Agrotechnology

Abstract

An Optimized dsRNA Production Protocol from Bacterial Cultures: A Case for Fusarium graminearum Genes

Binod Gyawali, Houshang Alizadeh and Mohsen Mohammadi*

Double-stranded RNA (dsRNA) triggers RNA interference (RNAi), leading to the directed silencing of specific genes. This mechanism has become significant and recognized as a sustainable tool in crop disease and pest- management applications. In vivo transcription of dsRNA in bacterial cells and in vitro transcription using kits are two methods of dsRNA production at laboratory scale. Design and testing of dsRNA at laboratory-scale requires efficient dsRNA production that yields large and high-quality dsRNA in a relatively lower price. In this report, we present an optimized protocol for production of double-stranded RNA (dsRNA) from bacterial cultures, achieving significantly higher yields compared to existing laboratory methods by five-fold (on average). Post extraction enzymatic digestion experiment revealed that the extracted dsRNA is pure and contains no trace amount of genomic DNA or single-stranded RNA (ssRNA) contamination. This study holds significant implications for RNAi-mediated gene silencing in agricultural biotechnology, particularly in crop protection.

Published Date: 2024-04-08; Received Date: 2024-03-07