All analyses were performed with SAS V8.2 statistics software. All means and standard errors

are presented as untransformed values. Besides tiller number, all other agronomic traits differed across cultivars (Table S1), with Kanlow displaying more biomass, leaf area and root surface area, and longer culms across all N deficiency treatments (Fig. 1). No significant difference was observed between Alamo and Kanlow in any traits but tiller number (Fig. 1). All cultivars of lowland ecotypes outperformed upland cultivars, and no significant difference was observed among upland cultivars for any trait (Fig. 2). There were significant cultivar-by-treatment and ecotype-by-treatment interactions for all agronomic traits except tiller number. Capmatinib supplier Tiller number showed only extremely strong responses to treatment (Table S1). Aside from tiller number and Veliparib nmr R:S, all other agronomic traits varied

across ecotypes (Table S1), with lowland cultivars producing 47% more biomass, 58% longer culms, 48% more leaf area, and 42% more root surface area than upland cultivars (Fig. 2). Nitrogen deficiency affected agronomic traits, and all traits showed large differences across the four treatments, with the control yielding an average of 168% more total biomass, 148% more aboveground biomass, 189% more belowground biomass, 53% more tillers, 127% more leaf area, 99% more root surface area, and 58% longer culms than the N deficiency treatments (Table 2). Clearly, cultivars performed best under the control conditions, followed

by moderate stress, and worst under extreme stress. No significance for R:S was observed between the control and N1 or N2. Tiller number, leaf area, root surface area, total biomass, aboveground biomass, and belowground biomass under the N2 treatment were significantly higher than under the N1. Height and belowground biomass did not differ between the N1 and N0 treatments (Table 2). Surprisingly, there were highly significant interactions between stress treatments and cultivars for all agronomic traits 3-mercaptopyruvate sulfurtransferase but tiller number (Table S1); response to N deficiency stress depended on cultivar. For Alamo, height showed no difference across the three stress levels (Fig. 3-A); for Pathfinder, height and aboveground biomass did not differ between the N1 and N2 treatments (Fig. 3-A, D). For both ecotypes, all the agronomic traits varied across N stress treatments (Fig. 3). According to Fig. 3, accumulation can also be calculated in height, leaf area, root surface area, aboveground biomass, belowground biomass and total biomass with decreasing N level for each cultivar (data not shown). Kanlow had the lowest overall response to decreasing N concentration for the agronomic traits in Fig. 3, immediately followed by Alamo. Kanlow also showed the best performance under the three N stress treatments for all the traits.