A Universal Expression/Silencing Vector in Plants

Abstract

A universal vector (IL-60 and auxiliary constructs), expressing or silencing genes in every plant tested to date, is described. Plants that have been successfully manipulated by the IL-60 system include hard-to-manipulate species such as wheat (Triticum duram), pepper (Capsicum annuum), grapevine (Vitis vinifera), citrus, and olive (Olea europaea). Expression or silencing develops within a few days in tomato (Solanum lycopersicum), wheat, and most herbaceous plants and in up to 3 weeks in woody trees. Expression, as tested in tomato, is durable and persists throughout the life span of the plant. The vector is, in fact, a disarmed form of Tomato yellow leaf curl virus, which is applied as a double-stranded DNA and replicates as such. However, the disarmed virus does not support rolling-circle replication, and therefore viral progeny single-stranded DNA is not produced. IL-60 does not integrate into the plant's genome, and the construct, including the expressed gene, is not heritable. IL-60 is not transmitted by the Tomato yellow leaf curl virus's natural insect vector. In addition, artificial satellites were constructed that require a helper virus for replication, movement, and expression. With IL-60 as the disarmed helper “virus,” transactivation occurs, resulting in an inducible expressing/silencing system. The system's potential is demonstrated by IL-60-derived suppression of a viral-silencing suppressor of Grapevine virus A, resulting in Grapevine virus A-resistant/tolerant plants.

Ubiquitination-Related MdBT Scaffold Proteins Target a bHLH Transcription Factor for Iron Homeostasis

“…The 'Gala' cultures and transgenic apple plantlets were grown on Murashige and Skoog medium containing 0.6 mg L 21 6-benzylaminopurine 35S-GFP/MYC vector, so that the genes were under the control of the CaMV 35S promoter. The viral vector pIR was used (Peretz et al, 2007). To construct the pIRMdCUL3-anti silencing vector, the 59-UTRs of either MdCUL3 or MdbHLH104 (up to 300 bp) were inserted into pIR.…”

Resources for Virus-Induced Gene Silencing in the Grasses

“…An interesting possibility is Tomato yellow leaf curl virus, which can infect and express reporter genes in a very wide range of dicot and monocot plants. This virus has been modified to serve as a VIGS vector, but has not yet been tested for silencing in a monocot (Peretz et al, 2007).…”

MdCOP1 Ubiquitin E3 Ligases Interact with MdMYB1 to Regulate Light-Induced Anthocyanin Biosynthesis and Red Fruit Coloration in Apple

“…To observe the effects of MdCOP1-1 and MdCOP1-2 overexpression or suppression in apple fruits, viral vectors were used as described by Ratcliff et al (2001) and Peretz et al (2007). The full-length cDNAs of MdCOP1-1 and MdCOP1-2 were amplified using the primer pairs BDP1-F/-R and BDP2-F/-R and then cloned into the pIR vector under the control of the 35S promoter (Supplemental Table S1).…”