Double-stranded RNA degrading nuclease affects RNAi efficiency in the melon fly, Zeugodacus cucurbitae

文献类型： 外文期刊

第一作者： Ahmad, Shakil

作者： Ahmad, Shakil;Jamil, Momana;Luo, Yanping;Jaworski, Coline C.;Jaworski, Coline C.

作者机构：

关键词： Zeugodacus cucurbitae; RNA interference (RNAi); DNA; RNA non-specific endonuclease (dsRNase); Co-silencing; Pest control

期刊名称：JOURNAL OF PEST SCIENCE （ 影响因子：4.8； 五年影响因子：5.0 ）

ISSN： 1612-4758

年卷期： 2023 年

页码：

收录情况： SCI

摘要： RNA interference (RNAi) is an efficient, species-specific technology to control invasive pests. However, its efficacy varies among insect species, notably due to the presence of double-stranded RNA degrading enzymes (dsRNases) in the midgut and hemolymph. These dsRNases degrade the gene-specific exogenous double-stranded ribonucleic acid (dsRNA) before it can bind to its target transcript. Here, we explored the role of dsRNases in RNAi efficacy in Zeugodacus cucurbitae, a major insect pest of cucurbit plants. Our RNAi gene target was the Coatomer subunit alpha (ZcCOPI-alpha), which plays a major role in insect development and survival. We identified two dsRNases (ZcdsRNase1, ZcdsRNase2) using a bioinformatics analysis. Using qRT-PCR, we showed that their expression level was highest in Z. cucurbitae's midgut and during the larval stage, compared to the lower levels measured in other tissue and other developmental stages. We therefore co-silenced the gene of each dsRNase together with ZcCOPI-alpha in third instar larvae. Co-silencing ZcdsRNase1 along with ZcCOPI-alpha led to 84% larval mortality, while co-silencing ZcdsRNase2 along with ZcCOPI-alpha caused 44% of larval mortality only, and silencing ZcCOPI-alpha alone caused 59% of larval mortality. Co-silencing ZcdsRNase1 reduced dsZcCOPI-alpha degradation and improved RNAi efficacy in Z. cucurbitae. We also showed that ZcCOPI-alpha knockdown severely affected the survival and development of Z. cucurbitae and caused ovarian abnormalities. The demonstrated co-silencing approach offers interesting avenues for the industrial development of RNAi-based pest control strategies targeting Z. cucurbitae.

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