CRISPRi in Xanthomonas demonstrates functional convergence of transcription activator-like effectors in two divergent pathogens
Functional analysis of large gene families in plant pathogens can be cumbersome using classical insertional mutagenesis. Additionally, Cas9 toxicity has limited the application of CRISPR–Cas9 for directed mutagenesis in bacteria. Here, we successfully applied a CRISPR interference strategy to investigate the cryptic role of the transcription activator-like effector (tale) multigene family in several plant-pathogenic Xanthomonas bacterial species, owing to their contribution to pathogen virulence. Single guide RNAs (sgRNAs) designed against Xanthomonas phaseoli pv manihotis tale conserved gene sequences efficiently silenced expression of all tales, with concomitant decrease in virulence and TALE-induced host gene expression. The system is readily translatable to other Xanthomonas species infecting rice, citrus, Brassica, and cassava, silencing up to 16 tales in a given strain using a single sgRNA. Complementation with plasmid-borne designer tales lacking the sgRNA-targeted sequence restored molecular and virulence phenotypes in all pathosystems. Our results evidenced that X. campestris pv campestris CN08 tales are relevant for symptom development in cauliflower. They also show that the MeSWEET10a sugar transporter is surprisingly targeted by the nonvascular cassava pathogen X. cassavae, highlighting a new example of TALE functional convergence between phylogenetically distant Xanthomonas. Overall, this novel technology provides a platform for discovery and rapid functional understanding of highly conserved gene families.
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dig-cirad-fr-608019 |
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Francia |
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Europa del Oeste |
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Biblioteca del CIRAD Francia |
language |
eng |
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H20 - Maladies des plantes Oryza sativa Xanthomonas expression des gènes phylogénie Xanthomonas oryzae Transcription génique virulence Manihot esculenta pouvoir pathogène Xanthomonas campestris manihotis Xanthomonas campestris phaseoli effecteur moléculaire maladie des plantes gène phénotype http://aims.fao.org/aos/agrovoc/c_5438 http://aims.fao.org/aos/agrovoc/c_8455 http://aims.fao.org/aos/agrovoc/c_27527 http://aims.fao.org/aos/agrovoc/c_13325 http://aims.fao.org/aos/agrovoc/c_24383 http://aims.fao.org/aos/agrovoc/c_35128 http://aims.fao.org/aos/agrovoc/c_8261 http://aims.fao.org/aos/agrovoc/c_4579 http://aims.fao.org/aos/agrovoc/c_5629 http://aims.fao.org/aos/agrovoc/c_36122 http://aims.fao.org/aos/agrovoc/c_36124 http://aims.fao.org/aos/agrovoc/c_27506 http://aims.fao.org/aos/agrovoc/c_5962 http://aims.fao.org/aos/agrovoc/c_3214 http://aims.fao.org/aos/agrovoc/c_5776 H20 - Maladies des plantes Oryza sativa Xanthomonas expression des gènes phylogénie Xanthomonas oryzae Transcription génique virulence Manihot esculenta pouvoir pathogène Xanthomonas campestris manihotis Xanthomonas campestris phaseoli effecteur moléculaire maladie des plantes gène phénotype http://aims.fao.org/aos/agrovoc/c_5438 http://aims.fao.org/aos/agrovoc/c_8455 http://aims.fao.org/aos/agrovoc/c_27527 http://aims.fao.org/aos/agrovoc/c_13325 http://aims.fao.org/aos/agrovoc/c_24383 http://aims.fao.org/aos/agrovoc/c_35128 http://aims.fao.org/aos/agrovoc/c_8261 http://aims.fao.org/aos/agrovoc/c_4579 http://aims.fao.org/aos/agrovoc/c_5629 http://aims.fao.org/aos/agrovoc/c_36122 http://aims.fao.org/aos/agrovoc/c_36124 http://aims.fao.org/aos/agrovoc/c_27506 http://aims.fao.org/aos/agrovoc/c_5962 http://aims.fao.org/aos/agrovoc/c_3214 http://aims.fao.org/aos/agrovoc/c_5776 |
spellingShingle |
H20 - Maladies des plantes Oryza sativa Xanthomonas expression des gènes phylogénie Xanthomonas oryzae Transcription génique virulence Manihot esculenta pouvoir pathogène Xanthomonas campestris manihotis Xanthomonas campestris phaseoli effecteur moléculaire maladie des plantes gène phénotype http://aims.fao.org/aos/agrovoc/c_5438 http://aims.fao.org/aos/agrovoc/c_8455 http://aims.fao.org/aos/agrovoc/c_27527 http://aims.fao.org/aos/agrovoc/c_13325 http://aims.fao.org/aos/agrovoc/c_24383 http://aims.fao.org/aos/agrovoc/c_35128 http://aims.fao.org/aos/agrovoc/c_8261 http://aims.fao.org/aos/agrovoc/c_4579 http://aims.fao.org/aos/agrovoc/c_5629 http://aims.fao.org/aos/agrovoc/c_36122 http://aims.fao.org/aos/agrovoc/c_36124 http://aims.fao.org/aos/agrovoc/c_27506 http://aims.fao.org/aos/agrovoc/c_5962 http://aims.fao.org/aos/agrovoc/c_3214 http://aims.fao.org/aos/agrovoc/c_5776 H20 - Maladies des plantes Oryza sativa Xanthomonas expression des gènes phylogénie Xanthomonas oryzae Transcription génique virulence Manihot esculenta pouvoir pathogène Xanthomonas campestris manihotis Xanthomonas campestris phaseoli effecteur moléculaire maladie des plantes gène phénotype http://aims.fao.org/aos/agrovoc/c_5438 http://aims.fao.org/aos/agrovoc/c_8455 http://aims.fao.org/aos/agrovoc/c_27527 http://aims.fao.org/aos/agrovoc/c_13325 http://aims.fao.org/aos/agrovoc/c_24383 http://aims.fao.org/aos/agrovoc/c_35128 http://aims.fao.org/aos/agrovoc/c_8261 http://aims.fao.org/aos/agrovoc/c_4579 http://aims.fao.org/aos/agrovoc/c_5629 http://aims.fao.org/aos/agrovoc/c_36122 http://aims.fao.org/aos/agrovoc/c_36124 http://aims.fao.org/aos/agrovoc/c_27506 http://aims.fao.org/aos/agrovoc/c_5962 http://aims.fao.org/aos/agrovoc/c_3214 http://aims.fao.org/aos/agrovoc/c_5776 Zárate-Chaves, Carlos Andrés Audran, Corinne Medina Culma, César Augusto Escalon, Aline Javegny, Stéphanie Gagnevin, Lionel Thomas, Emilie Pimparé, Léa-Lou López, Camilo Jacobs, Jonathan M. Noel, Laurent D. Koebnik, Ralf Bernal, Adriana Jimena Szurek, Boris CRISPRi in Xanthomonas demonstrates functional convergence of transcription activator-like effectors in two divergent pathogens |
description |
Functional analysis of large gene families in plant pathogens can be cumbersome using classical insertional mutagenesis. Additionally, Cas9 toxicity has limited the application of CRISPR–Cas9 for directed mutagenesis in bacteria. Here, we successfully applied a CRISPR interference strategy to investigate the cryptic role of the transcription activator-like effector (tale) multigene family in several plant-pathogenic Xanthomonas bacterial species, owing to their contribution to pathogen virulence. Single guide RNAs (sgRNAs) designed against Xanthomonas phaseoli pv manihotis tale conserved gene sequences efficiently silenced expression of all tales, with concomitant decrease in virulence and TALE-induced host gene expression. The system is readily translatable to other Xanthomonas species infecting rice, citrus, Brassica, and cassava, silencing up to 16 tales in a given strain using a single sgRNA. Complementation with plasmid-borne designer tales lacking the sgRNA-targeted sequence restored molecular and virulence phenotypes in all pathosystems. Our results evidenced that X. campestris pv campestris CN08 tales are relevant for symptom development in cauliflower. They also show that the MeSWEET10a sugar transporter is surprisingly targeted by the nonvascular cassava pathogen X. cassavae, highlighting a new example of TALE functional convergence between phylogenetically distant Xanthomonas. Overall, this novel technology provides a platform for discovery and rapid functional understanding of highly conserved gene families. |
format |
article |
topic_facet |
H20 - Maladies des plantes Oryza sativa Xanthomonas expression des gènes phylogénie Xanthomonas oryzae Transcription génique virulence Manihot esculenta pouvoir pathogène Xanthomonas campestris manihotis Xanthomonas campestris phaseoli effecteur moléculaire maladie des plantes gène phénotype http://aims.fao.org/aos/agrovoc/c_5438 http://aims.fao.org/aos/agrovoc/c_8455 http://aims.fao.org/aos/agrovoc/c_27527 http://aims.fao.org/aos/agrovoc/c_13325 http://aims.fao.org/aos/agrovoc/c_24383 http://aims.fao.org/aos/agrovoc/c_35128 http://aims.fao.org/aos/agrovoc/c_8261 http://aims.fao.org/aos/agrovoc/c_4579 http://aims.fao.org/aos/agrovoc/c_5629 http://aims.fao.org/aos/agrovoc/c_36122 http://aims.fao.org/aos/agrovoc/c_36124 http://aims.fao.org/aos/agrovoc/c_27506 http://aims.fao.org/aos/agrovoc/c_5962 http://aims.fao.org/aos/agrovoc/c_3214 http://aims.fao.org/aos/agrovoc/c_5776 |
author |
Zárate-Chaves, Carlos Andrés Audran, Corinne Medina Culma, César Augusto Escalon, Aline Javegny, Stéphanie Gagnevin, Lionel Thomas, Emilie Pimparé, Léa-Lou López, Camilo Jacobs, Jonathan M. Noel, Laurent D. Koebnik, Ralf Bernal, Adriana Jimena Szurek, Boris |
author_facet |
Zárate-Chaves, Carlos Andrés Audran, Corinne Medina Culma, César Augusto Escalon, Aline Javegny, Stéphanie Gagnevin, Lionel Thomas, Emilie Pimparé, Léa-Lou López, Camilo Jacobs, Jonathan M. Noel, Laurent D. Koebnik, Ralf Bernal, Adriana Jimena Szurek, Boris |
author_sort |
Zárate-Chaves, Carlos Andrés |
title |
CRISPRi in Xanthomonas demonstrates functional convergence of transcription activator-like effectors in two divergent pathogens |
title_short |
CRISPRi in Xanthomonas demonstrates functional convergence of transcription activator-like effectors in two divergent pathogens |
title_full |
CRISPRi in Xanthomonas demonstrates functional convergence of transcription activator-like effectors in two divergent pathogens |
title_fullStr |
CRISPRi in Xanthomonas demonstrates functional convergence of transcription activator-like effectors in two divergent pathogens |
title_full_unstemmed |
CRISPRi in Xanthomonas demonstrates functional convergence of transcription activator-like effectors in two divergent pathogens |
title_sort |
crispri in xanthomonas demonstrates functional convergence of transcription activator-like effectors in two divergent pathogens |
url |
http://agritrop.cirad.fr/608019/ http://agritrop.cirad.fr/608019/1/New%20Phytologist%20-%202023%20-%20Z%C3%A1rate%E2%80%90Chaves%20-%20CRISPRi%20in%20Xanthomonas%20demonstrates%20functional%20convergence%20of%20transcription.pdf |
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dig-cirad-fr-6080192024-01-30T08:06:10Z http://agritrop.cirad.fr/608019/ http://agritrop.cirad.fr/608019/ CRISPRi in Xanthomonas demonstrates functional convergence of transcription activator-like effectors in two divergent pathogens. Zárate-Chaves Carlos Andrés, Audran Corinne, Medina Culma César Augusto, Escalon Aline, Javegny Stéphanie, Gagnevin Lionel, Thomas Emilie, Pimparé Léa-Lou, López Camilo, Jacobs Jonathan M., Noel Laurent D., Koebnik Ralf, Bernal Adriana Jimena, Szurek Boris. 2023. New Phytologist, 238 (4) : 1593-1604.https://doi.org/10.1111/nph.18808 <https://doi.org/10.1111/nph.18808> CRISPRi in Xanthomonas demonstrates functional convergence of transcription activator-like effectors in two divergent pathogens Zárate-Chaves, Carlos Andrés Audran, Corinne Medina Culma, César Augusto Escalon, Aline Javegny, Stéphanie Gagnevin, Lionel Thomas, Emilie Pimparé, Léa-Lou López, Camilo Jacobs, Jonathan M. Noel, Laurent D. Koebnik, Ralf Bernal, Adriana Jimena Szurek, Boris eng 2023 New Phytologist H20 - Maladies des plantes Oryza sativa Xanthomonas expression des gènes phylogénie Xanthomonas oryzae Transcription génique virulence Manihot esculenta pouvoir pathogène Xanthomonas campestris manihotis Xanthomonas campestris phaseoli effecteur moléculaire maladie des plantes gène phénotype http://aims.fao.org/aos/agrovoc/c_5438 http://aims.fao.org/aos/agrovoc/c_8455 http://aims.fao.org/aos/agrovoc/c_27527 http://aims.fao.org/aos/agrovoc/c_13325 http://aims.fao.org/aos/agrovoc/c_24383 http://aims.fao.org/aos/agrovoc/c_35128 http://aims.fao.org/aos/agrovoc/c_8261 http://aims.fao.org/aos/agrovoc/c_4579 http://aims.fao.org/aos/agrovoc/c_5629 http://aims.fao.org/aos/agrovoc/c_36122 http://aims.fao.org/aos/agrovoc/c_36124 http://aims.fao.org/aos/agrovoc/c_27506 http://aims.fao.org/aos/agrovoc/c_5962 http://aims.fao.org/aos/agrovoc/c_3214 http://aims.fao.org/aos/agrovoc/c_5776 Functional analysis of large gene families in plant pathogens can be cumbersome using classical insertional mutagenesis. Additionally, Cas9 toxicity has limited the application of CRISPR–Cas9 for directed mutagenesis in bacteria. Here, we successfully applied a CRISPR interference strategy to investigate the cryptic role of the transcription activator-like effector (tale) multigene family in several plant-pathogenic Xanthomonas bacterial species, owing to their contribution to pathogen virulence. Single guide RNAs (sgRNAs) designed against Xanthomonas phaseoli pv manihotis tale conserved gene sequences efficiently silenced expression of all tales, with concomitant decrease in virulence and TALE-induced host gene expression. The system is readily translatable to other Xanthomonas species infecting rice, citrus, Brassica, and cassava, silencing up to 16 tales in a given strain using a single sgRNA. Complementation with plasmid-borne designer tales lacking the sgRNA-targeted sequence restored molecular and virulence phenotypes in all pathosystems. Our results evidenced that X. campestris pv campestris CN08 tales are relevant for symptom development in cauliflower. They also show that the MeSWEET10a sugar transporter is surprisingly targeted by the nonvascular cassava pathogen X. cassavae, highlighting a new example of TALE functional convergence between phylogenetically distant Xanthomonas. Overall, this novel technology provides a platform for discovery and rapid functional understanding of highly conserved gene families. article info:eu-repo/semantics/article Journal Article info:eu-repo/semantics/publishedVersion http://agritrop.cirad.fr/608019/1/New%20Phytologist%20-%202023%20-%20Z%C3%A1rate%E2%80%90Chaves%20-%20CRISPRi%20in%20Xanthomonas%20demonstrates%20functional%20convergence%20of%20transcription.pdf text cc_by info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by/4.0/ https://doi.org/10.1111/nph.18808 10.1111/nph.18808 info:eu-repo/semantics/altIdentifier/doi/10.1111/nph.18808 info:eu-repo/semantics/altIdentifier/purl/https://doi.org/10.1111/nph.18808 info:eu-repo/semantics/dataset/purl/https://www.ncbi.nlm.nih.gov/datasets/genome/GCF_000019585.2/ info:eu-repo/semantics/dataset/purl/https://www.ncbi.nlm.nih.gov/datasets/genome/GCF_003031365.1/ info:eu-repo/semantics/dataset/purl/https://www.ncbi.nlm.nih.gov/datasets/genome/GCF_000007165.1/ info:eu-repo/grantAgreement///ANR-18-EURE-0019//(FRA) The Toulouse-Perpignan/TULIP-GSR |