title: Enhanced production of microsclerotia in recalcitrant Verticillium dahliae isolates and its use for inoculation of olive plants.
abstract: OBJECTIVE
  The optimization of a simple protocol for the mass production of viable microsclerotia (MS) of Verticillium spp., even for recalcitrant isolates, to the inoculation of olive cuttings.
  
  
  RESULTS
  Four Verticillium spp. isolates were characterized by growth rate and morphology. Then, the production ability and the viability of MS over time were assessed in seven solid culture media and five aqueous media. The best culture medium, according to the quantity and the quality (size) of the MS produced, was the alkaline-modified sodium polipectate (AMSP) aqueous medium. The MS viability was higher in peat moss substrates. Finally, the MS obtained in this work were infective causing 100% incidence of Verticillium wilt (VW) disease in inoculated olive plants.
  
  
  CONCLUSIONS
  This study demonstrates that the modified sodium polipectate medium amended with 0·1% agar is the most suitable for the production of MS of Verticillium dahliae isolates that have lost the ability to produce MS in standard culture media.
  
  
  CONCLUSIONS
  Mass production of MS for artificial infestation of soil is critical to the study of epidemiological and control aspects of the VW. To overcome the failure in the production of MS in recalcitrant isolates, a culture media was optimized and a successful plant inoculation experiment was carried out with artificial MS.
authors.affiliation: Departamento de Agronomía, Universidad de Córdoba-Campus de Excelencia Internacional Agroalimentario ceiA3, Córdoba, Spain.
authors.forename: A
authors.identifier: http://orcid.org/0000-0002-6592-383X
authors.initials: A
authors.lastname: Varo
authors.affiliation: Departamento de Agronomía, Universidad de Córdoba-Campus de Excelencia Internacional Agroalimentario ceiA3, Córdoba, Spain.
authors.forename: M C
authors.initials: MC
authors.lastname: Raya-Ortega
authors.affiliation: Departamento de Agronomía, Universidad de Córdoba-Campus de Excelencia Internacional Agroalimentario ceiA3, Córdoba, Spain.
authors.forename: A
authors.identifier: http://orcid.org/0000-0003-3089-8301
authors.initials: A
authors.lastname: Trapero
mesh_terms: D003470:Culture Media / Q000737:chemistry / Q000378:metabolism*; D031658:Olea / Q000382:microbiology*; D010935:Plant Diseases / Q000382:microbiology*; D020172:Verticillium / Q000737:chemistry / Q000254:growth & development* / Q000378:metabolism
chemical_list: D003470:Culture Media
keywords: Olea europaea; Verticillium dahliae; mass production; microsclerotia; recalcitrant isolates