期刊
IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-PLANT
卷 53, 期 2, 页码 97-103出版社
SPRINGER
DOI: 10.1007/s11627-017-9806-9
关键词
Phytoene Desaturase gene; Spinach; Tobacco rattle virus; Virus-induced gene silencing
资金
- Cooperative Research Program for Agriculture Science & Technology Development [PJ011289032016]
- Rural Development Administration, Republic of Korea
- National Research Foundation of Korea (NRF)
- Ministry of Education [2009-0094059, 2016R1A6A1A03012862, 2016R1D1A1B02012307]
- Rural Development Administration (RDA), Republic of Korea [PJ011289032016] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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 similar to 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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