In Silico Analysis of Two Hard Tick P450s: Identification, Characterization, and Putative Metabolism of Cymbopogon citratus Essential Oil Constituents

利用计算机模拟分析两种硬蜱P450酶:香茅精油成分的鉴定、表征及推测代谢

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Abstract

The cytochrome p450 gene family is widely involved in various biological processes in arthropods. Tick p450s are often associated with chemical acaricides, but knowledge of their involvement in the metabolism of plant-derived essential oil components is limited. In this study, we identified the non-redundant number of p450 transcripts (NRNPTs) from Haemaphysalis longicornis and Hyalomma asiaticum under the Cymbopogon citratus essential oil (CCEO) and terpinolene stress using de novo transcriptome data, respectively. In this study, we identified and characterized the NRNPTs of Ha. longicornis and Hy. asiaticum. Their gene expression patterns and biological functions under CCEO and terpinolene stress were further analyzed. Finally, Hy. asiaticum NRNPTs (87) were more numerous than Ha. longicornis (58). Phylogenetic analyses showed that NRNPTs of both Hy. asiaticum and Ha. longicornis could be categorized in clan 2, clan 3, clan 4, and clan mito, this data comes from the NRNPTs. Phylogenetic analyses showed that NRNPTs of both Hy. asiaticum and Ha. longicornis could be categorized in clan 2, clan 3, clan 4, and clan mito. p450 members of both were most distributed in clan 3. In addition, one Hy. asiaticum NRNPT was identified as belonging to the new classification clan 20 (HyasCYP20A1). The biological functions and pathways of p450 family members enriched in Hy. asiaticum and Ha. longicornis under different exogenous substance stresses were different, and the expression patterns of these genes were inconsistent. Molecular docking results showed that Ha. longicornis p450 members (HaloCYP3A4 and HaloCYP4B1), which were significantly up-regulated under CCEO stress, as well as Hy. asiaticum HyasCYP24A1 and HyasCYP4V2 (the HaloCYP3A4 and HaloCYP4B1 homologous genes), encode proteins that differ in their ability to metabolize CCEO components, but they all bind well to Germacrene D and naphthalene. Our study enriches the knowledge of the involvement of p450 family members of different tick species in the metabolism of essential oil components of plants, and provides a theoretical basis for further in-depth studies on the function of tick p450 enzymes.

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