Virus-induced gene silencing in tomato.
We have previously demonstrated that a tobacco rattle virus (TRV)-based vector can be used in virus-induced gene silencing (VIGS) to study gene function in Nicotiana benthamiana. Here we show that recombinant TRV infects tomato plants and induces efficient gene silencing. Using this system, we suppressed the PDS, CTR1 and CTR2 genes in tomato. Suppression of CTR1 led to a constitutive ethylene response phenotype and up-regulation of an ethylene response gene, CHITINASE B. This phenotype is similar to Arabidopsis ctr1 mutant plants. We have constructed a modified TRV vector based on the GATEWAY recombination system, allowing restriction- and ligation-free cloning. Our results show that tomato expressed sequence tags (ESTs) can easily be cloned into this modified vector using a single set of primers. Using this vector, we have silenced RbcS and an endogenous gene homologous to the tomato EST cLED3L14. In the future, this modified vector system will facilitate large-scale functional analysis of tomato ESTs.
Liu
Yule
Y
Department of Molecular, Cellular and Developmental Biology, OML 451, Yale University, PO Box 208104, New Haven, CT 06520-8104, USA.
Schiff
Michael
M
Dinesh-Kumar
S P
SP
D002352:Carrier Proteins / Q000235:genetics; D027682:Cation Transport Proteins; D002688:Chitinases / Q000235:genetics; D003001:Cloning, Molecular; D000079923:Copper Transporter 1; D005030:Ethylenes / Q000494:pharmacology; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D005822:Genetic Vectors / Q000235:genetics; D018551:Solanum lycopersicum / Q000235:genetics* / Q000821:virology; D008565:Membrane Proteins / Q000235:genetics; D026901:Membrane Transport Proteins; D009154:Mutation; D010641:Phenotype; D010942:Plant Viruses / Q000235:genetics / Q000254:growth & development*; D010957:Plasmids / Q000235:genetics; D011494:Protein Kinases; D012231:Rhizobium / Q000235:genetics; D000076988:Sulfate Transporters; D014026:Nicotiana / Q000821:virology
D002352:Carrier Proteins; D027682:Cation Transport Proteins; D000079923:Copper Transporter 1; D005030:Ethylenes; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; C085609:SLC26A4 protein, human; C490079:SLC31A1 protein, human; D000076988:Sulfate Transporters; C036216:ethylene; D011494:Protein Kinases; C477753:CTR1 protein, Arabidopsis; D002688:Chitinases