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Methods in Molecular Biology 2020

Virus-Induced Gene Silencing in Olive Tree (Oleaceae)

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Konstantinos Koudounas
Margarita Thomopoulou
Elisavet Angeli
Dikran Tsitsekian
Stamatis Rigas
Polydefkis Hatzopoulos

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Abstrak

Research on gene functions in non-model tree species is hampered by a number of difficulties such as time-consuming genetic transformation protocols and extended period for the production of healthy transformed offspring, among others. Virus-induced gene silencing (VIGS) is an alternative approach to transiently knock out an endogenous gene of interest (GOI) by the introduction of viral sequences encompassing a fragment of the GOI and to exploit the posttranscriptional gene silencing (PTGS) mechanism of the plant, thus triggering silencing of the GOI. Here we describe the successful application of Tobacco rattle virus (TRV)-mediated VIGS through agroinoculation of olive plantlets. This methodology is expected to serve as a fast tracking and powerful tool enabling researchers from diversified fields to perform functional genomic analyses in the olive tree.

Keywords: Agrobacterium tumefaciens; Functional genomics; Olea europaea; Olive; Posttranscriptional gene silencing (PTGS ); RNA interference (RNAi); Tobacco rattle virus (TRV ); Virus-induced gene silencing (VIGS ).

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