Pesticide Science Lab, Agricultural University of Athens, 75 Iera Odos, 118 55 Athens, Greece; Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, GR-700 13 Heraklion, Crete, Greece.
Department of Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Coupure links 653, Ghent, Belgium.
Pestic Biochem Physiol. 2024 Sep;204:106031. doi: 10.1016/j.pestbp.2024.106031. Epub 2024 Jul 17.
The spider mite Tetranychus urticae is a major agricultural pest with a global distribution, extremely diverse host range and a remarkable ability to develop resistance to a wide variety of acaricides. P450 mono-oxygenases have been frequently associated with resistance development in this species. In particular enzymes of the CYP392A-subfamily were shown to metabolize a number of key acaricides, including abamectin, amitraz, fenpyroximate and the active metabolite of pyflubumide. However, transcriptomic studies comparing highly resistant and susceptible populations have often revealed high expression of members of the CYP392D-subfamily, but these have been only poorly studied. Here, we conducted a meta-analysis of gene expression data of 20 populations and identified two key enzymes of this family, CYP392D2 and CYP392D8, whose expression is associated with resistance. We subsequently functionally expressed these enzymes, together with CYP392A11 and CYP392A16 as known metabolizers, and compared their potential to accept a wide diversity of acaricides as substrate. This study overall confirms previous discovered substrates for CYP392A11 and CYP392A16, but also reveals unreported metabolic activity towards new acaricides. These include carbaryl, chlorpyrifos and etoxazole for CYP392A16 and carbaryl, chlorpyrifos and NNI-0711-NH pyflubumide for CYP392A11. For the newly studied CYP392D-family, we show that CYP392D2 metabolizes pyridaben, fenpyroximate, etoxazole and chlorpyrifos, while CYP392D8 metabolizes carbaryl, fenazaquin and tebufenpyrad. Last, we observed that both CYP392A- and CYP392D-subfamily enzymes activate chlorpyrifos to its corresponding oxon. Our study indicates that there is both overlap and specificity in the activity of A- and D-subfamily enzymes against acaricides and model substrates. With the recent advent of highly efficient CRISPR/Cas9 gene editing protocols in T. urticae, the way is now paved to conduct further genetic experiments revealing and quantifying the role of these enzymes in the resistance phenotype in field populations.
叶螨 Tetranychus urticae 是一种分布广泛的农业害虫，具有极其多样化的宿主范围和显著的抗药性发展能力。细胞色素 P450 单加氧酶经常与该物种的抗药性发展有关。特别是 CYP392A 亚家族的酶已被证明能代谢多种关键杀螨剂，包括阿维菌素、甲氰菊酯、唑螨酯和 pyflubumide 的活性代谢物。然而，比较高度抗性和敏感性种群的转录组学研究经常揭示 CYP392D 亚家族成员的高表达，但这些研究还很不完善。在这里，我们对 20 个种群的基因表达数据进行了荟萃分析，鉴定出两个关键酶，CYP392D2 和 CYP392D8，其表达与抗性有关。随后，我们功能性表达了这些酶，以及作为已知代谢物的 CYP392A11 和 CYP392A16，并比较了它们作为底物接受广泛多样性杀螨剂的潜力。这项研究总体上证实了以前发现的 CYP392A11 和 CYP392A16 的底物，但也揭示了对新杀螨剂的未报告的代谢活性。这些包括对 CYP392A16 的carbaryl、chlorpyrifos 和 etoxazole，以及对 CYP392A11 的 carbaryl、chlorpyrifos 和 NNI-0711-NH pyflubumide。对于新研究的 CYP392D 家族，我们表明 CYP392D2 代谢吡虫啉、唑螨酯、etoxazole 和 chlorpyrifos，而 CYP392D8 代谢 carbaryl、fenazaquin 和 tebufenpyrad。最后，我们观察到 CYP392A-和 CYP392D-亚家族酶都能将 chlorpyrifos 激活为其相应的 oxon。我们的研究表明，A-和 D-亚家族酶对杀螨剂和模型底物的活性既有重叠又有特异性。随着最近在 T. urticae 中高效的 CRISPR/Cas9 基因编辑协议的出现，现在为进行进一步的遗传实验铺平了道路，这些实验可以揭示和量化这些酶在田间种群抗性表型中的作用。
