Two-Fold Genes, Three-Fold Control Regions, lncRNA, and the "Missing" Found in the Mitogenomes of (Rhacophridae: ).

In prior research on the mitochondrial genome (mitogenome) of , the one copy of gene was translocated to the control region (CR) and the gene was not found. Gene loss is uncommon among vertebrates. However, in this study, we resequenced the mitogenomes of from different regions using a "primer bridging" approach with Sanger sequencing technologies, which revealed the "missing" gene in as well as three other previously published . The mitogenome of this species was found to contain two copies of the genes and three copies of the control regions. Furthermore, multiple tandem repeats were identified in the control regions. Notably, we observed that there was no correlation between genetic divergence and geographic distance. However, using the mitogenome, gene expression analysis was performed via RT-qPCR of liver samples and it was thus determined that , , , and were reduced to 0.64 ± 0.24, 0.55 ± 0.34, 0.44 ± 0.21 and 0.65 ± 0.17, respectively, under low-temperature stress (8 °C) as compared with controls ( < 0.05). Remarkably, the transcript of long non-coding RNA (lncRNA) between positions 8029 and 8612 decreased significantly with exposure to low-temperature stress (8 °C). Antisense gene expression showed a downward trend, but this was not significant. These results reveal that modulations of protein-coding mitochondrial genes and lncRNAs of play a crucial role in the molecular response to cold stress.