VIGS of CaNAC2c/d and CaHSFA5 in pepper plants
To silence CaNAC2c(d) or CaHSFA5 in pepper plants by virus
induced gene silencing (VIGS), GV3101 cells containing PYL192 and
PYL279-CaNAC2c (CaNAC2d , CaHSFA5 ), or PYL279 (as a
negative control) were mixed and resuspended in the induction medium at
1:1 ratio to OD600 = 0.6, which were infiltrated into
cotyledons of 2-week-old pepper plants. To simultaneously silenceCaNAC2c and CaNAC2d in pepper plants, GV3101 cells
containing PYL192 and PYL279-CaNAC2c and cells containing PYL192
and PYL279-CaNAC2d were mixed at ratio 1:1 and resuspended in the
induction medium to OD600 = 0.6. 100 μl were infiltrated
into cotyledons of 2-week-old pepper plants, which were placed in dark
at 16 ℃ for 56 h, and then moved to a growth room under conditions of 28
℃, 60–70 mmol photons m-2 s-1, a
relative humidity of 70%, and a 16-h light/8-h dark. At 15 days post
inoculation (dpi), the success and specificity of gene silencing were
assayed in pepper plants challenged with HTS by measuring the transcript
level of CaNAC2c (CaNAC2d , CaHSFA5 ) or the
transcript level of CaNAC2c and CaNAC2d .