The symbiotic effect of Piriformospora indica on induced resistance against bakanae disease in rice (Oryza sativa L.)

Document Type: Research Paper


1 Agricultural Biotechnology, University of Guilan, Iran

2 Department of Biotechnology, University of Guilan, Iran

3 Department of plant protection, Sari Agricultural Sciences and Natural Resources University, Iran

4 Agricultural biotechnology,University of Guilan, Iran


The root endophytic fungus, Piriformospora indica, colonizes roots of a large number of plant species including Cereals and Brasicaceae. There are several reports indicating that P. indica protects roots from different path- ogens. In the present study, rice plants were pre-inoculated with P. indica and were subsequently infected with Fusarium proliferatum, as the causal agent of root rot and crown rot (Bakanae) disease. The effects of P. indica on the protection of rice plants against F. proliferatum were evaluated. Transcription levels of some pathogenesis-related genes such as NPR1, PR1, PR4 and PR5, as well as two transcriptional factor genes namely WRKY62 and WRKY85 were quantified via quantitative real time PCR in rice roots. Pre-inoculated and challenged rice plants showed mild disease symptoms with two weeks delay compared to non P. indica colonized plants. Quantification of Fusarium in roots via q-PCR con- firmed the higher quantity of the fungi in non pre-inoculated plants. Expression analysis revealed a faster induction of pathogenesis related genes following F. proliferatum inoculation of pre-inoculated plants i.e. priming of these genes is a part of mechanism of P. indica induced systemic resistance.


Main Subjects

[1]      Achatz, B., Von Ruden, S., Andrade, D., Neumann, E., Pons-Kuhnemann, J., Kogel, K., Franken, P. and Waller, F. 2010. Root colonization by Piriformospora indica enhances grain yield in barley under diverse nutrient regimes by accelerating plant development. J Plant Soil, 333:59-70.

[2]      Agrawal, G. K., Rakwal, R., Jwa. N and Agrawal, V. P. 2001. Signalling molecules and blast pathogen attack activates rice OsPR1a and OsPR1b genes: A model illustrating components participating during defence/stress response. J Plant Physiol and Biochem, 39: 95–110.

[3]      Amatulli, M.T., Spadaro, D., Gullino, M.L. and Garibaldi, A. 2012. Conventional and real-time PCR for the identification of Fusarium fujikuroi and Fusarium proliferatum from diseased rice tissues and seeds. J Eur J Plant Pathol, 134:401– 408.

[4]      Balmer, A., Victoria, P., Jordi, G., Victor, F and Brigitte, M.M. 2015. The ‘prime-ome’: towards a holistic approach to priming. J Trends Plant Sci, 20: 443–452.

[5]      Cao, H., Glazebrook, J., Clarke, J., Volko, S. and Dong, X. 1997. The Arabidopsis NPR1 gene that controls systemic acquired resistance encodes a novel protein containing ankyrin repeats. J Cell, 88: 57–63.

[6]      Carter, L.L.A., Leslie, L.F. and Webster, R. K. 2008. Population structure of Fusarium fujikuroi from California rice and Water Grass. J Phytophatol, 9: 992–998.

[7]      Conrath, U., Beckers, G.J.M., Flors, V., Garcia, A.P., Jakab, G., Mauch, F., Newman, M.A., Pieterse, C.M.J., Poinssot, B., Pozo, M.J., Pugin, A., Schaffrath, U., Ton, J., Wendehenne, D., Zimmerli, L. and Mauch- Mani, B. 2006. Priming: getting ready for battle. J Mol Plant Microbe Interact, 19: 1062–1071.

[8]      Delaney, T.P., Friedrich, l. and Ryals, J.A. 1995. Arabidopsis signal transduction mutant defective in chemically and biologically induced disease resistance. J Proc Natl Acad Sci USA, 92(14): 6602–6606.

[9]      Deshmukh, S. D and Kogel, K.H. 2007. Piriformospora indica protects barley from root rot caused by Fusarium graminearum. J Plant Dis Prot, 114: 263–268.

[10]    Desjardins, A.E and Proctor, R.H. 2007. Molecular biology of fusarium mycotoxins. J International journal of food microbial, 119: 47–50.

[11]    Hejgaard, J., Jacobsen, S. and Svendsen, I. 1991. Two antifungal thaumatin like proteins from barley grain. J FEBS Lett, 291: 127-131.

[12]    Kogel, K.H., P. Franken and Huckelhoven, R. 2006. Endophyte or parasite-what decides? J Curr Opi Plant Bio, 9:358–363.

[13]    Kohler, A., Schwindling, S. and Conrath, U. 2002. Benzothiadiazole-induced priming for potentiated responses to pathogen infection, wounding, and infiltration of water into leaves requires the NPR1/NIM1 gene in Arabidopsis. J Plant Physiol, 128: 1046-1056.

[14]    Livak, K.J. and Schmittgen T.D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta CT) Method. J Methods, 25: 402-408.

[15]    Molitor, A., Zajic, D., Voll, L.M., Pons, K., Hnemann, J., Samans, B., Kogel. K.H. and Waller, F. 2011. Barley leaf transcriptome and metabolite analysis reveals new aspects of compatibility and Piriformospora indica-mediated systemic induced resistance to powdery mildew. J Mol Plant Microbe Interact, 24 (12): 1427-1439.

[16]    Mousavi, S.H., Babaeizad, V., Sharifnabi, B., Tajik, M.A., Massah, A and Alavi, S.M. 2014. Induction of Blast disease resistance in Rice plants by endophyte fungus Piriformospora indica. J plant dis, 3(50): 255-268.

[17]    Murray,M and Thompson,W,F, 1980. Rapid isolation of high molecular weight plant DNA.Nucl. Acids Res, 8: 4321-4325.

[18]    Nimchuk, Z., Eulgem, T., Holt, B.F. and Dangl, J.L. 2003. Recognition and response in the plant immune system. J Ann Rev Gen, 37: 579–609.

[19]    Peškan T., Shahollari, B., Giong, P.H., Hehl, S., Markert, S., Blanke, V., Kost, G., Varma, A. and Oelmüller, R. 2004. Association of Piriformospora indica with Arabidopsis thaliana roots represents a novel system to study beneficial plant–microbe interactions and involves early plant protein modifications in the endoplasmic reticulum and at the plasma membrane. J Physiol Plant, 122: 465–477.

[20]    Pozo, M.J. and Azcon-Aguilar, C. 2007. Unraveling mycorrhiza-induced resistance. J Curr Opi Plant Biol, 10:393–398.

[21]    Rasmussen, R. 2001. Quantification on the Light Cycler. In: Meuer, S., Wittwer, C. and Nakagawara, K.I., eds. Rapid Cycle Real-Time PCR, Springer press, Heidelberg, Pp: 21-34.

[22]    Schenk, P.M., Kazan, K., Wilson, I.W., Anderson, J.P., Richmond, T., Somerville, S.C. and Manners, J.M. 2000. Coordinated plant defense responses in Arabidopsis revealed by microarray analysis, J Proc Nat. Acad Sci U.S.A, 97: 11655–11660.

[23]    Song, F and Goodman, R. 2001. Molecular biology of disease resistance in rice. J Physiol Mol Plant Pathol, 59:1-11.

[24]    Souaze, F., Ntodou-Thome, A., Tran, C. Y., Rostene, W. and Forgez, P. 1996. Quantitative RT-PCR: limits and accuracy. J BioTechniques, 21:280–285.

[25]    Stein, E., Molitor, A., Kogel, K.H. and Waller, F. 2008. Systemic resistance in Arabidopsis conferred by the mycorrhizal fungus Piriformospora indica requires jasmonic acid signaling and the cytoplasmic function of NPR1. J Plant Cell Physiol, 11: 1747–1751.

[26]    Triant, D.A., Whitehead, A. 2009. Simultaneous extraction of high-quality RNA and DNA from small tissue samples. Journal of Heredity, 110: 246-250.

[27]    Van Loon, L.C., Rep, M. and Pieterse, C.M. 2006. Significance of inducible defense-related proteins in infected plants. J Annu Rev Phytopathol, 44, 135–162.

[28]    Van Wees, S.C., Van der Ent, S. and Pieterse, C.M. 2008. Plant immune responses triggered by beneficial microbes. J Curr Opi Plant Biol, 11: 443–448.

[29]    Varma, A., Verma, S., Sudha, N., Butehorn, B. and Franken, P. 1999. Piriformospora indica, a cultivable plant growth-promoting root endophyte. J Applied Environ Microbiol, 65:2741-2744.

[30]    Verhagen, B.W., Glazebrook, J., Zhu, T., Chang, H.S., van Loon, L.C. and Pieterse, C.M. 2004. The transcriptome of rhizobacteria-induced systemic resistance in Arabidopsis. J Mol Plant Microbe Interact, 17:895-908.

[31]    Vierheilig, h., Coughlan, A.P., Wyss, U. and Piche, Y. 1998. Ink and vinegar, a simple staining technique for arbuscular- mycorrhizal fungi. J Appl environ microbial. 64;5004-5007.

[32]    Waller, F., Achatz, H. Baltruschat, J. Fodor, K. Becker, M. Fischer, T. Heier, R. H. Ckelhoven, C., Neumann, D., Wettstein, P. F. and Kogel, K. H. 2005. The endophytic fungus Piriformospora indica reprograms barley to salt-stress tolerance, disease resistance, and higher yield. J Proc Nat. Acad Sci U.S.A, 102:13386–13391.

[33]    Yoshida, S., Forno A.D., Cook, J.H. and Gomes, K.A. 1976. Routine procedure for growing rice plants in culture solution: Laboratory Manual for Physiological Studies of Rice, 3rd ed. The International Rice Research Institute, Los Baños, Laguna, Philippines. Pp:61–65.

[34]    Zhang, X., Wang, C., Zhang, Y., Sun, Y. and Mou, Z. 2012. The Arabidopsis mediator complex subunit16 positively regulates salicylate-mediated systemic acquired resistance and jasmonate/ethylene induced defense pathways. J Plant Cell, 24: 4294–4309.