Construction and biological activities of the first infectious cDNA clones of the genus Foveavirus

• Published in: Virology (New York, N.Y.) N.Y.), 2013-01, Vol.435 (2), p.453-462
• Main Authors: Meng, Baozhong, Venkataraman, Srividhya, Li, Caihong, Wang, Weizhou, Dayan-Glick, Cathy, Mawassi, Munir
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; Agro-infiltration; BRASSICA; Cauliflower mosaic virus; Cloning, Molecular; Disseminated infection; DNA, Complementary - genetics; ELECTRON MICROSCOPY; Flexiviridae - genetics; Flexiviridae - pathogenicity; FLUORESCENCE; Fluorescence microscopy; Foveavirus; Green fluorescent protein; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Immuno electron microscopy; Infectious Disease; Microscopy, Fluorescence; Microscopy, Immunoelectron; movement protein; Necrosis; NICOTIANA; Nicotiana - virology; Nicotiana benthamiana; Plant Diseases - virology; Plant Stems - virology; Plant viruses; POLYMERASE CHAIN REACTION; Promoters; Replication; Reverse Transcriptase Polymerase Chain Reaction; RT-PCR; Sequence Analysis, DNA; Stem pitting; Transcription; Veins; Virion; Virions; VIRUSES; Vitaceae; Vitis - virology; Western blotting
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1016/j.virol.2012.09.045

Abstract Grapevine rupestris stem pitting-associated virus (GRSPaV, genus Foveavirus , family Betaflexiviridae) is one of the most prevalent viruses in grapevines and is associated with three distinct diseases: rupestris stem pitting, vein necrosis and Syrah decline. Little is known about the biology and pathological properties of GRSPaV. In this work, we engineered a full-length infectious cDNA clone for GRSPaV and a GFP-tagged variant, both under the transcriptional control of Cauliflower mosaic virus 35 S promoter. We demonstrated that these cDNA clones were infectious in grapevines and Nicotiana benthamiana through fluorescence microscopy, RT-PCR, Western blotting and immuno electron microscopy. Interestingly, GRSPaV does not cause systemic infection in four of the most commonly used herbaceous plants, even in the presence of the movement proteins of two other viruses which are known to complement numerous movement-defective viruses. These infectious clones are the first of members of Foveavirus which would allow further investigations into mechanisms governing different aspects of replication for GRSPaV and perhaps related viruses.