Labriocimbex sinicus Yan & Wei gen. et sp. nov. of Cimbicidae is described. The new genus is similar to Praia Andre and Trichiosoma Leach. A key to extant Holarctic genera of Cimbicinae is provided. To identify the phylogenetic placement of Cimbicidae, the mitochondrial genome of L. sinicus was annotated and characterized using high-throughput sequencing data. The complete mitochondrial genome of L. sinicus was obtained with a length of 15,405 bp (GenBank: MH136623; SRA: SRR8270383) and a typical set of 37 genes (22 tRNAs, 13 PCGs, and two rRNAs). The results demonstrated that all PCGs were initiated by ATN codon, and ended with TAA or T stop codons. The study reveals that all tRNA genes have a typical clover-leaf secondary structure, except for trnS1. Remarkably, the secondary structures of the rrnS and rrnL of L. sinicus were much different from those of Corynis lateralis. Phylogenetic analyses verified the monophyly and positions of the three Cimbicidae species within the superfamily Tenthredinoidea and demonstrated a relationship as (Tenthredinidae + Cimbicidae) + (Argidae + Pergidae) with strong nodal supports. Furthermore, we found that the generic relationships of Cimbicidae revealed by the phylogenetic analyses based on COI genes agree quite closely with the systematic arrangement of the genera based on the morphological characters. Phylogenetic tree based on two methods shows that L. sinicus is the sister group of Praia with high support values. We suggest that Labriocimbex belongs to the tribe Trichiosomini of Cimbicinae based on adult morphology and molecular data. Besides, we suggest to promote the subgenus Asitrichiosoma to be a valid genus.
The Asian potamid subfamily Potamiscinae is the most species-rich group of primary freshwater crabs in the world, and a large number of these (318 species in 52 genera) are found in China where they exhibit a remarkably high rate of endemism. However, the evolutionary history and phylogenetic relationships of this subfamily is still poorly understood and the hypothesis that the Chinese potamids originated in the Indochina Peninsula has yet to be tested using a modern phylogeographic reconstruction and molecular dating methods. Here we analyze the phylogenetic relationships and biogeographic history of the Chinese potamiscines based on whole mitogenomes of 72 species of Asian freshwater crabs representing 65% of the potamiscine genera found in China, including new mitogenome sequences for 57 species. The mitogenomic phylogeny generated four separate clades: Southwest China (SWC), Indochina-Southwest China (ISWC), Central China (CC), and South China-adjacent Islands (SCI). Our results suggest that the Chinese potamiscine fauna most likely originated in Indochina during the Eocene. The establishment of the East Asian monsoon during the Eocene/Oligocene resulted in major climate changes that coincide with the local diversification of the CC and SCI clades. In addition, diversification of two other clades (ISWC and SWC) may have been driven by past large scale environmental changes taking place at that time such as increased local precipitation and expanding freshwater habitats associated with the uplifting of the Hengduan Mountains. In addition, our phylogenies did not support monophyly for six currently recognized potamiscine genera, highlighting the need for further taxonomic revisions of this fauna. The broad taxonomic sampling and well-supported molecular phylogenies used here allow for the updating of our understanding of the evolutionary origins and diversification of the potamiscine freshwater crabs in China.
Single-cell genomics (SCG) is a recently developed tool to study the genomes of unculturable bacterial species. SCG relies on multiple-strand displacement amplification (MDA), PCR, and next-generation sequencing (NGS); however, obtaining sufficient amounts of high-quality DNA from samples is a major challenge when performing this technique. Here we present an improved bacterial cell lysing procedure that combines incubation in an alkaline buffer with a thermal shock (freezing/heating) treatment to yield highly intact genomic DNA with high efficiency. This procedure is more efficient in lysing Bacillus subtilis and Synechocystis cells compared with two other frequently used lysis methods. Furthermore, 16S ribosomal RNA gene and overall genome recovery were found to be improved by this method using single cells from a Utah desert soil community or Escherichia coli single cells, respectively. The efficiency of genome recovery for E. coli single cells using our procedure is comparable with that of the REPLI-g Single Cell (sc) Kit, but our method is much more economical. By providing high-quality genome templates suitable for downstream applications, our procedure will be a promising improvement for SCG research.
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