Kr-h1 Encoding Juvenile Hormone Transcription Factor Impacts Reproductive Functions in Coccinella septempunctata
Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Insects
2.2. Artificial Diets
2.3. Expression Profile of Kr-h1 in Adult Female Ladybugs
2.4. RNAi Methodology
2.4.1. Synthesis of Kr-h1-dsRNA
2.4.2. dsRNA Injection
2.4.3. Effects of RNAi on Female Adults
2.5. Data Analysis
3. Results
3.1. JH Affects Kr-h1 Expression in C. septempunctata
3.2. Kr-h1-dsRNA Affects Kr-h1 Expression in C. septempunctata
3.3. Impact of Kr-h1-dsRNA Injection on Ovary Development
3.4. Kr-h1-dsRNA Impacts on C. septempunctata Fecundity
4. Discussion
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
- Liu, Y.; Sheng, Z.T.; Liu, H.H.; Wen, D.; He, Q.Y.; Wang, S.; Shao, W.; Jiang, R.J.; An, S.H.; Sun, Y.N.; et al. Juvenile hormone counteracts the bHLH-PAS transcription factors MET and GCE to prevent caspase-dependent programmed cell death in Drosophila. Development 2009, 136, 2015–2025. [Google Scholar] [CrossRef] [Scilit] [PubMed]
- Santos, C.G.; Humann, F.C.; Hartfelder, K. Juvenile hormone signaling in insect oogenesis. Curr. Opin. Insect Sci. 2019, 31, 43–48. [Google Scholar] [CrossRef] [Scilit]
- Shpigler, H.Y.; Cohen, T.M.; Ben-Shimol, E.; Ben-Betzalel, R.; Levin, E. Juvenile hormone functions as a metabolic rate accelerator in bumble bees (Bombus terrestris). Horm. Behav. 2021, 136, 105073. [Google Scholar] [CrossRef] [Scilit] [PubMed]
- Zhang, X.S.; Li, S.; Liu, S.N. Juvenile hormone studies in Drosophila melanogaster. Front. Physiol. 2022, 12, 785320. [Google Scholar] [CrossRef] [Scilit]
- Liu, M.Y.; Wang, J.; Wang, M.Z.; Gao, F.; Zhang, H.Z.; Li, Y.Y.; Zang, L.S.; Zhang, L.S. Cloning and expression analysis of juvenile hormone epoxide hydrolase in Coccinella septempunctata. Plant Prot. 2019, 45, 156–162. [Google Scholar]
- Han, H.; Feng, Z.Y.; Zhang, S.; He, Y.Z. Effects of exogenous juvenile hormone on the ovarian development and transcription levels of the reproduction-related genes in Harmonia axyridis (Coleoptera: Coccinellidae). Acta Entomol. Sin. 2022, 65, 1090–1097. [Google Scholar]
- Jin, M.N.; Lin, X.D. Molecular mechanisms of the insect juvenile hormone. Acta Ecol. Sin. 2014, 34, 1361–1370. [Google Scholar] [CrossRef] [Scilit]
- Chen, J.X.; Geng, B.; He, J.Y. Advances in insect juvenile hormone receptor. J. Environ. Entomol. 2023, 45, 1167–1173. [Google Scholar]
- Li, D.; Muramatsu, N.; Numata, H.; Ito, C. Functional analysis of a juvenile hormone inducible transcription factor, Krüppel homolog 1, in the bean bug, Riptortus pedestris. Zool. Sci. 2022, 39, 562–569. [Google Scholar]
- Song, Z.C.; Gao, Y.Q.; Yan, Z.Q.; Zhao, Y.F.; Zhang, A.Y.; Liu, F.H.; Sun, J.H.; Xu, Y.Y.; Tian, H.G.; Li, S.; et al. Krüppel-homolog 1 is key in development and parthenogenetic reproduction in pea aphids. Entomol. Gen. 2025, 45, 205–215. [Google Scholar] [CrossRef] [Scilit]
- Wu, Y.K.; Yu, J.Q.; Jin, L.; Li, G.Q. Krüppel-homolog 1 is critical for larval metamorphosis in an herbivorous ladybird. J. Asia-Pac. Entomol. 2025, 28, 102447. [Google Scholar] [CrossRef] [Scilit]
- Yang, B.B.; Miao, S.Y.; Lu, Y.J.; Wang, S.S.; Wang, Z.Y.; Zhao, Y.R. Involvement of Methoprene-tolerant and Krüppel homolog 1 in juvenile hormone-mediated vitellogenesis of female Liposcelis entomophila (End.) (Psocoptera: Liposcelididae). Arch. Insect Biochem. Physiol. 2023, 112, e21973. [Google Scholar] [CrossRef] [Scilit] [PubMed]
- Aguilar, P.; Bourgeois, T.; Maria, A.; Couzi, P.; Demondion, E.; Bozzolan, F.; Gassias, E.; Force, E.; Debernard, S. Methoprene-tolerant and Krüppel homolog 1 are actors of juvenile hormone-signaling controlling the development of male sexual behavior in the moth Agrotis ipsilon. Horm. Behav. 2023, 150, 105330. [Google Scholar] [CrossRef] [Scilit]
- Fussnecker, B.; Grozinger, C. Dissecting the role of Kr-h1 brain gene expression in foraging behavior in honey bees (Apis mellifera). Insect Mol. Biol. 2008, 17, 515–522. [Google Scholar] [CrossRef] [Scilit]
- Fichelson, P.; Brigui, A.; Pichaud, F. Orthodenticle and Krüppel homolog 1 regulate Drosophila photoreceptor maturation. Proc. Natl. Acad. Sci. USA 2012, 109, 7893–7898. [Google Scholar] [CrossRef] [Scilit]
- Song, J.; Wu, Z.; Wang, Z.; Deng, S.; Zhou, S. Krüppel-homolog 1 mediates juvenile hormone action to promote vitellogenes is and oocyte maturation in the migratory locust. Insect Biochem. Mol. Biol. 2014, 52, 94–101. [Google Scholar] [CrossRef] [Scilit] [PubMed]
- Liu, W.; Guo, S.; Sun, D.; Zhu, L.; Zhu, F.; Lei, C.L.; Sheng, L.; Phelps, B.; Wang, X.P. Molecular characterization and juvenile hormone regulated transcription of the vitellogenin receptor in the cabbage beetle Colaphellus bowringi. Comp. Biochem. Physiol. Part A 2019, 229, 69–75. [Google Scholar] [CrossRef] [Scilit]
- Duportets, L.; Bozzolan, F.; Abrieux, A.; Maria, A.; Gadenne, C.; Debernard, S. The transcription factor Krüppel homolog 1 is linked to the juvenile hormone-dependent maturation of sexual behavior in the male moth, Agrotis ipsilon. Gen. Comp. Endocr. 2012, 176, 158–166. [Google Scholar] [CrossRef] [Scilit]
- Guan, C.X. Effects of exogenic juvenile hormone analogue on haemolymph vitellogenin content in Coccinella septempunctata L. Acta Entomol. Sin. 1989, 32, 6–11. [Google Scholar]
- Cheng, Y.; Zhou, Y.H.; Ran, H.Y.; Li, F.L. Effects of different hormones as dietary supplements on biological characteristics of Coccinella septempunctata L. J. Appl. Entomol. 2023, 147, 888–894. [Google Scholar] [CrossRef] [Scilit]
- Li, Z.Y.; Liu, Y.M.; Liang, Y.Q.; Pan, T.Q.; Liu, J.S. RNA interference-based pesticides: Mechanism, application, and commercialization in sustainable pest management. Pestic. Biochem. Physiol. 2026, 219, 107034. [Google Scholar] [CrossRef] [Scilit]
- Cheng, Y.; Zhou, Y.H.; Li, C.; Jin, J.X. Cloning and functional analysis of the juvenile hormone receptor gene CsMet in Coccinella septempunctata. J. Insect Sci. 2024, 24, ieae065. [Google Scholar] [CrossRef] [Scilit] [PubMed]
- Pfaffl, M.W. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001, 29, e45. [Google Scholar] [CrossRef] [Scilit]
- Tang, Q.Y.; Zhang, C.X. Data Processing System (DPS) software with experimental design, statistical analysis and data mining developed for use in entomological research. Insect Sci. 2013, 20, 254–260. [Google Scholar] [CrossRef] [Scilit]
- Cheng, Y.; Zhou, Y.H.; Li, F.L. Cloning and spatio-temporal expression of CsKr-h1 encoding the juvenile hormone response gene in Coccinella septempunctata L. Bull. Entomol. Res. 2024, 114, 99–106. [Google Scholar] [CrossRef] [Scilit]
- Minakuchi, C.; Zhou, X.; Riddiford, L.M. Krüppel homolog 1 (Kr-h1) mediates juvenile hormone action during metamorphosis of Drosophila melanogaster. Mech. Dev. 2008, 125, 91–105. [Google Scholar] [CrossRef] [Scilit] [PubMed]
- Zhang, T.L.; Song, W.; Li, Z.; Qian, W.L.; Wei, L.; Yang, Y.; Wang, W.N.; Zhou, X.; Meng, M.; Peng, J.; et al. Krüppel homolog 1 represses insect ecdysone biosynthesis by directly inhibiting the transcription of steroidogenic enzymes. Proc. Natl. Acad. Sci. USA 2018, 115, 3960–3965. [Google Scholar] [CrossRef] [Scilit] [PubMed]
- Li, Y.L. Functional Analysis of Juvenile Hormone Signaling Pathway Genes Met and Kr-h1 in Bactrocera dorsalis (Hendel) (Diptera: Tephritidae). Mater’s Thesis, Southwest University, Chongqing, China, 2016. [Google Scholar]
- Miao, L.J.; Zhang, N.; Jiang, H.; Dong, F.; Yang, X.M.; Xu, X.; Qian, K.; Meng, X.K.; Wang, J.J. Involvement of two paralogous methoprene-tolerant genes in the regulation of vitellogenin and vitellogenin receptor expression in the rice stem borer, Chilo suppressalis. Front. Genet. 2020, 11, 609–618. [Google Scholar] [CrossRef] [Scilit]
- Hu, K.; Tian, P.; Yang, L.; Qiu, L.; He, H.L.; Ding, W.B.; Li, Z.C.; Li, Y.Z. Knockdown of methoprene-tolerant arrests ovarian development in the Sogatella furcifera (Hemiptera: Delphacidae). J. Insect Sci. 2019, 19, 5. [Google Scholar] [CrossRef] [Scilit]
- Gijbels, M.; Lenaerts, C.; Broeck, J.V.; Marchal, E. Juvenile hormone receptor Met is essential for ovarian maturation in the desert locust, Schistocerca gregaria. Sci. Rep. 2019, 9, 10797. [Google Scholar] [CrossRef] [Scilit]
- Jin, M.N.; Xue, J.; Yao, Y.; Lin, X.D. Molecular characterization and functional analysis of Krüppel-homolog 1 (Kr-h1) in the Brown Planthopper, Nilaparvata lugens (Stl). J. Integr. Agr. 2014, 13, 1972–1981. [Google Scholar] [CrossRef] [Scilit]
- Yao, Y.; Wang, B.; Jiang, J.R.; Lin, X.D. Function of Methoprene-tolerant and Krüppel-homolog 1 genes in metamorphosis in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae). Acta Entomol. Sin. 2015, 58, 1151–1159. [Google Scholar]
- Liu, J.S.; He, Q.Y.; Lin, X.F.; Smagghe, G. Recent progress in nanoparticle-mediated RNA interference in insects: Unveiling new frontiers in pest control. J. Insect Physiol. 2025, 167, 104884. [Google Scholar] [CrossRef] [Scilit] [PubMed]
- Qiao, H.; Zong, S.M.; Zhao, J.; Xiao, L.B.; Zhu-Salzman, K.Y.; Xu, D.J.; Xu, G.C.; Tan, Y.A. High-efffciency green management to Spodoptera frugiperda by synergistic control of pH-responsive chitosan nanoparticle-mediated RNAi and pesticide. Chem. Eng. J. 2026, 528, 172175. [Google Scholar] [CrossRef] [Scilit]





| Primer Name | Primer Sequence (5′-3′) | Annealing Temperature |
|---|---|---|
| Kr-h1-F | AACCTTTCGAGTGCCCTGAAT | 58.3 |
| Kr-h1-R | ATGCCTCCTCCTGAACCTACT | |
| Kr-h1-dsRNA-F | taatacgactcactatagggAAGGATCTCACCACCGACAC | |
| Kr-h1-dsRNA-R | taatacgactcactatagggGGCTCCGTTTGTTCTGGTAA | |
| GFP-dsRNA-F | taatacgactcactatagggGCCAACACTTGTCACTACTT | |
| GFP-dsRNA-R | taatacgactcactatagggGGAGTATTTTGTTGATAATGGTCTG | |
| Actin-F | GATTCGCCATCCAGGACATCTC | 60.0 |
| Actin-R | TCCTTGCTCAGCTTGTTGTAGTC |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2026 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license.
Share and Cite
Cheng, Y.; Zhou, Y.; Li, C. Kr-h1 Encoding Juvenile Hormone Transcription Factor Impacts Reproductive Functions in Coccinella septempunctata. Insects 2026, 17, 577. https://doi.org/10.3390/insects17060577
Cheng Y, Zhou Y, Li C. Kr-h1 Encoding Juvenile Hormone Transcription Factor Impacts Reproductive Functions in Coccinella septempunctata. Insects. 2026; 17(6):577. https://doi.org/10.3390/insects17060577
Chicago/Turabian StyleCheng, Ying, Yuhang Zhou, and Cao Li. 2026. "Kr-h1 Encoding Juvenile Hormone Transcription Factor Impacts Reproductive Functions in Coccinella septempunctata" Insects 17, no. 6: 577. https://doi.org/10.3390/insects17060577
APA StyleCheng, Y., Zhou, Y., & Li, C. (2026). Kr-h1 Encoding Juvenile Hormone Transcription Factor Impacts Reproductive Functions in Coccinella septempunctata. Insects, 17(6), 577. https://doi.org/10.3390/insects17060577

