Skip to content
1887

Abstract

The São Paulo state citrus belt in Brazil is a major citrus production region. Since at least 1957, citrus plantations in this region have been affected by citrus canker, an economically damaging disease caused by subsp. (). For about 50 years, until 2017, a citrus canker eradication programme was carried out in this region. In this work, our aim was to investigate the effects of the eradication programme on genetic variability and evolution of . To this end, we sequenced and analysed 758 genomes sampled in the São Paulo citrus belt, together with 730 publicly available genomes from around the world. Our phylogenomic analyses show that these genomes can be grouped into seven major lineages and that in São Paulo, lineage L7 is dominant. Our time estimate for its appearance closely matches the date when citrus production expanded. L7 can be subdivided into lineages L7.1 and L7.2. In our samples, L7.2, which we estimate to have emerged around 1964, is by far the most abundant, showing that the eradication programme had little impact on strain diversification. On the other hand, oscillations in the estimated effective population size of L7.2 strains over time closely match the shifts in the eradication programme. In sum, we present a detailed view of the genomic diversity of in the world and in São Paulo, the largest such effort in terms of a number of genomes for a crop pathogen undertaken so far. The methods employed here can form the basis for active genomic surveillance of in major citrus production areas.

Funding
This study was supported by the:
  • Conselho Nacional de Desenvolvimento Científico e Tecnológico (Award 305961/2019-5)
    • Principal Award Recipient: JoaoCarlos Setubal
  • Conselho Nacional de Desenvolvimento Científico e Tecnológico (Award 307163/2020-2)
    • Principal Award Recipient: HenriqueFerreira
  • Conselho Nacional de Desenvolvimento Científico e Tecnológico (Award 302872/2020-5)
    • Principal Award Recipient: FranklinBehlau
  • Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Award Finance Code 001)
    • Principal Award Recipient: CaioFelipe Cavicchia Zamunér
  • Fundo de Defesa da Citricultura (Award 031686)
    • Principal Award Recipient: FranklinBehlau
  • Conselho Nacional de Desenvolvimento Científico e Tecnológico (Award 465440/2014-2)
    • Principal Award Recipient: HenriqueFerreira
  • Fundação de Amparo à Pesquisa do Estado de São Paulo (Award 2014/50880-0)
    • Principal Award Recipient: HenriqueFerreira
  • Fundação de Amparo à Pesquisa do Estado de São Paulo (Award 2021/10577-0)
    • Principal Award Recipient: HenriqueFerreira
  • Fundação de Amparo à Pesquisa do Estado de São Paulo (Award 2018/21164-5)
    • Principal Award Recipient: HenriqueFerreira
  • Fundação de Amparo à Pesquisa do Estado de São Paulo (Award 2017/50454-9)
    • Principal Award Recipient: HenriqueFerreira
  • Conselho Nacional das Fundações Estaduais de Amparo à Pesquisa (Award 2019/05497-7)
    • Principal Award Recipient: MarkC. Enright
  • Biotechnology and Biological Sciences Research Council (Award BB/S018891/1)
    • Principal Award Recipient: MarkC. Enright
  • Biotechnology and Biological Sciences Research Council (Award BB/R022720/1)
    • Principal Award Recipient: MarkC. Enright
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
Loading

Article metrics loading...

/content/journal/mgen/10.1099/mgen.0.001338
2025-01-16
2025-12-10

Metrics

Loading full text...

Full text loading...

/deliver/fulltext/mgen/11/1/mgen001338.html?itemId=/content/journal/mgen/10.1099/mgen.0.001338&mimeType=html&fmt=ahah

References

  1. Gottwald TR, Graham JH, Schubert TS. Citrus canker: the pathogen and its impact. Plant Health Progress. Plant Management Network 2002; 1993:48824 [View Article]
    [Google Scholar]
  2. Schaad NW, Postnikova E, Lacy G, Sechler A, Agarkova I et al. Emended classification of xanthomonad pathogens on citrus. Syst Appl Microbiol 2006; 29:690–695 [View Article] [PubMed]
    [Google Scholar]
  3. Shahbaz E, Ali M, Shafiq M, Atiq M, Hussain M et al. Citrus canker pathogen, its mechanism of infection, eradication, and impacts. Plants 2022; 12:123 [View Article] [PubMed]
    [Google Scholar]
  4. Ference CM, Gochez AM, Behlau F, Wang N, Graham JH et al. Recent advances in the understanding of Xanthomonas citri ssp. citri pathogenesis and citrus canker disease management. Mol Plant Pathol 2018; 19:1302–1318 [View Article] [PubMed]
    [Google Scholar]
  5. Behlau F. An overview of citrus canker in Brazil. Trop plant pathol 2021; 46:1–12 [View Article]
    [Google Scholar]
  6. Belasque Jr. J, Fernandes NG, Massari CA. The Success of erradication campaign os Citrus Canker in São Paulo States, Brazil. Summa phytopathol 2009; 35:91–92 [View Article]
    [Google Scholar]
  7. Raychaudhuri SP, Verma JP, Nariani TK, Sen B. The history of plant pathology in India. Annu Rev Phytopathol 1972; 10:21–36 [View Article]
    [Google Scholar]
  8. Bitancourt AA. O cancro cítrico. Biologico 1957; 23:101–111
    [Google Scholar]
  9. Behlau F, Fonseca AE, Belasque J. A comprehensive analysis of the Asiatic citrus canker eradication programme in São Paulo state, Brazil, from 1999 to 2009. Plant Pathol 2016; 65:1390–1399 [View Article]
    [Google Scholar]
  10. Behlau F, Belasque J. Cancro cítrico: a doença e seu controle: Fundecitrus; 2014 https://www.fundecitrus.com.br/comunicacao/manual_detalhes/cancro-citrico-a-doenca-e-seu-controle/49
  11. Gordon JL, Lefeuvre P, Escalon A, Barbe V, Cruveiller S et al. Comparative genomics of 43 strains of Xanthomonas citri pv. citri reveals the evolutionary events giving rise to pathotypes with different host ranges. BMC Genom 2015; 16:1098 [View Article] [PubMed]
    [Google Scholar]
  12. Zhang Y, Jalan N, Zhou X, Goss E, Jones JB et al. Positive selection is the main driving force for evolution of citrus canker-causing Xanthomonas. ISME J 2015; 9:2128–2138 [View Article] [PubMed]
    [Google Scholar]
  13. Patané JSL, Martins J Jr, Rangel LT, Belasque J, Digiampietri LA et al. Origin and diversification of Xanthomonas citri subsp. citri pathotypes revealed by inclusive phylogenomic, dating, and biogeographic analyses. BMC Genom 2019; 20:700 [View Article] [PubMed]
    [Google Scholar]
  14. Xu J, Zhang Y, Li J, Teper D, Sun X et al. Phylogenomic analysis of 343 Xanthomonas citri pv. citri strains unravels introduction history and dispersal paths. PLoS Pathog 2023; 19:e1011876 [View Article] [PubMed]
    [Google Scholar]
  15. Jaciani FJ, Ferro JA, Ferro MIT, Vernière C, Pruvost O et al. Genetic diversity of a Brazilian strain collection of Xanthomonas citri subsp. citri based on the type III effector protein genes. Plant Dis 2012; 96:193–203 [View Article] [PubMed]
    [Google Scholar]
  16. Carvalho FM de S, Caramori LPC, Leite Júnior RP. Genetic diversity of Xanthomonas axonopodis pv. citri based on plasmid profile and pulsed field gel electrophoresis. Genet Mol Biol 2005; 28:446–451 [View Article]
    [Google Scholar]
  17. Gonçalvez-Zuliani AMO, Zanutto C, Franco J, Cazetta A, Bock C et al. Genetic diversity of Xanthomonas citri subsp. citri in citrus orchards in northwest Paraná state, Brazil. Phytopahologia Mediterranea 2017; 56:40–49 [View Article]
    [Google Scholar]
  18. Behlau F, Jones JB, Myers ME, Graham JH. Monitoring for resistant populations of Xanthomonas citri subsp. citri and epiphytic bacteria on citrus trees treated with copper or streptomycin using a new semi-selective medium. Eur J Plant Pathol 2012; 132:259–270 [View Article]
    [Google Scholar]
  19. Coletta-Filho HD, Takita MA, Souza AA, Neto JR, Destefano SAL et al. Primers based on the rpf gene region provide improved detection of Xanthomonas axonopodis pv. citri in naturally and artificially infected citrus plants. J Appl Microbiol 2006; 100:279–285 [View Article]
    [Google Scholar]
  20. Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 2014; 30:2114–2120 [View Article] [PubMed]
    [Google Scholar]
  21. Wood DE, Lu J, Langmead B. Improved metagenomic analysis with Kraken 2. Genome Biol 2019; 20:257 [View Article] [PubMed]
    [Google Scholar]
  22. Bankevich A, Nurk S, Antipov D, Gurevich AA, Dvorkin M et al. SPAdes: a new genome assembly algorithm and its applications to single-cell sequencing. J Comput Biol 2012; 19:455–477 [View Article] [PubMed]
    [Google Scholar]
  23. Parks DH, Imelfort M, Skennerton CT, Hugenholtz P, Tyson GW. CheckM: assessing the quality of microbial genomes recovered from isolates, single cells, and metagenomes. Genome Res 2015; 25:1043–1055 [View Article] [PubMed]
    [Google Scholar]
  24. Richard D, Rieux A, Lefeuvre P, Hamza A, Lobin KK et al. Draft genome sequences of 284 Xanthomonas citri pv. citri strains causing Asiatic citrus canker. Microbiol Resour Announc 2021; 10:e01024-20 [View Article] [PubMed]
    [Google Scholar]
  25. Ragupathy R, Jolley KA, Zamuner C, Jones JB, Redfern J et al. Core-genome multilocus sequence typing for epidemiological and evolutionary analyses of phytopathogenic Xanthomonas citri. Appl Environ Microbiol 2023; 89:e0210122 [View Article] [PubMed]
    [Google Scholar]
  26. Seemann T. Prokka: rapid prokaryotic genome annotation. Bioinformatics 2014; 30:2068–2069 [View Article] [PubMed]
    [Google Scholar]
  27. Croucher NJ, Page AJ, Connor TR, Delaney AJ, Keane JA et al. Rapid phylogenetic analysis of large samples of recombinant bacterial whole genome sequences using Gubbins. Nucleic Acids Res 2015; 43:e15 [View Article] [PubMed]
    [Google Scholar]
  28. Page AJ, Taylor B, Delaney AJ, Soares J, Seemann T et al. SNP-sites: rapid efficient extraction of SNPs from multi-FASTA alignments. Microb Genom 2016; 2:e000056 [View Article] [PubMed]
    [Google Scholar]
  29. Minh BQ, Schmidt HA, Chernomor O, Schrempf D, Woodhams MD et al. IQ-TREE 2: new models and efficient methods for phylogenetic inference in the genomic era. Mol Biol Evol 2020; 37:1530–1534 [View Article] [PubMed]
    [Google Scholar]
  30. Yu G, Smith DK, Zhu H, Guan Y, Lam TTY et al. ggtree: an r package for visualization and annotation of phylogenetic trees with their covariates and other associated data. Methods Ecol Evol 2016; 8:28–36 [View Article]
    [Google Scholar]
  31. Tonkin-Hill G, Lees JA, Bentley SD, Frost SDW, Corander J. Fast hierarchical Bayesian analysis of population structure. Nucleic Acids Res 2019; 47:5539–5549 [View Article] [PubMed]
    [Google Scholar]
  32. Tonkin-Hill G, MacAlasdair N, Ruis C, Weimann A, Horesh G et al. Producing polished prokaryotic pangenomes with the Panaroo pipeline. Genome Biol 2020; 21:180 [View Article] [PubMed]
    [Google Scholar]
  33. Kolde R. Pheatmap: Pretty Heatmaps. Version 1.0. 12 edn R Package; 2019
    [Google Scholar]
  34. Wickham H. Ggplot2: Elegant Graphics for Data Analysis New York: Pringer-Verlag; 2016
    [Google Scholar]
  35. Sullivan MJ, Petty NK, Beatson SA. Easyfig: a genome comparison visualizer. Bioinformatics 2011; 27:1009–1010 [View Article] [PubMed]
    [Google Scholar]
  36. Alikhan NF, Petty NK, Ben Zakour NL, Beatson SA. BLAST Ring Image Generator (BRIG): simple prokaryote genome comparisons. BMC Genom 2011; 12:402 [View Article] [PubMed]
    [Google Scholar]
  37. Rambaut A, Lam TT, Max Carvalho L, Pybus OG. Exploring the temporal structure of heterochronous sequences using TempEst (formerly Path-O-Gen). Virus Evol 2016; 2:vew007 [View Article] [PubMed]
    [Google Scholar]
  38. Suchard MA, Lemey P, Baele G, Ayres DL, Drummond AJ et al. Bayesian phylogenetic and phylodynamic data integration using BEAST 1.10. Virus Evol 2018; 4:vey016 [View Article] [PubMed]
    [Google Scholar]
  39. Rambaut A, Drummond AJ, Xie D, Baele G, Suchard MA. Posterior summarization in Bayesian phylogenetics using tracer 1.7. Syst Biol 2018; 67:901–904 [View Article] [PubMed]
    [Google Scholar]
  40. Dellicour S, Gill MS, Faria NR, Rambaut A, Pybus OG et al. Relax, keep walking - a practical guide to continuous phylogeographic inference with BEAST. Mol Biol Evol 2021; 38:3486–3493 [View Article] [PubMed]
    [Google Scholar]
  41. Dellicour S, Rose R, Faria NR, Lemey P, Pybus OG. SERAPHIM: studying environmental rasters and phylogenetically informed movements. Bioinformatics 2016; 32:3204–3206 [View Article] [PubMed]
    [Google Scholar]
  42. Murrell B, Moola S, Mabona A, Weighill T, Sheward D et al. FUBAR: a fast, unconstrained bayesian approximation for inferring selection. Mol Biol Evol 2013; 30:1196–1205 [View Article] [PubMed]
    [Google Scholar]
  43. Zhou Y, Pan Q, Pires DEV, Rodrigues CHM, Ascher DB. DDMut: predicting effects of mutations on protein stability using deep learning. Nucleic Acids Res 2023; 51:W122–W128 [View Article] [PubMed]
    [Google Scholar]
  44. Cabrejos DAL, Alexandrino AV, Pereira CM, Mendonça DC, Pereira HD et al. Structural characterization of a pathogenicity-related superoxide dismutase codified by a probably essential gene in Xanthomonas citri subsp. citri. PLoS One 2019; 14:e0209988 [View Article] [PubMed]
    [Google Scholar]
  45. Campos PE, Pruvost O, Boyer K, Chiroleu F, Cao TT et al. Herbarium specimen sequencing allows precise dating of Xanthomonas citri pv. citri diversification history. Nat Commun 2023; 14:4306 [View Article] [PubMed]
    [Google Scholar]
  46. Tesson F, Hervé A, Mordret E, Touchon M, d’Humières C et al. Systematic and quantitative view of the antiviral arsenal of prokaryotes. Nat Commun 2022; 13:2561 [View Article] [PubMed]
    [Google Scholar]
  47. Richard D, Boyer C, Vernière C, Canteros BI, Lefeuvre P et al. Complete genome sequences of six copper-resistant Xanthomonas citri pv. citri strains causing Asiatic citrus canker, obtained using long-read technology. Genome Announc 2017; 5:e00010-17 [View Article] [PubMed]
    [Google Scholar]
  48. Behlau F, Hong JC, Jones JB, Graham JH. Evidence for acquisition of copper resistance genes from different sources in citrus-associated xanthomonads. Phytopathology 2013; 103:409–418 [View Article] [PubMed]
    [Google Scholar]
  49. Schmartz GP, Hartung A, Hirsch P, Kern F, Fehlmann T et al. PLSDB: advancing a comprehensive database of bacterial plasmids. Nucleic Acids Res 2022; 50:D273–D278 [View Article] [PubMed]
    [Google Scholar]
  50. Rensing C, Pribyl T, Nies DH. New functions for the three subunits of the CzcCBA cation-proton antiporter. J Bacteriol 1997; 179:6871–6879 [View Article] [PubMed]
    [Google Scholar]
  51. Smith SG, Wilson TJ, Dow JM, Daniels MJ. A gene for superoxide dismutase from Xanthomonas campestris pv. campestris and its expression during bacterial-plant interactions. Mol Plant Microbe Interact 1996; 9:584–593 [View Article] [PubMed]
    [Google Scholar]
  52. Artier J, da Silva Zandonadi F, de Souza Carvalho FM, Pauletti BA, Leme AFP et al. Comparative proteomic analysis of Xanthomonas citri ssp. citri periplasmic proteins reveals changes in cellular envelope metabolism during in vitro pathogenicity induction. Mol Plant Pathol 2018; 19:143–157 [View Article] [PubMed]
    [Google Scholar]
  53. Monteil CL, Yahara K, Studholme DJ, Mageiros L, Méric G et al. Population-genomic insights into emergence, crop adaptation and dissemination of Pseudomonas syringae pathogens. Microb Genom 2016; 2:e000089 [View Article] [PubMed]
    [Google Scholar]
  54. Straub C, Colombi E, McCann HC. Population genomics of bacterial plant pathogens. Phytopathology 2021; 111:23–31 [View Article] [PubMed]
    [Google Scholar]
  55. Jibrin MO, Sharma A, Mavian CN, Timilsina S, Kaur A et al. Phylodynamic insights into global emergence and diversification of the tomato pathogen Xanthomonas hortorum pv. gardneri. Mol Plant Microbe Interact 2024; 37:712–720 [View Article] [PubMed]
    [Google Scholar]
  56. Klein-Gordon JM, Timilsina S, Xing Y, Abrahamian P, Garrett KA et al. Whole genome sequences reveal the Xanthomonas perforans population is shaped by the tomato production system. ISME J 2022; 16:591–601 [View Article] [PubMed]
    [Google Scholar]
  57. Quibod IL, Atieza-Grande G, Oreiro EG, Palmos D, Nguyen MH et al. The green revolution shaped the population structure of the rice pathogen Xanthomonas oryzae pv. oryzae. ISME J 2020; 14:492–505 [View Article] [PubMed]
    [Google Scholar]
  58. Hawkey J, Paranagama K, Baker KS, Bengtsson RJ, Weill F-X et al. Global population structure and genotyping framework for genomic surveillance of the major dysentery pathogen, Shigella sonnei. Nat Commun 2021; 12:2684 [View Article] [PubMed]
    [Google Scholar]
  59. Coll F, McNerney R, Guerra-Assunção JA, Glynn JR, Perdigão J et al. A robust SNP barcode for typing Mycobacterium tuberculosis complex strains. Nat Commun 2014; 5:4812 [View Article]
    [Google Scholar]
  60. Oliveira PH, Touchon M, Rocha EPC. Regulation of genetic flux between bacteria by restriction-modification systems. Proc Natl Acad Sci U S A 2016; 113:5658–5663 [View Article] [PubMed]
    [Google Scholar]
  61. Dordet-Frisoni E, Vandecasteele C, Contarin R, Sagné E, Baranowski E et al. Impacts of Mycoplasma agalactiae restriction-modification systems on pan-epigenome dynamics and genome plasticity. Microb Genom 2022; 8:mgen000829 [View Article] [PubMed]
    [Google Scholar]
  62. Park S, Jung D, O’Brien B, Ruffini J, Dussault F et al. Comparative genomic analysis of Staphylococcus aureus isolates associated with either bovine intramammary infections or human infections demonstrates the importance of restriction-modification systems in host adaptation. Microb Genom 2022; 8:000779 [View Article] [PubMed]
    [Google Scholar]
  63. Nero TM, Dalia TN, Wang JC-Y, Kysela DT, Bochman ML et al. ComM is a hexameric helicase that promotes branch migration during natural transformation in diverse Gram-negative species. Nucleic Acids Res 2018; 46:6099–6111 [View Article] [PubMed]
    [Google Scholar]
  64. Douraghi M, Jasemi S, Kodori M, Rahbar M, Boroumand MA. Evidence of Interruption of the comM gene in a large series of clinical isolates of multidrug-resistant Acinetobacter baumannii. J Mol Microbiol Biotechnol 2016; 26:410–413 [View Article] [PubMed]
    [Google Scholar]
  65. Mazzamurro F, Chirakadavil JB, Durieux I, Poiré L, Plantade J et al. Intragenomic conflicts with plasmids and chromosomal mobile genetic elements drive the evolution of natural transformation within species. Genomics 2024 [View Article]
    [Google Scholar]
  66. Sheppard SK, Guttman DS, Fitzgerald JR. Population genomics of bacterial host adaptation. Nat Rev Genet 2018; 19:549–565 [View Article] [PubMed]
    [Google Scholar]
  67. Perez-Quintero AL, Ortiz-Castro M, Lang JM, Rieux A, Wu G et al. Genomic acquisitions in emerging populations of Xanthomonas vasicola pv. vasculorum Infecting corn in the United States and Argentina. Phytopathology 2020; 110:1161–1173 [View Article] [PubMed]
    [Google Scholar]
  68. Alvarez-Martinez CE, Sgro GG, Araujo GG, Paiva MRN, Matsuyama BY et al. Secrete or perish: the role of secretion systems in Xanthomonas biology. Comput Struct Biotechnol J 2021; 19:279–302 [View Article] [PubMed]
    [Google Scholar]
  69. Richard D, Ravigné V, Rieux A, Facon B, Boyer C et al. Adaptation of genetically monomorphic bacteria: evolution of copper resistance through multiple horizontal gene transfers of complex and versatile mobile genetic elements. Mol Ecol 2017; 26:2131–2149 [View Article] [PubMed]
    [Google Scholar]
  70. Richard D, Pruvost O, Balloux F, Boyer C, Rieux A et al. Time-calibrated genomic evolution of a monomorphic bacterium during its establishment as an endemic crop pathogen. Mol Ecol 2021; 30:1823–1835 [View Article] [PubMed]
    [Google Scholar]
  71. Gochez AM, Huguet-Tapia JC, Minsavage GV, Shantaraj D, Jalan N et al. Pacbio sequencing of copper-tolerant Xanthomonas citri reveals presence of a chimeric plasmid structure and provides insights into reassortment and shuffling of transcription activator-like effectors among X. citri strains. BMC Genom 2018; 19:16 [View Article] [PubMed]
    [Google Scholar]
  72. Fundecitrus. Levantamento da Incidência das Doenças dos Citros: Greening, CVC e Cancro Cítrico no Cinturão Citrícola de São Paulo e Triângulo/Sudoeste Mineiro. Araraquara; 2023
  73. Braga CAP, Silber SD. Brazilian Frozen Concentrated Orange Juice: The Folly of Unfair Trade Cases Washington: World Bank; 1991 p 46
    [Google Scholar]
  74. Fernandes W, Spreen T. The role of Brazil in the world orange Juice Market: a threat posed by Cvc. Paper presented at the American Agricultural Economics Association (New Name 2008: Agricultural and Applied Economics Association), 2002 Annual meeting
    [Google Scholar]
  75. Belasque Junior J, Barbosa JC, Bergamin Filho A, Massari CA. Prováveis consequências do abrandamento da metodologia de erradicação do cancro cítrico no Estado de São Paulo. Trop plant pathol 2010; 35:314–317 [View Article]
    [Google Scholar]
  76. Santos CFO. Cancro Cítirco: ocorrência no Brasil e seu combate. In Rodriguez O. eds Citricultura Brasileira. II, 2 edn. Campinas, SP: Fundação Cargill; 1991 pp 787–823
    [Google Scholar]
  77. Steunou AS, Babot M, Bourbon M-L, Tambosi R, Durand A et al. Additive effects of metal excess and superoxide, a highly toxic mixture in bacteria. Microb Biotechnol 2020; 13:1515–1529 [View Article] [PubMed]
    [Google Scholar]
/content/journal/mgen/10.1099/mgen.0.001338
Loading
/content/journal/mgen/10.1099/mgen.0.001338
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Supplementary material 2

EXCEL
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An error occurred
Approval was partially successful, following selected items could not be processed due to error