Phylogenetic Reconstruction Research Articles

The chloroplast genomes of Satureja are highly conserved but contain hypervariable loci like ndhF and ycf1. These loci provide powerful molecular tools for precise species identification, phylogenetic resolution, and future conservation efforts in this important genus. The genus Satureja (Lamiaceae) comprises a valuable number of aromatic herbs and shrubs with significant ecological, medicinal, and economic value, particularly in the Mediterranean and Southwest Asia. Satureja species are renowned for their therapeutic essential oils, yet their chloroplast (cp) genomic resources remain vastly understudied, with only S. montana complete cp genome available. To address this gap, we sequenced and analyzed the complete cp genomes of three taxonomically and ecologically distinct Satureja species (S. hortensis, S. khuzestanica, and S. montana subsp. variegata) and a new individual of S. montana, using Illumina sequencing and comparative genomic approaches. The assembled cp genomes exhibited a conserved quadripartite structure, with sizes ranging narrowly from 151,777 to 151,924bp, featuring typical large (LSC) and small (SSC) single-copy regions flanked by inverted repeats (IRs). Genome annotation revealed 128-130 genes, including core photosynthetic and ribosomal genes, with notable interspecific variation in tRNA copy numbers. Comparative analyses identified hypervariable loci (ndhF, ycf1, petN) as promising DNA barcodes for species discrimination, while codon use bias strongly favored A/T-ending codons. Simple sequence repeats (SSRs) were predominantly mononucleotide (A/T), and long repetitive sequences showed species-specific length polymorphisms. Phylogenetic reconstruction using the whole cp genomes strongly supported the monophyly of Satureja and resolved infrageneric relationships with high bootstrap confidence (> 90%), placing these species in a distinct clade sister to Thymus and Mentha. Our study provides the first comprehensive cp genomic resource for Satureja, elucidating evolutionary patterns, identifying molecular markers for taxonomy, and establishing a foundation for future conservation and biotechnological applications in this economically important genus.

Cryptotermes mobydicki sp. nov. is described from the soldier caste. The soldier is unique among all Cryptotermes species worldwide for its very narrow, elongate head capsule terminating with an extended frontal process. The new species constitutes the sixteenth Cryptotermes species for South America. Our phylogenetic reconstruction indicates that C. mobydicki sp. nov. is most closely related to the Neotropical and Central American C. mangoldi , C. parvifrons , C. cymatofrons , C. rotundiceps , and C. cavifrons .

The accurate phylogenetic distinction between Prochilodus magdalenae and Prochilodus reticulatus (Characiformes: Prochilodontidae) has been hindered by overlapping morphology and limited sequence data. Previous studies, relying on partial mitochondrial markers, have even suggested that Prochilodus magdalenae and Prochilodus reticulatus might be a single species. This study presents three annotated complete mitochondrial genomes for P. reticulatus and phylogenetic analyses that contribute to resolving uncertainty around these species’ boundaries. Our phylogenetic reconstructions, using both mitochondrial markers and complete mitogenomes, consistently support the segregation of P. magdalenae and P. reticulatus into distinct clades. Bayesian time-calibrated analysis estimates their divergence at approximately 6.9 mya (10.2–4.2 mya), coinciding with the Andean Eastern Cordillera’s final uplift. This study provides essential data for future taxonomic and conservation efforts. Our findings clarify the phylogenetic relationship between these species, emphasizing the utility of complete mitogenomes and demonstrating that sequence mislabeling, probably caused by the difficulty of accurately identifying these species based on morphological characteristics, has contributed to inconsistencies in previous phylogenetic studies within the genus Prochilodus .

ABSTRACT Aim We examine biogeography and speciation patterns in Miliusa Lesch. ex A. DC. (~65 species) distributed in tropical Asia to understand the uneven distribution of its extant diversity, with Indo‐Burma having twice the species richness of peninsular India (PI) and four times that of Wallacea and Sahul. Location Tropical Asia. Taxon Miliusa (Annonaceae). Methods Phylogenetic reconstruction was performed using six plastid markers across fifty‐two species using both ML and BI approaches. Divergence times were estimated using two fossil calibrations and an optimized relaxed clock, and ancestral areas were inferred with ‘BioGeoBEARS’. Speciation rates were examined using ClaDS and the DR statistic, and community structure was assessed using phylogenetic diversity metrics. Results Miliusa likely originated in the mid‐Miocene, with Indo‐Burma and PI as its ancestral range. Its extant diversity is primarily attributed to in‐situ speciation, with dispersal or vicariance playing limited but important roles in PI, and Wallacea and Sahul. Lineages in Indo‐Burma began accumulating in the mid‐Miocene, preceding those in PI (~10 Myr) and Wallacea and Sahul (~5 Myr). PI showed signs of saturation in lineage accumulation and had lower speciation rates compared to Wallacea and Sahul and Indo‐Burma, both of which had similar rates. All regions exhibited phylogenetic clustering, but Indo‐Burma and PI differed in their sensitivity to phylogenetic depths. Main Conclusions The uneven diversity of Miliusa is shaped by time for speciation, age and dispersal, although their relative influence varies across regions. In Indo‐Burma, long‐term geo‐climatic stability and greater niche availability likely facilitated the persistence of lineages, rapid speciation and dispersal, making it an evolutionary hotspot for Miliusa . In contrast, PI exhibited lower richness and speciation rates despite being old, likely due to the contraction of wet habitats in the Miocene that limited available niches for speciation. Lineages in Wallacea and Sahul show typical island‐like radiations, with speciation rates comparable to the larger and more geo‐climatically stable Indo‐Burma, despite their more recent origin. Overall, our results highlight the role of Miocene‐driven climatic vicariance and Pliocene–Pleistocene climatic fluctuations in shaping the diversification dynamics and regional diversity patterns across tropical Asia.

Accurate species identification within Vicia remains challenging due to high morphological variability, phenotypic plasticity, and frequent hybridization. This study presents the first molecular assessment of nine wild Vicia species from the mountainous area of Iraqi Kurdistan using the ITS region. A dataset of 32 ITS sequences (nine Iraqi accessions and 23 GenBank references) was analyzed for genetic divergence and species discrimination. Intraspecific K2P distances ranged from 0.002 to 0.018, while interspecific distances varied from 0.005 to 0.111, generally exceeding intraspecific divergence. Barcode gap analysis revealed a clear species-level separation in Vicia ervilia L., Vicia faba L., Vicia. narbonensis L., and Vicia sericocarpa Fenzl, whereas only partial overlap was observed in Vicia tenuifolia Roth., Vicia villosa Roth., Vicia palaestina Boiss., Vicia michauxii Spreng., and Vicia sativa L. The species identification tests (TaxonDNA) offered approximately 90% correct assignments, but the All-Species Barcode provided only 56% of correct assignments, reflecting the loss of resolution of closely related taxa. Maximum Likelihood phylogenetic inference confirmed that most Iraqi accessions clustered with conspecific references from neighboring countries. However, the weak support for some lineages raised questions about the resolving power of ITS for assessing recently diverged or hybridizing taxa. Generally, ITS barcodes were suitable for confirming the species identity and phylogenetic reconstruction of Iraqi Vicia but were insufficient to completely resolve the closely related lineages. This study provides the first molecular dataset for Vicia in Iraq and underscores the need for multilocus or genomic-scale approaches to solidify species delimitation and support regional plant genetic resource conservation.

Peach ( Prunus persica ) is an important temperate fruit crop and a model species for genomic research due to its diploid genome, short juvenile period, and relatively small genome size. Despite advances in next-generation sequencing (NGS), most peach genome-wide studies focused on a limited number of elite cultivars, and thus, the diversity of conserved germplasm is underrepresented. In Korea, a large number of peach genetic resources are maintained at the National Institute of Horticultural and Herbal Science (NIHHS), a branch of the Rural Development Administration (RDA), but no genome-scale core collection has been developed to date. This study aimed to perform whole-genome sequencing (WGS) on 445 peach accessions conserved in Korea between 2020 and 2025 using the Illumina NovaSeq 6000 platform, with the primary objective of constructing a representative genome-scale core collection and secondary objectives of identifying genome-wide single-nucleotide polymorphisms (SNPs) and assessing genetic diversity, population structure, and phylogenetic relationships. A total of 944,670 high-confidence SNPs were identified, with chromosomes 2 (G2) and 4 (G4) showing the highest variant density. Analyses using fastSTRUCTURE, principal component analysis (PCA), and phylogenetic reconstruction revealed a complex population structure and substantial genetic variation. From this data, a representative core collection was established, effectively capturing the majority of the genetic diversity present in the Korean peach germplasm. These results offer valuable genomic resources for peach improvement, marker development, pan-genome construction, and comparative genomics within the Rosaceae family.

ABSTRACT Tardigrades, or water bears, are microscopic invertebrates renowned for their extraordinary resilience and cryptic diversity. Their microscopic size, paucity of taxonomically distinctive morphological characters and limited molecular data have caused significant taxonomic deflation. Echiniscoides, the most speciose genus in the family Echiniscoididae, also harbours multiple undescribed species. In this study, we describe a new marine tardigrade species, Echiniscoides testudolapis sp. nov. collected from barnacles inhabiting the intertidal zone of Uljin, Korea. The new species was characterized using an integrative taxonomic approach combining detailed morphological, morphometric and molecular analyses, including DIC microscopy, SEM microscopy and molecular phylogenetic reconstructions based on mitochondrial COI sequences. Morphologically, E. testudolapis sp. nov. is distinguished by its dorsal cuticular sculpturing in the form of circular discs with central punctations (weakly elevated processes), the presence of sensory organs on all legs (papilliform on legs I–II, IV and spike-like on legs III) and a dominant claw formula of 9,9,9,8. Notably, our COI analyses revealed substantial intraspecific genetic variability, with 20 haplotypes identified among 28 individuals (p-distances = 0.1–2.8%). In contrast, the examined nuclear markers exhibited low to no genetic variation (p-distances are 0.05%, 0%, 0.2–0.5% and 0.2–0.4% for 18S, 28S, ITS1 and ITS2, respectively). Single nucleotide polymorphism (SNP) analyses indicated that the observed COI polymorphisms are likely functionally neutral. These findings highlight the value of an integrative taxonomic approach in revealing cryptic diversity and provide new insights into the evolutionary processes and biogeographic patterns shaping marine tardigrade diversity. https://zoobank.org/urn:lsid:zoobank.org:act:90D62D8D-E170-4CC2-AE46-1E8F8E593C9C

Background/Objectives: Leeches constitute a pharmacologically significant animal group in traditional medicine due to their antithrombotic peptides, which include numerous members of the hirustasin gene superfamily. However, a comparative expression profile of this pharmaceutically important family across different leech species is lacking. Methods: This study conducted a comparative transcriptomic analysis of hirustasin gene superfamily expression in the hematophagous leech Hirudinaria manillensis and the non-hematophagous leech Whitmania pigra. Results: The total expression of the hirustasin gene superfamily, quantified as transcripts per million (TPM), showed no significant difference (p = 0.237) between H. manillensis (11,802.60 ± 1596.59) and W. pigra (8623.12 ± 965.96). However, both species exhibited pronounced intergenic expression heterogeneity. Five dominantly expressed genes (TPM > 1000) in H. manillensis and three in W. pigra were identified, collectively comprising 81% and 62% of the total hirustasin gene superfamily expression per species, respectively. Critically, the dominantly expressed genes exhibited no phylogenetic correspondence between species. Integrating expression profiles with phylogenetic reconstruction identified five high-potential candidate genes: poecistasin_Hman2, hirustasin_like_Hman01, hirustasin_like_Hman11, guamerin_Wpig, and bdellastasin_Wpig. Population-level analysis revealed marked population-specific expression patterns in H. manillensis, contrasting with minimal inter-population divergence in W. pigra. Nevertheless, geographically distinct populations of both species showed significant variation in the expression of their respective dominantly expressed genes. Conclusions: These findings provide a set of high-priority candidate genes and insights into their expression characteristics, serving as a starting point for subsequent functional validation and, when integrated with other screening methods, for future antithrombotic drug discovery.

Crown rot is a major postharvest disease affecting bananas (Musa spp.), resulting in substantial economic losses by compromising fruit quality and limiting marketability. In 2023, symptoms of crown rot were observed on banana fruits sourced from ten states across the northern and northeastern regions of Brazil. Pathogen identification was carried out using morphological characteristics and multigene phylogenetic analysis based on the internal transcribed spacer (ITS), translation elongation factor 1-alpha (tef1-α), β-tubulin (tub2), and RNA polymerase II subunit (rpb2). A total of 116 fungal isolates recovered from diseased fruit exhibited morphological traits consistent with the genus Lasiodiplodia. Phylogenetic reconstruction based on concatenated multilocus data revealed that the new isolates belonged to the species Lasiodiplodia brasiliensis, Lasiodiplodia hormozganensis, Lasiodiplodia iraniensis, Lasiodiplodia laeliocattleyae, Lasiodiplodia pseudotheobromae, and Lasiodiplodia theobromae. Pathogenicity tests confirmed that all Lasiodiplodia species/isolates experimentally induced crown rot symptoms on banana fruits of cultivars 'Prata', 'Pacovan', and 'Terra'. Inoculated fungi were re-isolated from symptomatic tissues, which exhibited morphological characteristics identical to those of the original cultures, fulfilling Koch's postulates. This study represents the first large-scale and comprehensive survey on Lasiodiplodia species associated with banana crown rot in Brazil, revealing that a complex of at least six phylogenetically distinct Lasiodiplodia species are involved in the disease etiology.

Ephedra rhytidosperma belongs to the genus Ephedra (Ephedraceae) and plays a significant role in desertification control. In this study, we assembled the complete chloroplast genome of E. rhytidosperma for the first time, revealing a typical quadripartite structure. The chloroplast genome was 109,545 base pair (bp) long with a GC content of 36.8%, encoding 118 complete genes. Phylogenetic reconstruction resolved E. rhytidosperma, E. minuta, E. likiangensis, E. equisetina, Ephedra gerardiana, E. fedtschenkoae, and E. monosperma formed a monophyletic group, indicating their close relationship. The chloroplast genome of E. rhytidosperma establishes a good foundation for further genetic and genomic studies of the Ephedra.

As an old group that has diversified in South America over millions of years, the tinamous (Palaeognathae: Tinamidae) are of high interest for understanding the evolution of birds and the assembly of the Neotropical biota. However, there are currently no complete species-level phylogenies of this group. Most prior work has been based on either morphological data or a small number of molecular markers, each of which has limited capability for reconstructing the tinamou phylogeny. Therefore, the interrelationships of most tinamou species are uncertain. We analyzed 80 whole-genomes from a mix of historical study skins and frozen tissues, including all 46 recognized species of tinamous to (1) reconstruct their interrelationships, (2) estimate the timeframe of tinamou evolution, and (3) examine for the effects of incomplete lineage sorting (ILS) and ancestral introgression on genome evolution. We compared results for coding (BUSCO) and ultraconserved element (UCE) loci, as well as sex-linked and autosomal markers, and used fossil-calibrated tip-dating to estimate divergence times. Tinamous diverged from their sister-group, the extinct Moas, 50-60 mya, and their crown divergence occurred roughly 30-40 mya, followed by constant diversification rates until the present. Phylogenetic reconstructions were largely robust across methods and datasets. Only one clade in the genus Crypturellus displayed substantial species-tree discordance across the different data sets. To investigate the impacts of introgression on this discordance, we quantified introgression for 100kb non-overlapping windows across the genome, and identified pervasive genome-wide introgression. The distribution of this introgression across the genome was dependent on the assumed phylogeny applied to the f-branch model. When assuming one of these topologies in the f-branch model, patterns of introgression matched theoretical predictions about genome architecture. Overall, we present the most complete phylogeny for tinamous to date, identify an unrecognized species, and provide a case study for species-level phylogenomic analysis using whole-genomes.

The emergence of coronavirus variants that evade vaccine protection poses a serious threat to both human health and animal husbandry, including the novel SADS-CoV, an emerging bat-derived coronavirus. Herein, we introduce a "phylogenetic antigen reconstruction" strategy to design a trimeric ancestral spike (tAS) antigen sequence based on evolutionary spike protein analysis. Delivered via nucleoside-modified mRNA-LNP, the tAS mRNA vaccine induced robust and broad-spectrum immunity in mice and sows, conferring complete protection against lethal SADS-CoV challenges in their F1 offspring. Actively immunized piglets also exhibited full resistance to fatal infection. Importantly, the vaccine-elicited antibodies indicated efficient species cross-reactivity blocking SADS-CoV replication in human, monkey-derived cells, and chicken primary cells, and maintained efficacy for 140 days. This study highlights the potential of integrating ancestral antigen design with mRNA-LNP technology to address coronavirus evolution, offering a scalable solution for livestock protection and proactive control of emerging zoonotic coronaviruses.Graphical abstract

Phanerothecium acutum sp. n., the eighth described species of the genus Phanerothecium, is characterized by a unique combination of morphological and molecular traits. Morphologically, it differs from closely related species such as P. spinatus, P. deiropedeum, and P. macrosomum by features of the male copulatory organ (MCO), anchor configuration, and ventral bar structure. Notably, P. acutum sp. n. possesses an MCO armed with delicate spines, robust and truncated anchors with a slight elevation on the shaft, and a ventral bar with expanded extremities. Molecular analyses yielded three new 18S and two partial 28S rDNA sequences. A newly developed primer (DAC18F2) markedly improved PCR success and sequencing quality for 18S amplification. Genetic distance analyses based on 18S support P. acutum sp. n. as a distinct species, with interspecific divergence from most congeners ranging from 1.4-2.0%, and intraspecific variation of 0.0-0.2%. However, its genetic similarity to Phanerothecium sp. KX981456 (0.2-0.9%) highlights the need for additional data to clarify species boundaries. Phylogenetic reconstructions based on 18S and 28S confirmed the monophyly of Phanerothecium, placed within a strongly supported clade alongside viviparous gyrodactylideans, and reaffirmed the validity of the family Oogyrodactylidae. These findings underscore the importance of integrative approaches for resolving taxonomic relationships and suggest that the genus Phanerothecium, particularly among loricariid hosts, harbors greater diversity than currently recognized.

Grapes (Vitis spp.) are a globally significant fruit crop with a long history of cultivation and substantial cultivar diversity. Their high genetic differentiation and complex evolutionary history make them a valuable system for studying plant evolution. The chloroplast genome, known for its structural conservation and uniparental inheritance, offers a reliable molecular marker for phylogenetic reconstruction. In this study, we sequenced and assembled the complete chloroplast genomes of nine representative grape cultivars, analyzed their phylogenetic relationships, and compared structural variations. All chloroplast genomes displayed a typical quadripartite structure, with high conservation in genomic architecture, gene order and content, codon usage, and simple sequence repeats (SSRs). However, additional sequence comparisons revealed seven regions with high variation, including the genes rbcL and ndhF, and the intergenic regions rps16-trnQ, ndhC-trnV, accD-psaI, ndhF-rpl32, and trnL-ccsA. At the same time, seven natural variation sites were identified in the amino acid sequences of rbcL and ndhF. Additionally, the study’s maximum likelihood (ML) phylogenetic trees and photosynthetic index measurements suggest that developmental characteristics of grape photosynthesis may be related to the evolutionary origins of different populations. This phylogenetic classification not only elucidates the evolutionary origins of these germplasm resources but also provides a foundation for molecular-assisted breeding by identifying distinct genetic groups.

Robust termite phylogenies built using transposable element composition and insertion events.

BackgroundsFlies of the tribe Xylotini (Insecta, Diptera, Eristalinae) primarily inhabit dense forests or are frequently observed visiting flowers. Although many genera within this tribe are easily distinguishable and exhibit distinct characteristics, the scarcity of mitochondrial genome resources has limited phylogenetic research. This study addresses this gap by analyzing the mitochondrial genomes of seven newly sequenced species to investigate the tribe’s phylogeny and codon usage patterns.ResultsAll mitochondrial genomes exhibited typical Syrphid structure (37 genes, 15,663 − 16,341 bp) with conserved gene order and strong AT bias (78.56% overall, 93.21% at third codon positions). Codon usage analysis revealed differential evolutionary pressures: Brachypalpoides makiana and Chalcosyrphus curvaria (R² = 0.34–0.56) showed mutation-dominated bias, while other species displayed selection-driven patterns. ENC values (25.6-45.23) and RSCU analysis identified 14 preferred A/U-ending codons, notably UUA-Leu (RSCU = 3.20) and AGA-Arg (RSCU = 2.59). Phylogenetic reconstruction challenged the tribe’s monophyly, with Brachypalpoides and Xylota forming a well-supported clade (BS = 100%), while Milesia appeared polyphyletic. Additionally, C. curvaria showed closer affinity to M. ferruginosa than congenerics.ConclusionsThese findings provide novel insights into: (1) the dual influence of mutational pressure and natural selection on codon evolution, and (2) systematic relationships within this ecologically diverse dipteran group. The study establishes molecular foundations for re-evaluating Xylotini taxonomy and understanding mitochondrial genome evolution in hoverflies.

Introduction. The registration of sporadic cases of glanders in horses in Russia, caused by Burkholderia mallei, highlights the importance of developing genotyping algorithms for this pathogen. The MLVA method (multilocus-variable tandem repeat analysis), based on a comparative analysis of the number of variable tandem repeats (VNTRs), remains a promising genotyping tool. As the number of whole-genome sequences in international databases increases, the informative value of VNTR loci changes, necessitating a revisiting of existing typing schemes. The aim of this study was to assess the feasibility of including the VNTR locus BPSS1974#I in the MLVA-6 scheme for intraspecies differentiation of B. mallei. Materials and methods. The study of 64 strains of B. mallei was conducted in silico and in vitro using MLVA, differentiating region amplification, whole-genome sequencing, and bioinformatic analysis. Results. Genotyping B. mallei using the MLVA-6 scheme failed to determine in silico VNTR profiles of 13 strains from the GenBank NCBI database for one or more loci due to low read coverage of the corresponding genomic regions or their complete absence (null alleles). The effective number of alleles (ne) and the polymorphic information content (PIC) index for the MLVA-6 scheme loci ranged from 3.842–8.103 and 0.740–0.877, respectively. The potential for including the VNTR locus BPSS1974#I in this scheme was determined based on the molecular stability of the motif within it and a high values for ne and PIC, which were 4.299 and 0.767, respectively. VNTR profiles of the collection strains at locus BPSS1974#I were identical to the corresponding strains in the GenBank database. The results of the cluster analysis using a combined MLVA-6 scheme and the BPSS1974#I locus were consistent with the phylogenetic reconstructions obtained using other molecular genetic methods. Conclusion. The VNTR locus BPSS1974#I can be considered a marker, the inclusion of which in the MLVA-6 scheme will improve the accuracy of genotyping and the determination of the regions of origin of newly isolated B. mallei strains.

BackgroundVanessa butterflies, with 22 known species with local and cosmopolitan distributions, are good model organisms to study factors that contribute to the distribution and adaptation in the animal kingdom. Here, the complete mitochondrial genome of Vanessa cardui, a cosmopolitan member of Vanessa butterflies in Nymphalidae, was sequenced, assembled, annotated, and characterized. ResultsThe mitogenome was a midsize circular molecule with 15,299 bps containing regular 37 elements, including 13 protein-coding genes (PCGs), 2 rRNA genes, 22 tRNA genes, and an A + T-rich control region. The genes were organized by 11 intergenic and 13 overlapping spaces. The mitogenome showed a GC content of 19 % and a GC skew of -0.191724138, which was the lowest among the other sequenced Vanessa species. Out of 13 PCGs, only 7 start with ATG, and only 9 end with a TAA stop codon. Analysis of Relative Synonymous Codon Usage (RSCU) demonstrated that out of 66 codons, including stop codons, 25 codons were overrepresented, 7 were unbiased, and 34 were underrepresented. The tRNAs with the size range of 60 bps (tRNASer(AGN)) to 74 bps (tRNATrp) all showed typical cloverleaf secondary structures except for tRNASer(AGN), in which the D-loop was not seen. A Bayesian phylogenomic analysis of V. cardui's mitogenome and 42 other butterfly mitogenomes showed a well-resolved tree with 7 major clades.ConclusionsStudying Vanessa butterflies with different distribution ranges and their mitogenomes may provide insights into genetic factors influencing species distribution and adaptation. The complete mitogenome data provided better resolution in phylogenetic reconstructions for exploring the evolutionary history of the species. Supplementary InformationThe online version contains supplementary material available at 10.1186/s12864-025-12171-y.

Background: Thrushes (family Turdidae) are ecologically important passerine birds widely distributed across the Northern Hemisphere. However, the phylogenetic placement of several East Asian congeners, including Turdus pallidus, remains insufficiently resolved due to the limited resolution of partial mitochondrial or nuclear markers used in previous studies. Methods: In this work, we sequenced and annotated the complete mitochondrial genome of T. pallidus (16,739 bp) using high-throughput Illumina sequencing. The mitogenome exhibited the typical circular architecture and contained 37 genes (13 protein-coding genes, 22 tRNAs, and 2 rRNAs), with an overall GC content of 47.32%. Results: Most protein-coding genes initiated with the standard ATG codon, although lineage-specific deviations such as GTG in COX1 and ND2 were identified, and incomplete stop codons (T– or TA–) were observed, consistent with post-transcriptional polyadenylation. The 22 tRNA genes displayed typical cloverleaf secondary structures, except for trnS(AGN), which lacked a DHU arm, while rRNA genes were 977 bp (12S, 48.52% GC) and 1590 bp (16S, 44.65% GC), showing conserved stem regions but variable loop regions. Codon usage analysis revealed a strong bias toward A/T-ending codons, with a total of 3798 codons and an effective number of codons (ENC) of ~40, indicating moderate codon bias shaped by both mutational pressure and translational selection. Comparative analysis of evolutionary rates demonstrated that conserved genes such as COX1 and CYTB are suitable for resolving deeper relationships, whereas rapidly evolving genes like ATP8 provide resolution among closely related taxa. Conclusions: Phylogenetic reconstructions based on 13 mitochondrial protein-coding genes robustly supported the monophyly of Turdidae and recovered T. pallidus as most closely related to T. obscurus. Overall, this study provides a novel mitogenomic resource for T. pallidus, enhances phylogenetic resolution within Turdus, and underscores the value of complete mitochondrial genomes for molecular identification, conservation management, and avian evolutionary studies.

Entomopathogenic fungi play a crucial role in integrated pest management by targeting pests through specific infection mechanisms, offering both environmental compatibility and sustainability. In recent years, the growing challenge of pesticide resistance and the increasing demand for green agriculture have made the exploration of novel entomopathogenic fungal resources a major research focus in biological control. In this study, we employed a combination of traditional morphological characterization and multi-locus phylogenetic analyses to identify and describe three new species: Gongronella yichunensis , Gongronella shangraoensis , and Yunnania jiujiangensis . For the genus Gongronella , the internal transcribed spacer (ITS) and rRNA large subunit (LSU) regions were used for phylogenetic reconstruction, while the genus Yunnania was analyzed using ITS, LSU, β-tubulin (β-TUB), and translation elongation factor (TEF) gene sequences. Furthermore, preliminary bioactivity assessments revealed varying levels of pathogenicity of the new species against Ostrinia furnacalis . Gongronella yichunensis (strain JX09A02) demonstrated the highest virulence, with a median lethal time (LT 50 ) of 7.2 days and a median lethal concentration (LC 50 ) of 28.0 × 10 6 spores/mL. Gongronella shangraoensis (strain JX20B02) exhibited intermediate efficacy, showing an LT 50 of 8.4 days and an LC 50 of 92.3 × 10 6 spores/mL. Yunnania jiujiangensis (strain JX11B02) displayed relatively lower virulence, with an LT 50 of 9.5 days and an LC 50 of 243.8 × 10 6 spores/mL. These findings not only enrich the genetic resources of entomopathogenic fungi in China but also provide a valuable theoretical and germplasm foundation for developing novel biocontrol agents.