Development of a virus induced gene silencing vector from a legumes infecting tobamovirus.
Medicago truncatula, the model plant of legumes, is well characterized, but there is only a little knowledge about it as a viral host. Viral vectors can be used for expressing foreign genes or for virus-induced gene silencing (VIGS), what is a fast and powerful tool to determine gene functions in plants. Viral vectors effective on Nicotiana benthamiana have been constructed from a number of viruses, however, only few of them were effective in other plants. A Tobamovirus, Sunnhemp mosaic virus (SHMV) systemically infects Medicago truncatula without causing severe symptoms. To set up a viral vector for Medicago truncatula, we prepared an infectious cDNA clone of SHMV. We constructed two VIGS vectors differing in the promoter element to drive foreign gene expression. The vectors were effective both in the expression and in the silencing of a transgene Green Fluorescent Protein (GFP) and in silencing of an endogenous gene Phytoene desaturase (PDS) on N. benthamiana. Still only one of the vectors was able to successfully silence the endogenous Chlorata 42 gene in M. truncatula.
Agricultural Biotechnology Centre, Plant Biology Institute, Gödöllő, Hungary. varallya@abc.hu
Eva
E
Várallyay
Zsuzsanna
Z
Lichner
Judit
J
Sáfrány
Z
Z
Havelda
P
P
Salamon
Gy
G
Bisztray
J
J
Burgyán
D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D007887:Fabaceae / Q000235:genetics*; D005786:Gene Expression Regulation; D020868:Gene Silencing; D005821:Genetic Techniques; D005822:Genetic Vectors; D023281:Genomics / Q000379:methods; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D008957:Models, Genetic; D009841:Oligonucleotides / Q000235:genetics; D010088:Oxidoreductases / Q000737:chemistry; D017898:Tobamovirus / Q000235:genetics*
D018076:DNA, Complementary; D009841:Oligonucleotides; D049452:Green Fluorescent Proteins; D010088:Oxidoreductases; C022398:phytoene dehydrogenase