Russian Journal of Plant Physiology

, Volume 65, Issue 4, pp 588–597 | Cite as

Activities of Adenylate Cyclase and Changes in cAMP Concentration in Root Cells of Pea Seedlings Infected with Mutualists and Phytopathogens

  • L. A. LomovatskayaEmail author
  • O. V. Kuzakova
  • A. S. Romanenko
  • A. M. Goncharova
Research Papers


Work was carried out on pea (Pisum sativum L.) seedling roots to assess the attachment of the nitrogen-fixing symbiotic bacteria Rhizobium leguminosarum bv. vicea (Rlv) and the bacterial phytopathogens—specific Pseudomonas syringae pv. pisi (Psp) and nonspecific Clavibacter michiganensis ssp. sepedonicus (Cms). Different root zones were examined: (I) the meristem, 2 mm from the root tip; (II) the root hair-free zone, 27 mm; (III) the zone of root hair anlages, 712 mm; (IV) the young root hair zone, 1217 mm; and (V) the zone of root hair that completed the growth, 1722 mm. It was found earlier that the zones differed in their susceptibility to Rlv. In the present work, reactions of particular components of the adenylate cyclase signaling system (ACSS) were estimated, i.e., concentration of cAMP and activities of transmembrane adenylate cyclase (tAC) and soluble adenylate cyclase (sAC) in these zones after different times post inoculation (5, 15, 120, and 360 min). It was revealed that the degree of activation of particular components of ACSS did not depend on the sorption rate of differently specialized bacteria. Upon contact with Rlv, the character of changes in tAC and sAC activities was almost the same in different root zones and resembled the dynamics of the cAMP content. Inoculation with Psp changed the cAMP level similarly to that with Rlv, but the dynamics of tAC and sAC was opposite to each other in most cases. Inoculation with Cms, in spite of the absence of its attachment, elevated the cAMP content and activated tAC and sAC. It is suggested that the above-mentioned changes in ACSS is associated with exometabolites of Rlv, Psp, and Cms, which activate the PAMP-induced immunity of the pea seedling cells. The uniform dynamics of cAMP in different root zones upon the exposure to Rlv and Psp seems to reflect the specific reaction and, presumably, fulfills different functions—regulatory with Rlv and defensive with Psp. Upon short-term contact with Cms, the cAMP dynamics in the same root zones displayed a nonspecific character that might be related to the rate of adsorption of exopolysaccharides by the root hair. The systemic response of ACSS was observed in the hypocotyls of the seedlings exposed to any of the three organisms.


Pisum sativum Rhizobium leguminosarum bv. vicea Pseudomonas syringae pv. pisi Clavibacter michiganensis ssp. sepedonicus adenylate cyclase cAMP root hair zones sorption 



adenylate cyclase signaling system


colony-forming units


Clavibacter michiganensis ssp. sepedonicus




microbial-associated molecular patterns


Pseudomonas syringae pv. pisi


Rhizobium leguminosarum bv. vicea


soluble adenylate cyclase


transmembrane adenylate cyclase


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Makarova, L.E. and Nurminskii, V.N., Temperature impact on localization of “free” phenolic compounds in the root tissues and deformation of root hairs in pea seedlings inoculated by Rhizobium, Tsitologiya, 2005, vol. 47, pp. 519–525.Google Scholar
  2. 2.
    Glyan'ko, A.K., Immunity of a leguminous plant infected by nodular bacteria Rhizobium spp. F.: Review, Appl. Biochem. Microbiol., 2017, vol. 53, pp. 140–148.CrossRefGoogle Scholar
  3. 3.
    Granqvist, E., Sun, J., Camp, R.O., Pujic, P., Hill, L., Normand, P., Morris, R.J., Downie, A.J., Geurts, R., and Oldroyd, G.E.D., Bacterial-induced calcium oscillations are common to nitrogen-fixing associations of nodulating legumes and non-legumes, New Phytol., 2015, vol. 207, pp. 551–558.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Terakado, I., Fujihara, S., and Yoneyama, T., Changes in cyclic nucleotides during nodule formation, Soil Sci. Plant Nutr., 2003, vol. 49, pp. 459–462.CrossRefGoogle Scholar
  5. 5.
    Lomovatskaya, L.A., Romanenko, A.S., and Filinova, N.V., Plant adenylate cyclases: the effect of the biotic stressor on the kinetic parameters of the transmembrane and “soluble” forms of adenylate cyclase, Biol. Membr., 2014, vol. 31, pp. 129–136.Google Scholar
  6. 6.
    Lomovatskaya, L.A., Romanenko, A.S., Filinova, N.V., and Dudareva, L.V., Determination of cAMP in plant cells by a modified enzyme immunoassay method, Plant Cell Rep., 2011, vol. 30, pp. 125–132.CrossRefPubMedGoogle Scholar
  7. 7.
    Soto, M.J., Domínguez-Ferreras, A., Pérez-Mendoza, D., Sanjuán, J., and Olivares, J., Mutualism versus pathogenesis: the give-and-take in plant–bacteria interactions, Cell Microbiol., 2009, vol. 11, pp. 381–388.CrossRefPubMedGoogle Scholar
  8. 8.
    Jeandroz, S., Lamotte, O., Astier, J., Rasul, S., Trapet, P., Besson-Bard, A., Bourque, S., Nicolas-Frances, V., Berkowitz, G.A., and Wendehenne, D., There’s more to the picture than meets the eye: nitric cross talk with Ca2+ signaling, Plant Physiol., 2013, vol. 163, pp. 459–470.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Turnbull, G.A., Morgan, A.W., Whipps, J.M., and Saunders, J.R., The role of mobility in the in vitro attachment of Pseudomonas putida Pa W8 to wheat roots, FEMS Microbiol. Ecol., 2001, vol. 35, pp. 57–65.CrossRefPubMedGoogle Scholar
  10. 10.
    Shafikova, T.N. and Omelichkina, Yu.V., Molecular–genetic aspects of plant immunity to phytopathogenic bacteria and fungi, Russ. J. Plant Physiol., 2015, vol. 62, pp. 571–585.CrossRefGoogle Scholar
  11. 11.
    Jiang, J., Fan, L.W., and Wu, W.H., Evidences for involvement of endogenous cAMP in Arabidopsis defense responses to Verticillium toxins, Cell Res., 2005, vol. 15, pp. 585–592.CrossRefPubMedGoogle Scholar
  12. 12.
    Ma, W., Qi, Z., Smigel, A., Walker, R.K., Verma, R., and Berkowitz, G.A., Ca2+, cAMP, and transduction of non-self perception during plant immune responses, Proc. Natl. Acad. Sci. USA, 2009, vol. 106, pp. 20995–21000.CrossRefPubMedGoogle Scholar
  13. 13.
    Zhao, J., Guo, Y., Fujita, K., and Sakai, K., Involvement of cAMP signaling in elicitor-induced phytoalexin accumulation in Cupressus lusitanica cell cultures, New Phytol., 2003, vol. 161, pp. 723–731.CrossRefGoogle Scholar
  14. 14.
    Bindschedler, L.V., Minibayeva, F., Gardner, S.L., Gerrish, C., Davies, D.R., and Bolwell, G.P., Early signalling events in the apoplastic oxidative burst in suspension cultured French bean cells involve camp and Ca2+, New Phytol., 2001, vol. 151, pp. 185–194.CrossRefGoogle Scholar
  15. 15.
    Saeki, K., Rhizobial measures to evade host defense strategies and endogenous threats to persistent symbiotic nitrogen fixation: a focus on two legume rhizobium model systems, Cell. Mol. Life Sci., 2011, vol. 68, pp. 1327–1339.CrossRefPubMedGoogle Scholar
  16. 16.
    Tampakaki, A., Commonalities and differences of T3SSs in rhizobia and plant pathogenic bacteria, Front. Plant Sci., 2014, vol. 27, no. 5: 114. doi 10.3389/fpls.2014.00114Google Scholar
  17. 17.
    Gourion, B., Berrabah, F., Ratet, P., and Stacey, G., Rhizobium–legume symbioses: the crucial role of plant immunity, Trends Plant Sci., 2015, vol. 20, pp. 186–194.CrossRefPubMedGoogle Scholar
  18. 18.
    Fliegmann, J. and Bono, J.J., Lipo-chitooligosaccharidic nodulation factors and their perception by plant receptors, Glycoconj. J., 2015, vol. 32, pp. 455–464. doi 10.1007/s10719-015-9609-3CrossRefPubMedGoogle Scholar
  19. 19.
    Nelson, M.S. and Sadowsky, M.J., Secretion systems and signal exchange between nitrogen-fixing rhizobia and legumes, Front. Plant Sci., 2015, vol. 6: 491. doi 10.3389/fpls.2015.00491CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Steegborn, C., Structure, mechanism, and regulation of soluble adenylyl cyclases—similarities and differences to transmembrane adenylyl cyclases, Biochim. Biophys. Acta, 2014, vol. 1842, pp. 2535–2547.CrossRefPubMedGoogle Scholar
  21. 21.
    Shpakov, A.O., Plant chemosignal systems, Tsytologia, 2009, vol. 51, pp. 721–734.Google Scholar
  22. 22.
    Lomovatskaya, L.A., Romanenko, A.S., and Filinova, N.V., Adenilattsiklazy i ustoichivost' rastenii k stressam (Adenylate Cyclases and Plant Resistance to Stress), Irkutsk: Inst. Geogr. SO RAN, 2010.Google Scholar
  23. 23.
    Libault, M., Farmer, A., Brechenmacher, L., Drnevich, J., Landley, R.L., Bilgin, D.D., Radwan, O., Neece, D.J., Clough, S.J., May, G.D., and Stacey, G., Complete transcriptome of the soybean root hair cell, a single-cell model, and its alteration in response to Bradyrhizobium japonicum infection, Plant Physiol., 2010, vol. 152, pp. 541–552.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Lohar, D.P., Sharopova, N., Endre, G., Penuela, S., Samac, D., Town, C., Silverstein, K.A.T., and van den Bosch, K.A., Transcript analysis of early nodulation events in Medicago truncatula, Plant Physiol., 2006, vol. 140, pp. 221–234.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Ichinose, Y., Taguchi, F., and Mukaihara, T., Pathogenicity and virulence factors of Pseudomonas syringae, J. Gen. Plant Pathol., 2013, vol. 79, pp. 285–296.CrossRefGoogle Scholar
  26. 26.
    Zheng, X.Y., Spivey, N.W., Zeng, W., Liu, P.P., Fu, Q., Klessig, D.F., He, H.Y., and Dong, X., Coronatine promotes Pseudomonas syringae virulence in plants by activating a signaling cascade that inhibits salicylic acid accumulation, Cell Host Microbe, 2012, vol. 11, pp. 587–596.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Holtsmark, I., Takle, G.W., and Brurberg, M.B., Expression of putative virulence factors in the potato pathogen Clavibacter michiganensis subsp. sepedonicus during infection, Arch. Microbiol., 2008, vol. 189, pp. 131–139.CrossRefPubMedGoogle Scholar
  28. 28.
    Wroblewski, T., Caldwell, K.S., Piskurewicz, U., Cavanaugh, K.A., Xu, H., Kozik, A., Ochoa, O., McHale, L.K., Lahre, K., Jelenska, J., Castillo, J.A., Blumenthal, D., Vinatzer, B.A., Greenberg, J.T., and Michelmore, R.W., Comparative large-scale analysis of interactions between several crop species and the effector repertoires from multiple pathovars of Pseudomonas and Ralstonia, Plant Physiol., 2009, vol. 150, pp. 1733–1749. CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Omelichkina, Yu.V., Boyarkina, S.V., and Shafikova, T.N., Effector-activated immune responses in potato and tobacco cell cultures caused by phytopathogen Clavibacter michiganensis ssp. sepedonicus, Russ. J. Plant Physiol., 2017, vol. 64, pp. 423–430.CrossRefGoogle Scholar
  30. 30.
    Romanenko, A.C., Kustov, M.L., Graskova, I.A., Rifel’, A.A., Konenkina, T.A., and Salyaev, R.K., The role of pH, reception and endocytosis in nonspecific and specific immunity of plants, Dokl. Akad. Nauk, 1994, vol. 338, no. 2, pp. 275–277.Google Scholar

Copyright information

© Pleiades Publishing, Ltd. 2018

Authors and Affiliations

  • L. A. Lomovatskaya
    • 1
    Email author
  • O. V. Kuzakova
    • 1
  • A. S. Romanenko
    • 1
  • A. M. Goncharova
    • 1
  1. 1.Siberian Institute of Plant Physiology and Biochemistry, Siberian BranchRussian Academy of SciencesIrkutskRussia

Personalised recommendations