SILENCING OF F-BOX KELCH GENE FOR VIRUS RESISTANCE STUDIES IN MODEL PLANT NICOTIANA BENTHAMIANA
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Date
2019
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CSKHPKV, Palampur
Abstract
In the present investigation entitled ―Silencing of F-Box kelch gene for virus
resistance studies in model plant Nicotiana benthamiana‖, construct has been prepared for
generating the silent lines of the mentioned plant. F-Box kelch is an important protein that
associates with the viral proteins and that the protein is an important factor that helps in the
establishment of viral infection. A conserved portion of the gene was amplified in sense and
antisense orientation and in a sequential manner it was cloned into the silencing vector
pSAT4. The construct development was ascertained with the help of restriction digestion and
sequence determination. The construct is ready to be transformed into plants. For the
biochemical analysis, purified Cucumber mosaic virus (CMV) was inoculated at their 3-4 leaf
stage and infection was confined by symptom development. Both infected and healthy plants
were subjected to biochemical analysis of the phenolic compounds (total phenols, simple
phenols, Ortho-dihydric phenols and bound phenols), alkaloids and total chlorophyll and
carotenoids. It was observed that the total phenol content was increased in the infected plant
in comparison to the healthy leaves. However, total chlorophyll and carotenoids were reduced
in infected plants as compared to healthy leaves. This is expected as virus infection leads to
loss of chlorophyll as reflected by mosaic symptoms that the virus produces.
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