NR  Vol.6 No.4 , April 2015
Cross-Effects of Nitrogen and Sulphur Starvation in Chlorella sorokiniana 211/8K
ABSTRACT
Nitrogen (N) and sulphur (S), being essential macronutrients, have important roles in microalgae metabolism. Effects of N- or S-shortage were investigated in the green microalgae Chlorella sorokiniana subjected to 24 h of starvation, by measuring the glutamine synthetase (GS) and O-ace- tylserine(thiol)lyase (OASTL) activities, proteins and amino acids levels. To test possible metabolic impact related to carbon (C) metabolism in response to N- or S-deprivation, starch and total C, N and S contents were also determined. The growth of C. sorokiniana cells was affected by N or S availability. The algae cultured for 24 h in a medium deprived of nitrogen or sulphur showed a decrease in the growth rate and changes in the average volume cell. Nitrogen starvation affected proteins level in the algae cells more than S-deprivation did. The decline in the protein levels observed under S-deficient conditions was coupled with the accumulation of the amide glutamine and with OASTL activity increase; additionally, N-deficiency promoted a decrease in cysteine (Cys) levels (50%) and an increase in GS activity. Nevertheless, S-deprivation had negligible effects on GS activity, while N-deprivation significantly affected OASTL activity. Total C was also estimated in cells N- or S-deprived; nitrogen deprivation strongly affected total C content more than S-deprivation, which in addition reduced the content of C and N, but leaves intact their ratios. Our results support the hypothesis that in Chlorella sorokiniana cells a reciprocal influence of N, S and C assimilation occurs.

Cite this paper
Carfagna, S. , Salbitani, G. , Bottone, C. , Marco, A. and Vona, V. (2015) Cross-Effects of Nitrogen and Sulphur Starvation in Chlorella sorokiniana 211/8K. Natural Resources, 6, 221-229. doi: 10.4236/nr.2015.64020.
References
[1]   Cakmak, T., Angun, P., Ozkan, A.D., Cakmak, Z., Olmez, T.T. and Tekinay, T. (2012) Nitrogen and Sulfur Deprivation Deprivation Differentiate Lipid Accumulation Targets of Chlamydomonas reinhardtii. Bioengineered, 3, 343-346. http://dx.doi.org/10.4161/bioe.21427

[2]   Rigano, C., Di Martino-Rigano, V. and Vona, V. (1981) Nitrate Reductase and Glutamine Synthetase Activities, Nitrate and Ammonium Assimilation, in the Unicellular Alga Cyanidium caldarium. Archives of Microbiology, 129, 110-114. http://dx.doi.org/10.1007/BF00455343

[3]   Vona, V., Di Martino Rigano, V., Esposito, S., Carillo, P., Carfagna, S. and Rigano, C. (1999) Growth, Photosynthesis, and Respiration of Chlorella sorokiniana after N-Starvation. Interactions between Light, CO2 and NH4+ Supply. Physiologia Plantarum, 105, 288-293. http://dx.doi.org/10.1034/j.1399-3054.1999.105214.x

[4]   Ullrich, W.R. (1992) Transport of Nitrate and Ammonium through Plant Membranes. In: Mengel, K. and Pilbeam, D.J., Ed., Nitrogen Metabolism in Plants, Oxford University Press, Oxford, 121-137.

[5]   Hawkesford, M.J. and Belcher, A.R. (1991) Differential Protein Synthesis in Response to Sulfate and Phosphate Deprivation: Identification of Possible Components of Plasma-Membrane Transport Systems in Cultured Tobacco Roots. Planta, 185, 323-329. http://dx.doi.org/10.1007/BF00201051

[6]   Hell, R., Schwenn, J.D. and Bork, C. (1997) Light and Sulphur Sources Modulate mRNA Levels of Several Genes of Sulphate Assimilation. In: Cram, W.J., De Kok, L.J., Stulen, I., Brunold, C. and Rennenberg, H., Eds., Sulphur Metabolism in Higher Plants. Molecular, Ecophysiological and Nutritional Aspects, Backhuys Publishers, Leiden, 181- 185.

[7]   Carfagna, S., Vona, V., Di Martino-Rigano, V., Esposito, S. and Rigano, C. (2011) Nitrogen Assimilation and Cysteine Biosynthesis in Barley: Evidence for Root Sulphur Assimilation upon Recovery from N Deprivation. Environmental and Experimental Botany, 71, 18-24. http://dx.doi.org/10.1016/j.envexpbot.2010.10.008

[8]   Zhang, L., Happe, T. and Melis, A. (2002) Biochemical and Morphological Characterization of Sulfur-Deprived and H2-Producing Chlamydomonas reinhardtii (Green Alga). Planta, 214, 552-561. http://dx.doi.org/10.1007/s004250100660

[9]   Guan, M., Moller, I.S. and Schjoerring, J.K. (2014) Two Cytosolic Glutamine Synthetase Isoforms Play Specific Roles for Seed Germination and Seed Yield Structure in Arabidopsis. Journal of Experimental Botany, 66, 203-212. http://dx.doi.org/10.1093/jxb/eru411

[10]   Rigano, C., Di Martino-Rigano, V., Vona, V., Esposito, S. and Di Martino, C. (1993) Effect of Inhibitors on Ammonium Assimilation in Chlorella sorokiniana in Light and Darkness. Physiologia Plantarum, 89, 602-606. http://dx.doi.org/10.1111/j.1399-3054.1993.tb05221.x

[11]   Salbitani, G., Wirtz, M., Hell, R. and Carfagna, S. (2014) Affinity Purification of O-Acetylserine(thiol)lyase from Chlorella sorokiniana by Recombinant Proteins from Arabidopsis thaliana. Metabolites, 4, 629-639. http://dx.doi.org/10.3390/metabo4030629

[12]   Di Martino-Rigano, V., Vona, V., Carfagna, S., Esposito, S., Carillo, P. and Rigano, C. (2000) Effects of Sulfate- Starvation and Re-Supply on Growth, NH4+ Uptake and Starch Metabolism in Chlorella sorokiniana. Australian Journal of Plant Physiology, 27, 335-342. http://dx.doi.org/10.1071/PP99090

[13]   Gaitonde, M.K. (1967) A Spectrophotometric Method for the Direct Determination of Cysteine in the Presence of Other Naturally Occurring Amino Acids. Biochemical Journal, 104, 627-633.

[14]   Carfagna, S., Salbitani, G., Vona, V. and Esposito, S. (2011) Changes in Cysteine and O-Acetyl-L-serine Levels in the Microalga Chlorella sorokiniana in Response to the S-Nutritional Status. Journal of Plant Physiology, 168, 2188-2195. http://dx.doi.org/10.1016/j.jplph.2011.07.012

[15]   Bradford, M.M. (1976) Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding. Analytical Biochemistry, 72, 248-254. http://dx.doi.org/10.1016/0003-2697(76)90527-3

[16]   Zhang, Y.M., Chen, H., He, C.H. and Wang, Q. (2013) Nitrogen Starvation Induced Oxidative Stress in an Oil-Producing Green Alga Chlorella sorokiniana C3. PloS ONE, 8, e69225. http://dx.doi.org/10.1371/journal.pone.0069225

[17]   Juneja, A., Ceballos, R.M. and Murthy, G.S. (2013) Effects of Environmental Factors and Nutrient Availability on the Biochemical Composition of Algae for Biofuels Production: A Review. Energies, 6, 4607-4638. http://dx.doi.org/10.3390/en6094607

[18]   Hesse, H., Nikiforova, V., Gakiere, B. and Hoefgen, R. (2004) Molecular Analysis and Control of Cysteine Biosynthesis: Integration of Nitrose and Sulphur Metabolism. Experimental Botany, 55, 1283-1292. http://dx.doi.org/10.1093/jxb/erh136

[19]   Haas, F.H., Heeg, C., Queiroz, R., Bauer, A., Wirtz, M. and Hell, R. (2008) Mitochondrial Serine Acetyltransferase Functions as a Pacemaker of Cysteine Synthesis in Plant Cells. Plant Physiology, 148, 1055-1067. http://dx.doi.org/10.1104/pp.108.125237

[20]   Heeg, C., Kruse, C., Jost, R., Gutensohn, M., Ruppert, T., Wirtz, M. and Hell, R. (2008) Analysis of the Arabidopsis O-Acetylserine(thiol)lyase Gene Family Demonstrates Compartment-Specific Differences in the Regulation of Cysteine Synthesis. Plant Cell, 20, 168-185. http://dx.doi.org/10.1105/tpc.107.056747

[21]   Davidian, J.C. and Kopriva, S. (2010) Regulation of Sulfate Uptake and Assimilation—The Same or Not the Same. Molecular Plant, 3, 314-325. http://dx.doi.org/10.1093/mp/ssq001

[22]   Di Martino-Rigano, V., Vona, V., Esposito, S., Carillo, P., Carfagna, S. and Rigano, C. (1998) The Physiological Significance of Light and Dark NH4+ Metabolism in Chlorella sorokiniana. Phytochemistry, 47, 177-181. http://dx.doi.org/10.1016/S0031-9422(97)00569-4

[23]   Wykoff, D.D., Davies, J.P., Melis, A. and Grossman, A.R. (1998) The Regulation of Photosynthetic Electron Transport during Nutrient Deprivation in Chlamydomonas reinharditii. Plant Physiology, 117, 129-139. http://dx.doi.org/10.1104/pp.117.1.129

[24]   Ferreira, R.B.M. and Teixeira, A.R.N. (1992) A Preferential Degradation of Ribulose-Bisphosphate Carboxylase without Plant Death. Journal of Biological Chemistry, 267, 7253-7257.

[25]   Salbitani, G., Vona, V., Bottone, C., Petriccione, M. and Carfagna, S. (2015) Sulfur Deprivation Results in Oxidative Perturbation in Chlorella Sorokininana (211/8k). Plant and Cell Physiology, pii: pcv015. [Epub ahead of print]

 
 
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