Browsing by Subject "16K gene"

Virusinduced gene silencing (VIGS) vectors based on tobacco rattle virus (TRV) are now widely used for characterizing the function of plant genes. However, previous TRV vectors using RNA2 to carry the targeted gene sequence had difficulties to induce gene silencing on some plant species (Gerbera hybrida etc.) due to the obstacle of RNA2 movement. To achieve efficient gene silencing in those species, it is necessary to develop new TRV vectors, in which the targeted gene will be included in TRV RNA1 and the 16K gene will be replaced. Based on TRV RNA1, two new VIGS vectors M1 and M2 were developed through deletion part of 16K gene. Another mutant 16Kstop was also constructed to carry an early terminator in the 4th codon of 16K gene. The infectivity and gene silencing efficiency of the new constructs were assessed through a series of infection experiments. It was found that the infectivity of M1 and M2 was lower than wild TRV RNA1. M1 and M2 could induce PDS gene silencing on Nicotiana benthamiana, but their gene silencing efficiency was limited as compared with previous TRV VIGS vectors in which the PDS gene fragment was contained in RNA2. We also found that the 16K gene sequence, rather than the 16K protein, was required for efficient virus movement and accumulation.