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Virus-induced gene silencing (VIGS) for functional analysis of MYB80 gene involved in Solanum lycopersicum cold tolerance

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Abstract

Virus-induced gene silencing (VIGS) is a method for transiently silencing genes in dicot and monocot plants. To evaluate the effects of chilling injury on activity of the MYB80 gene in Solanum lycopersicum, an investigation was conducted by VIGS using TRV-GFP (a modified TRV vector) to silence the target gene. During the chilling treatment (4/12 °C, 8/16-h dark/light photoperiod, with 60% humidity), the leaves were collected to analyze the malondialdehyde (MDA) content, soluble sugar content, free proline levels, and relative electric conductivity (REC). Leaves collected 2 weeks after chilling treatment were used to detect the in situ accumulation of superoxide radical (O2−). Additionally, we collected leaves at selected time points for semi-quantitative reverse transcription-PCR (RT-PCR) analysis. Eventually, 20 days after chilling treatment, all plants were evaluated at 4 °C for 7 days to assess the chilling injury index. The results validated that the MYB80 gene was related to cold tolerance of tomato plants, and that silencing of the MYB80 gene reduced the cold resistance ability.

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Abbreviations

VIGS:

Virus-induced gene silencing

TRV:

Tobacco rattle virus

MDA:

Malondialdehyde

REC:

Relative electric conductivity

NBT:

Nitro blue tetrazolium

CI:

Chilling injury

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Funding

This study received grants from the Ministry of Science and Technology for “A comparative study on the diversification of the main vegetable gene resources between China and Russia” (2011DFR31180-3).

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Authors and Affiliations

  1. Beijing Key Laboratory of New Technology in Agricultural Application, Plant Science and Technology College, Beijing University of Agriculture, Beijing, 102206, China

    Xiuhua Chen, Xuefen Duan, Shuai Wang, Weiyuan Wu & Xichun Zhang

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  1. Xiuhua Chen
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  2. Xuefen Duan
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  3. Shuai Wang
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  4. Weiyuan Wu
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  5. Xichun Zhang
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Contributions

ZXC conceived and designed the research; CXH conducted the experiments; CXH and DXF analyzed the data; and CXH, DXF, WS, and WWY wrote the paper. All authors have read and approved the final manuscript.

Corresponding author

Correspondence to Xichun Zhang.

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The authors declare that they have no conflict of interest.

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Handling Editor: Burkhard Becker

Xiuhua Chen and Xuefen Duan are co-first authors.

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Chen, X., Duan, X., Wang, S. et al. Virus-induced gene silencing (VIGS) for functional analysis of MYB80 gene involved in Solanum lycopersicum cold tolerance. Protoplasma 256, 409–418 (2019). https://doi.org/10.1007/s00709-018-1302-5

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  • DOI: https://doi.org/10.1007/s00709-018-1302-5

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