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.
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 (DNAmbeta) associated with Tomato yellow leaf curl China virus (TYLCCNV). A cDNA fragment of the ferric chelate reductase gene (FROl) from tomato was inserted into the DNAmbeta vector. Tomato roots agro-inoculated with DNAmbeta 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.
He
XiuXia
X
Key Laboratory for Environmental and Ecosystem Health of the Ministry of Education, College of Environmental and Resource Science, Zhejiang University, Hangzhou 310029, China.
Jin
ChongWei
C
Li
GuiXin
G
You
GuangYi
G
Zhou
XuePing
X
Zheng
Shao-Jian
SJ
D001483:Base Sequence; D015139:Blotting, Southern; D017931:DNA Primers; D004276:DNA, Satellite / Q000235:genetics*; D038181:FMN Reductase / Q000235:genetics*; D020868:Gene Silencing; D017343:Genes, Plant; D018551:Solanum lycopersicum / Q000201:enzymology / Q000235:genetics*; D010957:Plasmids
D017931:DNA Primers; D004276:DNA, Satellite; D038181:FMN Reductase; C023903:ferric citrate iron reductase