Zhao Wei, Zou Di, Wang ZhengYang, Wang GuiQiang, Jiang HongBo, Smagghe Guy, Wang JinJun, Wei DanDan
Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing, China.
Key Laboratory of Agricultural Biosafety and Green Production of Upper Yangtze River (Ministry of Education), Academy of Agricultural Sciences, Southwest University, Chongqing, China.
Pest Manag Sci. 2025 Sep;81(9):5864-5875. doi: 10.1002/ps.8942. Epub 2025 Jun 2.
Bactrocera dorsalis (Diptera: Tephritidae) is a highly adaptable invasive pest that causes significant economic losses. Long-term chemical control strategies have led to high levels of insecticide resistance and severe environmental risks. C-type lectins (CTLs), key pattern recognition receptors in insect innate immunity, are named for their calcium-dependent carbohydrate recognition domains (CRDs) and they play a critical role in detecting and binding to foreign pathogens. This study aims to identify B. dorsalis CTLs (BdCTLs) and investigate their roles in pathogen defense.
A total of 42 BdCTL genes were identified. Based on the number of CRDs, 36 genes were classified into the S subfamily, while the remaining six were assigned to the X subfamily. Spatio-temporal expression analysis by qPCR revealed that BdCTL genes were expressed throughout all developmental stages with a higher expression observed in the midgut, Malpighian tubules, and fat body. The expression levels of BdCTL-S4, BdCTL-S12, and BdCTL-S31 were significantly upregulated under the infection of the entomopathogenic fungus Beauveria bassiana. Notably, BdCTL-S12 also exhibited upregulated expression patterns in response to four different pathogens bacteria with sustained expression over time. Using RNA interference (RNAi), we successfully knocked down BdCTL-S12 expression, achieving a silencing efficiency of 68%. Survival assays demonstrated a 10-30% reduction in the survival rate of B. dorsalis after pathogen infection following dsRNA injection compared to the control group. Further in vitro experiments confirmed that BdCTL-S12 can agglutinate and bind to pathogens in a calcium-dependent manner. This binding was competitively inhibited by polysaccharides, underscoring a key functional mechanism of BdCTL-S12 in pathogen recognition.
In summary, we identified 42 BdCTLs and demonstrated that BdCTL-S12 is critical for pathogen recognition and immune defense in B. dorsalis. These results advance our understanding of the molecular mechanisms underlying B. dorsalis immunity. Specifically, elucidating the role of CTLs may facilitate the development of microbial-based biological control strategies (e.g., entomopathogenic fungi or bacteria) against this invasive pest. Additionally, RNAi-mediated knockdown of CTL-encoding genes could weaken the immune system of Bactrocera flies, increasing their susceptibility to microbial infections and thereby enhancing the effectiveness of biological control. © 2025 Society of Chemical Industry.
橘小实蝇(双翅目:实蝇科)是一种适应性极强的入侵害虫,会造成重大经济损失。长期的化学防治策略导致了高水平的抗药性以及严重的环境风险。C型凝集素(CTLs)是昆虫先天免疫中的关键模式识别受体,因其钙依赖性碳水化合物识别结构域(CRDs)而得名,在检测和结合外来病原体方面发挥着关键作用。本研究旨在鉴定橘小实蝇的CTLs(BdCTLs)并研究它们在病原体防御中的作用。
共鉴定出42个BdCTL基因。根据CRDs的数量,36个基因被归类到S亚家族,其余6个被归入X亚家族。通过qPCR进行的时空表达分析表明,BdCTL基因在所有发育阶段均有表达,在中肠、马氏管和脂肪体中表达较高。在球孢白僵菌感染下,BdCTL-S4、BdCTL-S12和BdCTL-S31的表达水平显著上调。值得注意的是,BdCTL-S12在受到四种不同病原菌感染时也呈现上调表达模式,且随时间持续表达。利用RNA干扰(RNAi),我们成功敲低了BdCTL-S12的表达,沉默效率达到68%。存活试验表明,与对照组相比,注射dsRNA后,橘小实蝇在病原体感染后的存活率降低了10%-30%。进一步的体外实验证实,BdCTL-S12能够以钙依赖的方式凝集并结合病原体。这种结合受到多糖的竞争性抑制,突出了BdCTL-S12在病原体识别中的关键功能机制。
总之,我们鉴定出42个BdCTLs,并证明BdCTL-S12对橘小实蝇的病原体识别和免疫防御至关重要。这些结果推进了我们对橘小实蝇免疫分子机制的理解。具体而言,阐明CTLs的作用可能有助于开发针对这种入侵害虫的基于微生物的生物防治策略(如昆虫病原真菌或细菌)。此外,RNAi介导的CTL编码基因敲低可能会削弱橘小实蝇的免疫系统,增加它们对微生物感染的易感性,从而提高生物防治的效果。© 2025化学工业协会。