A fungal sRNA silences a host plant transcription factor to promote arbuscular mycorrhizal symbiosis

Cross-kingdom RNA interference (ckRNAi) is a mechanism of interspecies communication where small RNAs (sRNAs) are transported from one organism to another; these sRNAs silence target genes in trans by loading into host AGO proteins. In this work, we investigated the occurrence of ckRNAi in Arbuscula...

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Detalhes bibliográficos
Autores: Silvestri, Alessandro, Ledford, William Conrad, Fiorilli, Valentina, Votta, Cristina, Scerna, Alessia, Tucconi, Jacopo, Mocchetti, Antonio, Grasso, Gianluca, Balestrini, Raffaella, Jin, Hailing, Rubio-Somoza, Ignacio, Lanfranco, Luisa
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2025
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositório:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/388629
Acesso em linha:http://hdl.handle.net/10261/388629
https://api.elsevier.com/content/abstract/scopus_id/85209774160
Access Level:Acceso aberto
Palavra-chave:Medicago truncatula
Rhizophagus irregularis
Arbuscular mycorrhizal symbiosis
Cross‐kingdom RNA interference
Small RNA
Descrição
Resumo:Cross-kingdom RNA interference (ckRNAi) is a mechanism of interspecies communication where small RNAs (sRNAs) are transported from one organism to another; these sRNAs silence target genes in trans by loading into host AGO proteins. In this work, we investigated the occurrence of ckRNAi in Arbuscular Mycorrhizal Symbiosis (AMS). We used an in silico prediction analysis to identify a sRNA (Rir2216) from the AM fungus Rhizophagus irregularis and its putative plant gene target, the Medicago truncatula MtWRKY69 transcription factor. Heterologous co-expression assays in Nicotiana benthamiana, 5' RACE reactions and AGO1-immunoprecipitation assays from mycorrhizal roots were used to characterize the Rir2216-MtWRKY69 interaction. We further analyzed MtWRKY69 expression profile and the contribution of constitutive and conditional MtWRKY69 expression to AMS. We show that Rir2216 is loaded into an AGO1 silencing complex from the host plant M. truncatula, leading to cleavage of a host target transcript encoding for the MtWRKY69 transcription factor. MtWRKY69 is specifically downregulated in arbusculated cells in mycorrhizal roots and increased levels of MtWRKY69 expression led to a reduced AM colonization level. Our results indicate that MtWRKY69 silencing, mediated by a fungal sRNA, is relevant for AMS; we thus present the first experimental evidence of fungus to plant ckRNAi in AMS.