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Use of the modified viral satellite DNA vector to silence mineral nutrition-related genes in plants: silencing of the tomato ferric chelate reductase gene, FRO1, as an example
Authors:HE XiuXia  JIN ChongWei  LI GuiXin  YOU GuangYi  ZHOU XuePing &  ZHENG ShaoJian
Institution:(1) Key Laboratory for Environmental and Ecosystem Health of the Ministry of Education, College of Environmental and Resource Science, Zhejiang University, Hangzhou, 310029, China;(2) College of Life Science, Changchun University of Science and Technology, Changchun, 130022, China;(3) Institute of Biotechnology, Zhejiang University, Hangzhou, 310029, China;(4) State Key Laboratory of Plant Physiology and Biochemistry, College of Life Science, Zhejiang University, Hangzhou, 310058, China
Abstract:Virus-induced gene silencing (VIGS) is potentially an attractive reverse-genetics tool for studies of plant gene function, but whether it is effective in silencing mineral nutritional-related genes in roots has not been demonstrated. Here we report on an efficient VIGS system that functions in tomato roots using a modified viral satellite DNA (DNAmβ) associated with Tomato yellow leaf curl China virus (TYLCCNV). A cDNA fragment of the ferric chelate reductase gene (FRO1) from tomato was inserted into the DNAmβ vector. Tomato roots agro-inoculated with DNAmβ carrying both a fragment of FRO1 and TYLCCNV used as a helper virus exhibited a significant reduction at the FRO1 mRNA level. As a consequence, ferric chelate reductase activity, as determined by visualization of the pink FeBPDS3 complex was significantly decreased. Our results clearly demonstrated that VIGS system can be employed to investigate gene function associated with plant nutrient uptake in roots. Contributed equally to this work Supported by the National Natural Science Foundation of China (Grant No. 30625026) and Cultivation Fund of the Key Scientific and Technical Innovation Project, Ministry of Education of China (Grant No. 705025)
Keywords:virus-induced gene silencing  roots  ferric chelate reductase
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