National Academy of Agricultural Sciences (NAAS)
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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
VIGS is a powerful tool for assaying the effects of gene silencing in plants. Efficient silencing requires the fragments that are more than 90% homologous to the target gene. Here, we designed a set of universal primers to a region of the PDS gene that is highly conserved among species, and therefore allows to isolate a fragment of the homologous PDS gene from the species of interest i.e., tomato. We typically use silencing of PDS which is used as a marker for gene silencing in tomatoes involved in carotene biosynthesis which results in a characteristic photobleaching phenotype in the leaves. The purpose of this study was to investigate the possibility of tomato PDS gene silencing, using VIGS technique by TRV based VIGS vector containing tomato PDS. The pTRV2: PDS vector carried inserts derived from tomato PDS gene was transformed into DH5α E. coli strain and then Agrobacterium tumefaciens GV3101 strain cells were transformed by the vectors containing PDS genes. The construct was validated for physical integrity as well as for biological functionality i.e., PDS by agroinfiltration method in the experimental host plant. The construct shows gene silencing results in photobleaching phenotype occurred in tomato plant leaves. However, plant infiltration with pTRV1 and pTRV2 (without PDS gene as control) mixed culture did not show any photobleaching. This construct found structurally as well as functionally active under plant system and can be further modified into transgene construct to be used to study the function of other genes through VIGS in tomato.
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