Purification and Characterization of AsES Protein A Subtilisin Secreted by Acremonium Strictum is a Novel Plant Defense Elicitor.

Chalfoun, Nadia R.; Grellet Bournonville, Carlos F.; Martínez Zamora, Martín G.; Díaz Perales, Araceli; Castagnaro, Atilio y Díaz Ricci, Juan C. (2013). Purification and Characterization of AsES Protein A Subtilisin Secreted by Acremonium Strictum is a Novel Plant Defense Elicitor.. "JOURNAL OF BIOLOGICAL CHEMISTRY", v. 288 (n. 20); pp. 14098-14113. ISSN 0021-9258. https://doi.org/10.1074/jbc.M112.429423.

Descripción

Título: Purification and Characterization of AsES Protein A Subtilisin Secreted by Acremonium Strictum is a Novel Plant Defense Elicitor.
Autor/es:
  • Chalfoun, Nadia R.
  • Grellet Bournonville, Carlos F.
  • Martínez Zamora, Martín G.
  • Díaz Perales, Araceli
  • Castagnaro, Atilio
  • Díaz Ricci, Juan C.
Tipo de Documento: Artículo
Título de Revista/Publicación: JOURNAL OF BIOLOGICAL CHEMISTRY
Fecha: Mayo 2013
Volumen: 288
Materias:
Escuela: E.T.S.I. Agrónomos (UPM) [antigua denominación]
Departamento: Biotecnologia [hasta 2014]
Licencias Creative Commons: Reconocimiento - Sin obra derivada - No comercial

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Resumen

In this work, the purification and characterization of an extracellular elicitor protein, designated AsES, produced by an avirulent isolate of the strawberry pathogen Acremonium strictum, are reported. The defense eliciting activity present in culture filtrates was recovered and purified by ultrafiltration (cutoff, 30 kDa), anionic exchange (Q-Sepharose, pH 7.5), and hydrophobic interaction (phenyl-Sepharose) chromatographies. Two-dimensional SDS-PAGE of the purified active fraction revealed a single spot of 34 kDa and pI 8.8. HPLC (C2/C18) and MS/MS analysis confirmed purification to homogeneity. Foliar spray with AsES provided a total systemic protection against anthracnose disease in strawberry, accompanied by the expression of defense-related genes (i.e. PR1 and Chi2-1). Accumulation of reactive oxygen species (e.g. H2O2 and O2̇̄) and callose was also observed in Arabidopsis. By using degenerate primers designed from the partial amino acid sequences and rapid amplification reactions of cDNA ends, the complete AsES-coding cDNA of 1167 nucleotides was obtained. The deduced amino acid sequence showed significant identity with fungal serine proteinases of the subtilisin family, indicating that AsES is synthesized as a larger precursor containing a 15-residue secretory signal peptide and a 90-residue peptidase inhibitor I9 domain in addition to the 283-residue mature protein. AsES exhibited proteolytic activity in vitro, and its resistance eliciting activity was eliminated when inhibited with PMSF, suggesting that its proteolytic activity is required to induce the defense response. This is, to our knowledge, the first report of a fungal subtilisin that shows eliciting activity in plants. This finding could contribute to develop disease biocontrol strategies in plants by activating its innate immunity.

Más información

ID de Registro: 26288
Identificador DC: http://oa.upm.es/26288/
Identificador OAI: oai:oa.upm.es:26288
Identificador DOI: 10.1074/jbc.M112.429423
URL Oficial: http://www.jbc.org/content/288/20/14098.abstract
Depositado por: Memoria Investigacion
Depositado el: 24 Jun 2014 14:53
Ultima Modificación: 22 Sep 2014 11:40
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