| Summary: | We sequenced the mitochondrial genome of six colonial volvocine algae, namely: <i>Pandorina morum</i>, <i>Pandorina colemaniae</i>, <i>Volvulina compacta</i>, <i>Colemanosphaera angeleri</i>, <i>Colemanosphaera charkowiensi</i>, and <i>Yamagishiella unicocca</i>. Previous studies have typically reconstructed the phylogenetic relationship between colonial volvocine algae based on chloroplast or nuclear genes. Here, we explore the validity of phylogenetic analysis based on mitochondrial protein-coding genes. We found phylogenetic incongruence of the genera <i>Yamagishiella</i> and <i>Colemanosphaera</i>. In <i>Yamagishiella</i>, the stochastic error and linkage group formed by the mitochondrial protein-coding genes prevent phylogenetic analyses from reflecting the true relationship. In <i>Colemanosphaera</i>, a different reconstruction approach revealed a different phylogenetic relationship. This incongruence may be because of the influence of biological factors, such as incomplete lineage sorting or horizontal gene transfer. We also analyzed the substitution rates in the mitochondrial and chloroplast genomes between colonial volvocine algae. Our results showed that all volvocine species showed significantly higher substitution rates for the mitochondrial genome compared with the chloroplast genome. The nonsynonymous substitution (dN)/synonymous substitution (dS) ratio is similar in the genomes of both organelles in most volvocine species, suggesting that the two counterparts are under a similar selection pressure. We also identified a few chloroplast protein-coding genes that showed high dN/dS ratios in some species, resulting in a significant dN/dS ratio difference between the mitochondrial and chloroplast genomes.
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