Contents lists available at ScienceDirect International Journal of Food Microbiology journal homepage: www.elsevier.com/locate/ijfoodmicro Antifungal activity of proteolytic fraction (P1G10) from (Vasconcellea cundinamarcensis) latex inhibit cell growth and cell wall integrity in Botrytis cinerea María José Torres-Ossandón a,b,c , Antonio Vega-Gálvez a , Carlos E. Salas d , Julia Rubio e , Evelyn Silva-Moreno e , Luis Castillo b,c, ⁎ a Departamento de Ingeniería en Alimentos, Universidad de La Serena, La Serena, Chile b Laboratorio de Bioquímica y Biología Molecular, Departamento de Biología, Universidad de La Serena, La Serena, Chile c Millennium Nucleus for Fungal Integrative and Synthetic Biology (FISB), Chile d Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Belo Horizonte, Brazil e Instituto de Ciencias Biomédicas, Universidad Autónoma de Chile, Santiago, Chile ARTICLE INFO Keywords: Antifungal activity P1G10 Botrytis cinerea Vasconcellea cundinamarcencis Proteolytic enzymes ABSTRACT The aim of this study was to determine the antifungal activity of the proteolytic fraction P1G10 from Vasconcellea cundinamarcencis (ex-Carica candamarcensis) against Botrytis cinerea, the causative agent of pre- and postharvest damaging disease in fruit and vegetables. The survival of B. cinerea at different concentrations of P1G10 showed that 1 mg/mL inhibited 50% of mycelium growth after 72 h incubation. The kinetic of growth inhibition fits the Weibull distribution function, and the data was confirmed by the IC 50 survival assay. The study shows that P1G10 inhibits conidia germination and germ tube elongation of B. cinerea relative to untreated conidia. Hypersensitivity to cell wall-perturbing agents (Calcofluor white and Congo red) was observed in mycelium cells treated with P1G10. In addition, P1G10 exhibited inhibitory effect on the adhesion of conidia, provoked alterations in membrane integrity and induced production of reactive oxygen species accompanied by cellular damage. Our results highlight the effect of P1G10 on mycelium growth, cell wall alterations, membrane integrity and adhesion. P1G10 emerges as promising antifungal to control disease causing agents in the food agroindustry. 1. Introduction Botrytis cinerea is an ubiquitous pathogenic fungus, responsible for severe damage in more than 200 plant species worldwide, including grapes, stone-fruit, berries and vegetables, causing important economic losses pre- and post-harvest (Olmedo et al., 2016; Wang et al., 2013). The fungus infects either by direct penetration, or through wounds caused by farming practices and is responsible for severe loss during long-distance transport and storage of vegetables, especially when en- vironmental conditions are appropriate for fungal development (Soylu et al., 2010). Traditionally, B. cinerea is primarily controlled by syn- thetic fungicides. For example, the chemical control applied to table grapes is mainly based on different groups of fungicides (dicarbox- imides, anilinopyrimidines, phenylpyrroles, carboxamides and hydro- xyanilides), with applications programmed four to six times during the season (Aqueveque et al., 2016). However, use of fungicides is linked to hazardous effects including carcinogenicity, teratogenicity, high acute residual toxicity, and delayed degradation period, impacting human health (Kast-Hutcheson et al., 2001). Hence, human health hazards and the adverse environmental impact occasioned by conventional pesti- cides prompted a search for new antifungal agents to meet the needs of food industry (Martínez-Romero et al., 2007). Several less aggressive alternatives, including near-harvest spraying with biological controlling agents and postharvest application of bi- carbonates, or antagonistic microorganisms, sanitization and addition of natural products with antimicrobial properties have been in- vestigated to control fungal diseases. In the last decade these studies focused on plant extracts and essential oils with antimicrobial activity, and they emerge nowadays as good alternatives instead of conventional synthetic fungicides (Aqueveque et al., 2016; Burt, 2004; Olmedo et al., https://doi.org/10.1016/j.ijfoodmicro.2018.08.027 Received 23 December 2017; Received in revised form 22 August 2018; Accepted 24 August 2018 ⁎ Corresponding author at: Laboratorio de Bioquímica y Biología Molecular, Departamento de Biología, Facultad de Ciencias, Universidad de La Serena, La Serena, Chile. E-mail address: lcastillo@userena.cl (L. Castillo). International Journal of Food Microbiology 289 (2019) 7–16 Available online 28 August 2018 0168-1605/ © 2018 Published by Elsevier B.V. T