Phytotoxic Tryptoquialanines Produced In Vivo by Penicillium digitatum Are Exported in Extracellular Vesicles

Autor(es):Mostrar menos -

Costa, Jonas Henrique ^[1] ; Bazioli, Jaqueline Moraes ^{[2, 1]} ; Barbosa, Luidy Darllan ^[1] ; Theodoro dos Santos Junior, Pedro Luis ^[1] ; Reis, Flavia C. G. ^{[3, 4]} ; Klimeck, Tabata ^[3] ; Crnkovic, Camila Manoel ^[5] ; Berlinck, Roberto G. S. ^[6] ; Sussulini, Alessandra ^[1] ; Rodrigues, Marcio L. ^{[3, 7]} ; Fill, Taicia Pacheco ^[1]

Afiliação do(s) autor(es)

^[1] Univ Estadual Campinas, Inst Chem, CP 6154, Campinas, SP - Brazil

^[2] Univ Estadual Campinas, Fac Pharmaceut Sci, Campinas, SP - Brazil

^[3] Fundacao Oswaldo Cruz FIOCRUZ, Inst Carlos Chagas, Curitiba, Parana - Brazil

^[4] Fiocruz MS, Ctr Desenvolvimento Tecnol Saude CDTS, Rio De Janeiro - Brazil

^[5] Univ Sao Paulo, Sch Pharmaceut Sci, Dept Biochem & Pharmaceut Technol, Sao Paulo, SP - Brazil

^[6] Univ Sao Paulo, Inst Quim Sao Carlos, CP 780, Sao Carlos, SP - Brazil

^[7] Univ Fed Rio de Janeiro, Inst Microbiol Paulo Goes IMPG, Rio De Janeiro, RJ - Brazil

• Tipo de documento: Artigo Científico
• Fonte: MBIO; v. 12, n. 1 JAN-FEB 2021.
• Citações Web of Science: 4

Resumo

Penicillium digitatum is the most aggressive pathogen of citrus fruits. Tryptoquialanines are major indole alkaloids produced by P. digitatum. It is unknown if tryptoquialanines are involved in the damage of citrus fruits caused by P. digitatum. To investigate the pathogenic roles of tryptoquialanines, we initially asked if tryptoquialanines could affect the germination of Citrus sinensis seeds. Exposure of the citrus seeds to tryptoquialanine A resulted in a complete inhibition of germination and an altered metabolic response. Since this phytotoxic effect requires the extracellular export of tryptoquialanine A, we investigated the mechanisms of extracellular delivery of this alkaloid in P. digitatum. We detected extracellular vesicles (EVs) released by P. digitatum both in culture and during infection of citrus fruits. Compositional analysis of EVs produced during infection revealed the presence of a complex cargo, which included tryptoquialanines and the mycotoxin fungisporin. The EVs also presented phytotoxicity activity in vitro and caused damage to the tissues of citrus seeds. Through molecular networking, it was observed that the metabolites present in the P. digitatum EVs are produced in all of its possible hosts. Our results reveal a novel phytopathogenic role of P. digitatum EVs and tryptoquialanine A, implying that this alkaloid is exported in EVs during plant infection. IMPORTANCE During the postharvest period, citrus fruits can be affected by phytopathogens such as Penicillium digitatum, which causes green mold disease and is responsible for up to 90% of total citrus losses. Chemical fungicides are widely used to prevent green mold disease, leading to concerns about environmental and health risks. To develop safer alternatives to control phytopathogens, it is necessary to understand the molecular basis of infection during the host-pathogen interaction. In the P. digitatum model, the virulence strategies are poorly known. Here, we describe the production of phytotoxic extracellular vesicles (EVs) by P. digitatum during the infection of citrus fruits. We also characterized the secondary metabolites in the cargo of EVs and found in this set of molecules an inhibitor of seed germination. Since EVs and secondary metabolites have been related to virulence mechanisms in other host-pathogen interactions, our data are important for the comprehension of how P. digitatum causes damage to its primary hosts. (AU)

• Processo FAPESP: 17/24462-4 - Entendendo as estratégias de virulência do fungo Penicillium digitam contra seu hospedeiro citros
• Beneficiário: Taicia Pacheco Fill
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 13/50228-8 - Componentes da biodiversidade, e seus caracteres metabólicos, de ilhas do Brasil: uma abordagem integrada
• Beneficiário: Roberto Gomes de Souza Berlinck
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOTA - Temático
• Processo FAPESP: 19/06359-7 - Compreensão dos fatores químicos envolvidos no endofitismo x patogenicidade de diferentes isolados de Penicillium brasilianum
• Beneficiário: Taicia Pacheco Fill
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 15/01017-0 - EMU concedido no âmbito do projeto aprovado 2013/50228-8, nome do equipamento: uPLC-HRMS (cromatógrafo líquido de ultra resolução acoplado com espectrômetro de massas de alta resolução)
• Beneficiário: Roberto Gomes de Souza Berlinck
• Modalidade de apoio: Auxílio à Pesquisa - Programa Equipamentos Multiusuários
• Processo FAPESP: 19/11563-2 - Prospecção de metabólitos secundários com atividade anticâncer produzidos por fungos fitopatogênicos
• Beneficiário: Jaqueline Moraes Bazioli
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto