Nanoparticle-Shielded dsRNA Delivery for Enhancing RNAi Efficiency in Cotton Spotted Bollworm Earias vittella (Lepidoptera: Nolidae)

• Published in: International journal of molecular sciences Vol. 24; no. 11; p. 9161
• Main Authors: Sandal, Shelja, Singh, Satnam, Bansal, Gulshan, Kaur, Ramandeep, Mogilicherla, Kanakachari, Pandher, Suneet, Roy, Amit, Kaur, Gurmeet, Rathore, Pankaj, Kalia, Anu
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 23.05.2023; MDPI
• Subjects: Adenosine triphosphatase; Algorithms; Aminopeptidase; Cadherin; Chitin; Chitin synthase; Chitosan; Conjugates; Cotton; Double-stranded RNA; dsRNA stability; Earias vittella; Efficiency; Elongation; Gene expression; Gene silencing; Genes; Glyceraldehyde; H+-transporting ATPase; Hemolymph; Information processing; Insects; Juvenile hormones; Lepidoptera; Metabolism; nanoparticle encapsulated dsRNA; Nanoparticles; Nolidae; Okra; Pest control; Quantum dots; reference genes; RNA-mediated interference; Signal transduction; spotted bollworm; transcriptome; Transcriptomes; α-Amylase; reference genes; cotton; dsRNA stability; nanoparticle encapsulated dsRNA; transcriptome; spotted bollworm
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms24119161

The spotted bollworm (Lepidoptera: Nolidae) is a polyphagous pest with enormous economic significance, primarily affecting cotton and okra. However, the lack of gene sequence information on this pest has a significant constraint on molecular investigations and the formulation of superior pest management strategies. An RNA-seq-based transcriptome study was conducted to alleviate such limitations, and de novo assembly was performed to obtain transcript sequences of this pest. Reference gene identification across developmental stages and RNAi treatments were conducted using its sequence information, which resulted in identifying ( ), ( ), and ( ) as the most suitable reference genes for normalization in RT-qPCR-based gene expression studies. The present study also identified important developmental, RNAi pathway, and RNAi target genes and performed life-stage developmental expression analysis using RT-qPCR to select the optimal targets for RNAi. We found that naked dsRNA degradation in the hemolymph is the primary reason for poor RNAi. A total of six genes including ( ) ( ) ( ) ( ) ( ), and ( ) were selected and knocked down significantly with three different nanoparticles encapsulated dsRNA conjugates, i.e., Chitosan-dsRNA, carbon quantum dots-dsRNA (CQD-dsRNA), and Lipofectamine-dsRNA conjugate. These results demonstrate that feeding nanoparticle-shielded dsRNA silences target genes and suggests that nanoparticle-based RNAi can efficiently manage this pest.