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20 pages, 5823 KiB

Open AccessArticle

Gene Co-Expression Network Analysis Reveals Key Regulatory Genes in Metisa plana Hormone Pathways

by Vinothienii Vengatharajuloo, Hoe-Han Goh, Maizom Hassan, Nisha Govender, Suhaila Sulaiman, Nor Afiqah-Aleng, Sarahani Harun and Zeti-Azura Mohamed-Hussein

Insects 2023, 14(6), 503; https://doi.org/10.3390/insects14060503 - 30 May 2023

Cited by 1 | Viewed by 1584

Abstract

Metisa plana Walker (Lepidoptera: Psychidae) is a major oil palm pest species distributed across Southeast Asia. M. plana outbreaks are regarded as serious ongoing threats to the oil palm industry due to their ability to significantly reduce fruit yield and subsequent productivity. Currently, [...] Read more.

Metisa plana Walker (Lepidoptera: Psychidae) is a major oil palm pest species distributed across Southeast Asia. M. plana outbreaks are regarded as serious ongoing threats to the oil palm industry due to their ability to significantly reduce fruit yield and subsequent productivity. Currently, conventional pesticide overuses may harm non-target organisms and severely pollute the environment. This study aims to identify key regulatory genes involved in hormone pathways during the third instar larvae stage of M. plana gene co-expression network analysis. A weighted gene co-expression network analysis (WGCNA) was conducted on the M. plana transcriptomes to construct a gene co-expression network. The transcriptome datasets were obtained from different development stages of M. plana, i.e., egg, third instar larvae, pupa, and adult. The network was clustered using the DPClusO algorithm and validated using Fisher’s exact test and receiver operating characteristic (ROC) analysis. The clustering analysis was performed on the network and 20 potential regulatory genes (such as MTA1-like, Nub, Grn, and Usp) were identified from ten top-most significant clusters. Pathway enrichment analysis was performed to identify hormone signalling pathways and these pathways were identified, i.e., hormone-mediated signalling, steroid hormone-mediated signalling, and intracellular steroid hormone receptor signalling as well as six regulatory genes Hnf4, Hr4, MED14, Usp, Tai, and Trr. These key regulatory genes have a potential as important targets in future upstream applications and validation studies in the development of biorational pesticides against M. plana and the RNA interference (RNAi) gene silencing method. Full article

(This article belongs to the Special Issue Insect Bioinformatics)

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17 pages, 4660 KiB

Open AccessFeature PaperArticle

Identification and Validation of Reference Genes for Expression Analysis Using RT-qPCR in Leptocybe invasa Fisher and La Salle (Hymenoptera: Eulophidae)

by Ya Liu, Jing Zhou, Zhisong Qiu, Ping Hu, Xiao Chen and Zhende Yang

Insects 2023, 14(5), 456; https://doi.org/10.3390/insects14050456 - 12 May 2023

Cited by 3 | Viewed by 1705

Abstract

Leptocybe invasa (Hymenoptera: Eulophidae) is a globally intrusive pest. Despite extensive research into the physiological responses of this pest, our understanding of the molecular mechanisms still needs to be improved. We want to accurately investigate the expression of L. invasa’s target genes, [...] Read more.

Leptocybe invasa (Hymenoptera: Eulophidae) is a globally intrusive pest. Despite extensive research into the physiological responses of this pest, our understanding of the molecular mechanisms still needs to be improved. We want to accurately investigate the expression of L. invasa’s target genes, so it is imperative to select fitting reference genes. In this study, eight housekeeping genes’ stability (RPS30, ACTR, 18S rRNA, ACT, RPL18, GAPDH, 28S rRNA, and TUB) was tested under five different experimental conditions, including male or female adults, somites (head, thorax, and abdomen), temperatures (0 °C, 25 °C, and 40 °C), diets (starvation, clear water, 10% honey water, Eucalyptus sap), and pesticides (acetone was used as a control, imidacloprid, monosultap). Gene stability was calculated using RefFinder, which integrates four algorithms (the ∆Ct method, geNorm, NormFinder, and BestKeeper). The findings implied that ACT and ACTR were the most accurate when comparing sexes. For analyzing different somites, 28S rRNA and RPL18 were ideal; the 28S rRNA and RRS30 were perfect for analyzing at different temperatures. The combination of ACT and GAPDH helped to analyze gene expression in different diets, and GAPDH and 28S rRNA were suitable for various pesticide conditions. Overall, this research offers a complete list of reference genes from L. invasa for precise analysis of target gene expression, which can improve the trustworthiness of RT-qPCR and lay the foundation for further investigations into the gene function of this pest. Full article

(This article belongs to the Special Issue Insect Bioinformatics)

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15 pages, 1854 KiB

Open AccessArticle

Transcriptome Analysis Revealed Genes Related to γ-Irradiation Induced Emergence Failure in Third-Instar Larvae of Bactrocera dorsalis

by Chao Sun, Samina Shabbir, Wenxiang Wang, Yan Gao, Cuicui Ge and Qingsheng Lin

Insects 2022, 13(11), 1017; https://doi.org/10.3390/insects13111017 - 03 Nov 2022

Cited by 1 | Viewed by 1291

Abstract

The oriental fruit fly is a polyphagous and highly invasive economically important pest in the world. We proposed the hypothesis that radiation treatment influence RNA expression in the larvae and leads to emergence failure. Therefore, transcriptome analyses of third-instar larvae of B. dorsalis [...] Read more.

The oriental fruit fly is a polyphagous and highly invasive economically important pest in the world. We proposed the hypothesis that radiation treatment influence RNA expression in the larvae and leads to emergence failure. Therefore, transcriptome analyses of third-instar larvae of B. dorsalis ionizing, irradiated with ⁶⁰Co-γ at 116Gy, were conducted and compared with the controls; a total of 608 DEGs were identified, including 348 up-regulated genes and 260 down-regulated ones. In addition, 130 SNPs in 125 unigenes were identified. For the DEGs, the most significantly enriched GO item was hemolymph coagulation, and some of the enriched pathways were involved in digestive processes. The subsequent validation experiment confirmed the differential expression of six genes, including sqd, ENPEP, Jhe, mth, Notch, and Ugt. Additionally, the 3401:G->A SNP in the Notch gene was also successfully validated. According to previous research, this was the first comparative transcriptome study to discover the candidate genes involved in insect molt to pupae. These results not only deepen our understanding of the emerging mechanism of B. dorsalis but also provide new insights into the research of biomarkers for quarantine insect treatment with the appropriate dose of radiation. Full article

(This article belongs to the Special Issue Insect Bioinformatics)

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13 pages, 1888 KiB

Open AccessArticle

Analysis of Long Non-Coding RNA-Mediated Regulatory Networks of Plutella xylostella in Response to Metarhizium anisopliae Infection

by Junaid Zafar, Junlin Huang, Xiaoxia Xu and Fengliang Jin

Insects 2022, 13(10), 916; https://doi.org/10.3390/insects13100916 - 09 Oct 2022

Cited by 7 | Viewed by 1451

Abstract

Long non-coding RNAs (lncRNAs) represent a diverse class of RNAs that are structurally similar to messenger RNAs (mRNAs) but do not encode proteins. Growing evidence suggests that in response to biotic and abiotic stresses, the lncRNAs play crucial regulatory roles in plants and [...] Read more.

Long non-coding RNAs (lncRNAs) represent a diverse class of RNAs that are structurally similar to messenger RNAs (mRNAs) but do not encode proteins. Growing evidence suggests that in response to biotic and abiotic stresses, the lncRNAs play crucial regulatory roles in plants and animals. However, the potential role of lncRNAs during fungal infection has yet to be characterized in Plutella xylostella, a devastating pest of cruciferous crops. In the current study, we performed a strand-specific RNA sequencing of Metarhizium anisopliae-infected (Px36hT, Px72hT) and uninfected (Px36hCK, Px72hCK) P. xylostella fat body tissues. Comprehensive bioinformatic analysis revealed a total of 5665 and 4941 lncRNAs at 36 and 72-h post-infection (hpi), including 563 (Px36hT), 532 (Px72hT) known and 5102 (Px36hT), 4409 (Px72hT) novel lncRNA transcripts. These lncRNAs shared structural similarities with their counterparts in other species, including shorter exon and intron length, fewer exon numbers, and a lower expression profile than mRNAs. LncRNAs regulate the expression of neighboring protein-coding genes by acting in a cis and trans manner. Functional annotation and pathway analysis of cis-acting lncRNAs revealed their role in several immune-related genes, including Toll, serpin, transferrin, βGRP etc. Furthermore, we identified multiple lncRNAs acting as microRNA (miRNA) precursors. These miRNAs can potentially regulate the expression of mRNAs involved in immunity and development, suggesting a crucial lncRNA–miRNA-mRNA complex. Our findings will provide a genetic resource for future functional studies of lncRNAs involved in P. xylostella immune responses to M. anisopliae infection and shed light on understanding insect host–pathogen interactions. Full article

(This article belongs to the Special Issue Insect Bioinformatics)

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19 pages, 1701 KiB

Open AccessArticle

Meta-Analysis of Immune Induced Gene Expression Changes in Diverse Drosophila melanogaster Innate Immune Responses

by Ashley L. Waring, Joshua Hill, Brooke M. Allen, Nicholas M. Bretz, Nguyen Le, Pooja Kr, Dakota Fuss and Nathan T. Mortimer

Insects 2022, 13(5), 490; https://doi.org/10.3390/insects13050490 - 23 May 2022

Cited by 2 | Viewed by 2600

Abstract

Organisms are commonly infected by a diverse array of pathogens and mount functionally distinct responses to each of these varied immune challenges. Host immune responses are characterized by the induction of gene expression, however, the extent to which expression changes are shared among [...] Read more.

Organisms are commonly infected by a diverse array of pathogens and mount functionally distinct responses to each of these varied immune challenges. Host immune responses are characterized by the induction of gene expression, however, the extent to which expression changes are shared among responses to distinct pathogens is largely unknown. To examine this, we performed meta-analysis of gene expression data collected from Drosophila melanogaster following infection with a wide array of pathogens. We identified 62 genes that are significantly induced by infection. While many of these infection-induced genes encode known immune response factors, we also identified 21 genes that have not been previously associated with host immunity. Examination of the upstream flanking sequences of the infection-induced genes lead to the identification of two conserved enhancer sites. These sites correspond to conserved binding sites for GATA and nuclear factor κB (NFκB) family transcription factors and are associated with higher levels of transcript induction. We further identified 31 genes with predicted functions in metabolism and organismal development that are significantly downregulated following infection by diverse pathogens. Our study identifies conserved gene expression changes in Drosophila melanogaster following infection with varied pathogens, and transcription factor families that may regulate this immune induction. Full article

(This article belongs to the Special Issue Insect Bioinformatics)

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12 pages, 1060 KiB

Open AccessArticle

Analysis of Transcriptome Difference between Blood-Fed and Starved Tropical Bed Bug, Cimex hemipterus (F.) (Hemiptera: Cimicidae)

by Li Lim and Abdul Hafiz Ab Majid

Insects 2022, 13(4), 387; https://doi.org/10.3390/insects13040387 - 14 Apr 2022

Viewed by 2202

Abstract

The reference transcriptome for Cimex hemipterus (tropical bed bug) was assembled de novo in this study, and differential expression analysis was conducted between blood-fed and starved tropical bed bug. A total of 24,609 transcripts were assembled, with around 79% of them being annotated [...] Read more.

The reference transcriptome for Cimex hemipterus (tropical bed bug) was assembled de novo in this study, and differential expression analysis was conducted between blood-fed and starved tropical bed bug. A total of 24,609 transcripts were assembled, with around 79% of them being annotated against the Eukaryotic Orthologous Groups (KOG) database. The transcriptomic comparison revealed several differentially expressed genes between blood-fed and starved bed bugs, with 38 of them being identifiable. There were 20 and 18 genes significantly upregulated in blood-fed and starved bed bugs, respectively. Differentially expressed genes (DEGs) were revealed to be associated with regulation, metabolism, transport, motility, immune, and stress response; endocytosis; and signal transduction. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed an enrichment of genes encoding steroid biosynthesis, glycosaminoglycan biosynthesis, butanoate metabolism, and autophagy in both blood-fed and starved bed bugs. However, in blood-fed bed bugs, genes involved in histidine metabolism, caffeine metabolism, ubiquinone/terpenoid-quinone biosynthesis, and sulfur relay system were enriched. On the other hand, starvation activates genes related to nicotinate and nicotinamide metabolism, fatty acid elongation, terpenoid backbone biosynthesis, metabolism of xenobiotics by cytochrome P450, riboflavin metabolism, apoptosis, and protein export. The present study is the first to report a de novo transcriptomic analysis in C. hemipterus and demonstrated differential responses of bed bugs in facing blood-feeding and starvation. Full article

(This article belongs to the Special Issue Insect Bioinformatics)

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Review

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21 pages, 791 KiB

Open AccessReview

Current Status of Omics Studies Elucidating the Features of Reproductive Biology in Blood-Feeding Insects

by Aditi Kulkarni, Frida M. Delgadillo, Sharan Gayathrinathan, Brian I. Grajeda and Sourav Roy

Insects 2023, 14(10), 802; https://doi.org/10.3390/insects14100802 - 06 Oct 2023

Viewed by 1558

Abstract

Female insects belonging to the genera Anopheles, Aedes, Glossina, and Rhodnius account for the majority of global vector-borne disease mortality. In response to mating, these female insects undergo several molecular, physiological, and behavioral changes. Studying the dynamic post-mating molecular responses in these insects [...] Read more.

Female insects belonging to the genera Anopheles, Aedes, Glossina, and Rhodnius account for the majority of global vector-borne disease mortality. In response to mating, these female insects undergo several molecular, physiological, and behavioral changes. Studying the dynamic post-mating molecular responses in these insects that transmit human diseases can lead to the identification of potential targets for the development of novel vector control methods. With the continued advancements in bioinformatics tools, we now have the capability to delve into various physiological processes in these insects. Here, we discuss the availability of multiple datasets describing the reproductive physiology of the common blood-feeding insects at the molecular level. Additionally, we compare the male-derived triggers transferred during mating to females, examining both shared and species-specific factors. These triggers initiate post-mating genetic responses in female vectors, affecting not only their reproductive success but also disease transmission. Full article

(This article belongs to the Special Issue Insect Bioinformatics)

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14 pages, 2655 KiB

Open AccessReview

REDfly: An Integrated Knowledgebase for Insect Regulatory Genomics

by Soile V. E. Keränen, Angel Villahoz-Baleta, Andrew E. Bruno and Marc S. Halfon

Insects 2022, 13(7), 618; https://doi.org/10.3390/insects13070618 - 11 Jul 2022

Cited by 5 | Viewed by 2000

Abstract

We provide here an updated description of the REDfly (Regulatory Element Database for Fly) database of transcriptional regulatory elements, a unique resource that provides regulatory annotation for the genome of Drosophila and other insects. The genomic sequences regulating insect gene expression—transcriptional cis-regulatory [...] Read more.

We provide here an updated description of the REDfly (Regulatory Element Database for Fly) database of transcriptional regulatory elements, a unique resource that provides regulatory annotation for the genome of Drosophila and other insects. The genomic sequences regulating insect gene expression—transcriptional cis-regulatory modules (CRMs, e.g., “enhancers”) and transcription factor binding sites (TFBSs)—are not currently curated by any other major database resources. However, knowledge of such sequences is important, as CRMs play critical roles with respect to disease as well as normal development, phenotypic variation, and evolution. Characterized CRMs also provide useful tools for both basic and applied research, including developing methods for insect control. REDfly, which is the most detailed existing platform for metazoan regulatory-element annotation, includes over 40,000 experimentally verified CRMs and TFBSs along with their DNA sequences, their associated genes, and the expression patterns they direct. Here, we briefly describe REDfly’s contents and data model, with an emphasis on the new features implemented since 2020. We then provide an illustrated walk-through of several common REDfly search use cases. Full article

(This article belongs to the Special Issue Insect Bioinformatics)

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