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Identification of Botryticidal Proteins with Similarity to NBS–LRR Proteins in Rosemary Pepper (Lippia sidoides Cham.) Flowers

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Abstract

Heavy agricultural losses are closely related to attacks by insect-pests and phytopathogens such as bacteria and fungi. Among them, the fungus Botrytis cinerea can cause gray mold in more than 200 different species of plants, and is considered a challenging problem for agribusiness. Fungicides are commonly used to control this pathogen because they are fast-working and easy to apply. However, the continuous use of fungicides may promote the selection of resistant fungi and can also cause profound contamination in ecosystems. Aiming to find alternative strategies to solve these problems, several studies have focused on searching for plant proteins and peptides with antifungal activities (AFPs). With this in mind, this report shows the isolation and characterization of two novels antifungal proteins from flowers of rosemary pepper (Lippia sidoides Cham.) with 10 and 15 kDa. Isolation was performed by using an Octyl-Sepharose hydrophobic column. In vitro bioassays indicated that isolated proteins were able to inhibit B. cinerea development, but were not effective against all bacteria tested. Moreover, N-termini sequences indicate that both proteins showed sequence homology with NBS–LRR R proteins with a lower molecular mass, suggesting possible protein fragmentation. Data reported here could help in the development of biotechnological products for crop protection against phytopathogenic fungi in the near future.

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Abbreviations

AFPs:

Antifungal proteins

ATP:

Adenosine triphosphate

GTP:

Guanosine triphosphate

Ls-AMP1 and 2:

Lippia sidoides antimicrobial peptide 1 and 2

NBS–LRR:

Nucleotide binding site- leucine rich repeat protein

R-Genes:

Resistance genes

References

  1. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) J Mol Biol 215:403–410

    CAS  Google Scholar 

  2. Ashikawa I, Hayashi N, Yamane H, Kanamori H, Wu J, Matsumoto T, Ono K, Yano M (2008) Genetics 180:2267–2276

    Article  CAS  Google Scholar 

  3. Baser KH (2008) Curr Pharm Des 14:3106–3119

    Article  CAS  Google Scholar 

  4. Berger S, Sinha AK, Roitsch T (2007) J Exp Bot 58:4019–4026

    Article  CAS  Google Scholar 

  5. Blum H, Beier H, Gross H (1987) Electrophoresis 8:93–99

    Article  CAS  Google Scholar 

  6. Botelho MA, Nogueira NAP, Bastos GM, Fonseca SGC, Lemos TLG, Matos FJA, Montenegro D, Heukelbach J, Rao VS, Brito GAC (2007) Braz J Med Biol Res 40:349–356

    Article  CAS  Google Scholar 

  7. Choquer M, Fournier E, Kunz C, Levis C, Pradier JM, Simon A, Viaud M (2007) FEMS Microbiol Lett 277:1–10

    Article  CAS  Google Scholar 

  8. Collins N, Drake J, Ayliffe M, Sun Q, Ellis J, Hulbert S, Pryor T (1999) Plant Cell 11:1365–1376

    Article  CAS  Google Scholar 

  9. Costa SMO, Lemos TLG, Pessoa ODL, Pessoa C, Montenegro RC, Braz-Filho R (2001) J Nat Prod 64:792–795

    Article  CAS  Google Scholar 

  10. Costa JGM, Rodrigues FFG, Angélico EC, Silva MR, Mota ML, Santos NKA, Cardoso ALH, Lemos TLG (2005) Rev Bras Farmacog 15:304–309

    CAS  Google Scholar 

  11. de Maagd RA, Weemen-Hendriks M, Stiekema W, Bosch D (2000) Appl Environ Microb 66:1559–1563

    Article  Google Scholar 

  12. Elad Y, Williamson B, Tudzinski P, Delen N (2007) Springer, Dordrecht, the Netherlands:1–6

  13. Faris JD, Zhang Z, Lu HL, Reddy L, Cloutier S, Fellers JP, Meinhardt SW, Rasmussen JB, Xu SS, Oliver RP, Simons KJ, Friesen TL (2010) Proc Natl Acad Sci USA 107(30):13544–13549

    Article  CAS  Google Scholar 

  14. Fernández-Acero FJ, Jorge I, Calvo E, Vallejo E, Carbú M, Camafeita E, Garrido C, López JA, Jorrin J, Cantoral JM (2007) Arch Microbiol 187:207–215

    Article  Google Scholar 

  15. Finkina EL, Shramova EL, Tagaev AA, Ovchinnikova TV (2008) Biochem Biophys Res Comm 371:860–865

    Article  CAS  Google Scholar 

  16. Fontenelle ROS, Morais SM, Brito EHS, Kerntopf MR, Brilhante RSN, Cordeiro RA, Tomé AR, Queiroz MGR, Nascimento NRF, Sidrim JJC, Rocha MFG (2007) Cham J Antimicrob Chemoth 59:934–940

    Article  CAS  Google Scholar 

  17. Franco OL, Murad AM, Leite JR, Mendes PAM, Prates MV, Bloch C Jr (2006) FEBS J 273:3489–3497

    Article  CAS  Google Scholar 

  18. Gruber CW, Cemazar M, Anderson MA, Craik DJ (2007) Toxicon 49:561–575

    Article  CAS  Google Scholar 

  19. Jurgensen CW, Madsen A (2009) Ann Agric Environ Med 16:183–196

    Google Scholar 

  20. Kimura MK, Souza PE, Castro HA (2001) Cienc Agrotec 25:1150–1160

    CAS  Google Scholar 

  21. Laemmli UK (1970) Nature 15:680–685

    Article  Google Scholar 

  22. Lee JR, Park S-C, Kim J-Y, Lee SS, Park Y, Cheong G-W, Hahm K-S, Lee SY (2007) Biochem Biophys Res Comm 353:672–678

    Article  CAS  Google Scholar 

  23. Lee H, Churey JJ, Worabo RW (2008) J Appl Microbiol 105:663–673

    Article  CAS  Google Scholar 

  24. Lienqueo M, Salazar O, Henriquez K, Calado CRC, Fonseca LP, Cabral JMS (2007) J Chromatogr 1154:460–463

    Article  CAS  Google Scholar 

  25. Lin P, Ng TB (2008) Peptides 29:1664–1671

    Article  CAS  Google Scholar 

  26. Liu Z, Liu H, Liu X, Wu X (2008) Comp Biochem Phys B 149:557–561

    Article  Google Scholar 

  27. Lokossou AA, Park TH, van Arkel G, Arens M, Ruyter-Spira C, Morales J, Whisson SC, Birch PR, Visser RG, Jacobsen E, van der Vossen EA (2009) Mol Plant Microbe Int 22:630–641

    Article  CAS  Google Scholar 

  28. McHale L, Tan X, Koehl P, Michelmore RW (2006) Gen Biol 7:212

    Article  Google Scholar 

  29. Montesinos E (2007) FEMS Microbiol Lett 270:1–11

    Article  CAS  Google Scholar 

  30. Montesinos E, Barbají E (2008) Chem Biodivers 5:1225–1237

    Article  CAS  Google Scholar 

  31. Odintsova TI, Rogozhin EA, Baranov Y, Musolyamov AKH, Yalpani N, Egorov TA, Grishin EV (2008) Biochimie 90:1667–1673

    Article  CAS  Google Scholar 

  32. Park HC, Kang YH, Chun HJ, Koo JC, Cheong YH, Kim CY, Kim MC, Chung WS, Kim JC, Yoo JH, Koo YD, Koo SC, Lim CO, Lee SY, Cho MJ (2002) Plant Mol Biol 50:59–69

    Article  CAS  Google Scholar 

  33. Parish CA, Jiang H, Tokiwa Y, Berova N, Nakanishi K, Mccabe D, Zuckerman W, Xia MM, Gabay JE (2001) Bioorg Med Chem 9:377–382

    Article  CAS  Google Scholar 

  34. Pelegrini PB, Murad AM, Silva LP, Santos RCP, Costa FT, Tagliari PD, Bloch C Jr, Noronha EF, Miller RN, Franco OL (2008) Peptides 29:1271–1279

    Article  CAS  Google Scholar 

  35. Pellegrini A, Thomas U, Bramaz N, Hunziker P, Fellenberg RV (1999) Biochim Biophys Acta 1426:439–448

    CAS  Google Scholar 

  36. Pellegrini A, Hulsmeier AJ, Hunziker P, Thomas U (2004) Biochim Biophys Acta 1672:76–85

    CAS  Google Scholar 

  37. Shah P, Atwood JA III, Orlando R, Mubarek HE, Podila GK, Davis MR (2009) J Proteome Res 8:1123–1130

    Article  CAS  Google Scholar 

  38. Smith SM, Steinau M, Trick HN, Hulbert SH (2010) Mol Gen Genomics 283:591–602

    Article  CAS  Google Scholar 

  39. Tanaka T, Antonio BA, Kikuchi S, Matsumoto T, Nagamura Y, Numa H, Sakai H, Wu J, Itoh T, Sasaki T, Aono R, Fujii Y, Habara T, Harada E, Kanno M, Kawahara Y, Kawashima H, Kubooka H, Matsuya A, Nakaoka H, Saichi N, Sanbonmatsu R, Sato Y, Shinso Y, Suzuki M, Takeda J, Tanino M, Todokoro F, Yamaguchi K, Yamamoto N, Yamasaki C, Imanishi T, Okido T, Tada M, Ikeo K, Tateno Y, Gojobori T, Lin YC, Wei FJ, Hsing YI, Zhao Q, Han B, Kramer MR, McCombie RW, Lonsdale D, O’Donovan CC, Whitfield EJ, Apweiler R, Koyanagi KO, Khurana JP, Raghuvanshi S, Singh NK, Tyagi AK, Haberer G, Fujisawa M, Hosokawa S, Ito Y, Ikawa H, Shibata M, Yamamoto M, Bruskiewich RM, Hoen DR, Bureau TE, Namiki N, Ohyanagi H, Sakai Y, Nobushima S, Sakata K, Barrero RA, Sato Y, Souvorov A, Smith-White B, Tatusova T, An S, An G, OOta S, Fuks G, Fuks G, Messing J, Christie KR, Lieberherr D, Kim H, Zuccolo A, Wing RA, Nobuta K, Green PJ, Lu C, Meyers BC, Chaparro C, Piegu B, Panaud O, Echeverria M (2008) Nucleic Acids Res 36:1028–1033

    Google Scholar 

  40. Tavares LS, Santos MO, Viccini LF, Moreira JS, Miller RN, Franco OL (2008) Peptides 29:1842–1851

    Article  CAS  Google Scholar 

  41. Thennebelle T, Sahpaz S, Dermont C, Joseph H, Bailleul F (2006) Chem Biodivers 3:1116–1125

    Article  Google Scholar 

  42. Thevissen K, Kristensen H-H, Thomma BPHJ, Cammue BPA, François IEJA (2007) Drug Discov Today 12:966–971

    Article  CAS  Google Scholar 

  43. Thompson JD, Higgins DG, Gibson TJ (1994) Nucleic Acids Res 22:4673–4680

    Article  CAS  Google Scholar 

  44. Valero A, Sanchis V, Ramos AJ, Marín S (2008) Lett Appl Microbiol 47:327–332

    Article  CAS  Google Scholar 

  45. Velasco R, Zharkikh A, Troggio M, Cartwright DA, Cestaro A, Pruss D, Pindo M, Fitzgerald LM, Vezzulli S, Reid J, Malacarne G, Iliev D, Coppola G, Wardell B, Micheletti D, Macalma T, Facci M, Mitchell JT, Perazzolli M, Eldredge G, Gatto P, Oyzerski R, Moretto M, Gutin N, Stefanini M, Chen Y, Segala C, Davenport C, Demattè L, Mraz A, Battilana J, Stormo K, Costa F, Tao Q, Si-Ammour A, Harkins T, Lackey A, Perbost C, Taillon B, Stella A, Solovyev V, Fawcett JA, Sterck L, Vandepoele K, Grando SM, Toppo S, Moser C, Lanchbury J, Bogden R, Skolnick M, Sgaramella V, Bhatnagar SK, Fontana P, Gutin A, Van de Peer Y, Salamini F, Viola R (2007) PLoS ONE 2:E1326

    Article  Google Scholar 

  46. Wanderley-Nogueira AC, Soares-Cavalcanti NM, Morais DA, Belarmino LC, Barbosa-Silva A, Benko-Iseppon AM (2007) Genet Mol Res 6:866–889

    CAS  Google Scholar 

  47. Wang HX, Ng TB (2006) Peptides 28:547–552

    Article  Google Scholar 

  48. Wong JH, Ng TB, Cheung RCF, Ye XJ, Wang HX, Lam SK, Lin P, Chan YS, Chan Fang EF, Ngai PHK, Xia LX, Ye XY, Jiang Y, Liu F (2010) Appl Microbiol Biotechnol 87:1221–1235

    Article  CAS  Google Scholar 

  49. Wong JH, Ng TB (2005) Peptides 26:1120–1126

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by FAPEMIG, CAPES, CNPq and UCB. The authors thank Ludovico Migliolo for his valuable help in protein alignment.

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Authors and Affiliations

  1. Centro de Análises Proteômicas e Bioquímicas, Programa de Pós-Graduação em Ciências Genômicas e Biotecnologia, Universidade Católica de Brasília, SGAN Quadra 916, Módulo B, Av. W5 Norte 70. 790-160 Asa Norte, Brasília, DF-Brazil

    João S. Moreira, Renato G. Almeida, Simoni C. Dias & Octavio L. Franco

  2. Departamento de Biologia, Universidade Federal de Juiz de Fora, Juiz de Fora, Minas Gerais, Brazil

    João S. Moreira, Letícia S. Tavares, Marcelo O. Santos, Lyderson F. Viccini & Octavio L. Franco

  3. Departamento de Bioquímica e Biologia Celular, Laboratório de toxinas vegetais, Universidade Federal do Ceará, Fortaleza, Ceará, Brazil

    Ilka M. Vasconcelos & José T. A. Oliveira

  4. Faculdade de Farmácia, Universidade Federal de Juiz de Fora, Juiz de Fora, Minas Gerais, Brazil

    Nádia R. B. Raposo

Authors
  1. João S. Moreira
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  2. Renato G. Almeida
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  3. Letícia S. Tavares
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  4. Marcelo O. Santos
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  5. Lyderson F. Viccini
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  6. Ilka M. Vasconcelos
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  7. José T. A. Oliveira
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  8. Nádia R. B. Raposo
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  10. Octavio L. Franco
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Correspondence to Octavio L. Franco.

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Moreira, J.S., Almeida, R.G., Tavares, L.S. et al. Identification of Botryticidal Proteins with Similarity to NBS–LRR Proteins in Rosemary Pepper (Lippia sidoides Cham.) Flowers. Protein J 30, 32–38 (2011). https://doi.org/10.1007/s10930-010-9299-4

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  • DOI: https://doi.org/10.1007/s10930-010-9299-4

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