Virus‐induced gene silencing in tomato

TLDR

TRV‐based virus‑induced gene silencing has been successfully applied in Nicotiana benthamiana and is now shown to infect tomato plants and silence genes efficiently. The goal is to use a Gateway‑compatible TRV vector to enable large‑scale functional analysis of tomato expressed sequence tags. The authors constructed a restriction‑ and ligation‑free TRV vector that allows cloning of tomato ESTs with a single primer set and used it to silence PDS, CTR1, CTR2, RbcS, and a cLED3L14 homolog. TRV infection of tomato leads to efficient gene silencing, with CTR1 knockdown producing an ethylene‑responsive phenotype similar to Arabidopsis ctr1 mutants and successful silencing of RbcS and a cLED3L14 homolog.

Abstract

Abstract 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.