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Planta

, Volume 250, Issue 5, pp 1757–1772 | Cite as

Gene isolation and structural characterization of a legume tree defensin with a broad spectrum of antimicrobial activity

  • Susana Rodríguez-Decuadro
  • Pablo D. Dans
  • María Alejandra Borba
  • Ana Maria Benko-Iseppon
  • Gianna CecchettoEmail author
Original Article
  • 98 Downloads

Abstract

Main conclusion

The recombinant EcgDf1 defensin has an antimicrobial effect against both plant and human pathogens. In silico analyses predict that EcgDf1 is prone to form dimers capable of interacting with the membranes of microorganisms.

Abstract

Plant defensins comprise a large family of antimicrobial peptides (AMP) with a wide range of biological functions. They are cysteine-rich molecules, highly sequence diverse but with a conserved and stable structure. In this work, a defensin gene (EcgDf1) was isolated from Erythrina crista-galli, a legume tree native from South America. The predicted peptide presents eight cysteines, with a γ-core motif GXCX3-9C and six cysteines distributed like the typical defensin αβ motif. The mature EcgDf1 coding sequence was heterologously expressed in Escherichia coli strains and purified by affinity chromatography. Possible dimer and oligomers of EcgDf1 were visible in SDS electrophoresis. Moreover, its 3D structure, determined by homology modeling, docking, and molecular dynamics simulations, was found to be compatible with the formation of homodimers between the β3 and β1-loop-α1, leaving the β2-loop-β3 free to interact with lipid membranes. The purified recombinant peptide inhibited the growth of several critical plant and human pathogens, like the opportunistic fungi Candida albicans and Aspergillus niger and the plant pathogens Clavibacter michiganensis ssp. michiganensis, Penicillium expansum, Botrytis cinerea, and Alternaria alternata. EcgDf1 is a promising candidate for the development of antimicrobial products for use in agriculture and medicine.

Keywords

Antimicrobial peptides Dimerization Docking Erythrina crista-galli Heterologous expression Molecular dynamics simulation 

Abbreviations

AMP

Antimicrobial peptides

hpi

Hours post-induction

IC50

Peptide concentration at which 50% inhibitions was reached

Trx

Thioredoxin

Notes

Acknowledgements

The authors thank CSIC (Comisión Sectorial de Investigación Científica, Universidad de la República, Uruguay), PEDECIBA (Programa de Desarrollo de las Ciencias Básicas, Uruguay), CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Brazil), and CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico, Brazil). The authors thank Dr. Rosario Durán and Madelón Portela for MALDI-TOFF analysis and Dr. Magdalena Vaio for flow cytometry analysis. SRD was supported by a doctoral fellowship from CSIC. IRB Barcelona is the recipient of a Severo Ochoa Award of Excellence from MINECO (Ministerio de Economía, Industria y Competitividad. Government of Spain). PDD and GC are PEDECIBA and SNI (Sistema Nacional de Investigadores, Uruguay) researchers.

Funding

This research was funded by CSIC (Comisión Sectorial de Investigación Científica, Universidad de la República, Uruguay) (Grant #267).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

425_2019_3260_MOESM1_ESM.pdf (744 kb)
Supplementary material 1 (PDF 744 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Departamento de Biología Vegetal, Facultad de AgronomíaUniversidad de la RepúblicaMontevideoUruguay
  2. 2.Institute for Research in Biomedicine (IRB Barcelona)The Barcelona Institute of Science and TechnologyBarcelonaSpain
  3. 3.Joint BSC-IRB Research Program in Computational BiologyBarcelonaSpain
  4. 4.Departamento de Biociencias, Facultad de QuímicaUniversidad de la RepúblicaMontevideoUruguay
  5. 5.Universidade Federal de Pernambuco, Centro de BiociênciasRecifeBrazil
  6. 6.Instituto de Química Biológica, Facultad de Ciencias-Facultad de QuímicaUniversidad de la RepúblicaMontevideoUruguay

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