miR-125-3p and miR-276b-3p Regulate the Spermatogenesis of Bactrocera dorsalis by Targeting the orb2 Gene

Summar Sohail; Kaleem Tariq; Muhammad Sajid; Muhammad Waqar Ali; Wei Peng; Hongyu Zhang

doi:10.3390/genes13101861

miR-125-3p and miR-276b-3p Regulate the Spermatogenesis of Bactrocera dorsalis by Targeting the orb2 Gene

Genes (Basel). 2022 Oct 15;13(10):1861. doi: 10.3390/genes13101861.

Authors

Summar Sohail^{1

2}, Kaleem Tariq³, Muhammad Sajid⁴, Muhammad Waqar Ali¹, Wei Peng⁵, Hongyu Zhang¹

Affiliations

¹ Key Laboratory of Horticultural Plant Biology (MOE), China-Australia Joint Research Centre for Horticultural and Urban Pests, Institute of Urban and Horticultural Entomology, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
² Department of Environmental Sciences, Kohsar University Murree, Murree 47150, Pakistan.
³ Department of Entomology, Abdul Wali Khan University, Mardan 23200, Pakistan.
⁴ Department of Biotechnology, University of Okara, Okara 56300, Pakistan.
⁵ Hunan Provincial Key Laboratory of Animal Intestinal Function and Regulation, State Key Laboratory of Animal Intestinal Ecology and Health, Hunan Normal University, Changsha 410081, China.

Abstract

Bactrocera dorsalis is considered a major threat to horticultural crops. It has evolved resistance against insecticides. It is believed that development of new methods is highly desirable to control this destructive agricultural pest. Sterile insect technique is emerging as a potential tool to control this insect pest by reducing their reproductive ability. Here we report that orb2 has high expression in the testis of B. dorsalis which is the target of miR-125-3p and miR-276b-3p and plays a critical role in the spermatogenesis. Dual luciferase reporter assay using HEKT293 cells demonstrates that orb2 gene is downregulated by miR-125-3p and miR-276b-3p and is a common target of these miRNAs. Dietary treatment of adult male flies separately and in combination of agomir-125-3p (Ago-125-3p) and agomir-276b-3p (Ago-276b-3p) significantly downregulated the mRNA of orb2. The combined treatments of agomirs suppressed the level of mRNA of orb2 significantly more than any single treatment. Altered expression of miR-125-3p and miR-276b-3p significantly decreased the total and live spermatozoa in the testis which ultimately caused reduction in male fertility. Furthermore, we demonstrate that miR-125-3p, miR-276b-3p, and orb2 dsRNA are the novel agents that could be used in a genetic-based sterile insect technique (SIT) to control the B. dorsalis.

Keywords: Bactrocera dorsalis; miR-125-3p; miR-276b-3p; orb2.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Insecta / genetics
Insecticides*
Male
MicroRNAs* / genetics
MicroRNAs* / metabolism
RNA, Messenger / metabolism
Spermatogenesis / genetics
Tephritidae* / genetics

Substances

Insecticides
MicroRNAs
RNA, Messenger

Grants and funding

This research was supported by the National Key R&D Program of China (No. 2021YFC2600400), The Major Special Science and Technology Project of Yunnan Province (NO. 202102AE090054), The Earmarked Fund for CARS (CARS-26), and Higher Education Commission of Pakistan (Ref. No. 303/IPFP-II (Batch-I)/SRGP/NAHE/HEC/2020/296).