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To accumulate evidence of the phylogeny of Mileewinae and the relationships among Mileewa , Processina, and Ujna genera, we sequenced the complete mitochondrial genomes of four Mileewa spp., namely, Mileewa mira , Mileewa lamellata , Mileewa sharpa, and Mileewa amplimacula . The first complete mitogenome of the genus Processina ( P. sexmaculata ), established by Yang, Deitz & Li from China and comprising five species, was also sequenced in this study. Annotation showed that the five mitogenomes were 14787 –15436 bp in length, and all harbored 37 typical genes. The AT content of the five mitogenomes ranged from 78.3% to 80.2%, which was similar to that of other sequenced Mileewinae species. For protein-coding genes (PCGs), ATN was the start codon, while atp8 and nad5 genes were initiated with TTG, and a great majority of them used TAA or TAG as stop codons, whereas cox2 and nad1 ended with an incomplete codon T–. All tRNAs had a typical cloverleaf secondary structure, except for trnS1, which had a reduced dihydrouridine arm. We further used 59 Membracoidea species and two outgroups to reconstruct phylogenetic trees based on 13 PCGs under an independent partition model with Bayesian inference and Maximum-likelihood methods. Among these two trees, each of the subfamilies Cicadellinae, Typhlocybinae, and Mileewinae were recovered as a monophyletic group with high support values, suggesting that Typhlocybinae was more ancient than Mileewinae and Cicadellinae. Within the Mileewinae subfamily, all species maintained the same relationships and topologies according to both the BI and ML analyses (PP > 0.8, BS > 83) as follows: ( M. sharpa  + ( U. puerana  + (( M. ponta  + ( M. mira  +  M. lamellata )) + (( M. albovittata  + ( M. margheritae  +  M. amplimacula )) + ( M. rufivena  + ( P. sexmaculata  +  M. alara ))), and the monophyly of the genera Processina, Mileewa and Ujna were not supported. This study further enriches the Mileewinae mitogenome database and will contribute to future research on the systematics, evolution, and classification of this group.

The Cicadellinae subfamily of the Cicadellidae (Hemiptera: Auchenorrhyncha: Cicadelloidea) is quite large, with about 2400 species across 330 genera worldwide, of which 263 species in 23 genera occur in China. This work involved the sequencing of the mitochondrial genomes of ten species and nine genera within the Cicadellinae family: Anagonalia emeiensis , Anagonalia melichari, Anatkina vespertinula , Erragonalia choui , Gunungidia aurantiifasciata , Kolla paulula , Nanatka castenea , Paratkina nigrifasciana , Seasogonia rosea , and Stenatkina angustata. The acquired mitogenomes displays a significant AT bias, with the AT contents ranging from 76.1% to 81.7%. The lengths of the mitochondrial genomes range from 14,768 bp to 16,194 bp. TAG and a single T are less frequently used as stop codons, whereas TAA is the most frequently used one. Less PCGs begin with T/GTG, and the majority begin with the conventional ATN (ATA/T/G/C) codon. All tRNA genes in Cicadellinae mitogenomes fold into the standard secondary structure of a cloverleaf,except for trnS1, which lacks a stable dihydrouridine (DHU) stem and instead has s simple loop. Six phylogenetic trees constructed by maximum likelihood (ML) and Bayesian inference (BI) for the three mitochondrial datasets, respectively, consistently showed intergeneric and interspecific relationships within the Cicadellinae. Our results in particular shed light on the molecular and phylogenetic evolution of the pronotum in Cicadellinae.

Background Insect diapause is a critical adaptive strategy for surviving unfavorable environmental conditions. The predatory insect Aphidoletes aphidimyza , widely used in biological pest control, relies on diapause for commercial storage and application. Heat shock proteins (HSPs), particularly small HSPs (sHSPs), are known to play pivotal roles in stress tolerance and protein homeostasis maintenance during diapause. However, the regulatory mechanisms of the HSP gene superfamily, especially the sHSP gene family, in A.aphidimyza diapause remain poorly understood. This study aims to comprehensively analyze the HSP gene superfamily in A.aphidimyza and elucidate the functional and regulatory roles of sHSPs in diapause. Results Our findings reveal that A.aphidimyza has evolved a multi-layered regulatory mechanism through selective expansion of the sHSP gene family. At the transcriptional level, specific transcription factors such as Ftzf1 bind to sHSP gene promoters, enhancing their expression during diapause. Post-transcriptionally, Among the 34 members of the sHSP gene family, 32 have only one exon, the intronless, single-exon structure of sHSP genes facilitates rapid mRNA maturation, enabling swift protein synthesis. Post-translational modifications, such as phosphorylation, regulate the oligomeric state of sHSP proteins, allowing them to dissociate into functional dimers and protect cellular protein homeostasis under stress. Notably, a unique branch of the sHSP gene family (CladeX) exhibited expansion, higher expression during diapause, relatively conserved sequences, suggesting its critical role in environmental adaptation. Conclusions This study provides a comprehensive analysis of the A.aphidimyza HSP gene superfamily, highlighting the pivotal role of the sHSP gene family, particularly CladeX, in diapause regulation. The multi-level regulatory mechanisms—transcriptional, post-transcriptional, and post-translational—enable A.aphidimyza to rapidly respond to adverse conditions like low temperature and short photoperiod, ensuring survival triggering diapause. These insights not only deepen our understanding of insect diapause but also offer potential applications for improving the storage and commercial use of A.aphidimyza as a biological control agent.

The subfamily Mileewinae in China comprises one tribe (Mileewini), four genera (Ujna, Mileewa, Processina, Anzihelus), and 71 species, yet only 11 mitochondrial genomes have been published. This study aimed to elucidate ambiguous diagnostic traits in traditional taxonomy and examined phylogenetic relationships among genera by sequencing mitochondrial genomes from 16 species. The lengths of the mitochondrial genomes ranged from 14,532 to 15,280 bp, exhibiting an AT content of 77.2%–80.7%, indicative of AT bias. Each genome contains 13 protein‐coding genes (PCGs), 22 transfer RNA genes (tRNAs), and 2 ribosomal RNA genes (rRNAs). Notably, the genome of the Xizang endemic Mileewa digitata measures 15,280 bp, with an AT content of 80.7%, 10,941 bp of PCGs, and a 949 bp control region (CR) followed by a 19 bp poly‐A tail. Gene arrangement among the 17 genomes mirrors that of other Mileewinae species. Analyses of nucleotide diversity and evolutionary rates indicate purifying selection on PCGs, with atp8 exhibiting the greatest variability and evolutionary rate, whereas cox1 shows the least. Genetic distance studies display minimal separation among Mileewa species, specifically Mileewa nii with Mileewa margheritae and Mileewa rufivena with Mileewa ussurica, as well as the two morphs of Mileewa digitata. Phylogenetic analyses using Bayesian Inference (BI) and Maximum Likelihood (ML) generated six trees, further questioning the monophyly of the genera Mileewa, Ujna, and Processina. The reconstructed ancestral state based on the forewing patch position suggests that the common ancestor of Chinese Mileewini species was situated exclusively on the costal margin, prompting a proposed division of the tribe's species into two categories (genera). This research enriches the understanding of phylogenetic relationships within Mileewinae and contributes to the mitochondrial genome database for this group, paving the way for future taxonomic studies in China. In this study, we sequenced and comparatively analyzed 17 mitochondrial genomes from 16 species of Mileewinae, including the distinct mitochondrial genome of the unique species Mileewa digitata. Phylogenetic analyses employing Bayesian Inference (BI) and Maximum Likelihood (ML) methods produced six trees that further challenge the monophyly of the genera Mileewa, Ujna, and Processina. The reconstructed ancestral state based on the forewing patch position suggests that the common ancestor of Chinese Mileewini species was situated exclusively on the costal margin, prompting a proposed division of the tribe's species into two categories (genera).

A new leafhopper genus, Conjunctus Yao, Yu & Yang, gen. nov. , together with its type species, Conjunctus decussatus Yao, Yu & Yang, sp. nov. , and an additional new species, Inflatopina pilosa Yao, Yu & Yang, sp. nov. (Hemiptera, Auchenorrhyncha, Cicadellidae, Typhlocybinae, Empoascini), are described from China. An updated identification key to species of the genus Inflatopina Lu, Dietrich & Qin, 2017 is provided.

Tomicus minor (Hartig) and Tomicus yunnanensis Kirkendall and Faccoli are two sympatric species that infest Pinus yunnanensis (Franchet) in southwest China, contributing to growth losses. Accurate sampling plans are needed to make informed control decisions for these species. We investigated three pine forests within experimental sites in Yuxi, Yunnan province, China from 2016 to 2018. The spatial distribution patterns of two pine shoot beetles during the shoot-feeding phase were determined using Taylor’s power law. The optimum sample sizes and stop lines for precision levels of 0.25 and 0.10 were calculated. The model was validated using an additional 15 and 17 independent field datasets ranging in density from 0.06 to 1.90 beetles per tree. T. minor and T. yunnanensis adults showed aggregated spatial distributions. For T. minor, sample sizes of 41 and 259 trees were adequate for a D of 0.25 and 0.10, respectively, while for T. yunnanensis, a mean density of one individual per tree required sample sizes of 33 plants (D = 0.25) and 208 plants (D = 0.10). The software simulations of this sampling plan showed precision levels close to the desired levels. At a fixed-precision level of 0.25, sampling is easily achievable. This sampling program is useful for the integrated pest management (IPM) of two sympatric Tomicus species.

The mitogenome data of leafhopper species Mileewa alara was assembled and annotated in this study. The results shows that length of M. alara is 16020 bp, consist of 13 protein-coding genes (PCGs), 22 tRNA genes, 2 rRNA genes, and one control region. The A + T content in the mitogenome was 77.9%. Phylogenetic analysis based on 13 PCGs of four Mileewa species and other 29 Cicadellidae species, each subfamily species well separated. And M. alara clustered with M. ponta. This study also raised mitogenome of Mileewa number in GenBank to four.

The subfamily Typhlocybinae is a ubiquitous, highly diverse group of mostly tiny, delicate leafhoppers. The tribal classification has long been controversial and phylogenetic methods have only recently begun to test the phylogenetic status and relationships of tribes. To shed light on the evolution of Typhlocybinae, we performed phylogenetic analyses based on 28 newly sequenced and 19 previously sequenced mitochondrial genomes representing all currently recognized tribes. The results support the monophyly of the subfamily and its sister‐group relationship to Mileewinae. The tribe Zyginellini is polyphyletic with some included genera derived independently within Typhlocybini. Ancestral character state reconstruction suggests that some morphological characters traditionally considered important for diagnosing tribes (presence/absence of ocelli, development of hind wing submarginal vein) are homoplastic. Divergence time estimates indicate that the subfamily arose during the Middle Cretaceous and that the extant tribes arose during the Late Cretaceous. Phylogenetic results support establishment of a new genus, Subtilissimia Yan & Yang gen. nov., with two new species, Subtilissimia fulva Yan & Yang sp. nov. and Subtilissimia pellicula Yan & Yang sp. nov.; but indicate that two previously recognized species of Farynala distinguished only by the direction of curvature of the processes of the aedeagus are synonyms, that is, Farynala dextra Yan & Yang, 2017 equals Farynala sinistra Yan & Yang, 2017 syn. nov. A key to tribes of Typhlocybinae is provided. We studied the evolutional relationships of Typhlocybinae based on 47 typhlocybine mitogenomic dataset representing all currently recognized tribes. The Ancestral character state reconstruction suggests that some morphological characters (fore‐ and hind wing patterns) traditionally considered important for diagnosing typhlocybine tribes are homoplastic. The divergence time estimates indicate that the subfamily Typhlocybinae arose during the Middle Cretaceous and that the extant tribes arose during the Late Cretaceous.

The mitogenome of one Mileewinae species Mileewa ponta was sequenced and annotated according to close Cicadellidae species in this study. Annotation shows the mitogenome of M. ponta is 15999 bp in length, containing 13 protein-coding genes (PCGs), 22 tRNA genes, two rRNA genes, and a control region. The A + T content in the mitogenome was 79.9%. Besides, we further constructed a Bayesian Inference phylogenetic tree among M. ponta, M. albovittata, M. margheritae and other Cicadellidae species. Each subfamily species clustered together, and M. ponta clustered with M. albovittata, M. margheritae. This mitogenome will contribute to the further systematics and evolution study of subfamily Mileewinae.