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Fig. S1. Tick collection site used in this study.

Fig. S2. Phylogenetic analysis based on flaB of REP Borrelia spp. The phylogenetic tree was constructed based on NJ methods, and bootstrap tests were carried out according to the Kimura 2-parameter distances method. All positions containing alignment gaps and missing data were eliminated only in pairwise sequence comparisons (pairwise deletion option). There were a total of 462 positions in the final data set. The percentage of replicate trees in which the associated taxa were clustered together in the bootstrap test (1000 replicates) was calculated. The phylogenetic branches were supported in > 70% by the bootstrap analysis. The bar indicates the percentage of sequence divergence.

Fig. S3. Phylogenetic analysis based on gyrB of the genus Borrelia. The phylogenetic trees were constructed based on NJ methods, and bootstrap tests were carried out according to the Kimura 2-parameter distances method. All positions containing alignment gaps and missing data were eliminated only in pairwise sequence comparisons (pairwise deletion option). There were a total of 1932 positions in the final data set. The percentage of replicate trees in which the associated taxa were clustered together in the bootstrap test (1000 replicates) was calculated. The phylogenetic branches were supported in > 70% by the bootstrap analysis. The bar indicates the percentage of sequence divergence. Treponema pallidum (NC_010741), Clostridium botulinum (NC_009495), and Clostridium beijerinckii (NC_009617) were used as outgroups.

Fig. S4. Phylogenetic analysis based on glpQ of Borrelia spp. The phylogenetic tree was constructed based on NJ methods, and bootstrap tests were carried out according to the Kimura 2-parameter distances method. All positions containing alignment gaps and missing data were eliminated only in pairwise sequence comparisons (pairwise deletion option). There were a total of 1035 positions in the final data set. The percentage of replicate trees in which the associated taxa were clustered together in the bootstrap test (1000 replicates) was calculated. The phylogenetic branches were supported in > 70% by the bootstrap analysis. The bar indicates the percentage of sequence divergence.

Fig. S5. The gene organization of the IGS region of Borrelia sp. tAG strain tAG158M. Comparison of gene organization patterns among REP Borrelia spp. The sequences of strain tAG158M (a Borrelia sp. tAG strain isolated in this study) were updated at GenBank (AB529425). The genes shown are cof (hydrolase), hpt (hypoxanthine-guanine phosphoribosyltransferase), purA (adenylosuccinate synthetase), purB (adenylosuccinate lyase), mag (methylpurine-DNA glycosylase), ileT (tRNA-Ile), glvA (the maltose-6′-phosphate glucosidase), glvC, alaT (tRNA-Ala) and 16S rDNA. The nucleotide sequence similarity between strains is shown in the figure.

Fig. S6. Borrelial cells in A. geoemydae during blood-feeding. Borrelial cells were found in the midgut (A and B) and salivary gland (C and D) during blood-feeding. Borrelial cells (red) were detected by confocal laser scanning microscopy using goat anti-Borrelia sp. primary and Alexa Fluor 488-labelled rabbit anti-goat secondary antisera. DRAQ5 (green) was used as the counterstain.

Table S1.Borrelia strains used in this study.

Table S2. Number of Amblyomma geoemydae ticks used in this study.

Table S3. Number of REP Borrelia sp. (Borrelia sp. tAG) positive ticks after tick moulted.

Table S4. Primers used in this study.

Table S5. Primers used sequencing of IGS region of Borrelia sp. tAG158M.

Table S6. Oligonucleotide primers and probes used for quantitative real-time PCR assay.

Appendix S1. Supplemental materials.

FilenameFormatSizeDescription
EMI4_280_sm_supp_f1.tif3524KSupporting info item
EMI4_280_sm_supp_f2.tif17176KSupporting info item
EMI4_280_sm_supp_f3.tif6153KSupporting info item
EMI4_280_sm_supp_f4.tif7758KSupporting info item
EMI4_280_sm_supp_f5.tif8238KSupporting info item
EMI4_280_sm_supp_f6.tif15469KSupporting info item
EMI4_280_sm_supp_materials.doc209KSupporting info item
EMI4_280_sm_suppmaterials.doc111KSupporting info item

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