The main purpose of this work was studied the effect of TMV infection on plant physiology concerned active gradient photochemicals and protein expression. Impact of Tobacco mosaic tobamovirus (TMV) on active gradient photochemicals quantitative and qualitative was performed in Nicotiana tobaccum cv. white burly. TMV isolated from single local lesions infected leaves of N. glutinosa Leaves. Inoculated N. tobaccum cv. white burly with TMV that showed severe systemic mosaic symptoms and reduction of leave size. It was confirmed that these symptoms were due to the effect of TMV virus by Datura plant. Datura metel has been reported as a diagnostic plant (indicator) for mosaic virus because of the vast array of symptoms incited by viruses, the pant should be useful in comparing an unknown with a known. Analysis of TMV infected leaves by GC-mass detected biosynthesis of novel photochemicals (2-cyclopenten-1-one, Furfural, Indene, Pyrrole, Benzonitrile, Guaiacol and Oxime, methoxy-phenyl) that not detected in healthy ones. As well as decreased in nicotine with 56.17% related healthy ones. In response to TMV infection, increase of soluble protein expression contents was observed in infected leaves than healthy ones. Alterations in protein patterns were observed in N. tabaccum leaves in response to TMV infection by SDS PAGE. Plants, even common food and feed produce a broad range of bioactive chemical compounds via their so called secondary metabolism. Several of secondary bioactive are found to hold important functions in the living plants. For example, flavonoids can protect against free radicals generated during photosynthesis. Terpenoids may attract pollinators or seed dispersers, or inhibit competing plants. Alkaloids usually ward off herbivore animals or insect attacks (phytoalexins). Other secondary metabolites function as cellular signaling molecules or have other functions in the plants, so it was necessary studying the effect of virus on phytochemical components of plant.