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Evaluation of suitable reference genes for quantitative RT-PCR during development and abiotic stress in Panonychus citri (McGregor) (Acari: Tetranychidae)

Molecular Biology Reports volume 39, pages 5841–5849 (2012)Cite this article

Abstract

Quantitative real time reverse transcriptase polymerase chain reaction (RT-qPCR) is preferred for gene expression analysis in living organisms. Currently, it is a valuable tool for biological and ecological studies as it provides a relatively straightforward way to assess the relevance of transcriptional regulation under developmental and stress tolerance conditions. However, studies have shown that some commonly used reference genes varied among different experimental treatments, thus, systematic evaluation of reference genes is critical for gene expression profiling, which is often neglected in gene expression studies of arthropods. The aim of this study is to identify the suitable reference genes for RT-qPCR experiments involving various developmental stages and/or under abiotic stresses in citrus red mite Panonychus citri, a key pest in citrus orchards worldwide. GeNorm, NormFinder, and Bestkeeper software analysis indicates that elongation factor-1 alpha (ELF1A), RNA polymerase II largest subunit, alpha tublin, and glyceraldhyde-3-phosphate dehydrogenase (GAPDH) are the most stable reference genes in various developmental stages, meanwhile, ELF1A and GAPDH were the most stable reference genes under various abiotic stresses. Furthermore, this study will serve as a resource to screen reference genes for gene expression studies in any other spider mite species.

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Abbreviations

RT-qPCR:

Quantitative real time reverse transcriptase polymerase chain reaction

ACTB:

Beta actin

TUBA:

Alpha tublin

ELF1A:

Elongation factor-1 alpha

GAPDH:

Glyceraldhyde-3-phosphate dehydrogenase

RNAP II:

RNA polymerase II largest subunit

SDHA:

Succinate dehydrogenase complex subunit A

5.8S rRNA:

5.8 S ribosomal RNA

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Acknowledgments

This study was supported in part by a Special Fund for Agro-scientific Research in the Public Interest (201103020), the Program for Changjiang Scholars and Innovative Research Team in University (IRT0976), Natural Science Foundation of Chongqing (CSTC, 2009BA1042), and the earmarked fund for Modern Agro-industry (Citrus) Technology Research System of China (CARS-27-06B) to Jin-Jun Wang; as well as the Specialized Research Fund for the Doctoral Program of Higher Education (20100182120022) and the Fundamental Research Funds for the Central Universities (XDJK2009C112) of China to Wei Dou. The manuscript was critically read by Dr. Helen Hull-Sanders.

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Affiliations

  1. Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing, 400716, People’s Republic of China

    Jin-Zhi Niu, Wei Dou, Tian-Bo Ding, Li-Hong Yang, Guang-Mao Shen & Jin-Jun Wang

  2. Mianyang Normal University, Mianyang, 631000, Sichuan, People’s Republic of China

    Li-Hong Yang

Authors
  1. Jin-Zhi Niu
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  2. Wei Dou
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  3. Tian-Bo Ding
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  4. Li-Hong Yang
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  5. Guang-Mao Shen
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  6. Jin-Jun Wang
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Corresponding author

Correspondence to Jin-Jun Wang.

Additional information

Jin-Zhi Niu and Wei Dou contributed equally to this work.

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Niu, JZ., Dou, W., Ding, TB. et al. Evaluation of suitable reference genes for quantitative RT-PCR during development and abiotic stress in Panonychus citri (McGregor) (Acari: Tetranychidae). Mol Biol Rep 39, 5841–5849 (2012). https://doi.org/10.1007/s11033-011-1394-x

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  • DOI: https://doi.org/10.1007/s11033-011-1394-x

Keywords

  • RT-qPCR
  • Panonychus citri
  • Housekeeping genes
  • Gene expression
  • Spider mites