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5 results for “Single copy orthologous gene”
Single-copy orthologous genes used for Ricefish phylogeny
<p>Ortholog set</p> <p> We generated a reference set consisting of 8390 single-copy protein-coding genes derived from OrthoDB v.9.1 (Waterhouse et al., 2013) available for the following species: <em>Austrofundulus limnaeus, Centrocoris variegatus, Fundulus heteroclitus, Kryptolebias marmoratus, Nothobranchius furzeri, Oryzias latipes, O. melastigma, Poecilia formosa, P. latipinna ,P. mexicana, P. reticulata</em> and <em>Xiphophorus maculatus </em>(NCBI Accession numbers in Table S7). The hierarchical split was set to Actinopterygii (ID 7898). We used the script “make-ogs-corresponding.pl” to check for inconsistencies between the amino acid sequences and the corresponding nucleotide sequences and removed 96 problematic genes (Tab. S7). </p> <p>Identification of orthologs for transcripts and genome and alignment of single-copy genes</p> <p> Ortholog identification among 16 ricefish species and four outgroups (DS1, supplementary tables Tab. S1a) was carried out with Orthograph v0.7.1 (Petersen et al., 2017). Forward search for candidate transcript was left at default. Best reciprocal hit: Ortholog candidate genes needed at least one hit in either <em>O. latipes</em> or <em>O. melastigma </em>and we allowed concatenation of hits if they met the criteria and did not overlap. Max-blast-searches were set to 50, blast-max-hits were also set to 50. “U” in the amino acid sequences was changed to “X” to avoid issues in downstream analysis. The results of the orthology prediction were summarized for all species using a custom perl script coming with the orthograph package. Sequences of only those orthologs with all species present were aligned using MAFFT v7.221 with the L-INS-I algorithm on amino acid level (Katoh & Standley, 2013). 915 orthologs with outliers were identified according to Misof et al. 2014 and were subsequently removed from further analysis. We used the amino-acid alignments as blue print to generate corresponding nucleotide alignments with a modified version of Pal2Nal v14 (Misof et al., 2014; Suyama et al., 2006). To check each amino acid alignment for ambiguously aligned regions, we ran ALISCORE v2.0 with the maximal number of possible sequence selected pairs to analyze (-r) (Kück et al., 2010; Misof et al., 2014; Misof & Misof, 2009). Sites which needed masking were cut out using ALICUT v2.3 (Kück, 2009) from the amino acid alignments and correspondingly also from the nucleotide alignments. For further analyses we only proceeded with the data set on nucleotide level.</p>
Data from: Mining from transcriptomes: 315 single-copy orthologous genes concatenated for the phylogenetic analyses of Orchidaceae
Phylogenetic relationships are hotspots for orchid studies with controversial standpoints. Traditionally, the phylogenies of orchids are based on morphology and subjective factors. Although more reliable than classic phylogenic analyses, the current methods are based on a few gene markers and PCR amplification, which are labor intensive and cannot identify the placement of some species with degenerated plastid genomes. Therefore, a more efficient, labor-saving and reliable method is needed for phylogenic analysis. Here, we present a method of orchid phylogeny construction using transcriptomes. Ten representative species covering five subfamilies of Orchidaceae were selected, and 315 single-copy orthologous genes extracted from the transcriptomes of these organisms were applied to reconstruct a more robust phylogeny of orchids. This approach provided a rapid and reliable method of phylogeny construction for Orchidaceae, one of the most diversified family of angiosperms. We also showed the rigorous systematic position of holomycotrophic species, which has previously been difficult to determine because of the degenerated plastid genome. We concluded that the method presented in this study is more efficient and reliable than methods based on a few gene markers for phylogenic analyses, especially for the holomycotrophic species or those whose DNA sequences have been difficult to amplify. Meanwhile, a total of 315 single-copy orthologous genes of orchids are offered and more informative loci could be used in the future orchid phylogenetic studies.
Data from: Mining from transcriptomes: 315 single-copy orthologous genes concatenated for the phylogenetic analyses of Orchidaceae
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Data from: Identification and qualification of 500 nuclear, single-copy, orthologous genes for the Eupulmonata (Gastropoda) using transcriptome sequencing and exon capture
The qualification of orthology is a significant challenge when developing large, multiloci phylogenetic data sets from assembled transcripts. Transcriptome assemblies have various attributes, such as fragmentation, frameshifts and mis-indexing, which pose problems to automated methods of orthology assessment. Here, we identify a set of orthologous single-copy genes from transcriptome assemblies for the land snails and slugs (Eupulmonata) using a thorough approach to orthology determination involving manual alignment curation, gene tree assessment and sequencing from genomic DNA. We qualified the orthology of 500 nuclear, protein-coding genes from the transcriptome assemblies of 21 eupulmonate species to produce the most complete phylogenetic data matrix for a major molluscan lineage to date, both in terms of taxon and character completeness. Exon capture targeting 490 of the 500 genes (those with at least one exon >120 bp) from 22 species of Australian Camaenidae successfully captured sequences of 2825 exons (representing all targeted genes), with only a 3.7% reduction in the data matrix due to the presence of putative paralogs or pseudogenes. The automated pipeline Agalma retrieved the majority of the manually qualified 500 single-copy gene set and identified a further 375 putative single-copy genes, although it failed to account for fragmented transcripts resulting in lower data matrix completeness when considering the original 500 genes. This could potentially explain the minor inconsistencies we observed in the supported topologies for the 21 eupulmonate species between the manually curated and 'Agalma-equivalent' data set (sharing 458 genes). Overall, our study confirms the utility of the 500 gene set to resolve phylogenetic relationships at a range of evolutionary depths and highlights the importance of addressing fragmentation at the homolog alignment stage for probe design.
Data from: Identification and qualification of 500 nuclear, single-copy, orthologous genes for the Eupulmonata (Gastropoda) using transcriptome sequencing and exon capture
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