MBF2转录调控小菜蛾谷胱甘肽S-转移酶代谢氯虫苯甲酰胺的功能

doi:10.3864/j.issn.0578-1752.2023.04.006

Abstract

Abstract:

【Objective】The objective of this study is to explore the regulation function of transcription regulation factor MBF2 (multiprotein bridging factor 2) on glutathione S-transferase (GST) in Plutella xylostella and its role in resistance to chlorantraniliprole, and to provide a theoretical basis for clarifying the metabolic mechanism of P. xylostella to detoxify chlorantraniliprole. 【Method】The resistance level of P. xylostella to chlorantraniliprole in three populations, susceptible (SS), Lianzhou (LZ) and Tonghai (TH), was determined by leaf dipping method and the GST activity of three populations was measured by enzyme kinetic method. Real-time quantitative PCR (RT-qPCR) was used to analyze the expression difference of MBF2 in different P. xylostella populations. After induction for 12 h under the LC₅₀ of chlorantraniliprole, the induced expression level of MBF2 in different populations was analyzed. The regulatory role of MBF2 on specific GST genes was confirmed by RNA interference (RNAi) and the GST activity of MBF2-silenced P. xylostella was analyzed. Susceptibility variations of MBF2-silenced P. xylostella populations to chlorantraniliprole were conducted by leaf dipping method, which support that MBF2 influenced the resistance of P. xylostella to chlorantraniliprole. 【Result】The TH population was moderately resistant and LZ population was highly resistant to chlorantraniliprole when respectively compared to susceptible population (SS), with the resistance ratio of 54.27 and 289.58. The GST activity of TH and LZ populations was significantly higher than that of SS population. Compared with SS population, MBF2 was significantly up-regulated in resistant populations, and the expression level of MBF2 in the moderately resistant and highly resistant populations was 4.6 and 9.4 times that of susceptible population, respectively. When P. xylostella was exposed to chlorantraniliprole at LC₅₀ for a short time, the expression levels of MBF2 in susceptible, moderately resistant and highly resistant populations were significantly increased within 8 hours, and the highest expression was 10 times that of the control. After RNAi treatment for 24 hours, the expression levels of 9 GST genes in MBF2-silenced P. xylostella were significantly decreased, and GSTZ1 as well as GSTU1 decreased by more than 90%. Compared with the negative control (dsGFP), the GST activity in the treatment group (dsMBF2) decreased by 57.76%. After treatment with 75 mg·L^-1 chlorantraniliprole, the mortality in the treatment group increased by 23.34% compared with the control group. 【Conclusion】It is speculated that MBF2 improves the metabolism of chlorantraniliprole by regulating the transcription of GSTs, thereby participating in the detoxification effect of P. xylostella to chlorantraniliprole. The results can provide experimental basis for further analysis of P. xylostella resistance mechanisms to insecticides and research as well as development of new insecticides.

Key words: Plutella xylostella, transcriptional regulation, glutathione S-transferase (GST), chlorantraniliprole, insecticide resistance, MBF2

GE TianCheng, YIN Fei, HU QiongBo, PENG ZhengKe, LI ZhenYu. Function of MBF2 Transcriptionally Regulating Glutathione S-transferase Metabolizing Chlorantraniliprole in Plutella xylostella[J].Scientia Agricultura Sinica, 2023, 56(4): 665-673.

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References 43

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种群 Population	斜率 Slope (mean±SE)	LC₅₀95%置信区间 Confidence interval (mg·L^-1)	相关系数 Relevant coefficient	χ² (df)	抗性倍数 Resistance ratio
SS	2.60±0.34	0.26 (0.19-0.38)	0.995	0.59 (3)	-
TH	2.72±0.49	14.11 (10.23-18.74)	0.932	5.74 (3)	54.27
LZ	2.86±0.52	75.29 (55.50-108.41)	0.981	0.82 (3)	289.58

种群 Population	酶活性 Enzymatic activity (nmol·min^-1·mg^-1)
SS	134.83±7.50c
TH	217.79±24.34b
LZ	295.71±12.13a

Function of MBF2 Transcriptionally Regulating Glutathione S-transferase Metabolizing Chlorantraniliprole in Plutella xylostella

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