Spatio-Temporal Profiling of Metarhizium anisopliae—Responsive microRNAs Involved in Modulation of Plutella xylostella Immunity and Development

Abstract

Metarhizium anisopliae, a ubiquitous pathogenic fungus, regulates a wide array of the insect pest population. The fungus has been employed to control Plutella xylostella, an insecticide-resistant destructive lepidopteran pest, which causes substantial economic losses in crops worldwide. Integration of modern gene-silencing technologies in pest control strategies has become more crucial to counter pesticide-resistant insects. MicroRNAs (miRNA) play essential roles in the various biological process via post-transcriptional gene regulation. In the present study, RNA-seq analysis of control (CK36h, CK72h) and fungal-infected (T36h, T72h) midguts was performed to reveal underlying molecular mechanisms occurring in larval midgut at different time courses. We aimed at exploring M. anisopliae-responsive miRNAs and their target genes involved in development and immunity. After data filtration, a combined set of 170 miRNAs were identified from all libraries. Interestingly, miR-281, miR-263, miR-1, miR-6094 and miR-8 were listed among the most abundantly expressed conserved miRNAs. Furthermore, we experimentally studied the role of differentially expressed miR-11912-5p in regulating corresponding target trypsin-like serine proteinase (Px_TLSP). The luciferase assay (in vitro) revealed that miRNA-11912-5p significantly downregulated its target gene, suggesting it might play a crucial role in defense mechanism of P. xylostella against M.+ anisopliae infection. We used synthetic miRNA mimic/inhibitor (in vivo), to overexpress/silence miRNA, which showed harmful effects on larval duration, survival and adult fecundity. Additionally, fungal application in the presence of mimics revealed enhanced sensitivity of P. xylostella to infection. Our finding provides an insight into the relatively obscure molecular mechanisms involved in insect midgut during the fungal infection.

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

“…The potential lncRNAs identified in our study as miRNA precursors are presented in Tables S13 and S14 . We filtered these miRNAs through our previous studies to identify their potential immunity and development-related target genes [ 54 , 55 ] and identified several important genes, including trypsin , serpin B1 , catalase , cholinesterase , chymotrypsin, and cytochrome p450 . A schematic diagram of the lncRNA-miRNA-mRNA complex is presented in Figure 3 , particularly focusing on immune-related genes.…”

MicroRNA-Mediated Host Immune Genes Manipulation Benefits AcMNPV Proliferation in Spodoptera frugiperda

“…MiRNAs are considered to be vital players in insect host− pathogen interactions. 48 Insects, as well as viruses, exploit miRNAs for their specific benefits. Studies conducted on several insect species have shown the response of host miRNAs to pathogen stress and revealed the differential expression of a vast number of miRNAs.…”

“…To systematically identify the AcMNPV-induced DEGs in the S. f rugiperda larval midgut, whole-transcriptome sequencing was performed at different time points (6,48, and 72 h). Differential expression analysis was carried out to determine variations in gene expressions followed by AcMNPV infection.…”

Cyromazine Effects the Reproduction of Drosophila by Decreasing the Number of Germ Cells in the Female Adult Ovary

“…A total reaction mixture of 25 μL was used, while rp49 was used as an internal control to normalize the expression of ecdysone signaling-related genes by the 2 −ΔΔ CT method. The conditions used were: 95 °C for 2 min, 40 cycles each of 95 °C for 5 s, 60 °C for 10 s, and melting curve from 65–95 °C [ 25 , 26 ]. Further, 3 biological replicates were used and each biological replicate was repeated 3 times with 20 ovaries per replicate.…”