Background The main objectives of this study were to find the possible structural association between the activity of enzymatic antioxidants and the grain yield of triticale plants as well as identifying the genotypic variability which might be effective on this association. Accordingly, expression levels of superoxide dismutase (SOD) isozymes (Mn-SOD, Cu/Zn-SOD, and Fe-SOD) were appraised to distinguish any possible relationship between SOD expression and drought resistance of triticale. A novel analytical method for distinguishing elite genotypes based on measured features was proposed. Additionally, a new programing based on SAS-language (IML) was introduced to estimate the genetic parameters rooted from combined ANOVA model (linear mixed model), which is capable of being used in any field study other than the current one. Methods Thirty genotypes of triticale were studied under normal and drought stress conditions during 6 years (three different locations). Accordingly, based on the results of genetic variability, heatmap analysis, biplot graph, and clustering technique, two genotypes with the highest genetic distance were selected to appraise the differential expression profiling of three SOD isozyme in shoot and root organs. Results Field experiments and bioinformatics results showed that superoxide dismutase (SOD) was the most influential antioxidant in resistance of triticale to drought stress; therefore, it could be used as an indirect selection index in early stages to distinguish resistant genotypes to drought stress. Additionally, Mn-SOD and Fe-SOD showed roughly similar expression levels for both genotypes under drought stress. However, Cu/Zn-SOD expression level was higher in root and shoot of the tolerant genotype than the susceptible genotype. Conclusion Heatmap analysis that is applied for the first time to screen suitable genotypes, showed to be highly capable of distinguishing elite genotypes and pointing out the proper features for selection criteria. Bioinformatics results indicated that SOD is more important than other enzymatic antioxidant for being considered as selection criteria or candidate gene for transgenic purposes. Based on expressional results, Mn-SOD announced as a general isozyme that is probably highly expressed in most of the species, while, Cu/Zn-SOD was introduced as a genotype specific isozyme that is likely more expressed in tolerant genotypes

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Two separate experiments, one as drought stress condition and the other one as the normal irrigation condition based on randomized complete block design (RCBD) with three replications were implemented to examine 20 triticale (Triticoseclae wittamack X) genotypes of which 19 genotypes have recently been generated by crossing different parent lines and the other one was Javaniloo, common cultivar. Based on the results of different statistical techniques and comparing relationships among traits for normal irrigation and stress condition, enzymatic antioxidant could be used as criteria for screening tolerant genotypes of triticale. On the other hand, it is pointed out that superoxide dismutase (SOD) are the most important criteria to achieving higher tolerant genotypes through indirect selection. Furthermore, hydrogen peroxide (H2O2) and malondialdehyde (MDA) content with having highly relationship with rain yield are also different possible criteria for screening triticale genotypes for water stress conditions in which genotypes with lower content of these traits could be screened.