Comparison of T7 In Vitro Transcription and E. coli Expression Systems for RNAi-Based Control of Euproctis pseudoconspersa by Targeting EpCHSA
by
,
Linyuan Huang
1,
Fanhui Meng
1,
Jinxiu Yu
2,3,
Ying Luo
4,
Zhen Liu
5,
Wan Deng
2,3,
Mi Li
2,3,
Xiudan Wang
1,* and
Yifei Xie
2,3,* 1
Laboratory of Insect Behavior and Evolutionary Ecology, College of Life and Environmental Sciences, Central South University of Forestry and Technology, Changsha 410004, China
2
Institute of Forestry and Grassland Protection, Hunan Academy of Forestry, Changsha 410004, China
3
Yuelushan Laboratory, Changsha 410018, China
4
Hunan Forest & Grassland Fire Prevention Center, Hunan Forestry Bureau, Changsha 410004, China
5
College of Life Sciences, Hunan University of Arts and Science, Changde 415000, China
*
Authors to whom correspondence should be addressed.
Insects 2026, 17(5), 453; https://doi.org/10.3390/insects17050453
Submission received: 11 February 2026
/
Revised: 1 April 2026
/
Accepted: 14 April 2026
/
Published: 24 April 2026
(This article belongs to the Special Issue Insect Pathogens as Biocontrol Agents Against Pests)
Simple Summary
This study compared T7 in vitro transcription and E. coli expression systems for dsRNA production against Euproctis pseudoconspersa. The bacterial system offered a cost-effective alternative, achieving superior EpCHSA silencing efficiency. This led to increased mortality and severe developmental deformities, providing a potent strategy for RNAi-based biopesticide development.
Abstract
Euproctis pseudoconspersa is a devastating pest in Camellia oleifera plantations, necessitating the development of sustainable molecular intervention strategies. This study targeted the chitin synthase A gene (EpCHSA) to evaluate and compare the RNA interference (RNAi) efficacy of dsRNA synthesized via the T7 in vitro transcription system and the Escherichia coli HT115 (DE3) expression system. The EpCHSA gene (2199 bp ORF) was cloned and characterized, exhibiting peak expression during the fourth-instar stage, and predominantly in the head tissues of fifth-instar larvae. Bioassays demonstrated that larvae fed with 500 ng/μL in vitro synthesized dsRNA exhibited continuous gene silencing for five days, reaching a maximum efficiency of 68.1%. Conversely, treatment with 100× concentrated bacterial broth (5000 ng/μL) elicited a superior silencing effect of 79.3% within 24 h. Furthermore, the bacterial treatment group reached a 14-day mortality rate of 46.66%, significantly higher than the in vitro group (38.33%). Both methods induced severe phenotypic abnormalities, including molting failure and pupal malformation. These findings conclude that the E. coli expression system offers a cost-effective and highly potent platform for dsRNA production. This research provides a critical technical foundation and promising application prospects for the field-scale management of E. pseudoconspersa utilizing RNAi-based biopesticides.
