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Background <p>Chitinases and ecdysone are both crucial for insect growth and development, yet the regulatory interplay between them remains poorly understood. Our previous research demonstrated that the chitinase gene <i>HaCHT4</i> critically regulates the chitin content and surface structure of peritrophic membrane (PM) in <i>Helicoverpa armigera</i> (Hübner) (Lepidoptera: Noctuidae).</p> Results <p>Here, a 1286&#xa0;bp 5’-flanking region of <i>HaCHT4</i> was cloned to investigate its regulatory mechanism. Bioinformatics analysis predicted the presence of several cis-regulatory elements (CREs) within its 5’-flanking region, including Broad-Complex Zinc-Finger isoforms (BRCs), GAGA, Nuclear Factors of activated T cells (NFAT), and POU domain factor (POU). Notably, genome-wide identification and characterization revealed that the BRC gene of <i>H. armigera</i> shared the highest similarity with that of <i>Bombyx mori</i> (L.) (Lepidoptera: Bombycidae). Molecular docking predicted that HaBRC Z2 exhibits a favorable binding energy (ΔG = -12&#xa0;kcal/mol) with the sequence containing the BRC Z2 CRE, leading to its selection as the primary candidate for experimental validation. An electrophoretic mobility shift assay (EMSA) provided preliminary evidence that HaBRC Z2 might bind to the sequence containing BRC Z2 CRE of the <i>HaCHT4</i> promoter region in vitro. Expression analysis showed concomitantly upregulated of <i>HaBRC Z2</i> and <i>HaCHT4</i> during the late stages of 4th and 5th instar larvae, and were also strongly induced by 20E. In addition, a substantial decrease in the transcript levels of both <i>HaBRC Z2</i> and <i>HaCHT4</i> was observed after <i>HaBRC Z2</i> silencing for 24, 48, 72, and 96&#xa0;h, and resulting in growth delay and reduced body length and weight.</p> Conclusion <p>Our results preliminarily suggested that HaBRC Z2 might act as a potential upstream regulator of <i>HaCHT4</i>, thereby regulating the larval molting process in <i>H. armigera</i>. However, direct promoter activation and in vivo occupancy remain to be experimentally validated.</p>

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Broad complex Z2 functions as an upstream 20E-responsive regulator of the midgut chitinase gene HaCHT4 in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

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  • 发表日期：

    2026年4月10日

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  • Deqin Hu,
  • Yuan Li,
  • Jingang Xie,
  • Hongsheng Pan,
  • Xiaoning Liu

摘要

Background

Chitinases and ecdysone are both crucial for insect growth and development, yet the regulatory interplay between them remains poorly understood. Our previous research demonstrated that the chitinase gene HaCHT4 critically regulates the chitin content and surface structure of peritrophic membrane (PM) in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae).

Results

Here, a 1286 bp 5’-flanking region of HaCHT4 was cloned to investigate its regulatory mechanism. Bioinformatics analysis predicted the presence of several cis-regulatory elements (CREs) within its 5’-flanking region, including Broad-Complex Zinc-Finger isoforms (BRCs), GAGA, Nuclear Factors of activated T cells (NFAT), and POU domain factor (POU). Notably, genome-wide identification and characterization revealed that the BRC gene of H. armigera shared the highest similarity with that of Bombyx mori (L.) (Lepidoptera: Bombycidae). Molecular docking predicted that HaBRC Z2 exhibits a favorable binding energy (ΔG = -12 kcal/mol) with the sequence containing the BRC Z2 CRE, leading to its selection as the primary candidate for experimental validation. An electrophoretic mobility shift assay (EMSA) provided preliminary evidence that HaBRC Z2 might bind to the sequence containing BRC Z2 CRE of the HaCHT4 promoter region in vitro. Expression analysis showed concomitantly upregulated of HaBRC Z2 and HaCHT4 during the late stages of 4th and 5th instar larvae, and were also strongly induced by 20E. In addition, a substantial decrease in the transcript levels of both HaBRC Z2 and HaCHT4 was observed after HaBRC Z2 silencing for 24, 48, 72, and 96 h, and resulting in growth delay and reduced body length and weight.

Conclusion

Our results preliminarily suggested that HaBRC Z2 might act as a potential upstream regulator of HaCHT4, thereby regulating the larval molting process in H. armigera. However, direct promoter activation and in vivo occupancy remain to be experimentally validated.