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Insights on mechanical transmission of turnip mosaic virus (TuMV) mutants in Nicotiana benthamiana

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Title: Insights on mechanical transmission of turnip mosaic virus (TuMV) mutants in Nicotiana benthamiana
Author(s): Jyakhwa, Sarkal
Contributor: University of Helsinki, Faculty of Agriculture and Forestry
Degree program: Erasmus-Mundus Master's Programme in Plant Breeding (EMPLANT)
Specialisation: Plant production sciences
Language: English
Acceptance year: 2022
Turnip mosaic virus (TuMV) is an economically important plant RNA virus causing huge damage to wide range of arable and vegetable crops. A study was conducted in Nicotiana benthamiana to know if a TuMV mutant carrying a mutation in a thoroughly conserved WD-domain interacting motif and WG motif in HCPro protein can be mechanically transmitted to a healthy plant or not. HCPRoWD is a mutation in “AELPR” motif where glutamic acid and arginine are replaced by alanine. This mutated virus is here referred as TuMVWD. Similarly, in TuMVAG the tryptophan residue in the WG pair is changed to alanine and this mutated HCPro is called as TuMVAG. Four treatments, TuMVWT (positive control), Mock (negative control), TuMVWD and TuMVAG were made. Three plants per treatment were agroinfiltrated and five plants per treatment were used for mechanical inoculation experiment. Green fluorescent protein (GFP), a quantitative reporter of gene expression, was measured followed by qPCR for quantification of vRNA (viral RNA) accumulation. In agroinfiltrated plants, newly emerged leaves showed strong fluorescence in TuMVWT and TuMVAG by 14 dpi (days post inoculation), but TuMVWD showed poor GFP as compared to TuMVWT. During mechanical inoculation experiment, none of the treatments developed GFP in systemic leaves by six dpi but by 14 dpi GFP accumulation in the upper leaves of TuMVWT and TuMVAG was increased. TuMVWD was not used for 2nd mechanical experiment as it did not cause systemic infection during 1st mechanical inoculation experiment even by 14 dpi. Results from vRNA accumulation showed that mechanical transmission of virus was reduced with TuMVAG and not possible with TuMVWD. However, mutations had negative effect on vRNA accumulation.
Keyword(s): Keywords Potyvirus agroinfiltration mechanical inoculation vRNA qPCR systemic infection.

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