Molecular characterization and functional analysis of a beta-1,3-glucan recognition protein from oriental fruit moth Grapholita molesta (Lepidoptera: Tortricidae).

The beta-1,3-glucan recognition protein (BGRP) is an important pattern recognition protein (PRP), which plays an important role in immune recognition and signaling pathway of insect innate immunity. Herein, a BGRP gene was obtained from the transcriptome of Grapholita molesta and its expression was verified by PCR. The full cDNA of the GmBGRP gene was 1691 bp encoding 486 amino acid residues. The calculated molecular mass of the mature protein was 54.83 kDa with an estimated pI of 6.14. The amino acid sequence of GmBGRP was highly homologous to BGRPs of other lepidopterans including Leguminivora glycinivorella BGRP-3. Expression profile of GmBGRP at different developmental stages and different tissues of 5th instar larvae showed that the expression level of this gene tends to slightly increase and then decrease at the adult stage, with the highest at the pupa stage; and mainly expressed in the epidermis, fat body and hemocytes compared with other tissues. In addition, we investigated the transcription levels of other immune-related genes, such as Serine-1, Serine-2, Serine-3, Serpin, SRCB (scavenger receptor gene), Toll, PPO (prophenoloxidase) upon GmBGRP gene silencing, indicating that GmBGRP expression is associated with immune responses of G. molesta. This was further supported by the upregulation of the mRNA level of GmBGRP following fungal infection. Taken together, these results provide experimental evidence for the role of GmBGRP gene in immune defense in G. molesta larvae.

Molecular characterization and functional analysis of the ecdysone receptor isoform (EcR) from the oriental fruit moth Grapholita molesta (Lepidoptera: Tortricidae).

20-Hydroxyecdysone (20E) plays a vital role in a series of biological processes, via the nuclear receptors, EcR/USP by activating the ecdysone regulatory cascade. To clarify the role of EcR during the development of Grapholita molesta, the complementary DNA of ecdysone receptor isoform B1 (GmEcR-B1) was obtained from the transcriptome of G. molesta and verified by PCR. Alignment analysis revealed that the deduced protein sequence of GmEcR-B1 was highly homologous to EcR proteins identified in other lepidopteran species, especially the EcR-B1 isoform in Spodoptera litura. Quantitative real-time PCR showed that GmEcRs was expressed at all test developmental stages, and the expression level of GmEcRs was relatively higher during the period of the 3rd day of fifth instar larvae to 2nd of pupa than those in other stages. Moreover, the messenger RNA of GmEcRs was much more strongly expressed in the Malpighian tubule and epidermis than those in other tissues, which suggests that this gene may function in a tissue-specific manner during larval development. Silencing of GmEcRs could significantly downregulate the transcriptional level of ecdysone-inducible genes and result in increased mortality during metamorphosis and prolonged prepupal duration. Taken together, the present results indicate that GmEcRs may directly or indirectly affect the development of G. molesta.

Two Complete Mitochondrial Genomes From Leuctridae (Plecoptera: Nemouroidea): Implications for the Phylogenetic Relationships Among Stoneflies.

The family-level relationships within Plecoptera have been a focused area of research for a long time. Its higher classification remains unstable, and the phylogenetic relationships within Plecoptera should be re-examined. Here, we sequenced and analyzed two complete mitochondrial genomes (mitogenomes) of Paraleuctra cercia and Perlomyia isobeae of the family Leuctridae. We reconstructed the phylogeny of Plecoptera based on 13 protein-coding genes (PCGs) from published stoneflies. Our results showed that the Bayesian inference and maximum-likelihood tree had similar topological structures except for the positions of two families, Peltoperlidae and Scopuridae. The Plecoptera is divided into two clades, the suborder Antarctoperlaria and the suborder Arctoperlaria. The two suborders subsequently formed two groups, Eusthenioidea and Gripopterygoidea, and Euholognatha and Systellognatha, which is consistent with the results of morphological studies. In addition, the Leuctridae is the earliest branch within the superfamily Nemouroidea. But the monophyly of Perloidea and Pteronarcyoidea are still not well supported.

The phylogeny and evolutionary timescale of stoneflies (Insecta: Plecoptera) inferred from mitochondrial genomes.

Phylogenetic analysis based on mitochondrial genomic data from 25 stonefly species recovered a well-supported tree resolving higher-level relationships within Plecoptera (stoneflies). The monophyly of both currently recognized suborders was strongly supported, concordant with previous molecular analyses of Plecoptera. The southern hemisphere suborder Antarctoperlaria formed two clades: Eustheniidae + Diamphipnoidae and Austroperlidae + Gripopterygidae; consistent with relationships proposed based on morphology. The largely northern hemisphere suborder Arctoperlaria also divided into two groups, Euholognatha and Systellognatha, each composed of the five families traditionally assigned to each infraorder (the placement Scopuridae by mt genome data remains untested at this time). Within Euholognatha, strong support for the clade Nemouridae + Notonemouridae confirmed the northern origin of the currently southern hemisphere restricted Notonemouridae. Other family level relationships within the Arctoperlaria differ from those recovered by previous morphology and molecular based analyses. A fossil-calibrated divergence estimation suggests the formation of two suborders dates back to the Jurassic (181 Ma), with subsequent diversification of most stonefly families during the Cretaceous. This result confirms the hypothesis that initial divergence between the suborders was driven by the breakup of the supercontinent Pangaea into Laurasia and Gondwanaland (commencing 200 Ma and complete by 150 Ma).

Ship Classification in High-Resolution SAR Images via Transfer Learning with Small Training Dataset.

Synthetic aperture radar (SAR) as an all-weather method of the remote sensing, now it has been an important tool in oceanographic observations, object tracking, etc. Due to advances in neural networks (NN), researchers started to study SAR ship classification problems with deep learning (DL) in recent years. However, the limited labeled SAR ship data become a bottleneck to train a neural network. In this paper, convolutional neural networks (CNNs) are applied to ship classification by using SAR images with the small datasets. To solve the problem of over-fitting which often appeared in training small dataset, we proposed a new method of data augmentation and combined it with transfer learning. Based on experiments and tests, the performance is evaluated. The results show that the types of the ships can be classified in high accuracies and reveal the effectiveness of our proposed method.

Complete Mitochondrial Genome of Suwallia teleckojensis (Plecoptera: Chloroperlidae) and Implications for the Higher Phylogeny of Stoneflies.

Stoneflies comprise an ancient group of insects, but the phylogenetic position of Plecoptera and phylogenetic relations within Plecoptera have long been controversial, and more molecular data is required to reconstruct precise phylogeny. Herein, we present the complete mitogenome of a stonefly, Suwallia teleckojensis, which is 16146 bp in length and consists of 13 protein-coding genes (PCGs), 2 ribosomal RNAs (rRNAs), 22 transfer RNAs (tRNAs) and a control region (CR). Most PCGs initiate with the standard start codon ATN. However, ND5 and ND1 started with GTG and TTG. Typical termination codons TAA and TAG were found in eleven PCGs, and the remaining two PCGs (COII and ND5) have incomplete termination codons. All transfer RNA genes (tRNAs) have the classic cloverleaf secondary structures, with the exception of tRNASer(AGN), which lacks the dihydrouridine (DHU) arm. Secondary structures of the two ribosomal RNAs were shown referring to previous models. A large tandem repeat region, two potential stem-loop (SL) structures, Poly N structure (2 poly-A, 1 poly-T and 1 poly-C), and four conserved sequence blocks (CSBs) were detected in the control region. Finally, both maximum likelihood (ML) and Bayesian inference (BI) analyses suggested that the Capniidae was monophyletic, and the other five stonefly families form a monophyletic group. In this study, S. teleckojensis was closely related to Sweltsa longistyla, and Chloroperlidae and Perlidae were herein supported to be a sister group.

Nanoscale three-dimensional single particle tracking by light-sheet-based double-helix point spread function microscopy.

The double-helix point spread function (DH-PSF) microscopy has become an essential tool for nanoscale three-dimensional (3D) localization and tracking of single molecules in living cells. However, its localization precision is limited by fluorescent contrast in thick samples because the signal-to-noise ratio of the system is low due to the inherent low transfer function efficiency and background fluorescence. Here we combine DH-PSF microscopy with light-sheet illumination to eliminate out-of-focus background fluorescence for high-precision 3D single particle tracking. To demonstrate the capability of the method, we obtain the single fluorescent bead image with light-sheet illumination, with three-dimensional localization accuracy better than that of epi-illumination. We also show that the single fluorescent beads in agarose solution can be tracked, which demonstrates the possibility of our method for the study of dynamic processes in complex biological specimens.

Review of the genus Sweltsa (Plecoptera: Chloroperlidae) in China.

The species of the stonefly genus Sweltsa known from China are reviewed. A new species, Sweltsa baiyunshana, sp. n. is described, and its relationship to similar species is discussed. Redescriptions of Sweltsa longistyla Wu and diagnostic notes on the other Sweltsa species are also presented.

Species of Neoperla (Plecoptera: Perlidae) from Yunnan Province, China.

Two new species of the genus Neoperla are described from Yunnan Province, China: Neoperla lihuae Li & Murányi, sp. nov. and Neoperla yanlii Li & Wang, sp. nov. They are compared with related congeners. Brief comments on the taxonomy of N. cavaleriei (Navás, 1922), N. limbatella Navás, 1933, N. obscurofulva Wu, 1962 and N. lui Du, 2004 are also presented.

A new species of Sphaeronemoura (Plecoptera: Nemouridae) from Henan Province of China, with additions to generic characters of the female and larva.

A new species of the genus Sphaeronemoura (Plecoptera: Nemouridae: Amphinemurinae), S. separata sp. n. is described from the male, female, and larvae collected in Henan Province of China. The internal genitalia of the female adult is described for the first time for the genus, and additional generic characters for the larva are also given. The known distributions of all Sphaeronemoura species is presented.

The mitochondrial genome of Neoperla bimaculata (Li et al. 2021) (plecoptera: perlidae) from Tibet of southwest China and its phylogenetic analysis.

The complete mitochondrial genome (mitogenome) of Neoperla bimaculata was sequenced and annotated in this study. We found that the mitogenome of N. bimaculata is 15,774 bp in length with an A + T content of 64.3%. It exhibits the classic structure of a mitogenome. Most protein-coding genes (PCGs) of the mitogenome initiate with the standard start codon ATN. Ten PCGs use the standard stop codon TAA/TAG, while the COI, COII, and ND5 genes terminate with a single T nucleotide. Phylogenetic analyses suggested that N. bimaculata, along with two unpublished Neoperla species, formed a cluster within the phylogenetic tree. Our results indicated that the genus Neoperla and Neoperlops were sister groups. Meanwhile, the monophyly of Perlinae and Acroneuriinae was supported in the mitochondrial phylogeny.

Additional records of Neoperla (Plecoptera: Perlidae) from Yunnan Province of China, with description of a new species.

A new species of the genus Neoperla, N. donglaii sp. nov., is described from Yunnan Province of southwestern China. The new species is assigned to the N. lushana-complex of the N. (Formosita) subgenus due to the presence of paired, ventral spiny lobes of the aedeagus. The new species was compared with related species in the subgenus. Neoperla hamata Jewett, 1975 and N. idella Stark & Sivec, 2008 are newly recorded for China and N. quadrata Wu & Claassen, 1934 specifically from Yunnan Province, China. Neoperla quadrata had not been collected for 90 years.

Identity on Filchneria heteroptera (Wu, 1938) (Plecoptera: Perlodidae) with a new record of Paraleuctra cercia (Okamoto, 1922) from China.

Filchneria heteroptera (Wu, 1938) is rediscovered from Heilongjiang Province of northeastern China. A supplementary description and illustrations including aedeagal structures based on newly collected specimens are provided. Filchneria dongruihangi Chen, 2020 is considered here as a junior synonym of F. heteroptera (Wu, 1938). Paraleuctra cercia (Okamoto, 1922) is reported from Liaoning and Heilongjiang provinces, China for the first time.

A new species and three records of Chloroperlidae (Plecoptera) from China.

A new species of the genus Suwallia Ricker, 1943, S. rostrata sp. nov., from Xizang of southwestern China is described, illustrated and compared with similar species. Alloperla kurentzovi Zhiltzova & Zapekina-Dulkeit, 1977, A. kurilensis Zhiltzova, 1978, A. teleckojensis mal, 1939 are reported for the first time from China. The diagnostic features of these species are illustrated with color images.

Mitochondrial genomes provide insights into the Euholognatha (Insecta: Plecoptera).

BACKGROUND: Euholognatha is a monophyletic group within stoneflies comprised by a superfamily Nemouroidea and a family Scopuridae. Based on morphological data, the family-level phylogenetic relationships within Euholognatha are widely accepted, but there is still controversy among different molecular studies. To better understand the phylogeny of all six extant euholognathan families, we sequenced and analyzed seven euholognathan mitogenomes. RESULTS: The sequence heterogeneity analysis observed a low degree of compositional heterogeneity in euholognathan mitogenomes. Meanwhile, leuctrid mitogenomes were more heterogeneous than other euholognathan families, which may affect the phylogenetic reconstruction. Phylogenetic analyses with various datasets generated three topologies. The Leuctridae was recovered as the earliest branching lineage, and the sister relationship of Capniidae and Taeniopterygidae was supported by most tree topologies and FcLM analyses. When separately excluding sparsely sampled Scopuridae or high heterogeneity leuctrid taxa, phylogenetic analyses under the same methods generated more stable and consistent tree topologies. Finally, based on the results of this study, we reconstructed the relationships within Euholognatha as: Leuctridae + (Scopuridae + ((Taeniopterygidae + Capniidae) + (Nemouridae + Notonemouridae))). CONCLUSION: Our research shows the potential of data optimizing strategies in reconstructing phylogeny within Euholognatha and provides new insight into the phylogeny of this group.

Two new species of stoneflies (Plecoptera: Nemouridae) from Northeastern India with a checklist of the family in the Indian Subcontinent.

A new species of the genus Amphinemura (Plecoptera: Nemouridae: Amphinemurinae), A. baumanni sp. n. is described on the basis of five males collected in the Darjeeling District of West Bengal, India. A new species of the genus Sphaeronemoura (Amphinemurinae), S. siveci sp. n. is described on the basis of a single male collected in the West Garo Hills District of Meghalaya, India. A checklist of the family Nemouridae from the Indian Subcontinent is given with zoogeographical comments.

Haploperla choui sp. n. (Plecoptera: Chloroperlidae), a remarkable new stonefly from Qinling Mountains of China.

A new chloroperline stonefly species, Haploperla choui from the Qinling Mountains, Shaanxi Province, China is described. It is compared to other species of the genus known from Asia.

Species of the family Nemouridae (Plecoptera) from Taihang Mountains of northern China, with description of Nemoura taihangshana sp. n.

A distinctive new species of the genus Nemoura is described from the Taihang Mountains of northern China, N. taihangshana. The new species is compared with similar species. Amphinemura sinensis (Wu, 1926) is redescribed and the first record of N. geei Wu, 1929 from Henan Province is also given.

Discovery of the genus Paraleuctra (Plecoptera: Leuctridae) from Palearctic China, with description of P. qilianshana sp. nov.

A new species of the genus Paraleuctra, P. qilianshana sp. nov., is described from Mt. Qilianshan, Gansu Province, China. The new species is the first representative of Paraleuctra discovered in the Palearctic Region of China. The new species is compared with its regional congeners.

Review of Neoperla (Plecoptera: Perlidae) from Zhejiang Province, China.

The species of the perlid genus Neoperla are reviewed from eastern coastal Zhejiang Province, China. Neoperla xuanson- gae sp. nov. is described and compared with related congeners. Neoperla yangae Du is considered a junior synonym of N. anjiensis Yang & Yang and the first record of N. flagellata Li & Murányi is given for the Province. Several additional species are redescribed or diagnosed. A provisional key to the known species for Zhejiang Province is also presented.