Introducing targeted epigenetic changes in Solanum tuberosum and Nicotiana benthamiana by plant viruses

In this thesis we have focused on the induction of targeted epigenetic changes using modified Potato Virus X vector (PVX, Potato virus X). The gene coding sequence phytoene desaturase (PDS) was isloated from Solanum tuberosum and Nicotiana benthamiana plants using PCR. Primers were specifically designed as universal for both plant species and their Tm temperatures was in the range of 5 °C. The primers also added flanking restriction sites XhoI and ClaI to simplify cloning of PDS PCR products into full length PVX viral vector in AS orientation. The PVX viral vector pGR106 was later introduced into the plants using Agrobacterium tumefaciens. Infection led to the degradation of the target transcript and to efficient gene silencing (PTGS posttranscriptional gene silencing). The process has been manifested by complete or partial fading of a green plant tissue phenotype.. Some of the plants were aslso inoculated in in vitro conditions, for which a novel technique has been implemented