Multiple sequence alignments: Detection and isolation of a new member of Burkholderiaceae‑related endofungal bacteria from Saksenaea boninensis sp. nov., a new thermotolerant fungus in Mucorales
<p><strong>Methods:</strong></p><p>Nucleotide sequences were aligned independently for each region using MAFFT v7.212 (Katoh and Standley, 2013). The obtained alignment blocks were subject to Gblocks 0.91b (Castresana, 2000) to remove poorly aligned positions with the relaxed selection setting described in Talavera & Castresana (2007) using the following parameters (-t = d -b2 = 9 -b3 = 10 -b4 = 5 -b5 = h). After automatically removing gaps, the alignment blocks were viewed using MEGA 6.06 software (Tamura et al., 2013) and poorly aligned positions at either end of the alignments were removed manually. Pairwise distances of the nucleotide sequences (ITS2, ITS1-5.8S-ITS2, LSU, and tef1) of the ex-type strains of seven <i>Saksenaea</i> spp. and the representative isolate <i>S. boninensis</i> Sak4 were calculated by MEGA 6.06 software (Tamura et al. 2013). Multiple sequence alignment of 16S rRNA gene of the family <i>Burkholderiaceae</i> was prepared for the phylogeny of a bacterial endosymbiont. Multiple sequence alignments of ITS, LSU, and tef1 genes of <i>Saksenaea</i> spp. (Mucorales) were separately prepared for the phylogeny of a fungal host. Concatenated dataset of these genes were also prepared. All nucleotide sequences were retrieved from GenBank (See "Sequence_ID.csv" and taxon names of each alignment). </p><p> </p><p><strong>Description of files:</strong></p><p><strong>A. Phylogeny of the family </strong><i><strong>Burkholderiaceae</strong></i><strong> (Bacterial endosymbiont):</strong></p><p>1. Burkholderiaceae_16S_RAW.fasta</p><p>Non-aligned dataset of 16S rRNA gene of the family <i>Burkholderiaceae</i>.</p><p> </p><p>2. Burkholderiaceae_16S_aligned.fasta</p><p>Aligned dataset of 16S rRNA gene of the family <i>Burkholderiaceae</i>.</p><p> </p><p><strong>B. Phylogenies of </strong><i><strong>Saksenaea</strong></i><strong> spp. (Fungal host):</strong></p><p>1. Sequence_ID_v2.csv</p><p>Taxon names, accession numbers, and sequence ID for the concatenated multiple sequence alignment are listed.</p><p> </p><p>2. Saksenaea_ITS_RAW_v2.fasta</p><p>Non-aligned dataset of ITS1-5.8S-ITS2 region of <i>Saksenaea</i> spp. </p><p> </p><p>3. Saksenaea_ITS_aligned_v2.fasta</p><p>Aligned dataset of ITS1-5.8S-ITS2 region of <i>Saksenaea</i> spp. Only used for ITS1-5.8S-ITS2 phylogeny.</p><p> </p><p>4. Saksenaea_LSU_RAW_v2.fasta</p><p>Non-aligned dataset of LSU gene region of <i>Saksenaea</i> spp. </p><p> </p><p>5. Saksenaea_LSU_aligned_v2.fasta</p><p>Aligned dataset of LSU gene region of <i>Saksenaea</i> spp. Only used for LSU phylogeny.</p><p> </p><p>6. Saksenaea_tef1_RAW_v2.fasta</p><p>Non-aligned dataset of tef1 gene region of <i>Saksenaea</i> spp. </p><p> </p><p>7. Saksenaea_tef1_aligned_v2.fasta</p><p>Aligned dataset of tef1 gene region of <i>Saksenaea</i> spp. Only used for tef1 phylogeny.</p><p> </p><p><strong><Concatenated dataset 1 (ITS2, LSU, tef1)></strong></p><p>8. Saksenaea_ITS2_for_concatenated_RAW_v2.fasta</p><p>Non-aligned dataset of ITS2 region of <i>Saksenaea</i> spp. used for preparation of a concatenated dataset 1.</p><p> </p><p>9. Saksenaea_ITS2_for_concatenated_aligned_v2.fasta</p><p>Aligned dataset of ITS2 region of <i>Saksenaea</i> spp. used for preparation of a concatenated dataset 1.</p><p> </p><p>10. Saksenaea_LSU_for_concatenated_RAW_v2.fasta</p><p>Non-aligned dataset of LSU gene region of <i>Saksenaea</i> spp. used for preparation of concatenated datasets 1 and 2.</p><p> </p><p>11. Saksenaea_LSU_for_concatenated_aligned_v2.fasta</p><p>Aligned dataset of LSU gene region of <i>Saksenaea</i> spp. used for preparation of concatenated datasets 1 and 2.</p><p> </p><p>12. Saksenaea_tef1_for_concatenated_RAW_v2.fasta</p><p>Non-aligned dataset of tef1 gene region of <i>Saksenaea</i> spp. used for preparation of concatenated datasets 1 and 2.</p><p> </p><p>13. Saksenaea_tef1_for_concatenated_aligned_v2.fasta</p><p>Aligned dataset of tef1 gene region of <i>Saksenaea</i> spp. used for preparation of concatenated datasets 1 and 2.</p><p> </p><p>14. Saksenaea_ITS2_LSU_tef1_concatenated_dataset1.fasta</p><p>Concatenated dataset of three multiple sequence alignments (9, 11, and 13). This concatenated dataset was used for the main phylogeny of <i>Saksenaea</i> spp.</p><p> </p><p><strong><Concatenated dataset 2 (ITS1-5.8S-ITS2, LSU, tef1)></strong></p><p>15. Saksenaea_ITS_for_concatenated_RAW_v2.fasta</p><p>Non-aligned dataset of ITS1-5.8S-ITS2 region of <i>Saksenaea</i> spp. used for preparation of a concatenated dataset 2.</p><p> </p><p>16. Saksenaea_ITS_for_concatenated_aligned_v2.fasta</p><p>Aligned dataset of ITS1-5.8S-ITS2 region of <i>Saksenaea</i> spp. used for preparation of a concatenated dataset 2.</p><p>Blank sequences were inserted for five isolates of <i>Saksenaea longicolla</i> after the alignment.</p><p> </p><p>17.Saksenaea_ITS_LSU_tef1_concatenated_dataset2.fasta</p><p>Concatenated dataset of three multiple sequence alignments (15, 11, and 13). This concatenated dataset was used for the main phylogeny of <i>Saksenaea</i> spp.</p><p> </p><p><strong>C. Pairwise distances of the ex-type strains of </strong><i><strong>Saksenaea</strong></i><strong> spp.</strong></p><p>1. Saksenaea_ITS_type_RAW.fasta</p><p>Non-aligned dataset of ITS1-5.8S-ITS2 region of the ex-type strains of <i>Saksenaea</i> spp.</p><p> </p><p>2. Saksenaea_ITS2_type_aligned.fasta</p><p>Aligned dataset of ITS2 region of the ex-type strains of <i>Saksenaea</i> spp.</p><p> </p><p>3.Saksenaea_ITS_type_aligned.fasta</p><p>Aligned dataset of ITS1-5.8S-ITS2 region of the ex-type strains of <i>Saksenaea</i> spp. without <i>Saksenaea longicolla</i>.</p><p> </p><p>4. Saksenaea_LSU_type_RAW.fasta</p><p>Non-aligned dataset of LSU gene region of the ex-type strains of <i>Saksenaea </i>spp.</p><p> </p><p>5. Saksenaea_LSU_type_aligned.fasta</p><p>Aligned dataset of LSU gene region of the ex-type strains of <i>Saksenaea</i> spp.</p><p> </p><p>6. Saksenaea_tef1_type_RAW.fasta</p><p>Non-aligned dataset of tef1 gene region of the ex-type strains of <i>Saksenaea</i> spp.</p><p> </p><p>7. Saksenaea_tef1_type_aligned.fasta</p><p>Aligned dataset of tef1 gene region of the ex-type strains of <i>Saksenaea</i> spp.</p>
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