Based on morphological comparisons and molecular phylogenetic analyses, a new species of the genus Tylototriton from Wufeng County, Hubei Province, China, is described. Phylogenetic analyses of mitochondrial 16S rRNA and ND2 gene sequences supported the new species as an independent clade, sister to a clade containing Tylototriton dabienicus and T. gaowangjienensis. The uncorrected p-distances of the 16S rRNA and ND2 genes between the new species and its closest congeners were 0.7% and 1.7%, respectively. Divergence time estimates indicated separation from T. dabienicus and T. gaowangjienensis at 2.53 million years ago (Mya; 95% highest posterior density [HPD]: 1.53–3.72 Mya). Bayesian species delimitation (BFD and bPTP analyses) strongly supported the new taxon as a distinct species. Morphologically, it is distinguished by: (1) moderate body size (SVL 63.7–77.2 mm in males, 74.9–88.1 mm in females); (2) head width exceeding head length; (3) presence of a gular fold; (4) tail shorter than snout–vent length; (5) orange distal digits, cloacal periphery, and tail margin; (6) limb tips meeting when fore- and hindlimbs are pressed along the trunk; (7) fingertips extending beyond the snout when forelimbs are stretched forward; and (8) continuous, indistinct, nodule-like warts on the body sides.

Anopheles mosquitoes serve as a significant vector for the transmission of malaria and arboviruses holding great medical importance, but their phylogenetic relationships have not yet been clearly elucidated. This study was the first to sequence and analyze the mitochondrial genomic features of An. hyrcanus and An. messeae, and to incorporate them into a phylogenetic analysis with 102 other species within the genus Anopheles. Both genomes contain 37 genes, including 22 tRNAs, 13 protein-coding genes and 2 rRNAs, as well as one control region. The genome characteristics, codon usage, and tRNA secondary structure of the two species align with those observed in mitochondrial genomes of other mosquito species. Phylogenetic analysis revealed that the relationship among six subgenera was Lophopodomyia + ((Kerteszia + Stethomyia) + ((Cellia + Anopheles) + Nyssorhynchus)), providing strong support for their monophyly. Nevertheless, the phylogenetic relationships within subgenus Cellia were inconsistent with previous findings. The findings of this study will enrich the biological information repository for Anopheles mosquitoes, lay the foundation for further research on systematic classification, and hold significant implications for guiding the research, prevention and control of mosquito-borne diseases.

Surveying biodiversity has taken a quantum leap with environmental DNA (eDNA) metabarcoding, an immensely powerful approach lauded for its efficiency, sensitivity, and non-invasiveness. This approach emerges as a game-changer for the elusive realm of endangered and rare species-think nocturnal, environmentally elusive amphibians. Here, we have established a framework for constructing a reliable metabarcoding pipeline for amphibians, covering primer design, performance evaluation, laboratory validation, and field validation processes. The Am250 primer, located on the mitochondrial 16S gene, was optimal for the eDNA monitoring of amphibians, which demonstrated higher taxonomic resolution, smaller species amplification bias, and more extraordinary detection ability compared to the other primers tested. Am250 primer exhibit an 83.8% species amplification rate and 75.4% accurate species identification rate for Chinese amphibians in the in silico PCR and successfully amplified all tested species of the standard samples in the in vitro assay. Furthermore, the field-based mesocosm experiment showed that DNA can still be detected by metabarcoding even days to weeks after organisms have been removed from the mesocosm. Moreover, field mesocosm findings indicate that eDNA metabarcoding primers exhibit different read abundances, which can affect the relative biomass of species. Thus, appropriate primers should be screened and evaluated by three experimental approaches: in silico PCR simulation, target DNA amplification, and mesocosm eDNA validation. The selection of a single primer set or multiple primers' combination should be based on the monitoring groups to improve the species detection rate and the credibility of results.

Phylogenetic studies in the phylogenomics era have demonstrated that reticulate evolution greatly impedes the accuracy of phylogenetic inference, and consequently can obscure taxonomic treatments. However, the sys-tematics community lacks a broadly applicable strategy for taxonomic delimitation in groups characterized by pervasive reticulate evolution. The red-fruit genus, Stranvaesia, provides an ideal model to examine the influence of reticulation on generic circumscription, particularly where hybridization and allopolyploidy dominate the evolutionary history. In this study, we conducted phylogenomic analyses integrating data from hundreds of single-copy nuclear (SCN) genes and plastomes, and interrogated nuclear paralogs to clarify the inter/intra-generic relationship of Stranvaesia and its allies in the framework of Maleae. Analyses of phylogenomic discord and phylogenetic networks showed that allopolyploidization and introgression promoted the origin and diversification of the Stranvaesia clade, a conclusion further bolstered by cytonuclear and gene tree discordance. With a well-inferred phylogenetic backbone, we propose an updated generic delimitation of Stranvaesia and introduce a new genus, Weniomeles. This new genus is distinguished by its purple-black fruits, thorns trunk and/ or branches, and a distinctive fruit core anatomy characterized by multilocular separated by a layer of sclereids and a cluster of sclereids at the top of the locules. Through this study, we highlight a broadly-applicable workflow that underscores the significance of reticulate evolution analyses in shaping taxonomic revisions from phylogenomic data.

The type species of Filchneria Klapálek, 1908, F. mongolica (Klapálek, 1901), is based on a single female collected from Mongolia, but it was considered the same as another species, F. songi from Qinling, China, when the genus Filchneria was proposed. This study narrates the story of these two species, which have been confused for a century. Until now, the distribution of F. mongolica has been confirmed only in Mongolia and Russia, and we recently recorded it for the first time in Inner Mongolia as a new species record in China. Additionally, the genus Sinoperlodes is a junior synonym of Filchneria, as demonstrated by both the morphological and molecular analysis. Phylogenetic analysis based on the subfamily Perlodinae is provided, along with morphological and biogeographic comparisons of Filchneria and its relatives.

Mitochondrial metagenomics (MMG) using Illumina sequencers for mixed-species samples provides a promising tool for evolutionary and ecological studies using mitogenomes. However, the traditional assembly procedure is still computationally intensive and expensive. Here, a novel MMG pipeline was applied to different DNA extractions, one per species, and their sequence as a mixed sample for rapid mitogenome assembly is presented. Our method integrated a faster and more accurate read mapper for filtering non-mitochondrial reads. A seed-and-extend assembler for species-specific mitogenomes that detects ‘noisy species/sequences’ was also assessed. The MMG pipeline for each dataset was completed in a few hours on desktop PCs, maintaining high accuracy and completeness (COI divergence >10%), except for some very closely related taxa. Particularly for closely related species, the exclusion of ‘noisy reads’ (including chimera of non-targeted species) improved the target assembly. In addition, we observed that short barcodes used as references had almost identical detection power compared with mitogenomes but required greater sequencing depth. We tested our MMG pipeline on two real and one simulated dataset to validate its high efficiency in mixed-species sample mitogenome assembly.

A nov. during field 2021. The new species can be well distinguished from its congeners by the combination of the following characteristics: mouth shallow arc-shaped, width of cutting edge on upper jaw equal to half mouth width, central portion of anterior papillae arranged in one row with 6–10 well-developed papillae of equal size, medial pad on lower lip bisected into two squircle-shaped pads, and grooved; midventral region completely scaled and thorax scaleless, lateral line scales 36–38, pre-dorsal scales 8–10; barbel length 26.7%–31.4% of head length; eye diameter 30.9%–36.9% of head length, interorbital width 21.3%–27.6% of head length; posterior chamber of air-bladder weak, length smaller than half eye diameter. Bayesian inference and maximum-likelihood phylogenetic analyses based on the cyt b gene sequence and species delineation also the specimens as a distinct species, sister to M. brevirostris (Günther, 1868). Project of Biodiversity Survey and Conservation in and Sino BON-Inland Fish Diversity

DEAR EDITOR, A new gobionine species,Microphysogobio oujiangensis sp.nov.,was collected from the Oujiang River Basin during field work in Lishui City,Zhejiang Province in 2021.The new species can be well distinguished from its congeners by the combination of the following characteristics:mouth shallow arc-shaped,width of cutting edge on upper jaw equal to half mouth width,central portion of anterior papillae arranged in one row with 6-10 well-developed papillae of equal size,medial pad on lower lip bisected into two squircle-shaped pads,and grooved;midventral region completely scaled and thorax scaleless,lateral line scales 36-38,pre-dorsal scales 8-10;barbel length 26.7％-31.4％of head length;eye diameter 30.9％-36.9％of head length,interorbital width 21.3％-27.6％of head length;posterior chamber of air-bladder weak,length smaller than half eye diameter.Bayesian inference and maximum-likelihood phylogenetic analyses based on the cyt b gene sequence and species delineation also supported the specimens as a distinct species,sister to M.brevirostris(Gunther,1868).

A new species of the frog genus Rana sensu lato from Wuyi Mountain, Fujian Province, China is described. Molecular phylogenetic analyses clustered the new species into the R.johnsi group and indicated that it was genetically divergent from its closely related species. The new species could be distinguished from its congeners by a combination of the following characters: body size medium, SVL 41.4-45.6 mm (42.9 ± 1.9 mm, n = 4) in adult males and 47.6-50.3 mm (n = 2) in adult females; adult male with a pair of internal subgular vocal sacs; lateroventral grooves present on tip of toes; webbing on fourth toes reaching the tip of toe; transverse skin ridges distinctly present on the dorsal surface of thigh and tibia, the number large (mean 26.5 ± 2.7, range 22-29, n = 6); breeding males possess creamy white nuptial pad with tiny velvety spines on the dorsal surface of the first finger, divided into three parts.

A new species of the genusMegophrysfrom Zhejiang Province, China is described. Molecular phylogenetic analyses supported the new taxon as an independent clade nested into theMegophrysclade and sister toM. lishuiensis. The new species could be distinguished from its congeners by a combination of the following morphological characteristics: (1) small size (SVL 31.0–36.3 mm in male and 41.6 mm in female); (2) vomerine ridge present and vomerine teeth absent; (3) tongue not notched behind; (4) a small horn-like tubercle at the edge of each upper eyelid; (5) tympanum distinctly visible, rounded; (6) two metacarpal tubercles in hand; (7) relative finger lengths: II < I < IV < III; (8) toes with rudimentary webbing at bases; (9) heels overlapping when thighs are positioned at right angles to the body; (10) tibiotarsal articulation reaching tympanum to eye when leg stretched forward; (11) an internal single subgular vocal sac in male; (12) in breeding male, the nuptial pads with black nuptial spines on the dorsal bases of the first and second fingers.

2018年6月,在位于云南省文山壮族苗族自治州的文山国家级自然保护区(23°21′41″N,104°5026″E,海拔1 300 m)采集到无尾目(Anura)姬蛙科(Microhylidae)狭口蛙属(Kaloula)标本3号,包括雄性成体标本1号和雌性成体标本2号,并对相应形态特征进行了测量,雄性头体长为35.79mm,雌性头体长为48.01mm和42.78mm.根据形态特征比较发现,该种群的标本皮肤光滑,背部呈橄榄绿色,有不规则的深绿色斑纹,指端平切且具两排小乳突,雄性胸部有小柠檬黄色斑点,雄性全蹼,雌性为1/3蹼,与采集自弄岗国家级自然保护区的弄岗狭口蛙(K.nonggangensis)地模标本(海拔186m)的形态鉴定特征基本一致,仅体色和体型大小稍有差异.基于16S rRNA基因片段进行系统发育分析表明,这些标本与弄岗狭口蛙模式产地标本聚在一起,共享同一单倍型.可以确定所采集的3号标本均为弄岗狭口蛙,为云南省两栖纲分布新记录物种.新发现的种群分布于中高海拔处,较地模标本的低海拔分布记录有较大的拓展.该物种的发现说明,云南东南部与广西西南部的喀斯特地区具有相似的物种组成,对该区域将来的生物地理学研究具有重要意义.

A new species of the Asian horned toad genus Megophrys is described from Zhejiang Province, China, based on multiple data. Molecular phylogenetic analyses based on mitochondrial DNA indicated the new species as an independent clade deeply clustered into the Megophrys clade. The new species is identified from its congeners by a combination of the following characters: body size small (SVL 28.4-32.4 mm in males); vomerine teeth absent; tongue not notched behind; tympanum distinctly visible, oval; a small horn-like tubercle present at the edge of each upper eyelid; two metacarpal tubercles distinctly visible in hand; toes without webbing; heels overlapped when thighs are positioned at right angles to the body; tibiotarsal articulation reaching the level to middle of eye when leg stretched forward; an internal single subgular vocal sac in male; in breeding male, the nuptial pads present on the dorsal base of the first two fingers.