Genome-Wide Analysis Provides Evidence on the Genetic Relatedness of the Emergent Xylella fastidiosa Genotype in Italy to Isolates from Central America.

Xylella fastidiosa is a plant-pathogenic bacterium recently introduced in Europe that is causing decline in olive trees in the South of Italy. Genetic studies have consistently shown that the bacterial genotype recovered from infected olive trees belongs to the sequence type ST53 within subspecies pauca. This genotype, ST53, has also been reported to occur in Costa Rica. The ancestry of ST53 was recently clarified, showing it contains alleles that are monophyletic with those of subsp. pauca in South America. To more robustly determine the phylogenetic placement of ST53 within X. fastidiosa, we performed a comparative analysis based on single nucleotide polymorphisms (SNPs) and the study of the pan-genome of the 27 currently public available whole genome sequences of X. fastidiosa. The resulting maximum-parsimony and maximum likelihood trees constructed using the SNPs and the pan-genome analysis are consistent with previously described X. fastidiosa taxonomy, distinguishing the subsp. fastidiosa, multiplex, pauca, sandyi, and morus. Within the subsp. pauca, the Italian and three Costa Rican isolates, all belonging to ST53, formed a compact phylotype in a clade divergent from the South American pauca isolates, also distinct from the recently described coffee isolate CFBP8072 imported into Europe from Ecuador. These findings were also supported by the gene characterization of a conjugative plasmid shared by all the four ST53 isolates. Furthermore, isolates of the ST53 clade possess an exclusive locus encoding a putative ATP-binding protein belonging to the family of histidine kinase-like ATPase gene, which is not present in isolates from the subspecies multiplex, sandyi, and pauca, but was detected in ST21 isolates of the subspecies fastidiosa from Costa Rica. The clustering and distinctiveness of the ST53 isolates supports the hypothesis of their common origin, and the limited genetic diversity among these isolates suggests this is an emerging clade within subsp. pauca.

[1]  M. Sagot,et al.  Pierce's Disease of Grapevines: A Review of Control Strategies and an Outline of an Epidemiological Model , 2018, Front. Microbiol..

[2]  F. Palmisano,et al.  Isolation and pathogenicity of Xylella fastidiosa associated to the olive quick decline syndrome in southern Italy , 2017, Scientific Reports.

[3]  S. Marcelletti,et al.  Xylella fastidiosa CoDiRO strain associated with the olive quick decline syndrome in southern Italy belongs to a clonal complex of the subspecies pauca that evolved in Central America. , 2016, Microbiology.

[4]  M. Muthuramalingam,et al.  Toxin-Antitoxin Modules Are Pliable Switches Activated by Multiple Protease Pathways , 2016, Toxins.

[5]  Sara S. K. Koenig,et al.  Whole Genome Sequencing for Genomics-Guided Investigations of Escherichia coli O157:H7 Outbreaks , 2016, Front. Microbiol..

[6]  M. Takita,et al.  The MqsRA Toxin-Antitoxin System from Xylella fastidiosa Plays a Key Role in Bacterial Fitness, Pathogenicity, and Persister Cell Formation , 2016, Front. Microbiol..

[7]  David S. Wishart,et al.  PHASTER: a better, faster version of the PHAST phage search tool , 2016, Nucleic Acids Res..

[8]  D. Stenger,et al.  Draft Genome Sequence of Xylella fastidiosa subsp. fastidiosa Strain Stag’s Leap , 2016, Genome Announcements.

[9]  Sudhir Kumar,et al.  MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. , 2016, Molecular biology and evolution.

[10]  G. Martelli,et al.  Intercepted isolates of Xylella fastidiosa in Europe reveal novel genetic diversity , 2016, European Journal of Plant Pathology.

[11]  J. West,et al.  Statement on diversity of Xylella fastidiosa subsp. pauca in Apulia , 2016 .

[12]  D. Crouzillat,et al.  New Coffee Plant-Infecting Xylella fastidiosa Variants Derived via Homologous Recombination , 2015, Applied and Environmental Microbiology.

[13]  G. Martelli,et al.  Draft Genome Sequence of CO33, a Coffee-Infecting Isolate of Xylella fastidiosa , 2015, Genome Announcements.

[14]  R. Almeida,et al.  How Do Plant Diseases Caused by Xylella fastidiosa Emerge? , 2015, Plant disease.

[15]  Tom Slezak,et al.  kSNP3.0: SNP detection and phylogenetic analysis of genomes without genome alignment or reference genome , 2015, Bioinform..

[16]  A. Italiano,et al.  Draft Genome Sequence of the Xylella fastidiosa CoDiRO Strain , 2015, Genome Announcements.

[17]  D. Makowski,et al.  Scientific Opinion on the risk to plant health posed by Xylella fastidiosa in the EU territory, with the identification and evaluation of risk reduction options , 2015 .

[18]  L. Nunney,et al.  The Complex Biogeography of the Plant Pathogen Xylella fastidiosa: Genetic Evidence of Introductions and Subspecific Introgression in Central America , 2014, PloS one.

[19]  J. Shao,et al.  Genome Sequence of a Xylella fastidiosa Strain Causing Sycamore Leaf Scorch Disease in Virginia , 2014, Genome Announcements.

[20]  M. Scally,et al.  Large-Scale Intersubspecific Recombination in the Plant-Pathogenic Bacterium Xylella fastidiosa Is Associated with the Host Shift to Mulberry , 2014, Applied and Environmental Microbiology.

[21]  J. Shao,et al.  Genome Sequence of a Xylella fastidiosa Strain Causing Mulberry Leaf Scorch Disease in Maryland , 2014, Genome Announcements.

[22]  L. R. Nunes,et al.  Genomic Sequencing of Two Coffee-Infecting Strains of Xylella fastidiosa Isolated from Brazil , 2014, Genome Announcements.

[23]  Fangfang Xia,et al.  The SEED and the Rapid Annotation of microbial genomes using Subsystems Technology (RAST) , 2013, Nucleic Acids Res..

[24]  D. Hopkins,et al.  Intersubspecific Recombination in Xylella fastidiosa Strains Native to the United States: Infection of Novel Hosts Associated with an Unsuccessful Invasion , 2013, Applied and Environmental Microbiology.

[25]  G. P. Martelli,et al.  IDENTIFICATION OF DNA SEQUENCES RELATED TO XYLELLA FASTIDIOSA IN OLEANDER, ALMOND AND OLIVE TREES EXHIBITING LEAF SCORCH SYMPTOMS IN APULIA (SOUTHERN ITALY) , 2013 .

[26]  D. Stenger,et al.  Draft Genome Sequence of Xylella fastidiosa subsp. multiplex Strain Griffin-1 from Quercus rubra in Georgia , 2013, Genome Announcements.

[27]  B. Contreras-Moreira,et al.  GET_HOMOLOGUES, a Versatile Software Package for Scalable and Robust Microbial Pangenome Analysis , 2013, Applied and Environmental Microbiology.

[28]  F. Jan,et al.  Pierce's Disease of Grapevines in Taiwan: Isolation, Cultivation and Pathogenicity of Xylella fastidiosa , 2013 .

[29]  D. Stenger,et al.  A Conjugative 38 kB Plasmid Is Present in Multiple Subspecies of Xylella fastidiosa , 2012, PloS one.

[30]  Michael T Laub,et al.  Evolution of two-component signal transduction systems. , 2012, Annual review of microbiology.

[31]  L. T. Kishi,et al.  Global Expression Profile of Biofilm Resistance to Antimicrobial Compounds in the Plant-Pathogenic Bacterium Xylella fastidiosa Reveals Evidence of Persister Cells , 2012, Journal of bacteriology.

[32]  D. Hopkins,et al.  The Xylella fastidiosa Biocontrol Strain EB92-1 Genome Is Very Similar and Syntenic to Pierce's Disease Strains , 2011, Journal of bacteriology.

[33]  Geeta Shakya,et al.  Population Genetics of Vibrio cholerae from Nepal in 2010: Evidence on the Origin of the Haitian Outbreak , 2011, mBio.

[34]  Nicola K. Petty,et al.  BLAST Ring Image Generator (BRIG): simple prokaryote genome comparisons , 2011, BMC Genomics.

[35]  J. Hartung,et al.  Population Genomic Analysis of a Bacterial Plant Pathogen: Novel Insight into the Origin of Pierce's Disease of Grapevine in the U.S. , 2010, PloS one.

[36]  G. Xie,et al.  Whole Genome Sequences of Two Xylella fastidiosa Strains (M12 and M23) Causing Almond Leaf Scorch Disease in California , 2010, Journal of bacteriology.

[37]  Lavanya Kannan,et al.  A low-polynomial algorithm for assembling clusters of orthologous groups from intergenomic symmetric best matches , 2010, Bioinform..

[38]  P. Christie,et al.  Biological Diversity of Prokaryotic Type IV Secretion Systems , 2009, Microbiology and Molecular Biology Reviews.

[39]  A. Oskooi Molecular Evolution and Phylogenetics , 2008 .

[40]  Rick L. Stevens,et al.  The RAST Server: Rapid Annotations using Subsystems Technology , 2008, BMC Genomics.

[41]  Jiqiang Yao,et al.  Analysis of the genome-wide variations among multiple strains of the plant pathogenic bacterium Xylella fastidiosa , 2006, BMC Genomics.

[42]  Jaideep P. Sundaram,et al.  Genome analysis of multiple pathogenic isolates of Streptococcus agalactiae: implications for the microbial "pan-genome". , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[43]  O. Pellegrini,et al.  The Bacillus subtilis ydcDE operon encodes an endoribonuclease of the MazF/PemK family and its inhibitor , 2005, Molecular microbiology.

[44]  C. Stoeckert,et al.  OrthoMCL: identification of ortholog groups for eukaryotic genomes. , 2003, Genome research.

[45]  S. Lopes,et al.  Microarray analyses of Xylella fastidiosa provide evidence of coordinated transcription control of laterally transferred elements. , 2003, Genome research.

[46]  G H Goldman,et al.  Comparative Analyses of the Complete Genome Sequences of Pierce's Disease and Citrus Variegated Chlorosis Strains of Xylella fastidiosa , 2003, Journal of bacteriology.

[47]  D. A. Palmieri,et al.  The genome sequence of the plant pathogen Xylella fastidiosa , 2000, Nature.

[48]  D. Helinski,et al.  The parDE operon of the broad-host-range plasmid RK2 specifies growth inhibition associated with plasmid loss. , 1994, Journal of molecular biology.