Assessment of resistance to Oidium neolycopersici ( On ) in RNAi transformants of NIL-Ol-1 . (A), Segregation for phenotypic traits in T2 plants obtained after selfing RNAi-ALS1 + 2 transformant 216, compared with the NIL-Ol-1 untransformed plant. PCR analysis of the 35S promoter of the silencing construct showed co-segregation of altered phenotype with presence of the expected PCR product. (B) On symptoms on MM, NIL-Ol-1 and NIL-Ol-1 transformant obtained with the ALS1 + 2 RNAi construct (T2 plant). (C), On fungal biomass and (D), transcript levels of ALS1, ALS2, and ALS3 in RNAi-ALS1 + 2 T2 plants of family 216 showing an altered phenotype, compared with that in NIL-Ol-1 plants. (E), On fungal biomass and (F), transcript levels of ALS1, ALS2, and ALS3 in NPTII-containing T2 plants in which three ALS genes were targeted individually by RNAi (RNAi-ALS1, RNAi-ALS2, and RNAi-ALS3). For each plant RNA was isolated from pooled 3rd and 4th leaves. Values were normalized relative to EF, and calibrated to the levels in untransformed NIL-Ol-1 plants. Error bars represent standard deviation of three biological replicates. Asterisks indicate significant difference from the controls according to independent-samples t-test and one way analysis of variance (P <0.05).