Comparative Transcriptomic Analysis of Riptortus pedestris (Hemiptera: Alydidae) to Characterize Wing Formation across All Developmental Stages

Abstract

Riptortus pedestris (Hemiptera: Alydidae) is a major agricultural pest in East Asia that causes considerable economic losses to the soybean crop each year. However, the molecular mechanisms governing the growth and development of R. pedestris have not been fully elucidated. In this study, the Illumina HiSeq6000 platform was employed to perform de novo transcriptome assembly and determine the gene expression profiles of this species across all developmental stages, including eggs, first-, second-, third-, fourth-, and fifth-instar nymphs, and adults. In this study, a total of 60,058 unigenes were assembled from numerous raw reads, exhibiting an N50 length of 2126 bp and an average length of 1199 bp, and the unigenes were annotated and classified with various databases, such as the Kyoto Encyclopedia of Genes and Genomes (KEGG), Clusters of Orthologous Groups (COG), and Gene Ontology (GO). Furthermore, various numbers of differentially expressed genes (DEGs) were calculated through pairwise comparisons of all life stages, and some of these DEGs were associated with immunity, metabolism, and development by GO and KEGG enrichment. In addition, 35,158 simple sequence repeats (SSRs) and 715,604 potential single nucleotide polymorphisms (SNPs) were identified from the seven transcriptome libraries of R. pedestris. Finally, we identified and summarized ten wing formation-related signaling pathways, and the molecular properties and expression levels of five wing development-related genes were analyzed using quantitative real-time PCR for all developmental stages of R. pedestris. Taken together, the results of this study may establish a foundation for future research investigating developmental processes and wing formation in hemimetabolous insects and may provide valuable data for pest control efforts attempting to reduce the economic damage caused by this pest.

Identification and expression patterns of somatic piRNAs and PIWI genes in Riptortus pedestris (Hemiptera: Alydidae)

“…China, and reared in climatic chambers at a temperature of 25 ± 2°C and 60% ± 5% relative humidity (RH) under a long‐day photoperiod cycle (16:8 h light/dark). The methods for sample treatment and tissue dissection for WTS and RNA transcriptome sequencing were described in our previous reports (Fu et al, 2021; Ren, Chen, Fu, et al, 2023). In brief, newly hatched eggs from the above parents were reared until successful incubation of 1st‐instar nymphs, which were randomly divided into three groups: (1) feeding soybean seeds and distilled water containing 0.05% (w/v) ascorbic acid (DWA) from 1st‐instar nymph to adult (Blank group; B); (2) feeding potted soybean plants and DWA from 1st‐instar nymph to adult (Control group; C); (3) alternatively fed soybean seeds and DWA or only distilled water containing 50 μg/mL tetracycline from 2nd‐instar nymph to adult on the following day (Antibiotic group; A).…”

The ncRNA-mediated regulatory networks of defensins and lysozymes in Riptortus pedestris: involvement in response to gut bacterial disturbances

“…The quantitative real-time PCR (qRT-PCR) assay was executed with a Green qPCR SuperMix Kit (TransGen, Beijing, China) on a CFX96 real-time PCR detection system (Bio-Rad Laboratories, Hercules, CA) following the manufacturer's instructions. The PCR procedures were performed as described previously (Fu et al, 2021;Ren et al, 2023). The EF-1α gene was used as an internal control for normalization of the expression levels, and specific primers of selected DE AMPs and ncRNAs were designed by Primer Premier 5 software (Supplementary Table S5).…”

Elucidating the ecophysiology of soybean pod-sucking stinkbug Riptortus pedestris (Hemiptera: Alydidae) based on de novo genome assembly and transcriptome analysis

“…Investigating this aspect may provide a better understanding of soybean-pest interaction, and ways of utilizing this knowledge for HPR offers a potential and an appealing target for mitigating R. pedestris infestation [ 17 , 25 , 27 – 31 ]. Interestingly, secreted proteins from R. pedestris that instigate plant cell death have been identified [ 32 , 33 ], and cross-kingdom species interactions through plant–insect microRNAs have also been reported [ 34 ]. However, how interaction occurs between soybeans and stinkbugs remains unknown, and profiling the gene expression changes as a result of this interaction is thus necessary.…”