Journal Article

RNAi-mediated silencing of vitellogenin gene curtails oogenesis in the almond moth Cadra cautella

Aldawood, Abdulrahman Saad, Alwaneen, Waleed Saleh, Husain, Mureed, Tufail, Muhammad, Rasool, Khawaja Ghulam

2021

Overview

Vitellogenins, major yolk protein precursors, play an essential role in the reproduction and spread of all oviparous species, including insects. To investigate reproductive strategies of the warehouse moth Cadra cautella at the molecular level, a partial transcript of the C . cautella vitellogenin ( CcVg ) gene was extended through the rapid amplification of cDNA ends PCR and sequenced. The complete CcVg mRNA transcript was 5,334 bp long, which encoded a protein of 1,778 amino acids, including the first 14 amino acids of the signal peptide. The deduced CcVg protein contained a putative cleavage site (RTRR) at the amino-terminal side, similar to several other insect species. DGQR and GI/LCG motifs were present at the CcVg gene C-terminus, followed by nine cysteine residues. CcVg harbored 131 putative phosphorylation sites, numbering 84, 19, and 28 sites for serine, threonine, and tyrosine, respectively. The transcript showed a great resemblance with other lepidopteran Vgs. CcVg protein analysis revealed three conserved regions: 1) vitellogenin-N domain, 2) DUF 1943 (domain of unknown function), and 3) a von Willebrand factor type D domain. Additionally, sex, stage-specific, and developmental expression profiles of the CcVg gene were determined through RT-PCR. The Vg was first expressed in 22-day-old female larvae, and its expression increased with growth. The phylogenetic analysis based on different insect Vgs revealed that the CcVg exhibited close ancestry with lepidopterans. The CcVg -based RNAi experiments were performed, and the effects were critically evaluated. The qRT-PCR results showed that CcVg -based dsRNA suppressed the Vg gene expression up to 90% at 48 h post-injection. Moreover, CcVg -based RNAi effects resulted in low fecundity and egg hatchability in the CcVg -based dsRNA-treated females. The females laid eggs, but because of insufficient yolk protein availability the eggs could not succeed to hatch. The significant difference in the fecundity and hatchability unveils the importance of CcVg gene silencing and confirmed that the Vg gene plays a key role in C . cautella reproduction and it has the potential to be used as a target for RNAi-mediated control of this warehouse pest.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Aedes aegypti / Agricultural commodities / Agricultural sciences / Agricultural technology / Agriculture / Amino Acid Sequence / Animals / Apis mellifera / Biological research / Biology and Life Sciences / Biology, Experimental / Blattidae / Bombyx mori / Bromides / Butterflies & moths / Computer and Information Sciences / Computer programs / Cotton / Data analysis / Dictyoptera / Drafting software / Economic analysis / Editing / Embryos / Entomology / Environmental management / Farms / Female / Food / Freezing / Freezing effects / Fruits / Fumigants / Gene expression / Gene Expression Regulation, Developmental - genetics / Genetic aspects / Genetic research / Genomics / Heat treatment / Heat treatments / Hemiptera / Insects / Laboratories / Lepidoptera / Methyl bromide / Mosquitoes / Moths - genetics / Moths - growth & development / Moths - physiology / Nilaparvata lugens / Noctuidae / Oogenesis / Oogenesis - genetics / Pests / Phosphine / Phosphines / Plant protection / Proteins / Research and Analysis Methods / Reviews / RNA Interference / RNA-mediated interference / Science and technology / Sex Characteristics / Silkworms / Software / Spodoptera exigua / Technology / Vector-borne diseases / Vitellogenin / Vitellogenins - chemistry / Vitellogenins - deficiency / Vitellogenins - genetics / Vitellogenins - metabolism / Yellow fever