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Inoculation of Nicotiana tabacum with recombinant potato virus X induces RNA interference in the solenopsis mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae)

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Abstract

Objectives

The chitin synthase 1 (CHS1) gene in Phenacoccus solenopsis (PsCHS1) was evaluated as a potential target of RNA interference (RNAi) by using Potato virus X (PVX) as a vector (recombinant PVX) for expressing RNAi triggering elements in Nicotiana tabacum L.

Results

RT-PCR analysis confirmed the expression of PsCHS1 in N. tabacum inoculated with recombinant-PVX–PsCHS1 (treated). RT- and multiplex-PCR further showed a reduction in mRNA levels of the target gene in mealybugs feeding on treated plants. Mortality in parent adults and emerging nymphs (21 and 29 %) exposed to the treated plants was significantly higher (P < 0.05) than those exposed to uninoculated (−ve control) or inoculated with non-recombinant PVX (PVX-control). The number of surviving adults and the combined number of adults and nymphs (47 and 60 %) was significantly (P < 0.05) lower on the treated plants than the −ve (76 %) or PVX (74 %) control. The visual observations verified the physical deformities in mealybugs exposed to the treated plants.

Conclusion

chitin synthase 1 is a potential RNAi target in P. solenopsis and the recombinant PVX can be used as a tool to evaluate candidate RNAi triggering elements in plants.

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Acknowledgments

This study was supported by the Higher Education Commission of Pakistan.

Conflict of interest

The authors declare no financial or commercial conflict of interest.

Supporting information

Supplementary Table 1−Primers and PCR conditions used in the study.

Author information

Authors and Affiliations

  1. National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan

    Arif Muhammad Khan, Muhammad Ashfaq, Akhtar Rasool & Shahid Mansoor

  2. Department of Biotechnology, University of Sargodha, Sargodha, Pakistan

    Arif Muhammad Khan

  3. Biodiversity Institute of Ontario, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada

    Muhammad Ashfaq

  4. College of Agriculture, Bahauddin Zakariya University, Bahadur Campus Layyah, Multan, Pakistan

    Azhar Abbas Khan

  5. School of Life Sciences, Beijing Institute of Technology, Beijing, People’s Republic of China

    Javed Iqbal

Authors
  1. Arif Muhammad Khan
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  2. Muhammad Ashfaq
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  3. Azhar Abbas Khan
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  4. Akhtar Rasool
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Corresponding author

Correspondence to Muhammad Ashfaq.

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Supplementary material 1 (DOCX 16 kb)

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Khan, A.M., Ashfaq, M., Khan, A.A. et al. Inoculation of Nicotiana tabacum with recombinant potato virus X induces RNA interference in the solenopsis mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae). Biotechnol Lett 37, 2083–2090 (2015). https://doi.org/10.1007/s10529-015-1880-7

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  • DOI: https://doi.org/10.1007/s10529-015-1880-7

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