Agroinoculation of Beet necrotic yellow vein virus cDNA clones results in plant systemic infection and efficient Polymyxa betae transmission

• Published in: Molecular plant pathology Vol. 14; no. 4; pp. 422 - 428
• Main Authors: Delbianco, Alice, Lanzoni, Chiara, Klein, Elodie, Rubies Autonell, Concepcion, Gilmer, David, Ratti, Claudio
• Format: Journal Article
• Language: English
• Published: Oxford Blackwell Publishing Ltd 01-05-2013; Blackwell; Wiley; John Wiley and Sons Inc
• Subjects: Agrobacterium tumefaciens - metabolism; Animals; Biochemistry, Molecular Biology; Biological and medical sciences; Blotting, Northern; Blotting, Western; Cellular Biology; DNA, Complementary - genetics; Fundamental and applied biological sciences. Psychology; Genes, Reporter; Genetic Techniques; Life Sciences; Molecular biology; Nicotiana - parasitology; Nicotiana - virology; Phytopathology. Animal pests. Plant and forest protection; Plant Diseases - parasitology; Plant Diseases - virology; Plant Leaves - parasitology; Plant Leaves - virology; Plant Viruses - genetics; Plasmodiophorida - physiology; Technical Advance; Virus; Infection; Protozoa; Plant; Plant pathogen; Complementary DNA; Polymyxa betae; Systemic; Benyvirus; Mycetozoa; Clone; Beet necrotic yellow vein virus
• ISSN: 1464-6722; 1364-3703
• DOI: 10.1111/mpp.12018

Summary
Agroinoculation is a quick and easy method for the infection of plants with viruses. This method involves the infiltration of tissue with a suspension of Agrobacterium tumefaciens carrying binary plasmids harbouring full‐length cDNA copies of viral genome components. When transferred into host cells, transcription of the cDNA produces RNA copies of the viral genome that initiate infection. We produced full‐length cDNA corresponding to Beet necrotic yellow vein virus (BNYVV) RNAs and derived replicon vectors expressing viral and fluorescent proteins in pJL89 binary plasmid under the control of the Cauliflower mosaic virus 35S promoter. We infected Nicotiana benthamiana and Beta macrocarpa plants with BNYVV by leaf agroinfiltration of combinations of agrobacteria carrying full‐length cDNA clones of BNYVV RNAs. We validated the ability of agroclones to reproduce a complete viral cycle, from replication to cell‐to‐cell and systemic movement and, finally, plant‐to‐plant transmission by its plasmodiophorid vector. We also showed successful root agroinfection of B. vulgaris, a new tool for the assay of resistance to rhizomania, the sugar beet disease caused by BNYVV.