Plant Physiol 87:745–750
Guy CL (2003) Freezing tolerance of plants: current understanding and selected emerging concepts. Can J Bot 81:1216–1223
Horváth I, Van Hasselt PR (1985) Inhibition of chilling-induced photooxidative damage to leaves of Cucumis sativus L. by treatment with amino alcohols. Planta 164:83–88
Huner NPA, Oquist G, Hurry VM, Krol M, Falk S, Griffith M (1993) Photosynthesis, photoinhibition and low temperature acclimation tolerant plants in cold. Photosynth Res 37:19–39
Ilker R, Warring AJ, Lyons JM, Breidenbach RW (1976) The cytological responses of tomato seedling cotyledons to chilling and the influence of membrane modifications upon these responses. Protoplasma 90:229–252
Jones PG, Inouye M (1994) The cold shock response: a hot topic. Mol Microbiol 11:811–818
Kamal-Eldin A, Määttä K, Toivo J, Lampi A-M, Piironen V (1998) Acid-catalyzed isomerization of fucosterol and Δ5-avenasterol. Lipids 33:1073–1077
Kariola T, Brader G, Li J, Palva ET (2005) Chlorophyllase 1, a damage control enzyme, affects the balance between defense pathways in plants. Plant Cell 17:282–294
Karpinski S, Gabrys H, Mateo A, Karpinska B, Mullineaux PM (2003) Light perception in plant disease defense signalling. Curr Opin Plant Biol 6:390–396
Lin C, Thomashow MF (1992) A cold-regulated Arabidopsis gene encodes a polypeptide having potent cryoprotective activity. Biochem Biophys Res Commun 183:1103–1108
McConn M, Hugly S, Browse J, Somerville C (1994) A mutation at the fad8 locus of Arabidopsis identifies a second chloroplast ω-3 desaturase. Plant Physiol 106:1609–1614
Metting B, Zimmerman WJ, Crouch T, van-Staden J (1990) Agronomic uses of seaweed and microalgae. In: Akatsuka I (ed) Introduction of applied phycology. SPB Academic, Hague, pp 589–627
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15:473–497
Nabati DA (1991) Response of two grass species to plant growth regulators, fertilizer N, chelated Fe, salinity and water stress. Ph.D. dissertation. Virginia Polytechnic Institute and State University, Blacksburg
Nabati DA, Schmidt RE, Parrish DJ (1994) Alleviation of salinity stress in Kentucky bluegrass by plant growth regulators and iron. Crop Sci 34:198–202
Nakayama K, Okawa K, Kakizaki T, Honma T, Itoh H, Inaba T (2007) Arabidopsis COR15A is a chloroplast stromal protein that has cryoprotective activity and forms oligomers. Plant Physiol 144:513–523
Nishida I, Murata N (1996) Chilling sensitivity in plants and Cyanobacteria: the crucial contribution of membrane lipid. Annu Rev Plant Physiol Plant Mol Biol 47:541–568
Rate DN, Cuenca JV, Bowman GR, Guttman DS, Greenberg JT (1999) The gain-of-function Arabidopsis acd6 mutant reveals novel regulation and function of the salicylic acid signaling pathway in controlling cell death, defenses, and cell growth. Plant Cell 11:1695–1708
Sharma P, Sharma N, Deswal R (2005) The molecular biology of the low-temperature response in plants. Bioessays 27:1048–1059
Spurlock BO, Skinner MS, Kattine AA (1966) A simple rapid method for staining epoxy- embedded specimens for light microscopy with the polychromatic stain paragon-1301. Am J Clin Path 46:252
Spurr AR (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26:31
Steponkus PL, Uemura M, Joseph RA, Gilmour SJ, Thomashow MF (1998) Mode of action of the COR15A gene on the freezing tolerance of Arabidopsis thaliana. Proc Natl Acad Sci USA 95:14570–14575
Stockinger EJ, Gilmour SJ, Thomashow MF (1997) Arabidopsis thaliana CBF1 encodes an AP2 domain-containing transcription activator that binds to the C-repeat/DRE, a cis-acting DNA regulatory element that stimulates transcription in response to low temperature and water deficit. Proc Natl Acad Sci USA 94:1035–1040
Temple WD, Bomke AA (1989) Effects of Kelp (Macrocystis integrifolia and Eklonia maxima) foliar applications on bean crop growth. Plant Soil 117:85–92
Thomashow MF (1994) Arabidopsis thaliana as a model for studying mechanisms of plant cold tolerance. In: Meyerowitz E, Somerville C (eds) Arabidopsis. Cold Spring Harbor Laboratory Press, New York, pp 807–834
Thomashow MF (1999) Plant cold acclimation: freezing tolerance genes and regulatory mechanisms. Annu Rev Plant Physiol Plant Mol Biol 50:571–599
Upchurch RG (2008) Fatty acid unsaturation, mobilization, and regulation in the response of plants to stress. Biotechnol Lett 30:967–977
Wilson S (2001) Frost management in cool climate vineyards. In: University of Tasmania Research Report—UT 99/1. Grape and Wine Research and Development Corporation
Xin Z, Browse B (1998) Eskimo1 mutants of Arabidopsis are constitutively freezing-tolerant. Proc Natl Acad USA 95:7799–7804
Yamaguchi-Shinozaki K, Shinozaki K (1993) Arabidopsis DNA encoding two desiccation-responsive rd29 genes. Plant Physiol 101:1119–1120
Yan J (1993) Influence of plant growth regulators on turfgrass polar lipid composition, tolerance to drought and salinity stresses, and nutrient efficiency. Ph.D. dissertation. Virginia Polytechnic Institute and State University, Blacksburg
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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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)
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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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DOI: https://doi.org/10.1007/s00425-009-0920-8