Limoniidae, the most speciose family in the superfamily Tipuloidea, consists of four subfamilies and more than 11,000 species. However, mitochondrial (mt) genome sequences, which have been widely used for phylogenetic study, are available for only 11 species across three subfamilies. Thus, a larger variety of mt genome sequences in Limoniidae are required to improve our understanding of tipuloid phylogeny and genomic evolution. Here we present mt genomes of Elephantomyia (Elephantomyia) inulta Alexander, 1938 and Helius (Helius) pluto Alexander, 1932, representing the first mt genomes of the tribe Elephantomyiini (Limoniidae). The two mt genomes are typical circular DNA molecules and show similar gene order, nucleotide composition and codon usage. Standard ATN start and TAR stop codons are present in most protein-coding genes. All transfer RNA (tRNA) genes exhibited the cloverleaf secondary structure typical for metazoans except in tRNASer(AGN), which lacks the dihydrouridine arm. Phylogenetic analyses were performed based on four nucleotide matrixes for the currently sequenced species of Tipuloidea using Bayesian inference and maximum likelihood methods. Four-cluster likelihood mapping was used to study incongruent signals between different topologies. Pediciidae is supported as the earliest lineage in Tipuloidea, and the sister-group relationship between Cylindrotomidae and Tipulidae is also supported, but the monophyly of Limoniidae is not supported. Our study also supports the monophyly of Elephantomyiini (Elephantomyia + Helius), as one of origins of flower-visiting in Limoniidae. Although Elephantomyiini is sister to Limoniinae + Epiphragma (Limnophilinae) in our study, a more precise understanding of its phylogenetic position in Tipuloidea will require additional studies that include a broader species sample.
A remarkable new genus and species of Nemourinae, Sinonemura balangshana gen. et sp. n., is described from Balang Mountains, Sichuan, southwestern China. The description is based on morphology and molecular data. The Nemourinae genera related to the new taxon are re-evaluated on the basis of comparative functional morphology of male epiproct. Notes on the Asian distribution of the Nemourinae are also given.
Two new complete mitochondrial genomes (mitogenomes) of the subfamily Chloroperlinae, Haploperla japonica Kohno, 1946 and Sweltsa sp., were sequenced. The two species showed similar gene order, nucleotide composition, and codon usage. The Sweltsa sp. and H. japonica mitogenomes were typical circular DNA molecules, with lengths of 15,893 bp and 16,012 bp, respectively. Standard ATN start and TAN stop codons were present in most PCGs. All tRNA genes exhibited the cloverleaf secondary structure typical for metazoans except the tRNASer(AGN), which lacked the dihydrouridine arm. In both species, the secondary structure of lrRNA contained five structural domains, while the srRNA included three domains. The A+T-rich regions contained different repeat regions in each species. Phylogenetic analyses using Bayesian inference (BI) and maximum likelihood methods (ML) showed identical results. The family Chloroperlidae was sister to the Perlodidae. Our analyses inferred relationships between six of the seven Systellognatha families: (((Chloroperlidae + Perlodidae) + Perlidae) + (Styloperlidae + Pteronarcyidae)) + Peltoperlidae.
Dicranoptychini is a tribe in the subfamily Limoniinae (Diptera, Tipuloidea, and Limoniidae) and includes only the genus Dicranoptycha Osten Sacken, 1860. However, the species diversity of the tribe in China was seriously underestimated, and the taxonomic status of Dicranoptycha has long been controversial. In this study, types of Chinese Dicranoptycha species and specimens collected from several localities in China were examined, and the first mitochondrial (mt) genome of the tribe Dicranoptychini is presented. Two Dicranoptycha species, D. jiufengshana sp. nov. and D. shandongensis sp. nov., from China, are described and illustrated as new to science. A Palaearctic species, D. prolongata Alexander, 1938, is recorded in China for the first time. In addition, the complete mt genome of D. shandongensis sp. nov. is sequenced and annotated, indicating that it is a typical circular DNA molecule with a length of 16,157 bp and shows a similar gene order, nucleotide composition, and codon usage to mt genomes of other Tipuloidea species. The two pairs of repeat elements are found in its control region. Phylogenetic results confirm the sister-group relationship between Cylindrotomidae and Tipulidae, question the position of the genus Epiphragma Osten Sacken, 1860 in Limoniidae, and indicate that Dicranoptychini may be a basal lineage within Limoniinae.
Three new species of the genus Nemoura Latreille, 1796: Nemoura alticalcaneum Mo, Wang & Li, sp. n., Nemoura ganeum Mo, Yang & Li, sp. n. and Nemoura tauripitis Mo, Li & Murányi, sp. n., are described from Guangxi Zhuang Autonomous Region of China. Eight new province records from Guangxi, two new province records from Guangdong and one new province record from Hunan are given, among them one new for China as well. The newly recorded species are depicted and complementary descriptions provided, including the first female description of seven species and first egg description of two species. The paper is completed with an updated checklist of Guangxi stoneflies, distribution of species richness across prefectural cities in Guangxi, with distribution maps for each species.
The genus Helius Lepeletier & Serville, 1828 is recorded in Guangxi, China for the first time with the following three species belonging to the subgenus H. (Helius): H. (H.) damingshanus sp. nov., H. (H.) nipponensis (Alexander, 1913) and H. (H.) stenorhynchus stenorhynchus Alexander, 1954. Among them, H. (H.) stenorhynchus stenorhynchus is also a new record in China. Their descriptions and illustrations, as well as the first key to Chinese H. (Helius) crane flies, are presented.