First report of Xanthomonas prunicola causing bacterial leaf streaks on wheat in Argentina
Since 2018, bacterial-like symptoms, such as leaf streaks were observed on wheat plants (Triticum aestivum L.) in Córdoba province in Argentina, with 1 to 5% of disease incidence. Samples of wheat stem and spike collected in a trial of varieties for summer/autumn sowing in the experimental field of...
| Main Authors: | , , , , , , , , , , , |
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| Format: | info:ar-repo/semantics/artículo |
| Language: | Inglés |
| Published: |
American Phytopathological Society
2022
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| Subjects: | |
| Online Access: | http://hdl.handle.net/20.500.12123/13392 https://apsjournals.apsnet.org/doi/10.1094/PDIS-04-22-0886-PDN https://doi.org/10.1094/PDIS-04-22-0886-PDN |
| _version_ | 1855036928657719296 |
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| author | Martino, Julia Andrea Fernandez, Franco Daniel Pozzi, Elizabeth Alicia Alberione, Enrique Javier Bainotti, Carlos Tomas Marquez, Nathalie Tolocka, Patricia Salines, Nicolas Gomez, Dionisio Tomas Donaire, Guillermo Manuel Conci, Luis Rogelio Alemandri, Vanina Maria |
| author_browse | Alberione, Enrique Javier Alemandri, Vanina Maria Bainotti, Carlos Tomas Conci, Luis Rogelio Donaire, Guillermo Manuel Fernandez, Franco Daniel Gomez, Dionisio Tomas Marquez, Nathalie Martino, Julia Andrea Pozzi, Elizabeth Alicia Salines, Nicolas Tolocka, Patricia |
| author_facet | Martino, Julia Andrea Fernandez, Franco Daniel Pozzi, Elizabeth Alicia Alberione, Enrique Javier Bainotti, Carlos Tomas Marquez, Nathalie Tolocka, Patricia Salines, Nicolas Gomez, Dionisio Tomas Donaire, Guillermo Manuel Conci, Luis Rogelio Alemandri, Vanina Maria |
| author_sort | Martino, Julia Andrea |
| collection | INTA Digital |
| description | Since 2018, bacterial-like symptoms, such as leaf streaks were observed on wheat plants (Triticum aestivum L.) in Córdoba province in Argentina, with 1 to 5% of disease incidence. Samples of wheat stem and spike collected in a trial of varieties for summer/autumn sowing in the experimental field of the INTA Marcos Juárez were disinfected, washed and macerated in mortars with sterile distilled water and extracts were streaked on Luria-Bertani (LB) agar. After 48 h incubation at 28 °C, circular, mucoid, convex, and cream colonies were observed and pure cultures were transferred to LB medium for further identification tests. Biochemical tests corroborated the detection of a Gram-negative bacillus. Conventional PCR was performed using DNA isolate from pure cultures and general primers for various species of genera Xanthomonas (Maes 1993) and Pseudomonas (Mulet et al. 2010). An isolate (Arg-1), with cream colored colonies was positive using general primers for Xanthomonas sp (amplified fragment of 444 bp). A bacterial suspension containing 108 CFU mL−1 grown for 48 h on LB medium at 28 °C was injected into three-week-old leaves of wheat plants to fulfill Koch’s postulates. After 5 days, plants showed symptoms of chlorosis, streaks and then necrosis on the leaves. The bacteria were re-isolated from the inoculated plants, showing same symptoms observed in the original plants. Negative control plants, inoculated with sterile water remained without symptoms. The amplified 444 bp fragment described above was sequenced by the Sanger method (GenBank accession OM972662), as well as another 757 bp fragment amplified with universal primers that amplify the partial 16S rDNA gene (GenBank accession OM972661). Analyses of these sequences, as well as the protein profile of the isolate obtained by matrix assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOF MS) Bruker Biotyper, allowed to identify only the genus Xanthomonas. With the purpose of determine the species status, the complete genome of isolate Arg-1 was sequenced using Oxford Nanopore Technologies (ONT). Total gDNA was isolate from pure cultures using a commercial kit (Wizard Genomic DNA Purification Kit, Promega). gDNA library was constructed using Ligation Sequencing Kit (SQK-LSK109) and sequenced using ONT platform on a MinION 1kb device. Raw basecalled sequences were filtered using Filtlong and assembled using Trycycler. The genome was assembled in a single contig comprising 5.410.641 bp with 4740 predicted CDSs and 63.9% GC content. Genome sequence was deposited in GenBank under accession number CP094827 and SRA data SRX14635308. Whole-genome Average Nucleotide Identity (ANI) analysis showed values of ~ 97% against the reference genomes of Xanthomonas prunicola (PHKX01.1, PHKV01.1 and PHKW01.1) and 100% in complete 16S rRNA gene sequences (1547 bp). These findings suggest that a new wheat pathogen within the genus Xanthomonas is present in Argentina, as well as was reported in Uruguay and USA (Clavijo et al. 2021). To our knowledge, this is the first report of X. prunicola affecting wheat in Argentina and the first complete genome registered for this specie. Accurate and specific diagnostics are required for the detection of X. prunicola in wheat crops to implement correct prevention and control strategies to this disease, avoiding the dissemination in lots where it has not yet been found. |
| format | info:ar-repo/semantics/artículo |
| id | INTA13392 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2022 |
| publishDateRange | 2022 |
| publishDateSort | 2022 |
| publisher | American Phytopathological Society |
| publisherStr | American Phytopathological Society |
| record_format | dspace |
| spelling | INTA133922022-11-11T18:08:33Z First report of Xanthomonas prunicola causing bacterial leaf streaks on wheat in Argentina Martino, Julia Andrea Fernandez, Franco Daniel Pozzi, Elizabeth Alicia Alberione, Enrique Javier Bainotti, Carlos Tomas Marquez, Nathalie Tolocka, Patricia Salines, Nicolas Gomez, Dionisio Tomas Donaire, Guillermo Manuel Conci, Luis Rogelio Alemandri, Vanina Maria Argentina Triticum Aestivum Trigo Wheat Plant Diseases Enfermedades de las Plantas Xanthomonas Bacterial Leaf Streak of Wheat Pathogen Detection Xanthomonas Prunicola Since 2018, bacterial-like symptoms, such as leaf streaks were observed on wheat plants (Triticum aestivum L.) in Córdoba province in Argentina, with 1 to 5% of disease incidence. Samples of wheat stem and spike collected in a trial of varieties for summer/autumn sowing in the experimental field of the INTA Marcos Juárez were disinfected, washed and macerated in mortars with sterile distilled water and extracts were streaked on Luria-Bertani (LB) agar. After 48 h incubation at 28 °C, circular, mucoid, convex, and cream colonies were observed and pure cultures were transferred to LB medium for further identification tests. Biochemical tests corroborated the detection of a Gram-negative bacillus. Conventional PCR was performed using DNA isolate from pure cultures and general primers for various species of genera Xanthomonas (Maes 1993) and Pseudomonas (Mulet et al. 2010). An isolate (Arg-1), with cream colored colonies was positive using general primers for Xanthomonas sp (amplified fragment of 444 bp). A bacterial suspension containing 108 CFU mL−1 grown for 48 h on LB medium at 28 °C was injected into three-week-old leaves of wheat plants to fulfill Koch’s postulates. After 5 days, plants showed symptoms of chlorosis, streaks and then necrosis on the leaves. The bacteria were re-isolated from the inoculated plants, showing same symptoms observed in the original plants. Negative control plants, inoculated with sterile water remained without symptoms. The amplified 444 bp fragment described above was sequenced by the Sanger method (GenBank accession OM972662), as well as another 757 bp fragment amplified with universal primers that amplify the partial 16S rDNA gene (GenBank accession OM972661). Analyses of these sequences, as well as the protein profile of the isolate obtained by matrix assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOF MS) Bruker Biotyper, allowed to identify only the genus Xanthomonas. With the purpose of determine the species status, the complete genome of isolate Arg-1 was sequenced using Oxford Nanopore Technologies (ONT). Total gDNA was isolate from pure cultures using a commercial kit (Wizard Genomic DNA Purification Kit, Promega). gDNA library was constructed using Ligation Sequencing Kit (SQK-LSK109) and sequenced using ONT platform on a MinION 1kb device. Raw basecalled sequences were filtered using Filtlong and assembled using Trycycler. The genome was assembled in a single contig comprising 5.410.641 bp with 4740 predicted CDSs and 63.9% GC content. Genome sequence was deposited in GenBank under accession number CP094827 and SRA data SRX14635308. Whole-genome Average Nucleotide Identity (ANI) analysis showed values of ~ 97% against the reference genomes of Xanthomonas prunicola (PHKX01.1, PHKV01.1 and PHKW01.1) and 100% in complete 16S rRNA gene sequences (1547 bp). These findings suggest that a new wheat pathogen within the genus Xanthomonas is present in Argentina, as well as was reported in Uruguay and USA (Clavijo et al. 2021). To our knowledge, this is the first report of X. prunicola affecting wheat in Argentina and the first complete genome registered for this specie. Accurate and specific diagnostics are required for the detection of X. prunicola in wheat crops to implement correct prevention and control strategies to this disease, avoiding the dissemination in lots where it has not yet been found. Instituto de Patología Vegetal Fil: Martino, Julia Andrea. Fondo para la Investigación Científica y Tecnológica (FONCYT); Argentina Fil: Martino, Julia Andrea. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina Fil: Fernandez, Franco Daniel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina Fil: Fernandez, Franco Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Fitopatología y Modelización Agrícola (UFyMA); Argentina Fil: Pozzi, Elizabeth Alicia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; Argentina Fil: Alberione, Enrique Javier. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; Argentina Fil: Bainotti, Carlos Tomas. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; Argentina Fil: Marquez, Nathalie. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina Fil: Marquez, Nathalie. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Fitopatología y Modelización Agrícola (UFyMA); Argentina Fil: Tolocka, Patricia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina Fil: Tolocka, Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Fitopatología y Modelización Agrícola (UFyMA); Argentina Fil: Salines, Nicolas. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; Argentina Fil: Gomez, Dionisio Tomas. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; Argentina Fil: Donaire, Guillermo Manuel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Marcos Juárez; Argentina Fil: Conci, Luis Rogelio. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina Fil: Conci, Luis Rogelio. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Fitopatología y Modelización Agrícola (UFyMA); Argentina Fil: Alemandri, Vanina Maria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina Fil: Alemandri, Vanina Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Fitopatología y Modelización Agrícola (UFyMA); Argentina 2022-11-11T17:59:33Z 2022-11-11T17:59:33Z 2022-06-13 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/13392 https://apsjournals.apsnet.org/doi/10.1094/PDIS-04-22-0886-PDN 0191-2917 https://doi.org/10.1094/PDIS-04-22-0886-PDN eng info:eu-repograntAgreement/INTA/2019-PD-E4-I090-001/2019-PD-E4-I090-001/AR./Análisis de patosistemas en cultivos agrícolas y especies forestales. Caracterización de sus componentes info:eu-repograntAgreement/INTA/2019-PE-E6-I126-001/2019-PE-E6-I126-001/AR./Mejoramiento genético de trigo pan (Triticum aestivum L.) y trigo candeal (Triticum turgidum ssp. durum Desf.) info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf American Phytopathological Society Plant Disease (Published Online:13 Jun 2022) |
| spellingShingle | Argentina Triticum Aestivum Trigo Wheat Plant Diseases Enfermedades de las Plantas Xanthomonas Bacterial Leaf Streak of Wheat Pathogen Detection Xanthomonas Prunicola Martino, Julia Andrea Fernandez, Franco Daniel Pozzi, Elizabeth Alicia Alberione, Enrique Javier Bainotti, Carlos Tomas Marquez, Nathalie Tolocka, Patricia Salines, Nicolas Gomez, Dionisio Tomas Donaire, Guillermo Manuel Conci, Luis Rogelio Alemandri, Vanina Maria First report of Xanthomonas prunicola causing bacterial leaf streaks on wheat in Argentina |
| title | First report of Xanthomonas prunicola causing bacterial leaf streaks on wheat in Argentina |
| title_full | First report of Xanthomonas prunicola causing bacterial leaf streaks on wheat in Argentina |
| title_fullStr | First report of Xanthomonas prunicola causing bacterial leaf streaks on wheat in Argentina |
| title_full_unstemmed | First report of Xanthomonas prunicola causing bacterial leaf streaks on wheat in Argentina |
| title_short | First report of Xanthomonas prunicola causing bacterial leaf streaks on wheat in Argentina |
| title_sort | first report of xanthomonas prunicola causing bacterial leaf streaks on wheat in argentina |
| topic | Argentina Triticum Aestivum Trigo Wheat Plant Diseases Enfermedades de las Plantas Xanthomonas Bacterial Leaf Streak of Wheat Pathogen Detection Xanthomonas Prunicola |
| url | http://hdl.handle.net/20.500.12123/13392 https://apsjournals.apsnet.org/doi/10.1094/PDIS-04-22-0886-PDN https://doi.org/10.1094/PDIS-04-22-0886-PDN |
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