Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (21): 4542-4549.doi: 10.3864/j.issn.0578-1752.2013.21.017

• PLANT PROTECTION • Previous Articles     Next Articles

Selection of Reference Genes and Study of the Expression Levels of Detoxifying Enzymes of Tetranychus urticae (Acari: Tetranychidae)

 YUE  Xiu-Li-1, GAO  Xin-Ju-1, WANG  Jin-Jun-2, 吕Juan-Juan-1 , SHEN  Hui-Min-1   

  1. 1.College of Prataculture, Gansu Agricultural University, Lanzhou 730070
    2.College of Plant Protection, Southwest University, Chongqing 400716)
  • Received:2013-04-15 Online:2013-11-01 Published:2013-06-05

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Abstract: 【Objective】The objective of this study is to select suitable reference genes for studying the expression levels of detoxifying enzymes on fenpropathrin susceptible (SS) and resistant strains (Fe-R) of Tetranychus urticae by using quantitative real-time PCR (RT-qPCR).【Method】The mRNA expression stability of eight candidate reference genes 5.8S rRNA, α-tubulin, β-actin, ELF, GAPDH, RPL13a, SDHA and TBP of T. urticae were detected by RT-qPCR, the relative expression of the main detoxifying enzymes CCEs, GSTs and P450s were analyzed. GeNorm and NormFinder softwares were used to analyze and evaluate the data for reference genes and comparison of Ct value method was used for the calculation of relative expression levels.【Result】α-tubulin was one of the most stable genes among the eight reference genes. When α-tubulin was used as a reference gene, the expression levels of TuGSTo1, TuGSTd1and CYP3D2 in Fe-R strain were significantly higher than that in SS strain while TuCCE1 was downregulated.【Conclusion】The gene α-tubulin can be used as an ideal reference in both SS and Fe-R strains of T. urticae. Futhermore, the elevated relative expression levels of TuGSTo1, TuGSTd1 and CYP3D2 may be the molecular mechanism of T. urticae that with high resistance to fenpropathrin.

Key words: reference gene , RT-qPCR , CCEs , GSTs , P450s , Tetranychus urticae

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