Abstract
We present here the draft genome sequences of bacterial pathogens of the Araceae family, Xanthomonas axonopodis pv. dieffenbachiae LMG 695 and Xanthomonas campestris pv. syngonii LMG 9055, differing in host range. A comparison between genome sequences will help understand the mechanisms involved in tissue specificity and adaptation to host plants.
GENOME ANNOUNCEMENT
Among the different members of the aroids, the tropical flower anthurium (Anthurium andreanum Linden ex André) is an economically important crop cultivated in tropical and temperate areas. Production of anthurium in the world is threatened by Anthurium bacterial blight (ABB), which is caused by Xanthomonas axonopodis pv. dieffenbachiae (1). Strains of X. axonopodis pv. dieffenbachiae isolated from anthurium can infect other aroids, and particularly different ornemental species of Dieffenbachia, Caladium, Philodendron, Syngonium, or edible aroids, such as Colocasia species. Other Xanthomonas strains, first classified as Xanthomonas campestris pv. syngonii (2), have a narrower host range and infect Syngonium plants causing the bacterial leaf blight of Syngonium. Based on rep-PCR and multilocus sequence analysis (MLSA), the two strains LMG 695 and LMG 9055 grouped in the X. axonopodis 9.4 cluster (3, 4). Hybridization values (<70%) between LMG 695 or LMG 9055 and the type strain of X. axonopodis LMG 982 did not support their inclusion in X. axonopodis (5). A polyphasic taxonomic approach based on MLSA, DNA/DNA hybridization, average nucleotide identity (ANI) values, and phenotypic analyses confirmed these results and proposed to reclassify this cluster as Xanthomonas phaseoli (6). In addition to differences in pathogenicity, these data also supported the separation of the two categories of strains represented by LMG 695 and LMG 9055 in distinct pathovars named dieffenbachiae and syngonii, respectively.
The genomes of both strains were sequenced using the Illumina HiSeq 2000 platform (GATC Biotech, Germany). Shotgun sequencing yielded 25,320,564 read pairs (12,846,409 100-bp paired-end reads with an insert size of 250 bp, and 12,474,156 50-bp mate-pair reads with an insert size of 3 kb) and 30,521,960 read pairs (17,529,832 100-bp paired-end reads with an insert size of 250 bp and 12,992,128 50-bp mate-pair reads with an insert size of 3 kb) for LMG 695 and LMG 9055, respectively.
A combination of Velvet (7), SOAPdenovo, and SOAP GapCloser (8) yielded 10 contigs >500 bp (N50, 1,343,781 bp), with the largest contig being 1,543,007 bp, for a total assembly size of 5,035,943 bp for strain LMG 695, and 33 contigs >500 bp (N50, 414,753 bp), with the largest contig being 751,399 bp, for a total assembly size of 5,000,894 bp for strain LMG 9055. Genomic contigs were annotated using the EuGene-P annotation pipeline to identify RNAs and protein-coding genes (9). The draft genomes were predicted to contain 4,423 and 4,665 coding sequences (CDSs) for strains LMG 695 and LMG 9055, respectively.
The two genomes have an average nucleotide identity (ANI) of 97.33% (10). The two genomes share >3,700 CDSs, and each contains approximately 200 CDSs, with no orthologs in other Xanthomonas species (11). Nineteen and 22 predicted type 3 effector (T3E) genes are present in the LMG 695 and LMG 9055 genomes, respectively. At least 13 of these T3E genes are shared by the two strains.
Accession number(s).
These whole-genome shotgun projects have been deposited in GenBank under the accession no. CP014347 for strain LMG 695 and LSLD00000000 for strain LMG 9055. The versions described in this paper are the first versions.
ACKNOWLEDGMENTS
This research was funded by the French Agence Nationale de la Recherche, the European Regional Development Fund 2007–2014 (grant 2007FR161PO004), the Conseil Régional de la Réunion, and CIRAD.
Footnotes
Citation Robène I, Bolot S, Pruvost O, Arlat M, Noël LD, Carrère S, Jacques M-A, Koebnik R, Gagnevin L. 2016. High-quality draft genome sequences of two Xanthomonas pathotype strains infecting aroid plants. Genome Announc 4(5):e00902-16. doi:10.1128/genomeA.00902-16.
REFERENCES
- 1.Alvarez AM, Toves PJ, Vowell TS. 2006. Bacterial blight of anthuriums: Hawaii’s experience with a global disease. APSNet Features doi: 10.1094/APSnetFeature-2006-0206. [DOI] [Google Scholar]
- 2.Dickey RS, Zumoff CH. 1987. Bacterial leaf blight of Syngonium caused by a pathovar of Xanthomonas campestris. Phytopathology 77:1257–1262. doi: 10.1094/Phyto-77-1257. [DOI] [Google Scholar]
- 3.Rademaker JL, Louws FJ, Schultz MH, Rossbach U, Vauterin L, Swings J, De Bruijn FJ. 2005. A comprehensive species to strain taxonomic framework for Xanthomonas. Phytopathology 95:1098–1111. doi: 10.1094/PHYTO-95-1098. [DOI] [PubMed] [Google Scholar]
- 4.Bui Thi Ngoc L, Vernière C, Jouen E, Ah-You N, Lefeuvre P, Chiroleu F, Gagnevin L, Pruvost O. 2010. Amplified fragment length polymorphism and multilocus sequence analysis-based genotypic relatedness among pathogenic variants of Xanthomonas citri pv. citri and Xanthomonas campestris pv. bilvae. Int J Syst Evol Microbiol 60:515–525. doi: 10.1099/ijs.0.009514-0. [DOI] [PubMed] [Google Scholar]
- 5.Donahoo RS, Jones JB, Lacy GH, Stromberg VK, Norman DJ. 2013. Genetic analyses of Xanthomonas axonopodis pv. dieffenbachiae strains reveal distinct phylogenetic groups. Phytopathology 103:237–244. doi: 10.1094/PHYTO-08-12-0191-R. [DOI] [PubMed] [Google Scholar]
- 6.Constantin EC, Cleenwerck I, Maes M, Baeyen S, Van Malderghem C, De Vos P, Cottyn B. 2015. Genetic characterisation of strains named as Xanthomonas axonopodis pv. dieffenbachiae leads to a taxonomic revision of the X. axonopodis species complex. Plant Pathol 65:792–806. doi: 10.1111/ppa.12461. [DOI] [Google Scholar]
- 7.Zerbino DR, Birney E. 2008. Velvet: algorithms for de novo short read assembly using de Bruijn graphs. Genome Res 18:821–829. doi: 10.1101/gr.074492.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8.Luo R, Liu B, Xie Y, Li Z, Huang W, Yuan J, He G, Chen Y, Pan Q, Liu Y, Tang J, Wu G, Zhang H, Shi Y, Liu Y, Yu C, Wang B, Lu Y, Han C, Cheung DW, Yiu SM, Peng S, Xiaoqian Z, Liu G, Liao X, Li Y, Yang H, Wang J, Lam TW, Wang J. 2012. SOAPdenovo2: an empirically improved memory-efficient short-read de novo assembler. GigaScience 1:18. doi: 10.1186/2047-217X-1-18. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 9.Sallet E, Roux B, Sauviac L, Jardinaud MF, Carrère S, Faraut T, de Carvalho-Niebel F, Gouzy J, Gamas P, Capela D, Bruand C, Schiex T. 2013. Next-generation annotation of prokaryotic genomes with EuGene-P: application to Sinorhizobium meliloti 2011. DNA Res 20:339–354. doi: 10.1093/dnares/dst014. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10.Goris J, Konstantinidis KT, Klappenbach JA, Coenye T, Vandamme P, Tiedje JM. 2007. DNA-DNA hybridization values and their relationship to whole-genome sequence similarities. Int J Syst Evol Microbiol 57:81–91. doi: 10.1099/ijs.0.64483-0. [DOI] [PubMed] [Google Scholar]
- 11.Blom J, Albaum SP, Doppmeier D, Pühler A, Vorhölter FJ, Zakrzewski M, Goesmann A. 2009. Edgar: a software framework for the comparative analysis of prokaryotic genomes. BMC Bioinformatics 10:154. doi: 10.1186/1471-2105-10-154. [DOI] [PMC free article] [PubMed] [Google Scholar]