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In this study, the mitochondrial (mt) genomes of Siphluriscus chinensis (Ephemeroptera: Siphluriscidae) were evaluated in specimens collected from two sites in China: Niutou Mountain, Zhejiang Province (S. chinensis NTS) and Leigong Mountain, Guizhou Province (S. chinensis LGS) and were successfully sequenced. The lengths of the mt genomes of S. chinensis NTS and S. chinensis LGS were 15,904 bp (ON729390) and 15,212 bp (ON729391), respectively. However, an in-depth comparison of the two mt genomes showed significant differences between the specimens collected from the two sites. A detailed analysis of the genetic distance between S. chinensis NTS and S. chinensis LGS was undertaken to further achieve an accurate delimitation of S. chinensis. The genetic distance between S. chinensis NTS and the other three species within Siphluriscidae was a high value, above 12.2%. The two mt genomes were used to reconstruct phylogenetic relationships and estimate divergence time. The results demonstrated robust differences between S. chinensis NTS and S. chinensis LGS, which revealed that a kind of cryptic species existed. Maximum likelihood (ML) and Bayesian inference (BI) analyses produced well-supported phylogenetic trees that showed evolutionary relationships between Siphluriscidae (((S. chinensis HQ875717 + S. chinensis MF352165) + S. chinensis LGS) + S. chinensis NTS). The most recent common ancestor (MRCA) of four species within Siphluriscidae began to diversify during the Neogene [11.80 million years ago (Mya); 95% highest posterior densities (HPD) = 6.17–19.28 Mya], and S. chinensis NTS was first to diverge from the branches of S. chinensis LGS. In short, based on mitochondrial genomes, our results showed that the specimens collected from Leigong Mountain, Guizhou Province (S. chinensis LGS) belonged to S. chinensis, and the specimens collected from Niutou Mountain, Zhejiang Province (S. chinensis NTS) were a cryptic species of S. chinensis. [ABSTRACT FROM AUTHOR]

Simple Summary: The family Vietnamellidae (Ephemeroptera) is one of the oldest insect families in the world. However, there are still controversies about the phylogenetic relationships among Vietnamellidae, Ephemerellidae, and Teloganodidae. The mitochondrial (mt) genome can be used to discuss phylogenetic relationships and cryptic species. We sequenced and annotated three complete mt genomes of Vietnamella sinensis from three different populations, identifying a cryptic species of V. sinensis and discuss the phylogenetic relationships of Vietnamellidae. Based on the genetic distance of the whole mt genomes, the phylogenetic relationship of three populations was uncovered and the divergence time of V. sinensis QY indicated that it was a cryptic species of V. sinensis. Ephemeroptera (Insecta: Pterygota) are widely distributed all over the world with more than 3500 species. During the last decade, the phylogenetic relationships within Ephemeroptera have been a hot topic of research, especially regarding the phylogenetic relationships among Vietnamellidae. In this study, three mitochondrial genomes from three populations of Vienamella sinensis collected from Tonglu (V. sinensis TL), Chun'an (V. sinensis CN), and Qingyuan (V. sinensis QY) in Zhejiang Province, China were compared to discuss the potential existence of cryptic species. We also established their phylogenetic relationship by combining the mt genomes of 69 Ephemeroptera downloaded from NCBI. The mt genomes of V. sinensis TL, V. sinensis CN, and V. sinensis QY showed the same gene arrangement with lengths of 15,674 bp, 15,674 bp, and 15,610 bp, respectively. Comprehensive analyses of these three mt genomes revealed significant differences in mt genome organization, genetic distance, and divergence time. Our results showed that the specimens collected from Chun'an and Tonglu in Zhejiang Province, China belonged to V. sinensis, and the specimens collected from Qingyuan, Zhejiang Province, China were a cryptic species of V. sinensis. In maximum likelihood (ML) and Bayesian inference (BI) phylogenetic trees, the monophyly of the family Vietnamellidae was supported and Vietnamellidae has a close relationship with Ephemerellidae. [ABSTRACT FROM AUTHOR]

The relationship of Matrona and Atrocalopteryx (Odonata: Calopterygidae) is still unclear. To better understand the phylogenetic relationship of Matrona and Atrocalopteryx, we sequenced and annotated two complete mitochondrial genomes of Matrona basilaris sampled from two different locations. The length of the two complete mitochondrial genomes of M. basilaris is 16,149 bp and 15,893 bp for the specimens collected in Jinhua, Zhejiang Province and Tianmushan, Zhejiang Province, China, respectively. The two mitochondrial genomes include the typical invertebrate set of 37 genes: 13 protein-coding genes (PGCs), 22 tRNA genes, and 2 rRNA genes. The nucleotide composition of the mitogenome is similar to other odonates with high content of A + T (68.9%) and all PCGs use ATN as the start codon. Tandem repeats were detected in the control regions of the two M. basilaris samples that accounted for the different sequence lengths of the mitochondrial genomes from the two locations. Finally, BI and ML phylogenetic analysis based on the concatenated nucleotide sequences of the 13 PCGs supported the conclusion that M. basilaris is a sister clade to Atrocalopteryx melli. [ABSTRACT FROM AUTHOR]