Skip to main content

Nitrogen metabolism-related enzymes in Mesembryanthemum crystallinum after Botrytis cinerea infection


We compared C3 and CAM (crassulacean acid metabolism) states in Mesembryanthemum crystallinum, a facultative CAM species, with respect to the involvement of phosphoenolpyruvate carboxylase (PEPC) and nitrogen metabolismrelated enzymes in plant response to Botrytis cinerea infection. The enzyme activities were monitored both in pathogeninoculated 2nd leaf pair and non-inoculated 3rd leaf pair. The control activities of most studied enzymes were dependent on the mode of photosynthesis. Compared to C3 plants, those performing CAM exhibited higher PEPC, nitrate reductase (NR), and deaminating glutamate dehydrogenase (NAD-GDH) activities but lower glutamine synthetase (GS) and alanine aminotransferase (ALT) activities. Regardless of the mode of photosynthetic carbon assimilation, the plants responded to infection with enhancement of PEPC and inhibition of NR activities in the inoculated leaves. Whereas the activity of GS remained unaffected, those of all glutamate-yielding enzymes, namely ferredoxin-dependent glutamate synthase (Fd-GOGAT), aspartate aminotransferase (AST), ALT, and aminating glutamate dehydrogenase (NADHGDH) were altered after infection. However, the time-course and extent of the observed changes differed in C3 and CAM plants. In general, CAM plants responded to infection with an earlier increase in PEPC and Fd-GOGAT activities as well as later inhibition of NR activity. Contrary to C3 plants, in those performing CAM the activities of PEPC, Fd-GOGAT, NADH-GDH, and AST in the non-inoculated 3rd leaf pair were similarly influenced by infection as in leaves directly inoculated with the pathogen. This implies that the local infection induced an alteration of carbon/nitrogen status in healthy upper leaves. This reprogramming resulting from changes in PEPC and nitrogen metabolism-related enzymes was C3- and CAM-specific.

This is a preview of subscription content, access via your institution.



alanine aminotransferase


aspartate aminotransferase


crassulacean acid metabolism


day after inoculation


ethylenediaminetetraacetic acid


ferredoxin-dependent glutamate synthase


glutamine synthetase


NAD-dependent glutamate dehydrogenase


NADH-dependent glutamate dehydrogenase


NADH-dependent glutamate synthase


nitrite reductase


nitrate reductase


phosphoenolpyruvate carboxylase

TCA cycle:

tricarboxylic acid cycle


  • Aubry, S., Brown, N.J., Hibberd, J.M.: The role of proteins in C(3) plants prior to their recruitment into the C(4) pathway. - J. exp. Bot. 62: 3049–3059, 2011.

    Article  PubMed  CAS  Google Scholar 

  • Bilgin, D.D., Zavala, J., Zhu, J., Clough, S.J., Or, D.R., DeLucia, E.H.: Biotic stress globally downregulates photosynthesis genes. - Plant Cell Environ. 33: 1597–1613, 2010.

    Article  PubMed  CAS  Google Scholar 

  • Bolton, M.D., Thomma, B.P.H.J.: The complexity of nitrogen metabolism and nitrogen-regulated gene expression in plant pathogenic fungi. - Physiol. mol. Plant Pathol. 72: 104–110, 2008.

    Article  CAS  Google Scholar 

  • Botrel, A., Kaiser, W.M.: Nitrate reductase activation state in barley roots in relation to the energy and carbohydrate status. - Planta 201: 496–501, 1997.

    Article  PubMed  CAS  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. 72: 248–254, 1976.

    Article  PubMed  CAS  Google Scholar 

  • Cebeci, O., Kokturk, B., Ergen, N., Ozturk, L., Cakmak, I., Budak, H.: Differential expression of wheat transcriptomes in response to varying cadmium concentrations. - Biol. Plant. 52: 703–708, 2008.

    Article  CAS  Google Scholar 

  • Champigny, M.L.: Integration of photosynthetic carbon and nitrogen metabolism in higher plants. - Photosynth. Res. 46: 117–127, 1995.

    Article  PubMed  CAS  Google Scholar 

  • Chang, Y.-M., Liu, W.-Y., Shih, A.C.-C., Shen, M.-N., Lu, C.-H., Lu, M.-Y.J., Yang, H.-W., Wang, T.-Y., Chen, S.C.-C., Chen, S.M., Li, W.-H., Ku, M.S.B.: Characterizing regulatory and functional differentiation between maize mesophyll and bundle sheath cells by transcriptomic analysis. - Plant Physiol. 160: 165–177, 2012.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Coruzzi, G.M., Zhou, L.: Carbon and nitrogen sensing and signaling in plants: emerging “matrix effects”. - Curr. Opin. Plant Biol. 4: 247–253, 2001.

    Article  PubMed  CAS  Google Scholar 

  • Delgado-Alvarado, A., Walker, R.P., Leegood, R.C.: Phosphoenolpyruvate carboxykinase in developing pea seeds is associated with tissues involved in solute transport and is nitrogen-responsive. - Plant Cell Environ. 30: 225–235, 2007.

    Article  PubMed  CAS  Google Scholar 

  • Doubnerová, V., Ryšlavá, H.: What can enzymes of C4 photosynthesis do for C3 plants under stress? - Plant Sci. 180: 575–583, 2011.

    Article  PubMed  CAS  Google Scholar 

  • Foyer, C.H., Noctor, G.: Photosynthetic nitrogen assimilation: inter-pathway control and signaling. - In: Foyer, C.H., Noctor, G., (ed.): Photosynthetic Nitrogen Assimilation and Associated Carbon and Respiratory Metabolism, Advances in Photosynthesis and Respiration. Pp. 1–22. Kluwer Academic Publishers, Dordrecht 2002.

    Google Scholar 

  • Gabara, B., Kuźniak, E., Skłodowska, M., Surówka, E., Miszalski, Z.: Ultrastructural and metabolic modifications at the plant-pathogen interface in Mesembryanthemum crystallinum leaves infected by Botrytis cinerea. - Environ. exp. Bot. 77: 33–43, 2012.

    Article  CAS  Google Scholar 

  • Gajewska, E., Niewiadomska, E., Tokarz, K., Słaba, M., Skłodowska, M.: Nickel-induced changes in carbon metabolism in wheat shoots. - J. Plant Physiol. 170: 369–377, 2013.

    Article  PubMed  CAS  Google Scholar 

  • Gajewska, E., Skłodowska, M.: Nickel-induced changes in nitrogen metabolism in wheat shoots. - J. Plant Physiol. 166: 1034–1044, 2009.

    Article  PubMed  CAS  Google Scholar 

  • Gao, S., Liu, K.-T., Chung, T.-W., Chen, F.: The effects of NaCl stress on Jatropha cotyledon growth and nitrogen metabolism. - J. Soil Sci. Plant Nutr. 13: 99–113, 2013.

    Google Scholar 

  • Gawronska, K., Niewiadomska, E.: Participation of citric acid and isocitric acid in the diurnal cycle of carboxylation and decarboxylation in the common ice plant. - Acta Physiol. Plant. 37: 61 doi:10.1007/s11738-015-1807-x, 2015.

    Article  CAS  Google Scholar 

  • Kuźniak, E., Gabara, B., Skłodowska, M., Libik-Konieczny, M., Miszalski, Z.: Effects of NaCl on the response of the halophyte Mesembryanthemum crystallinum callus to Botrytis cinerea infection. - Biol. Plant. 55: 423–430, 2011.

    Article  CAS  Google Scholar 

  • Kuźniak, E., Kaźmierczak, A., Wielanek, M., Głowacki, R., Kornas, A.: Involvement of salicylic acid, glutathione and protein S-thiolation in plant cell death-mediated defence response of Mesembryanthemum crystallinum against Botrytis cinerea. - Plant Physiol. Biochem. 63: 30–38, 2013.

    Article  PubMed  CAS  Google Scholar 

  • Kuźniak, E., Kornas, A., Gabara, B., Ullrich, C., Skłodowska, M., Miszalski, Z.: Interaction of Botrytis cinerea with the intermediate C3-CAM plant Mesembryanthemum crystallinum. - Environ. exp. Bot. 69: 137–147, 2010.

    Article  CAS  Google Scholar 

  • Kuźniak, E., Kornas, A., Kaźmierczak, A., Rozpądek, P., Nosek, M., Kocurek, M., Zellnig, G., Müller, M., Miszalski, Z.: Photosynthesis-related characteristics of the midrib and the interveinal lamina in leaves of C3-CAM intermediate plant Mesembryanthemum crystallinum. - Ann. Bot. 117: 1141–1151, 2016.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Lea, P.J., Miflin, B.J.: Glutamate synthase and the synthesis of glutamate in plants. - Plant Physiol. Biochem. 41: 555–564, 2003.

    Article  CAS  Google Scholar 

  • Leegood, R.C., Walker, R.P.: Regulation and roles of phosphoenolpyruvate carboxykinase in plants. - Arch. Biochem. Biophys. 414: 204–210, 2003.

    Article  PubMed  CAS  Google Scholar 

  • Libik-Konieczny, M., Surówka, E., Kuźniak, E., Nosek, M., Miszalski, Z.: Photosynthetic metabolism influences the direction of changes in antioxidant system activity in Mesembryanthemum crystallinum leaves inoculated with Botrytis cinerea or Pseudomonas syringae. - J. Plant Physiol. 168: 1052–1059, 2011.

    Article  PubMed  CAS  Google Scholar 

  • Liu, G., Ji, Y., Bhuiyan, N.H., Pilot, G., Selvaraj, G., Zou, J., Wei, Y.: Amino acid homeostasis modulates salicylic acidassociated redox status and defense responses in Arabidopsis. - Plant Cell 22: 3845–3863, 2010.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Masclaux-Daubresse, C., Daniel-Vedele, F., Dechorgnat, J., Chardon, F., Gaufichon, L., Suzuki, A.: Nitrogen uptake, assimilation and remobilization in plants: challenges for sustainable and productive agriculture. - Ann. Bot. 105: 1141–1157, 2010.

    Article  PubMed  PubMed Central  Google Scholar 

  • Miyashita, Y., Good, A.G.: NAD(H)-dependent glutamate dehydrogenase is essential for the survival of Arabidopsis thaliana during dark-induced carbon starvation. - J. exp. Bot. 59: 667–680, 2008.

    Article  PubMed  CAS  Google Scholar 

  • Müller, C., Scheible, W., Stitt, M., Krapp, A.: Influence of malate and 2-oxoglutarate on the NIA transcript level and nitrate reductase activity in tobacco leaves. - Plant Cell Environ. 24: 191–203, 2001.

    Article  Google Scholar 

  • Nunes-Nesi, A., Fernie, A.R., Stitt, M.: Metabolic and signaling aspects underpinning the regulation of plant carbon nitrogen interactions. - Mol. Plant 3: 973–996, 2010.

    Article  PubMed  CAS  Google Scholar 

  • Rojas, C.M., Senthil-Kumar, M., Tzin, V., Mysore, K.S.: Regulation of primary plant metabolism during plantpathogen interactions and its contribution to plant defense. - Front. Plant Sci. 5: 17. doi:10.3389/fpls.2014.00017, 2014.

    Article  PubMed  PubMed Central  Google Scholar 

  • Ryšlavá, H., Müller, K., Semorádová, Š., Synková, H., Čeřovská, N.: Photosynthesis and activity of phosphoenolpyruvate carboxylase in Nicotiana tabacum L. leaves infected by Potato virus A and Potato virus Y. - Photosynthetica 41: 357–363, 2003.

    Article  Google Scholar 

  • Sánchez, R., Flores, A., Cejudo, F.J.: Arabidopsis phosphoenolpyruvate carboxylase genes encode immunologically unrelated polypeptides and are differentially expressed in response to drought and salt stress. - Planta 223: 901–909, 2006.

    Article  PubMed  CAS  Google Scholar 

  • Scharte, J., Schön, H., Weis, E.: Photosynthesis and carbohydrate metabolism in tobacco leaves during an incompatible interaction with Phytophthora nicotianae. - Plant Cell Environ. 28: 1421–1435, 2005.

    Article  CAS  Google Scholar 

  • Schultz, J.C., Appel, H.M., Ferrieri, A.P., Arnold, T.M.: Flexible resource allocation during plant defense responses. Front. Plant Sci. 4: 324, 2013.

    Article  PubMed  PubMed Central  Google Scholar 

  • Seifi, H.S., Van Bockhaven, J., Angenon, G., Höfte, M.: Glutamate metabolism in plant disease and defense: friend or foe? - Mol. Plant Microbe Interact. 26: 475–485, 2013.

    Article  PubMed  CAS  Google Scholar 

  • Skopelitis, D.S., Paranychianakis, N.V., Paschalidis, K.A., Pliakonis, E.D., Delis, I.D., Yakoumakis, D.I., Kouvarakis, A., Papadakis, A.K., Stephanou, E.G., Roubelakis- Angelakis, K.A.: Abiotic stress generates ROS that signal expression of anionic glutamate dehydrogenases to form glutamate for proline synthesis in tobacco and grapevine. - Plant Cell 18: 2767–2781, 2006.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Snoeijers, S.S., Pérez-García, A., Joosten, M.H.A.J., De Wit, P.J.G.M.: The effect of nitrogen on disease development and gene expression in bacterial and fungal plant pathogens. - Eur. J. Plant Pathol. 106: 493–506, 2000.

    Article  CAS  Google Scholar 

  • Song, J.T., Lu, H., McDowell, J.M., Greenberg, J.T.: A key role for ALD1 in activation of local and systemic defenses in Arabidopsis. - Plant J. 40: 200–212, 2004.

    Article  PubMed  CAS  Google Scholar 

  • Stewart, G.R., Rhodes, D.: Nitrogen metabolism of halophytes. - New Phytol. 80: 307–316, 1978.

    Article  CAS  Google Scholar 

  • Sun, W., Huang, A., Sang, Y., Fu, Y., Yang, Z.: Carbonnitrogen interaction modulates plant growth and expression of metabolic genes in rice. - J. Plant Growth Regul. 32: 575–584, 2013.

    Article  CAS  Google Scholar 

  • Vlot, A.C., Klessig, D.F., Park, S.-W.: Systemic acquired resistance: the elusive signal(s). - Curr. Opin. Plant Biol. 11: 436–442, 2008.

    Article  PubMed  CAS  Google Scholar 

  • Zeier, J.: New insights into the regulation of plant immunity by amino acid metabolic pathways. - Plant Cell Environ. 36: 2085–2103, 2013.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations


Corresponding author

Correspondence to E. Gajewska.

Additional information

Acknowledgments: This work was supported by the Ministry of Science and Higher Education (grant 830/1/N-COST/2010/11).

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Gajewska, E., Surówka, E., Kornas, A. et al. Nitrogen metabolism-related enzymes in Mesembryanthemum crystallinum after Botrytis cinerea infection. Biol Plant 62, 579–587 (2018).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

Additional key words

  • crassulacean acid metabolism
  • glutamate dehydrogenase
  • grey mould
  • ice plant
  • nitrate reductase
  • phosphoenolpyruvate carboxylase