Abstract
One-year-old seedlings of Bruguiera gymnorrhiza (L) Savingay were exposed to 500 mM NaCl for 6d under hydroponic culture condition to characterize the changes in leaf and thylakoid protein profiles in response to short-term salt exposures. Significant changes in leaf dry mass, chlorophylls and soluble leaf proteins were observed in short term of salt exposures, as it happens under tidal situations in nature. Chlorophyll a/b ratio showed decrease of light harvesting efficiency in salt treatment. Total soluble proteins in leaves were extracted from control and NaCl-treated plants at 2d intervals and were analyzed by SDS-PAGE. Intensity of several protein bands of different molecular mass of leaf protein profile ranging from 10 to 86 kDa (10, 16, 23, 33, 37, 42, 44, 50 and 86 kDa) were decreased due to high salt treatment. Out of these, 16, 23 and 33 kDa protein bands decreased dramatically from 1–3 fold but recovered in 7d growth, except the 33 kDa band. SDSPAGE profile of thylakoid protein revealed that both number and the intensity of several protein bands got altered by salt concentration. However, 33 kDa protein band of thylakoid reappeared in recovery that might not be of the same characteristics with same molecular mass as shown in total leaf protein profile. The numbers of major bands found in SDS-PAGE were reduced when analyzed in urea-SDS-PAGE to minimize protein aggregations by high salt. It was noted that 47 kDa disappeared while some proteins of apparent molecular mass like 23 kDa, 33 kDa, 37 kDa and 50 kDa degraded to minor bands. Partial restoration of protein bands occurred when the salt-treated plants were brought back to initial growth condition. These results clearly demonstrate that short term high salt concentration could cause major alterations to photosynthetic apparatus of a true non salt-secreting tree mangrove Bruguiera gymnorrhiza and adapted against fluctuation of salinity by altering leaf protein pool relatively more than the thylakoid proteins.
Key words: Bruguiera gymnorrhiza, Mangrove, Polypeptides, Salt shock, Sodium chloride, Thylakoid
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Abbreviations
- kDa
kilo Dalton
- SDS
Sodium dodecyl sulfate
- PAGE
Polyacrylamide gel electrophoresis
- MW
Molecular weight
- Tris
Hydroxymethyl amino methane
- TEMED
Tetra ethylene methylene diamine
- Tricine
N-[tris-(hydroxymethyl)-methyl] glycine
- PMSF
Phenylmethylsulphonyl fluoride
Contributor Information
Anath Bandhu Das, Phone: 00-91-674-2557925, FAX: 00-91-674-2550274, Email: a_b_das@hotmail.com.
Prasanna Mohanty, Email: photosis@rediffmail.com.
References
- Agastiana P., Kingsley S.J., Vivekanandan M. Effect of salinity on photosynthesis and biochemical characteristics in mulberry genotypes. Photosynthetica. 2000;38:287–290. doi: 10.1023/A:1007266932623. [DOI] [Google Scholar]
- Behera B., Das A.B. Response of Antioxident defense system of Bruguiera gymnorrhiza under short-term exposure to high salt. In: Gupta N., Mohapatra A.K., editors. Proceedings of the symposium on Wetland and Mangrove Biodiversity in Orissa Coast. Bhubaneswar: RPRC; 2008. pp. 44–48. [Google Scholar]
- Bradford M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 1976;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Cheeseman J.M. Mechanism of salinity tolerance in plants. Plant Physiol. 1988;87:547–550. doi: 10.1104/pp.87.3.547. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Das A.B., Mishra S., Mohanty P. Physiology of mangroves: strategies on stress adaptation with special emphasis on tolerance to high salinity. In: Hemantaranjan A., editor. Advances in Plant Physiology. Jodhpur: Scientific Publishers; 2006. pp. 101–154. [Google Scholar]
- Gadallah M.A.A. Effects of proline and glycinebetaine on Vicia faba response to salt stress. Biol. Plant. 1999;42:249–257. doi: 10.1023/A:1002164719609. [DOI] [Google Scholar]
- Greenway H., Munns R. Mechanisms of salttolerance in non-halophytes. Ann. Rev. Plant Physiology. 1980;31:149–190. doi: 10.1146/annurev.pp.31.060180.001053. [DOI] [Google Scholar]
- Hernandez J., Jimenez A., Mullineaux P., Seville F. Tolerance of pea plants (Pisum sativum) to long-term salt stress is associated with induction of antioxidant defense. Plant Cell Environ. 2001;23:853–862. doi: 10.1046/j.1365-3040.2000.00602.x. [DOI] [Google Scholar]
- Hogarth P.J. The Biology of mangroves. London: Academic Press; 1999. [Google Scholar]
- Hoagland D.R., Arnon D.I. Crop production in artificial solutions and in soil with special reference to factors influencing yields and absorption of inorganic nutrients. Soil Science. 1940;50:463–471. [Google Scholar]
- Jitesh M.N., Prashanth S.R., Sivaprakash K.R., Parida A.K. Anioxidant response mechanisms in halophytes: their role in stress defense. J. Genet. 2006;85:237–254. doi: 10.1007/BF02935340. [DOI] [PubMed] [Google Scholar]
- Kathiresan K., Moorthy P. Influence of different irradiance on growth and photosynthetic characteristics in seedlings of Rhizophora species. Photosynthetica. 1994;29:143–146. [Google Scholar]
- Khan M.A., Ungar I.A., Showwalter A.M. The effects of salinity on growth, water status and ion content of a leaf succulent perennial halophyte Sueada fruticosa (L.) Forssk. Journal of Aried Environment. 2000;45:73–84. doi: 10.1006/jare.1999.0617. [DOI] [Google Scholar]
- Laemmli U.K. Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature. 1970;227:680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Lichtenthaler H.K. Chlorophylls and caratinoidspigments of photosynthetic biomembranes. In: Clockwise S.P., Kaplan N.O., editors. Methods in Enzymology. San Diego-New York-Bereley-Boston-London-Sydney-Tokyo-Toronto: Academic Press; 1987. pp. 350–382. [Google Scholar]
- Lowry O.H., Rosebrought N.J., Farr A.L., Randall R.J. Protein measurement with the Folin Phenol reagent. J. Biol. Chem. 1951;193:256–257. [PubMed] [Google Scholar]
- Mitra B., Das A.B., Das T.K., Parida A., Mishra S., Mohanty P. A salt responsive leaf protein, SSP23 in tree mangrove Bruguiera: Localization in leaf tissue by immunogold labeling. J. Plant. Biol. 2006;33(1&2):135–138. [Google Scholar]
- Morales M.A., SancheoBlanco M.J., Olmos E., Torrechlas A., Alarcon J.J. Changes in the growth, leaf water relations and all ultrastructure in Argyranthemum coronopifolium plants under saline conditions. J. Plant Physiol. 1998;153:174–180. [Google Scholar]
- Nakatani H.Y., Barber J. An improved method for isolating chloroplast retaining their outer membranes. Biochim. Biophys. Acta. 1977;416:510–512. [PubMed] [Google Scholar]
- Nandy P., Ghose M. Estimation of osmotic potential and free amino acids in some mangroves of Sundarbans, India. Acta Botanica Croatica. 2003;62(1):37–45. [Google Scholar]
- Nandy P., Das S., Ghose M. Relation of leaf micromorphology with photosynthesis and water efflux in some Indian mangroves. Acta Botanica Croatica. 2005;64(2):331–340. [Google Scholar]
- Parida A., Das A.B., Das P. NaCl stress causes changes in photosynthetic pigments, proteins and other metabolic components in the leaves of a true mangrove, Bruguiera parviflora, in hydroponic cultures. J. Plant Biol. 2002;45:28–36. doi: 10.1007/BF03030429. [DOI] [Google Scholar]
- Parida A., Das A.B., Mohanty P. Deffense potentials to NaCl in a mangrove, Bruguiera parviflora: Differential changes of isoforms of some antioxidative enzymes. J. Plant Physiol. 2004;161:531–542. doi: 10.1078/0176-1617-01084. [DOI] [PubMed] [Google Scholar]
- Parida A.K., Das A.B. Salt tolerance and salinity effects on plants: a review. Ecotoxicology and Environmental Safety. 2005;60:324–349. doi: 10.1016/j.ecoenv.2004.06.010. [DOI] [PubMed] [Google Scholar]
- Parida A., Mittra B., Das A.B., Das T.K., Mohanty P. High salinity reduces the content of abundant 23-kDa protein of the mangrove Bruguiera parviflora. Planta. 2005;221:135–140. doi: 10.1007/s00425-004-1415-2. [DOI] [PubMed] [Google Scholar]
- Selvam V., Gnanappazham L., Navamuniyammal M., Ravichandran K.K., Karunagaran V.M. Atlas of mangrove wetlands of India Part I Tamil Nadu, M.S. Swaminathan Research Foundation, Chennai, India. Chromepet, Chennai: TTK Healthcare ltd.; 2002. pp. 1–100. [Google Scholar]
- Sokal R.R., Rohlf F.J. Biometry: The Principles and Practice of Statistics in Biological Research. 3. New York: W.H. Freeman and Company; 1995. pp. 321–356. [Google Scholar]
- Sugihara K., Hanagata N., Dubinsky Z., Baba S., Karube J. Characterization of cDNA encoding oxygen evolving enhancer protein 1 increased by salt treatment in the mangrove Bruguiera gymnorrhiza. Plant Cell Physiol. 2000;41:1279–1285. doi: 10.1093/pcp/pcd061. [DOI] [PubMed] [Google Scholar]
- Takemura T., Hanagata N., Sugihara K., Baba S., Karube I., Dubinsky Z. Physiological and biochemical responses to salt stress in the mangrove. Bruguiera gymnorrhiza. Aquat. Bot. 2000;68:15–28. doi: 10.1016/S0304-3770(00)00106-6. [DOI] [Google Scholar]
- Wang Y., Nil N. Changes in chlorophyll, ribulose biphosphate carboxylase-oxygenase, glycine betaine content, photosynthesis and transpiration in Amaranthus tricolor leaves during salt stress. J. Hort. Sci. Biotech. 2000;75:632–627. [Google Scholar]
