Complete genome analysis of a novel chuvirus from a southern green stink bug (Nezara viridula).

A novel chuvirus from a southern green stink bug (Nezara viridula) was identified by RNA sequencing in this study and was tentatively named "Ningbo southern green stink bug chuvirus 1" (NBSGSBV-1). The complete genome sequence of NBSGSBV-1 consists of 11,375 nucleotides, and the genome was found to be circular by 'around-the-genome' reverse transcription polymerase chain reaction (RT-PCR) and Sanger sequencing. Three open reading frames (ORFs) were predicted in the NBSGSBV-1 genome, encoding a large polymerase protein (L protein), a glycoprotein (G protein), and a nucleocapsid protein (N protein). A phylogenetic tree was constructed based on all of the currently available RNA-dependent RNA polymerase amino acid sequences of viruses of the family Chuviridae, and NBSGSBV-1 was found to cluster together with Sanya chuvirus 2 and Hubei odonate virus 11, indicating that NBSGSBV-1 might belong to the genus Odonatavirus. Five conserved sites were identified in the L proteins of NBSGSBV-1 and other chuviruses. The abundance and characteristics of the NBSGSBV-1-derived small interfering RNAs suggested that NBSGSBV-1 actively replicates in the host insect. To the best of our knowledge, this is the first report of a chuvirus identified in a member of the insect family Pentatomidae. The discovery and characterization of NBSGSBV-1 will help us to understand the diversity of chuviruses in insects.

The evolution and functional divergence of 10 Apolipoprotein D-like genes in Nilaparvata lugens.

Apolipoprotein D (ApoD), a member of the lipocalin superfamily of proteins, is involved in lipid transport and stress resistance. Whereas only a single copy of the ApoD gene is found in humans and some other vertebrates, there are typically several ApoD-like genes in insects. To date, there have been relatively few studies that have examined the evolution and functional differentiation of ApoD-like genes in insects, particularly hemi-metabolous insects. In this study, we identified 10 ApoD-like genes (NlApoD1-10) with distinct spatiotemporal expression patterns in Nilaparvata lugens (BPH), which is an important pest of rice. NlApoD1-10 were found to be distributed on 3 chromosomes in a tandem array of NlApoD1/2, NlApoD3-5, and NlApoD7/8, and show sequence and gene structural divergence in the coding regions, indicating that multiple gene duplication events occurred during evolution. Phylogenetic analysis revealed that NlApoD1-10 can be clustered into 5 clades, with NlApoD3-5 and NlApoD7/8 potentially evolving exclusively in the Delphacidae family. Functional screening using an RNA interference approach revealed that only NlApoD2 was essential for BPH development and survival, whereas NlApoD4/5 are highly expressed in testes, and might play roles in reproduction. Moreover, stress response analysis revealed that NlApoD3-5/9, NlApoD3-5, and NlApoD9 were up-regulated after treatment with lipopolysaccharide, H2 O2 , and ultraviolet-C, respectively, indicating their potential roles in stress resistance.

Identification and Characterization of Three Novel Solemo-like Viruses in the White-Backed Planthopper, Sogatella furcifera.

Agricultural insects play a crucial role in transmitting plant viruses and host a considerable number of insect-specific viruses (ISVs). Among these insects, the white-backed planthoppers (WBPH; Sogatella furcifera, Hemiptera: Delphacidae) are noteworthy rice pests and are responsible for disseminating the southern rice black-streaked dwarf virus (SRBSDV), a significant rice virus. In this study, we analyzed WBPH transcriptome data from public sources and identified three novel viruses. These newly discovered viruses belong to the plant-associated viral family Solemoviridae and were tentatively named Sogatella furcifera solemo-like virus 1-3 (SFSolV1-3). Among them, SFSolV1 exhibited a prevalent existence in different laboratory populations, and its complete genome sequence was obtained using rapid amplification of cDNA ends (RACE) approaches. To investigate the antiviral RNA interference (RNAi) response in WBPH, we conducted an analysis of virus-derived small interfering RNAs (vsiRNAs). The vsiRNAs of SFSolV1 and -2 exhibited typical patterns associated with the host's siRNA-mediated antiviral immunity, with a preference for 21- and 22-nt vsiRNAs derived equally from both the sense and antisense genomic strands. Furthermore, we examined SFSolV1 infection and distribution in WBPH, revealing a significantly higher viral load of SFSolV1 in nymphs' hemolymph compared to other tissues. Additionally, in adult insects, SFSolV1 exhibited higher abundance in male adults than in female adults.

RNAi-mediated silencing of an egg-specific gene Nllet1 results in hatch failure in the brown planthopper.

BACKGROUND: The brown planthopper (BPH) is one of the most destructive agricultural pests in Asia. RNA interference (RNAi)-mediated pest management has been under development for years, and the selection of appropriate target genes is important for pest-targeted RNAi. C-type lectins (CTLs) are a class of genes that perform a variety of functions, such as the regulation of growth and development. RESULTS: A CTL-S protein named Nllet1, containing a single calcium ion (Ca2+ )-dependent carbohydrate-binding domain (CRD) with a conserved triplet motif QPD was identified and functionally characterized in BPH. Expression profiles at both the transcriptional and translational levels show that Nllet1 accumulates during the serosal cuticle (SC) formation period. Immunofluorescence and immunogold labeling further demonstrated that Nllet1 is located in the serosal endocuticle (en-SC). Maternal RNAi-mediated silencing of Nllet1 disrupted the SC structure, accompanied by a loss of the outward barrier and 100% embryo mortality. Injection of 10 ng dsNllet1 or dsNllet1' per female adult BPH resulted in a total failure of egg hatching. CONCLUSION: Nllet1 is essential for SC formation and embryonic development in BPH, which helps us understand the important roles of CTL-Ss. Additionally, BPH eggs show high sensitivity to the depletion of Nllet1. This study indicates that Nllet1 is a promising candidate gene that can be used to develop RNAi-based control strategies at the BPH egg stage, and it can also be used as a target for developing novel ovicides. © 2022 Society of Chemical Industry.

Exploration of the mechanism of Zisheng Shenqi decoction against gout arthritis using network pharmacology.

BACKGROUND: In this study, the network pharmacological methods were used to predict the target of effective components of compounds in Zisheng Shenqi Decoction (ZSD, or Nourishing Kidney Qi Decoction) in the treatment of gouty arthritis (GA). METHOD: The main effective components and corresponding key targets of herbs in the ZSD were discerned through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis (TCMSP), Bioinformatics Analysis Tool for Molecular mechanism of Traditional Chinese Medicine (BATMAN-TCM) database. UniProt database and Swiss Target Prediction (STP) database was used to rectify and unify the target names and supply the target information. The targets related to GA were obtained by using GeneCards database. After we discovered the potential common targets between ZSD and GA, the interaction network diagram of "ZSD-component-GA-target" was constructed by Cytoscape software (Version 3.7.1). Subsequently, the Protein-protein interaction (PPI) network of ZSD effective components-targets and GA-related targets was constructed by Search Tool for the Retrieval of Interacting Genes Database (STRING). Bioconductor package "org.Hs.eg.db" and "cluster profiler" package were installed in R software (Version 3.6.0) which used for Gene Ontology analysis and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analysis. RESULTS: 146 components and 613 targets of 11 herbal medicines in the ZSD were got from TCMSP database and BATMAN-TCM database. 987 targets of GA were obtained from GeneCards database. After intersected and removed duplications, 132 common targets between ZSD and GA were screened out by Cytoscape software (Version 3.7.1). These common targets derived from 81 effective components of 146 components, such as quercetin, stigmasterol and kaempferol. They were closely related to anti-inflammatory, analgesic and anti oxidative stress and the principal targets comprised of Purinergic receptor P2X, ligand-gated ion channel 7 (P2x7R), Nod-like receptor protein 3 (NLRP3) and IL-1ß. GO enrichment analysis and KEGG pathway enrichment analysis by R software (Version 3.6.0) showed that the key target genes had close relationship with oxidative stress, reactive oxygen species (ROS) metabolic process and leukocyte migration in aspects of biological process, cell components and molecular function. It also indicated that ZSD could decrease inflammatory reaction, alleviate ROS accumulation and attenuate pain by regulating P2 × 7R and NOD like receptor signaling pathway of inflammatory reaction. CONCLUSION: A total of 81 effective components and 132 common target genes between ZSD and GA were screened by network pharmacology. The PPI network, GO enrichment analysis and KEGG pathway enrichment analysis suggested that ZSD can exerte anti-inflammatory and analgesic effects on the treatment of GA by reducing decreasing inflammatory reaction, alleviating ROS accumulation, and attenuating pain. The possible molecular mechanism of it mainly involved multiple components, multiple targets and multiple signaling pathways, which provided a comprehensive understanding for further study. In general, the network pharmacological method applied in this study provides an alternative strategy for the mechanism of ZSD in the treatment of GA.

Long non-coding RNA UCA1 contributes to the progression of prostate cancer and regulates proliferation through KLF4-KRT6/13 signaling pathway.

Long non-coding RNAs (lncRNAs) UCA1 have been shown to paly an important regulatory roles in cancer biology, and UCA1 dysfunction is related to TNM stage, metastasis and postoperative survival in several cancers. However, the biological role and clinical significance of UCA1 in the carcinogenesis of prostate cancer (PC) remain largely unclear. Herein, we found that UCA1 was abnormally upregulated in tumor tissues from PC patients, and patients with high UCA1 levels had a significantly poorer prognosis. Intriguingly, the mRNA and protein levels of KLF4 were significantly increased in tumor tissues, which was highly correlated to UCA1 levels. Moreover, UCA1 depletion inhibited the growth and induced apoptosis in PC3 and LNCaP cell lines. In addition, UCA1 loss-of-function could decrease KLF4 expression, subsequently, the downregulation of KRT6 and KRT13. Taken together, our study indicated that UCA1 had a crucial role in the tumorigenesis of PC. Moreover, UCA1 loss-of-function inhibited cell proliferation and induced cell apoptosis, at least partially, through inactivation KLF4-KRT6/13 cascade.