High-level expression of alternative oxidase protein sequences enhances the spread of viral vectors in resistant and susceptible plants.
The alternative oxidase (AOX) is the terminal oxidase of the cyanide-resistant alternative respiratory pathway in plants and has been implicated in resistance to viruses. When tobacco mosaic virus (TMV) vectors were used to drive very high levels of expression of either AOX or AOX mutated in its active site (AOX-E), virus spread was enhanced. This was visualized as the induction of larger hypersensitive-response lesions after inoculation onto NN-genotype tobacco than those produced by vectors bearing sequences of comparable length [the green fluorescent protein (gfp) gene sequence or antisense aox] or the 'empty' viral vector. Also, in the highly susceptible host Nicotiana benthamiana, systemic movement of TMV vectors expressing AOX or AOX-E was faster than that of TMV constructs bearing gfp or antisense aox sequences. Notably, in N. benthamiana, TMV.AOX and TMV.AOX-E induced symptoms that were severe and ultimately included cell death, whereas the empty vector, TMV.GFP and the TMV vector expressing antisense aox sequences never induced necrosis. The results show that, if expressed at sufficiently high levels, active and inactive AOX proteins can affect virus spread and symptomology in plants.
Murphy
Alex M
AM
Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK.
Gilliland
Androulla
A
Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK.
York
Caroline J
CJ
Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK.
Hyman
Belinda
B
Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK.
Carr
John P
JP
Department of Plant Sciences, University of Cambridge, Cambridge CB2 3EA, UK.
D017209:Apoptosis; D001665:Binding Sites; D005822:Genetic Vectors / Q000502:physiology*; D024101:Mitochondrial Proteins; D009336:Necrosis; D010088:Oxidoreductases / Q000235:genetics / Q000502:physiology*; D010101:Oxygen Consumption; D010940:Plant Proteins; D014026:Nicotiana / Q000821:virology*; D014027:Tobacco Mosaic Virus / Q000201:enzymology* / Q000502:physiology; D000099188:Alternative Oxidase
D024101:Mitochondrial Proteins; D010088:Oxidoreductases; D010940:Plant Proteins; D000099188:Alternative Oxidase