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Cochliobolus lunatus down-regulates proteome at late stage of colonization and transiently alters StNPR1 expression in Solanum tuberosum L.


Cochliobolus lunatus abundantly produces four-celled conidia at high temperatures (>30 °C) and under suitable conditions; the fungus colonizes potato (Solanum tuberosum L.) cultivars by adopting different invasion strategies at the microscopic level. Long-lasting defence during infection requires an upsurge in proteome changes particularly pathogenesis-related proteins chiefly under the control of nonexpresser of pathogenesis-related proteins. In order to gain molecular insights, we profiled the changes in proteome and potato nonexpresser of pathogenesis-related proteins (StNPR1) during the infection process. It is found that C. lunatus significantly (P < 0.05) suppressed the host functional proteome by 96 h after infection (hai), principally, affecting the expression of ribulose bisphosphate carboxylase enzyme, plastidic aldolase enzyme, alcohol dehydrogenase 2 and photosystem II protein prior to the formation of brown-to-black leaf spot disease. Strongest host response was observed at 24 hai hallmarked by 307 differentially expressed peptide spots concurring with the active phase of production of penetrating hyphae. Additionally, C. lunatus differentially down-regulated StNPR1 transcript by 8.19 fold by 24 hai. This study is the first to elucidate that C. lunatus transiently down-regulates the expression of StNPR1 at the onset of infection, and as a whole, infection negatively affects the expression of proteome components involved in photosynthesis, carbon fixation and light assimilation. This study contributes towards better understanding of the mechanism underlining the invasion strategies of C. lunatus.

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  1. Agrios GN (2005) Plant pathology, 5th edn. Academic Press, Amsterdam, p 395p

    Google Scholar 

  2. Bengyella L, Waikhom SD, Roy P, Bhardwaj PK, Sharma CK, Singh MW, Talukdar NC (2014) Host-range dynamics of Cochliobolus lunatus: from a biocontrol agent to a severe environmental threat. BioMed Res Int, Article ID 378372, p 9. Doi: 10.1155/2014/378372

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

    CAS  Article  PubMed  Google Scholar 

  4. Dai FC, Gao WD, Wu RJ, Jin XH (1995) A noticeable corn disease: Curvularia leaf spot. Acta Phytopathol Sin 25:330

    Google Scholar 

  5. Dong X (2004) NPR1, all things considered. Curr Opin Plant Biol 7:547–552

    CAS  Article  PubMed  Google Scholar 

  6. Elias JE, Haas W, Faherty BK, Gygi SP (2005) Comparative evaluation of mass spectrometry platforms used in large-scale proteomics investigations. Nat Methods 2:667–675

    CAS  Article  PubMed  Google Scholar 

  7. Gao S, Liu T, Li Y, Wu Q, Fu K, Chen J (2012) Understanding resistant germplasm induced virulence variation through analysis of proteomics and suppression subtractive hybridisation in a maize pathogen Curvularia lunata. Proteomics 12:3524–3535

    CAS  Article  PubMed  Google Scholar 

  8. Goa J-X, Liu T, Chen J (2014) Insertional mutagenesis and cloning of the gene required for the biosynthesis of the non-host specific toxin in Cochliobolus lunatus that causes maize leaf spot. Phytopathology 104:332–339

    Article  Google Scholar 

  9. Huang X, Liu L, Zhai Y, Liu T, Chen C (2009) Proteomic comparison of four maize inbred lines with different levels of resistance to Culvularia lunata (wakker) boed infection. Prog Nat Sci 19:353–358

    Article  Google Scholar 

  10. Iftikhar A, Shazia I, Cullum J (2006) Genetic variability and aggressiveness in Curvularia lunata associated with rice-wheat cropping areas of Pakistan. Pak J Bot 38(2):475–485

    Google Scholar 

  11. Louis B, Roy P, Waikhom SD, Talukdar NC (2013) Report of foliar necrosis of potato caused by Cochliobolus lunatus in India. Afr J Biotechnol 12:833–835

    CAS  Google Scholar 

  12. Louis B, Waikhom SD, Roy P, Bhardwaj PK, Singh MW, Goyari S, Sharma CK, Talukdar NC (2014) Secretome weaponries of Cochliobolus lunatus interacting with potato leaf at different temperature regimes reveal a CL[xxxx]LHM-motif. BMC Genom 15:213

    Article  Google Scholar 

  13. Louis B, Waikhom SD, Jose RC, Goyari S, Talukdar NC, Roy P (2015) Cochliobolus lunatus colonizes potato by adopting different invasion strategies on cultivars: new insights on temperature-dependent virulence. Microb Pathog 87(2015):30–39

    Article  PubMed  Google Scholar 

  14. Maier T, Güell M, Serrano L (2009) Correlation of mRNA and protein in complex biological samples. FEBS Lett 583(24):3966–3973

    CAS  Article  PubMed  Google Scholar 

  15. Pechanova O, Pechan T (2015) Maize-pathogen interactions: an ongoing combat from a proteomics perspective. Int J Mol Sci 16:28429–28448

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  16. Rochon A, Boyle P, Wignes T, Fobert PR, Despres C (2006) The co-activator function of Arabidopsis NPR1 requires the core of its BTB/POZ domain and the oxidation of C-terminal cysteines. Plant Cell 18:3670–3685

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  17. Scheffer RP (1997) The nature of disease in plants. University Press, Cambridge

    Google Scholar 

  18. Shevchenko A, Tomas H, Havis J, Olsen JV, Mann M (2006) In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nat Protoc 1:2856–2860

    CAS  Article  PubMed  Google Scholar 

  19. Sugihara K, Hanagata N, Dubinsky Z, Baba S, Karube I (2000) Molecular characterization of cDNA encoding oxygen evolving enhancer protein 1 increased by salt treatment in the mangrove Bruguiera gymnorrhiza. Plant Cell Physiol 41:1279–1285

    CAS  Article  PubMed  Google Scholar 

  20. Trouvelot S, Varnier AL, Allergre M, Mercier L, Baillieuil F et al (2008) A beta-1, 3 glucan sulphate induces resistance in grapevine against Plasmopara viticola through priming of defense responses, including HR-Like cell death. Mol Plant Microbe Interact 21:232–243

    CAS  Article  PubMed  Google Scholar 

  21. Ullstrup AJ (1972) The impact of the southern corn leaf blight epidemics of 1970-71. Annu Rev Phytopathol 10:37–50

    Article  Google Scholar 

  22. Waikhom SD, Louis B, Pranab R, Talukdar NC (2015) Insights on predominant edible bamboo shoot proteins. Afr J Biotechnol 14(17):1511–1518

    CAS  Article  Google Scholar 

  23. Williams ME, Torabinejad J, Cohik E, Parker K, Drake EJ, Thompson JE, Hortter M, Dewald DB (2005) Mutations in the Arabidopsis phosphoinositide phosphatase gene SAC9 lead to over-accumulation of ptdIns(4,5)P2 and constitutive expression of the stress-response pathway. Plant Physiol 138:686–700

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  24. Wu L, Han Z, Wang S, Wang X, Sun A, Zu X, Chen Y (2013) Comparative proteomic analysis of the plant-virus interaction in resistant and susceptible ecotypes of maize infected with sugarcane mosaic virus. J Proteome 89:124–140

    Article  Google Scholar 

  25. Xu S, Chen J, Liu L, Wang X, Huang X, Zhai Y (2007) Proteomics associated with virulence differentiation of Curvularia lunata in maize in China. J Integr Plant Biol 49(2007):487–496

    CAS  Article  Google Scholar 

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This research was jointly supported by The World Academy of Sciences (TWAS), Trieste, Italy and the Department of Biotechnology, Government of India (DBT/TWAS PG fellowship No. 3240223450) and Alexander von Humbolt (AvH) foundation. The authors thank DK Hore for proofreading the manuscript.

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Corresponding authors

Correspondence to Narayan C. Talukdar or Pranab Roy.

Additional information

Communicated by Erko Stackebrandt.

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Below is the link to the electronic supplementary material.

Figure S1

A 2-D proteome map of potato leaves at 24 hai of C. lunatus. Spots of interest with fold expression >3 are indicated with arrows. The immobilized pH gradient scale and Precision Plus Protein™ WesternC™ Standards are shown (TIFF 227 kb)

Figure S2

A 2-D proteome map of potato leaves at 72 hai of C. lunatus. Spots of interest with fold expression >3 are indicated with arrows. The immobilized pH gradient scale and Precision Plus Protein™ WesternC™ Standards are shown (TIFF 219 kb)

Figure S3

Reverse transcriptase PCR for 26S rRNA endogenous control following cDNA synthesis from total RNA. The reaction mixture consisted of 0.5 μl of 10 mM dNTPs, 19.8 μl DEPC H2O, 2.5 μl of 10X buffer, 0.5 μl of 20 pmol/μl forward, 0.5 μl of 20 pmol/μl reverse primers, 0.2 μl of 5U/ul TaqPol, and 1 μl of cDNA. The PCR conditions were: 94 °C for 3 min initial denaturation, 35 cycles of 94 °C for 30 s, 54 °C for 40 s, 72 °C for 1 min and a final extension at 72 °C for 7 min. Agarose gel electrophoresis showing a 500 bp of 26S rDNA amplicon for samples extracted at different time points. A24, A48, A72 and A96 are cDNA for plants treated with sterile water. Ctlr is control plant not treated with any substance. EA24, EA48, EA72 and EA96 are cDNA for plants challenged with C. lunatus (TIFF 78 kb)

Supplementary Table S1

Primers set for genes that encode peptide spots, used for validating the expression of significantly alter peptide spots (XLSX 10 kb)

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Louis, B., Waikhom, S.D., Jose, R.C. et al. Cochliobolus lunatus down-regulates proteome at late stage of colonization and transiently alters StNPR1 expression in Solanum tuberosum L.. Arch Microbiol 199, 237–246 (2017).

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  • Proteome
  • StNPR1
  • Two-dimensional electrophoresis
  • qPCR
  • Western blotting