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

Summary 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 A...

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Veröffentlicht in:The New phytologist Jg. 246; H. 3; S. 924 - 935
Hauptverfasser: 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
Format: Journal Article
Sprache:Englisch
Veröffentlicht: England Wiley Subscription Services, Inc 01.05.2025
John Wiley and Sons Inc
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ISSN:0028-646X, 1469-8137, 1469-8137
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Abstract Summary 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. See also the Commentary on this article by Xie & Fan, 246: 814–817.
AbstractList Summary 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. See also the Commentary on this article by Xie & Fan, 246: 814–817.
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.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.
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. See also the Commentary on this article by Xie & Fan, 246: 814–817.
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.
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.
Author Mocchetti, Antonio
Votta, Cristina
Scerna, Alessia
Silvestri, Alessandro
Tucconi, Jacopo
Ledford, William Conrad
Balestrini, Raffaella
Rubio‐Somoza, Ignacio
Jin, Hailing
Lanfranco, Luisa
Fiorilli, Valentina
Grasso, Gianluca
AuthorAffiliation 4 Department of Microbiology and Plant Pathology, Center for Plant Cell Biology, Institute for Integrative Genome Biology University of California 900 University Avenue Riverside CA 92521 USA
3 Institute of Biosciences and Bioresources, CNR via Amendola 165/A 70126 Bari Italy
1 Department of Life Sciences and Systems Biology University of Torino Viale Mattioli 25 10125 Turin Italy
2 Molecular Reprogramming and Evolution (MoRE) Lab Centre for Research in Agricultural Genomics (CRAG) Carrer de la Vall Moronta, 08193 Cerdanyola del Vallès Barcelona Spain
AuthorAffiliation_xml – name: 3 Institute of Biosciences and Bioresources, CNR via Amendola 165/A 70126 Bari Italy
– name: 2 Molecular Reprogramming and Evolution (MoRE) Lab Centre for Research in Agricultural Genomics (CRAG) Carrer de la Vall Moronta, 08193 Cerdanyola del Vallès Barcelona Spain
– name: 4 Department of Microbiology and Plant Pathology, Center for Plant Cell Biology, Institute for Integrative Genome Biology University of California 900 University Avenue Riverside CA 92521 USA
– name: 1 Department of Life Sciences and Systems Biology University of Torino Viale Mattioli 25 10125 Turin Italy
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  surname: Grasso
  fullname: Grasso, Gianluca
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  orcidid: 0000-0001-5778-5193
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  surname: Rubio‐Somoza
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  email: luisa.lanfranco@unito.it
  organization: University of Torino
BackLink https://www.ncbi.nlm.nih.gov/pubmed/39555692$$D View this record in MEDLINE/PubMed
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IsOpenAccess true
IsPeerReviewed true
IsScholarly true
Issue 3
Keywords Medicago truncatula
cross‐kingdom RNA interference
small RNA
arbuscular mycorrhizal symbiosis
Rhizophagus irregularis
Language English
License Attribution
2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.
This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
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Notes These authors contributed equally to this work.
814–817
Xie & Fan
246
See also the Commentary on this article by
.
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 14
content type line 23
See also the Commentary on this article by Xie & Fan, 246: 814–817.
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Snippet Summary Cross‐kingdom RNA interference (ckRNAi) is a mechanism of interspecies communication where small RNAs (sRNAs) are transported from one organism to...
Cross‐kingdom RNA interference (ckRNAi) is a mechanism of interspecies communication where small RNAs (sRNAs) are transported from one organism to another;...
Cross-kingdom RNA interference (ckRNAi) is a mechanism of interspecies communication where small RNAs (sRNAs) are transported from one organism to another;...
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SubjectTerms Alfalfa
arbuscular mycorrhizal symbiosis
Arbuscular mycorrhizas
Argonaute Proteins - metabolism
computer simulation
cross‐kingdom RNA interference
Fungi
Gene Expression Regulation, Plant
Gene Silencing
genes
Glomeromycota - genetics
Glomeromycota - physiology
Host plants
Immunoprecipitation
Medicago truncatula
Medicago truncatula - genetics
Medicago truncatula - microbiology
Mycorrhizae - genetics
Mycorrhizae - physiology
mycorrhizal fungi
Nicotiana - genetics
Nicotiana - microbiology
Nicotiana benthamiana
Plant Proteins - genetics
Plant Proteins - metabolism
Plant Roots - microbiology
Plants
prediction
Priority Report
Rhizophagus irregularis
RNA interference
RNA, Fungal - genetics
RNA, Fungal - metabolism
RNA, Small Untranslated - genetics
RNA, Small Untranslated - metabolism
RNA-mediated interference
Roots
small RNA
Symbiosis
Symbiosis - genetics
Transcription factors
Transcription Factors - genetics
Transcription Factors - metabolism
vesicular arbuscular mycorrhizae
Title A fungal sRNA silences a host plant transcription factor to promote arbuscular mycorrhizal symbiosis
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