Expression characteristics of two novel cytochrome P450 genes involved in fenpropathrin resistance in Tetranychus cinnabarinus (Boisduval)

Abstract

The carmine spider mite, Tetranychus cinnabarinus, which is also considered as the red form of Tetranychus urticae, is one of the most serious mite pests on crops. It is capable of rapidly developing resistance to acaricides, and has caused difficulty in controlling. However, the resistance mechanism of this mite remains unclear at molecular level. As a member of main detoxification enzymes, cytochrome P450 monooxygenases (CYPs or P450s) play important roles in the development of acaricide resistance in arthropods. In this study, two novel P450 genes (CYP389B1 and CYP392A26) were identified and characterized from T. cinnabarinus. The opening reading frames (ORFs) of CYP389B1 and CYP392A26 contained 1545 and 1488 nucleotides, which encode 514 and 495 amino acids, respectively. Phylogenetic analysis showed that CYP389B1 and CYP392A26 were most closely related to CYP389B1 and CYP392A4 from T. urticae, respectively. When treated with piperonyl butoxide (PBO), an inhibitor of P450s, the resistance ratio of fenpropathrin-resistant (FeR) strain decreased from 101- to 75-fold, which suggested a correlation between P450 and fenpropathrin-resistance in T. cinnabarinus. Furthermore, constitutive over-expressions of CYP389B1 and CYP392A26 were detected in FeR strain. Meanwhile, the expressions of CYP389B1 and CYP392A26 were inducible in FeR strain after treatment in 6, 12 and 24 h with LC30 of fenpropathrin; especially, the expression of CYP392A26 increased to a markedly high level (20.88-fold higher than in the control) after treatment in 6 h. However, there was no significant difference between treatment and control in susceptible strain. Furthermore, stage specific expression profiles of these two genes did not show significant difference among developing stages, except for eggs, in which the mRNA levels were quite low. The results indicate that CYP389B1 and CYP392A26 were involved in the fenpropathrin-resistance in T. cinnabarinus, and the expression of CYP392A26 was more sensitive to fenpropathrin stress. These findings provide clues for further elucidating the function and regulation mechanism of these two cytochrome P450 genes in T. cinnabarinus.

Keywords: Cytochrome P450; Fenpropathrin resistance; Gene expression; Tetranychus cinnabarinus.