Characterization of tobacco mosaic virus isolated from tomato in India

Tomato mosaic tobamovirus (ToMV) differs from the type strain of tobacco mosaic virus (TMV) in producing local lesions instead of systemic infection on Nicotiana sylvestris. An isolate collected from Kolar district of Karnataka which produced this differential host reaction was propagated in the greenhouse on N. tabacum cp. Samsun and purified. The virus is a rigid rod shaped particle with a coat protein of molecular weight 18 kDa and genomic RNA of size 6.3 kb. A cDNA library was constructed using a specific primer designed based on the conserved nucleotide sequence at the 3 non coding region of tobamoviruses. The cDNA library was screened for recombinant Clones and, the recombinant clone 82 with an insert of size 1.04 kb was sequenced in both directions. This sequence was compared with the genomic sequence of TMV and ToMV which showed 93.1 and 73.7 per cent identity, respectively. The sequence encompassed the 3 non coding region, the complete coat protein ORF and 467 nucleotides of the, movement protein. The deduced amino acid sequence of the coat protein was compared with that of TMV and ToMV. This-sequence was nearly identical to TMV with nine amino acid changes whereas thirty-two changes were observed with ToMV. This suggests that the virus under study is a strain of TMV and therefore we have named it as tobacco mosaic virus tomato strain from Karnataka, India (TMV(Tom-K))

Engineering resistance against physalis mottle tymovirus by expression of the coat protein and 3' noncoding region

A 748 nucleotides cDNA fragment corresponding to the 3' terminal of physalis mottle virus, PhMV (formerly known as belladonna mottle virus) ($\#$Y16104) genomic RNA encompassing the tymobox, coat protein ORF and 3' noncoding region was cloned into the binary vector pKYLX 71 35 $S^2$ and introduced into N. tabacum cv. Havana plants using Agrobacterium-mediated transformation. The R0 transgenic plants showed accumulation of coat protein which self-assembled into capsids in vivo. The transgenic R1 and R2 plants showed delay in symptom expression and virus accumulation upon challenge with PhMV. 55 and 65% of the plants showed no detectable symptoms in the R1 and R2 transgenic plants respectively, when challenged with 10 $\mu$g/ml virus. Further, no detectable symptoms were observed in 75% and 25% of the R1 and R2 transgenic plants respectively, after 50 days of post infection when challenged with 10 $\mu$g/ml RNA. Thus the expression of PhMV coat protein and 3' noncoding sequence confers a high level of resistance against PhMV infection

Vitamin E TPGS and Its Applications in Nutraceuticals

Vitamin E TPGS (d-α-tocopheryl polyethylene glycol 1000 succinate), the water-soluble derivative of the naturally occurring d-a-tocopherol, was used initially to overcome malabsorption and correct severe vitamin E deficiency in cholestatic children. The observation that its administration increased the absorption of vitamin D focused attention on its amphiphilic properties as nonionic surfactant and applications in solubilizing lipophilic and poorly soluble compounds and enhancing their absorption and bioavailability. Further research has evaluated its safety and efficacy which combines solubilization and formation of micelle-like particles with inhibition of P-glycoprotein-mediated efflux, the key mechanism to the development of drug resistance. These properties expanded its applications in pharmaceuticals and dietary, disease-specific, supplements for malabsorbing patients. Emerging research on major nutraceuticals and the developing field of cannabinoids have shown that poor water solubility and extensive first-pass metabolism cause poor absorption and bioavailability. This chapter describes the properties, safety, and efficacy of vitamin E TPGS with a focus on its applications in nutraceuticals and cannabinoids