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Degrading enzymes and phytotoxins in Monilinia spp

  • C. Garcia-Benitez
  • P. Melgarejo
  • P. Sandin-España
  • B. Sevilla-Morán
  • A. De CalEmail author
Article
  • 59 Downloads

Abstract

Necrotrophic phytopathogenic fungi secrete degrading enzymes and phytotoxins to penetrate and colonise plant tissues. Since knowledge on the pathogenic processes of brown rot in fruit which becomes infected by Monilinia spp. is limited, we undertook an investigation whose aims were to detect: (i) the degrading enzymes of Monilinia spp. when cultured on media amended with different substrates, and (ii) the phytotoxins which are secreted by Monilinia fructicola, M. fructigena and M. laxa in brown rot-infected fruit. The sporulation and growth of 10 M. fructicola isolates, 9 M. fructigena isolates, and 10 M. laxa isolates plates, which contained 25 mL 2% (w/v) bacteriological agar, 0.67% (w/v) yeast nitrogen base, and 1% (w/v) of one of the following substrates: (1) β1–3 glucan (callose), (2) casein hydrolysate, (3) cutin, (4) methylcellulose, (5) apple pectin, (6) polygalacturonic acid sodium salt, (7) D(+)-sucrose, and (8) xylan were determined in order to detect the degrading enzymes. To look for the phytotoxins, the juice of nectarines that were inoculated with conidial suspensions M. fructicola, M. fructigena and M. laxa were subjected to liquid-liquid extraction with different organic solvents. The extracts were tested for their ability to cause necrosis on nectarine discs and the ones with a positive response were analysed by high performance liquid chromatography with DAD and MS spectrometer detectors and different fractions were collected to delimit metabolites. We found that Monilinia spp. secrete phytotoxins with a molar mass between 329 and 387 g mol−1 and degrading enzymes (cutinases, α-glucosidases, pectin lyases, polygalacturonases, proteases, and xylanases) for penetrating the fruit surface and invading and colonising fruit tissues.

Keywords

Brown rot Monilinia fructicola Monilinia fructigena Monilinia laxa Phytotoxins Degrading enzymes 

Notes

Acknowledgements

We thank M.T. Morales Clemente and R. Castillo for their technical collaboration. The authors would also like to acknowledge Dr. Arieh Bomzon, ConsulWrite (www.consulwrite.com) for his editorial assistance in preparing this manuscript.

Funding

This study was funded by grants AGL2011–30472-CO2–02 and AGL2014–55287-CO2–01 from the Spanish Ministry of Science and Innovation. C. Garcia-Benitez received a scholarship (BES-2012-053796) from the Spanish Ministry of Science and Innovation.

Compliance with ethical standards

Conflict of interest

Authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

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Copyright information

© Koninklijke Nederlandse Planteziektenkundige Vereniging 2018

Authors and Affiliations

  • C. Garcia-Benitez
    • 1
  • P. Melgarejo
    • 1
  • P. Sandin-España
    • 2
  • B. Sevilla-Morán
    • 2
  • A. De Cal
    • 1
    Email author
  1. 1.Department of Plant ProtectionInstituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA)MadridSpain
  2. 2.Unit Plant Protection Products, DTEVPFMadridSpain

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