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Development of a virus-induced gene silencing (VIGS) system for Spinacia oleracea L.

  • Functional Genomics
  • Published:
In Vitro Cellular & Developmental Biology - Plant Aims and scope Submit manuscript

Abstract

Virus-induced gene silencing (VIGS) is known as a rapid and efficient system for studying functions of interesting genes in plants. Tobacco rattle virus (TRV) is widely applied for the gene silencing of many plants. Although spinach is a TRV-susceptible plant, a TRV-based VIGS system has not yet been developed for spinach. In this study, we established a TRV-based VIGS system for spinach. To evaluate the functionality of the TRV-based VIGS system, the phytoene desaturase gene (SoPDS) was first isolated from spinach as a marker gene. Then, the VIGS vector pTRV2 was combined with the partial fragment of SoPDS gene in sense or antisense orientation. Using the Agrobacterium infiltration method, we introduced the pTRV2-SoPDS clone to silence the SoPDS gene in spinach. SoPDS was efficiently silenced, and consequently, greater than 90% of newly emerging leaves exhibited severe chlorosis symptoms in the treated plants. Levels of chlorosis symptoms were similar in both plants infected with pTRV2 vectors harboring sense (SoPDS_S) or antisense (SoPDS_A) gene fragments. Quantitative analysis of SoPDS gene expression by qRT-PCR revealed that gene expression was reduced by greater than 90% in both SoPDS_S and SoPDS_A VIGS plants. Chlorosis on leaves was prolonged up to 4~5 wk after Agrobacterium infiltration. The TRV-based VIGS system was effective in silencing the SoPDS gene in spinach, suggesting that it can be a useful reverse genetics tool for the functional study of spinach genes.

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Acknowledgments

This work was performed with the support of the “Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ011289032016),” the Rural Development Administration, Republic of Korea, and the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2009-0094059, 2016R1A6A1A03012862, and 2016R1D1A1B02012307).

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Authors and Affiliations

  1. Department of Biotechnology, College of Applied Life Science (SARI), Jeju National University, Jeju, 63243, Republic of Korea

    Jungmin Lee, Dang-Viet Cao, Jiwon Kim, Reniel S. Pamplona, Key-Zung Riu & Kyung-Hwan Boo

  2. Woojung Institute of Life Sciences, Woojung BSC, Suwon, Gyeonggi, 16229, Republic of Korea

    Jongcheol Ahn

  3. Department of Plant Biology and Biotechnology, Faculty of Life Science, University of Copenhagen, Thovaldsensvej 40, Frederiksberg, 1871, Copenhagen, Denmark

    Seok-Keun Cho & Seong-Wook Yang

  4. Subtropical/tropical Organism Gene Bank, Jeju National University, Jeju, 63243, Republic of Korea

    Key-Zung Riu & Kyung-Hwan Boo

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  1. Jungmin Lee
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  2. Dang-Viet Cao
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  3. Jiwon Kim
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Correspondence to Kyung-Hwan Boo.

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Editor: Ewen Mullins

Jungmin Lee and Dang-Viet Cao contributed equally to this work.

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Lee, J., Cao, DV., Kim, J. et al. Development of a virus-induced gene silencing (VIGS) system for Spinacia oleracea L.. In Vitro Cell.Dev.Biol.-Plant 53, 97–103 (2017). https://doi.org/10.1007/s11627-017-9806-9

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