Skip to main content

Advertisement

SpringerLink
Log in

Functional analysis of polygalacturonase gene RsPG2 from Rhizoctonia solani, the pathogen of rice sheath blight

  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript

Abstract

Rice sheath blight, which is caused by Rhizoctonia solani, is a devastating disease in many areas of the world where rice is cultivated. The role of polygalacturonase (PG) in the infection process of R. solani was investigated in the present study by cloning and investigating the function of a new PG-encoding gene, which was designated RsPG2. Bioinformatics showed that RsPG2 consisted of a 1,633 bp open reading frame (ORF) with six introns; the deduced protein contained 436 amino acids with a predicted mass of 45.88 kDa after cleavage of the predicted 17-amino acid signal sequence. Culture supernatants obtained from Pichia pastoris producing recombinant RsPG2 degraded polygalacturonic acid in vitro, thus confirming that RsPG2 encodes a PG gene. Purified recombinant RsPG2 also degraded rice tissue and elicited obvious necrotic symptoms 48 h after inoculation. Real-time qRT-PCR indicated that the expression of RsPG2 was strongly induced during the infection of rice by R. solani. These data unequivocally demonstrate that RsPG2 plays an important role in R. solani infection and provide further insights for elucidating the function of pg genes in the virulence of R. solani.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Akhgari, A. B., & Motallebi, Z. M. R. (2012). Bean PG-inhibiting protein expressed in transgenic Brassica napus inhibits PG from its fungal pathogen Rhizoctonia solani. Plant Protection Science, 48, 1–9.

    CAS  Google Scholar 

  • Arnold, K., Bordoli, L., Kopp, J., & Schwede, T. (2006). The SWISS-MODEL workspace: A web-based environment for protein structure homology modelling. Bioinformatics, 21, 195–201.

    Article  Google Scholar 

  • Baiswar, P., Bag, T .K., Basumatary, R., Chandra, S., & Ngachan, S. V. (2012). Molecular evidence reveals presence of Rhizoctonia solani AG1-IB on Tagetes patula in India. Australian Plant Disease Notes 7, 63–66.

  • Bendtsen, J. D., Nielsen, H., Heijne, G. V., & Brunak, S. (2004). Improved prediction of signal peptides: Signal P 3.0. Journal of Molecular Biology, 340, 783–795.

    Article  PubMed  Google Scholar 

  • Benedetti, M., Andreani, F., Leggio, C., Galantini, L., Di Matteo, A., Favel, N. V., De Lorenzo, G., Cervone, F., Federici, L., & Sicilia, F. (2013). A single amino-acid substitution allows endo-polygalacturonase of Fusarium verticillioides to acquire recognition by PGIP2 from Phaseolus vulgaris. PloS One, 8, 1–11.

    Article  Google Scholar 

  • Cabanne, C., & Doneche, B. (2002). Polygalacturonase isozymes produced during infection of the grape berry by Botrytis cinerea. Journal of Grape Research, 41, 129–132.

    CAS  Google Scholar 

  • Catharina, S., Vakhrushev, S. Y., Joshi, H. J., Kong, Y., Vester-Christensen, M. B., Schjoldager, K. T.-B. G., Lavrsen, K., Dabelsteen, S., Pedersen, N. B., Marcos-Silva, L., Gupta, R., Bennett, E. P., Mandel, U., Brunak, S., Wandall, H. H., Levery, S. B., & Clausen, H. (2013). Precision mapping of the human O-GalNAc glycoproteome through SimpleCell technology. EMBO Journal, 32, 1478–1488.

    Article  Google Scholar 

  • Chen, X. J., Zhang, H., Xu, J. Y., Tong, Y. H., & Ji, Z. L. (2006). Cell wall degrading enzymes produced by Rhizoctonia solani and their pathogenicity to rice plant (in Chinese). Jiangsu Journal of Agricultural Sciences, 22, 24–28.

    Google Scholar 

  • Chen, X. J., Xu, Y., Tong, Y. H., Meng, L. J., Ji, Z. L., & Xu, J. Y. (2009). Pathogenic mechanism of phytotoxin produced by Rhizoctonia solani, the causal pathogen of rice sheath blight (in Chinese). Acta Phytopathologica Sinica, 39, 439–443.

    Google Scholar 

  • Chen, X. J., Wang, Y. D., Zuo, S. M., Tong, Y. H., Pan, X. B., & Xu, J. Y. (2010). Isolation, purification and characterization of polygalacturonase (PG) from Rhizoctonia solani, the pathogen of rice sheath blight (in Chinese). Acta Phytopathologica Sinica, 40, 276–281.

    Google Scholar 

  • Chen, X. J., Chen, Y., Zhang, L. N., Xu, B., Zhang, J. H., Chen, Z. X., Tong, Y. H., Zuo, S. M., & Xu, J. Y. (2016). Overexpression of OsPGIP1 enhances rice resistance to sheath blight. Plant Disease, 100, 388–395.

    Article  CAS  Google Scholar 

  • Combet, C., Blanchet, C., Geourjon, C., & Deleage, G. (2000). NPS@: Network protein sequence analysis. Trends in Biochemical Sciences, 25, 147–150.

    Article  CAS  PubMed  Google Scholar 

  • Contreras, E. J. C., Hours, R. A., Voget, C. E., & Mignone, C. F. (1999). Aspergillus kawachii produces an acidic pectin releasing enzyme activity. Journal of Bioscience and Bioengineering, 88, 48–52.

    Article  Google Scholar 

  • Cserzo, M., Eisenhaber, F., Eisenhaber, B., & Simon, I. (2002). On filtering false positive transmembrane protein predictions. Protein Engineering, 15, 745–752.

    Article  CAS  PubMed  Google Scholar 

  • De Lorenzo, G., D’Ovidio, R., & Cervone, F. (2001). The role of PG-inhibiting proteins (PGIPs) in defense against pathogenic fungi. Annual Review of Phytopathology, 39, 313–335.

    Article  CAS  PubMed  Google Scholar 

  • Diaz, A. A., Tomba, E., Lennarson, R., Richard, R., Baqajewicz, M. J., & Harrison, R. G. (2009). Prediction of protein solubility in Escherichia coli using logistic regression. Biotechnology and Bioengineering, 105, 374–383.

    Article  Google Scholar 

  • Dong, Z. Y., & Wang, Z. Z. (2015). Isolation and heterologous expression of a PG produced by Fusarium oxysporum f. Sp. cubense race 1 and 4. International Journal of Molecular Science, 16, 7595–7607.

    Article  CAS  Google Scholar 

  • Eken, C., & Demirci, E. (2003). Identification and pathogenicity of Rhizoctonia solani and binucleate Rhizoctonia anastomosis groups isolated from forage legumes in Erzurum Turkey. Phytoparasitica, 31, 76–80.

    Article  Google Scholar 

  • Federici, L., Caprari, C., Mattei, B., Savino, C., Di Matteo, A., De Lorenzo, G., Cervone, F., & Tsernoglou, D. (2001). Structural requirements of endopolygalacturonase form interaction with PGIP (polygalacturonase-inhibiting protein). Proceedings of the National Academy of Science of the United States of America, 98, 13425–13430.

    Article  CAS  Google Scholar 

  • Ferrari, S., Galletti, R., Pontiggia, D., Manfredini, C., Lionetti, V., Bellincampi, D., Cervone, F., & De Lorenzo, G. (2008). Transgenic expression of a fungal endo-PG increases plant disease resistance to pathogens and reduces auxin sensitivity. Plant Physiology, 146, 669–681.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ferrari, S., Sella, L., Janni, M., De, L. G., & Favaron, F. (2012). Transgenic expression of PG-inhibiting protein in Arabidopsis and wheat increase resistance to the flower pathogen Fusarium graminearum. Plant Biology 14. Supplemental, 1, 31–38.

    Google Scholar 

  • Gainvors, A., Nedjaoum, N., Gognies, S., Muzart, M., Nedjma, M., & Belarbi, A. (2000). Purification and characterization of acidic endo-polygalacturonase encoded by the PGL1-1 gene from Saccharomyces cerevisiae. FEMS Microbiology Letters, 183, 131–135.

    Article  CAS  PubMed  Google Scholar 

  • Gasteiger, E., Hoogland, C., Gattiker, A., Duvaud, S., Wilkins, R., Appel, R. D., & Bairoch, A. (2005). Protein identification and analysis tools on the ExPASY server. (In J. M. Walker (Ed.) The proteomics protocols handbook (pp. 571–607). Totowa, New Jersey: Humana Press).

  • Hassan Dar, G. H., Beig, M. A., Ahanger, F. A., & Ganai, N. A. (2011). Management of root rot caused by Rhizoctonia solani and Fusarium oxysporum in blue pine (Pinus wallichiana) through use of fungal antagonists. Asian Journal of Plant Pathology, 5, 1–13.

    Article  Google Scholar 

  • Hwang, B. H., Bae, H. H., Lim, H. S., Kin, K. B., Kim, S. J., Im, M. H., Park, B. S., Kim, D. S., & Kim, J. K. (2010). Overexpression of polygalacturonase-inhibiting protein 2 (PGIP2) of Chinese cabbage (Brassica rapa ssp. pekinensis) increased resistance to the bacterial pathogen Pectobacterium carotovorum Ssp. carotovorum. Plant Cell Tissue and Organ Culture, 103, 293–305.

    Article  CAS  Google Scholar 

  • Joubert, D. A., Slaughter, A. R., Kemp, G., Becker, J. V., Krooshof, G. H., Bergmann, C., Benen, J., Pretorius, I. S., & Vivier, M. A. (2006). The grapevine polygalacturonase-inhibiting protein (VvPGIP1) reduces Botrytis cinerea susceptibility in transgenic tobacco and differentially inhibits fungal polygalacturonases. Transgenic Research, 15, 687–702.

    Article  CAS  PubMed  Google Scholar 

  • Kant, S., Vohra, A., & Gupta, R. (2013). Purification and physicochemical properties of polygalacturonase from Aspergillus niger MTCC 3323. Protein Expression and Purification, 7, 11–16.

    Article  Google Scholar 

  • Kumar, G. M., Mamidala, P., & Podile, A. R. (2009). Regulation of PG-inhibitory proteins in plants is highly dependent on stress and light responsive elements. Plant Omics, 2, 238–249.

    Google Scholar 

  • Lemanczyk, G., & Kwasna, H. (2013). Effects of sharp eyespot (Rhizoctonia cerealis) on yield and grain quality of winter wheat. European Journal of Plant Pathology, 135, 187–200.

    Article  Google Scholar 

  • Markovic, O., & Janecek, S. (2001). Pectin degrading glycoside hydrolases of family 28: Sequence-structural features, specificities and evolution. Protein Engineering., 14, 615–631.

    Article  CAS  PubMed  Google Scholar 

  • Martins, E. D. S., Leite, R. S., da Silva, R., & Gomes, E. (2013). Purification and properties of polygalacturonase produced by thermophilic fungus Thermoascus aurantiacus CBMAI-756 on solid-state fermentation. Enzyme Research, 2, 1–7.

    Article  Google Scholar 

  • Misawa, T., & Kuninaga, S. (2013). First report of white leaf rot on Chinese chives caused by Rhizoctonia solani AG-2-1. Journal of General Plant Pathology, 79, 280–283.

    Article  CAS  Google Scholar 

  • Niture, S. K. (2008). Comparative biochemical and structural characterizations of fungal polygalacturonases. Biologia, 63, 1–19.

    Article  CAS  Google Scholar 

  • Oeser, B., Heidrich, P. M., Muller, U., Tudzynski, P., & Tenberge, K. B. (2002). Polygalacturonase is a pathogenicity factor in the Claviceps purpurea/rye interaction. Fungal Genetics and Biology, 36, 176–186.

    Article  CAS  PubMed  Google Scholar 

  • Pan, X., Li, K., Ma, R., Shi, P. G., Huang, H. Q., Yang, P. L., Meng, K., & Yao, B. (2015). Biochemical characterization of three distinct polygalacturonase from Neosartorya fischeri P1. Food Chemistry, 188, 569–575.

    Article  CAS  PubMed  Google Scholar 

  • Rivera, M. C., Wright, E. R., Asciutto, K., & Gasoni, L. (2003). Occurrence of damping-off and wilt of Violax x witterockiana caused by Rhizoctonia solani in Argentina. Australasian Plant Pathology, 32, 561–562.

    Article  Google Scholar 

  • Rosa, D. D., Ohto, C. T., Basseto, M. A., Furtadoa, E. L., & Souzaa, N. L. D. (2008). First report of Rhizoctonia solani AG4 HG-II attacking Gazunia rigens plants in Brazil. Australasian Plant Disease Notes, 3, 1–2.

    Article  Google Scholar 

  • van Santen, Y., Benen, J. A. E., Schroter, K. H., Kalk, K. H., Armand, S., Visser, J., & Dijkstra, B. W. (1999). 1.68-Å crystal structure of endopolygalacturonase II from Aspergillus niger and identification of active site residues by site-directed mutagenesis. The Journal of Biological Chemistry, 274, 30474–30480.

    Article  PubMed  Google Scholar 

  • Sella, L., Castiglioni, C., Roberti, S., D’Ovidio, R., & Favaron, F. (2004). An endo-polygalacturonase (PG) of Fusarium moniliforme escaping inhibition by plant polygalacturonase-inhibiting proteins (PGIPs) provides new insight into the PG-PGIP interaction. FEMS Microbiology Letters, 240, 117–124.

    Article  CAS  PubMed  Google Scholar 

  • Shieh, M. T., Brown, R. L., Whitehead, M. P., Cary, J. W., Cotty, P. J., Cleveland, T. E., & Dean, R. A. (1997). Molecular genetic evidence for the involvement of a specific PG, P2c, in the invasion and spread of Aspergillus flavus in cotton bolls. Applied and Environmental Microbiology, 63, 3549–3552.

    Google Scholar 

  • Taylor, R. J., & Secor, G. A. (1988). An improved diffusion assay for quantifying the polygalacturonase content of Erwinia culture filtrates. Phytopathology, 78, 1101–1103.

    Article  CAS  Google Scholar 

  • Xu, J. Y., Zhang, H. D., Zhang, H., Tong, Y. H., Xu, Y., Chen, X. J., & Ji, Z. L. (2004). Toxin produced by Rhizoctonia solani and its relationship with pathogenicity of the fungus (in Chinese). Journal of Yangzhou University (Agricultural and Life Science Edition), 25, 61–64.

    Google Scholar 

  • Yang, J., Luo, H. Y., Li, J., Wang, K., Cheng, H. P., Bai, Y. G., Yuan, T. Z., Fan, Y. L., & Yao, B. (2011). Cloning, expression and characterization of an acidic endo-polygalacturonase from Bispora sp. MEY-1 and its potential application in juice clarification. Process Biochemistry, 46, 272–277.

    Article  Google Scholar 

  • Yang, Y. Q., Yang, M., Li, M. H., & Zhou, E. X. (2012). Cloning and functional analysis of an endo-PG-encoding gene Rrspg1 of Rhizoctonia solani, the causal agent of rice sheath blight. Canadian Journal of Plant Pathology, 34, 436–447.

    Article  CAS  Google Scholar 

  • Zhang, H., Chen, X. J., Tong, Y. H., Ji, Z. L., & Xu, J. Y. (2005). Damage of cell wall degrading enzymes produced by Rhizoctonia solani to rice tissue and cells (in Chinese). Journal of Yangzhou University (Agricultural and Life Science Edition), 26, 83–86.

    Google Scholar 

  • Zuo, S. M., Yin, Y. J., Pan, C. H., Chen, Z. X., Zhang, Y. F., Gu, S. L., Zhu, L. H., & Pan, X. B. (2013). Fine mapping of qSB-11 LE, the QTL that confers partial resistance to rice sheath blight. Theoretical and Applied Genetics, 126, 1257–1272.

    Article  PubMed  Google Scholar 

  • Zuo, S. M., Chen, X. J., Chen, H. Q., Xu, Y., Zhang, J. H., Chen, Y., Chen, Z. X., Tong, Y. H., Xu, J. Y., & Pan, X. B. (2014). Defense response and physiological difference of rice cultivars with different sheath blight resistance levels to the toxin produced by Rhizoctonia solani (in Chinese). Chinese Journal of Rice Sciences, 28, 551–558.

    CAS  Google Scholar 

Download references

Acknowledgments

This work was partially supported by the Science and Technology Project of Jiangsu Province (BE2015342), the National Transgenic Key Project (2014ZX0800103B), the key project of Jiangsu Natural Science Foundation for Colleges (14KJA210003), the Open Project of the Key Laboratory of Plant Functional Genomics, Jiangsu Province (K13004), the key plan of Yangzhou agriculture (YZ2015028) and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, respectively.

Author information

Authors and Affiliations

  1. Horticulture and Plant Protection College, Yangzhou University, Yangzhou, 225009, China

    Xijun Chen, Li Lili, Yun Zhang, Jiahao Zhang, Shouqiang Ouyang, Qingxia Zhang, Yunhui Tong & Jingyou Xu

  2. Key Laboratory of Crop Genetics and Physiology/Co-Innovation Center for Modern Production Technology of Grain Crops, Key Laboratory of Plant Functional Genomics of the Ministry of Education, Yangzhou University, Yangzhou, 225009, China

    Xijun Chen & Shimin Zuo

Authors
  1. Xijun Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Li Lili
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yun Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jiahao Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Shouqiang Ouyang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Qingxia Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yunhui Tong
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jingyou Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Shimin Zuo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shimin Zuo.

Electronic supplementary material

ESM 1

(DOCX 64 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chen, X., Lili, L., Zhang, Y. et al. Functional analysis of polygalacturonase gene RsPG2 from Rhizoctonia solani, the pathogen of rice sheath blight. Eur J Plant Pathol 149, 491–502 (2017). https://doi.org/10.1007/s10658-017-1198-5

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10658-017-1198-5

Keywords

Search

Navigation