Supplementary Figure 8: RNAi-mediated HisR silencing in NIL R-43
. a, Disease symptoms 15 dai of T0 transgenic plants #1 and #2 and the control R-43. Only one spike was used in inoculation. b, Semiquantitative RT-PCR of His-RNAi in the T0 transformants. M, DNA size standard in bp. The experiments were repeated independently three times with similar results. c, HisR expression levels in the two T0 transgenic plants relative to HisR in R-43 based on quantitative RT-PCR. Values are represented as the means ± s.d. (n = 3 technical replicates). In a–c, R43 indicates transgene-negative plants from the transformation experiment. d, Representative disease symptoms 15 dai of the derived T1 transgenic plants expressing the transgenes together with the resistance control R-43 and susceptible control PH691. e, HisR expression levels in the T1 transgenic lines relative to R-43. Values are represented as the means ± s.d. The number of biologically independent samples (n) and the P values for two-tailed Student’s t-test for comparison of HisR expression in the transgenic lines to that in R-43 are shown. f, NDS of the T1 lines. The number of inoculated spikes (n) and the P values for two-tailed Student’s t-test for comparison of the transgenic segregants to the transgene-negative segregants in each progenies are shown. g, Resistance phenotypes 15 dai of four transgenic T1 segregants without obvious HisR silencing. One to three spikes per plant were evaluated. h, Semiquantitative RT-PCR of the His-RNAi construct in the four plants mentioned in g. M, DNA size standard in bp. The experiments were repeated independently three times with similar results. i, HisR expression levels in the four plants mentioned in g relative to HisR expression in R-43, (n = 2 biologically independent experiments, each with three technical replicates). j, Disease symptoms 15 dai of two homozygous T2 transgenic families together with R-43. k, l, NDS (k) and LDR (l) of the two homozygous T2 transgenic families and R-43, with the box-and-whisker plots showing the medians (orange line), upper and lower quartiles (box edges), 1.5 × interquartile range (whiskers), and outliers (dots). The number of inoculated spikes (n) and the P values for two-tailed Student’s t-test for comparison of the transgenic plants to R-43 in each family are shown.
