A universal expression/silencing vector in plants.
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.
Peretz
Yuval
Y
Hebrew University of Jerusalem, Faculty of Agricultural, Food and Environmental Quality Sciences, Institute for Plant Sciences and Genetics, Rehovot 76100, Israel.
Mozes-Koch
Rita
R
Akad
Fuad
F
Tanne
Edna
E
Czosnek
Henryk
H
Sela
Ilan
I
D000818:Animals; D004276:DNA, Satellite; D004277:DNA, Single-Stranded; D004926:Escherichia coli / Q000235:genetics; D052677:Flexiviridae; D017791:Geminiviridae / Q000235:genetics*; D015870:Gene Expression; D020868:Gene Silencing; D005818:Genetic Engineering; D005822:Genetic Vectors; D018745:Genome, Plant; D006430:Hemiptera / Q000821:virology; D010944:Plants / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D010957:Plasmids; D012330:RNA, Double-Stranded; D014026:Nicotiana / Q000821:virology; D015533:Transcriptional Activation
D004276:DNA, Satellite; D004277:DNA, Single-Stranded; D012330:RNA, Double-Stranded