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Ailsa Craig. FIGURE 4 The effect of reduced NO production on Botrytis http://www.selleckchem.com/products/DAPT-GSI-IX.html cinerea infections of sitiens. (A) Representative B. cinerea lesion phenotypes in cv. Ailsa Craig and sitiens sprayed with either water or 5 mM Nitro-L-arginine methyl ester (L-NAME); an inhibitor of NO ... ABA Suppresses B. cinerea NO and ROS Production As sitiens is deficient in ABA, it was hypothesized that exogenous application of ABA could reduce NO production. Thus, sitiens plants as well as cv. Ailsa Craig were sprayed with 100 mM ABA and then after 1 h, we challenged with B. cinerea. Examining the effect of ABA treatment on lesion development in sitiens suggested that lesion sizes at 72 hpi tended to be greater compared to untreated controls. The phenotypic effects of ABA on cv. Ailsa Craig were more difficult to assess visually (Figure ?Figure5A5A). This slight effect of ABA on B. cinerea infected on of cv. Ailsa Craig was also suggested from estimations of fungal biomass at 72 hpi (Figure ?Figure5B5B). Although there was a trend toward increased fungal development with ABA treatment differences were not significant compared to controls. This was in contrast to sitiens where the application of ABA significantly increased fungal biomass suggesting that resistance was being reduced. FIGURE 5 Nitric oxide production in B. cinerea challenged sitiens treated with abscisic acid lazabemide (ABA). (A) cv. Ailsa Craig or sitiens plants were sprayed with either 100 mL ABA in 0.2% Silwet (v/v) or with water [+0.2% Silwet (v/v)] and then multiply spot-inoculated ... The impact on NO on ABA in sitiens was measured using the QCL-based system (Figure ?Figure5C5C). Logistical constraints limited the number of samples that could be measured so only the effect of ABA on sitiens was determined following challenge with B. cinerea. NO production in the control plant was in line with that previously observed (e.g., Figure ?Figure3A3A). However, rates of NO production were reduced by ?half in two ABA treated sitiens plants (Figure ?Figure5C5C). In a seminal paper, Asselbergh et al., (2007) linked resistance in sitiens to elevated H2O2 production. We assessed if NO could also be contributing to H2O2 generation in cv. Ailsa Craig and sitiens in cores from tomato leaves at 24 h after inoculation with B. cinerea and stained with DAB (Figure ?Figure6A6A). Lesions with brown Veliparib molecular weight DAB staining were clearly observed in cv. Ailsa Craig and this appeared to be reduced by treatment with L-NAME and ABA. Greater staining was observed in sitiens compared to cv. Ailsa Craig and this was also reduced by L-NAME and ABA. This was further suggested when the extent of staining was measured using image analysis software (Figure ?Figure6B6B). Although a semi-quantitative measure at best, the data suggested significant reductions (P