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Chemical measures of antioxidant activity within the plant, such as the oxygen radical absorbance capacity (ORAC) assay, have been reported for many plant-based foods. However, the extent to which chemical measures relate to cellular measures of oxidative stress is unclear. The natural variation in the phytochemical content of 22 broccoli genotypes was used to determine correlations among chemical composition (carotenoids, tocopherols and polyphenolics), chemical antioxidant activity (ORAC), and measures of cellular antioxidation [prevention of DNA oxidative damage and of oxidation of the biomarker dichlorofluorescein (DCFH) in HepG2 cells] using hydrophilic and lipophilic extracts of broccoli. For lipophilic extracts, ORAC (ORAC-L) correlated with inhibition of cellular oxidation of DCFH (DCFH-L, r = 0.596, p = 0.006). Also, DNA damage in the presence of the lipophilic extract was negatively correlated with both chemical and cellular measures of antioxidant activity as measured by ORAC-L (r = -0.705, p = 0.015) and DCFH-L (r = -0.671, p = 0.048), respectively. However, no correlations were observed for hydrophilic (-H) extracts, except between polyphenol content and ORAC (ORAC-H; r = 0.778, p < 0.001). Inhibition of cellular oxidation by hydrophilic extracts (DCFH-H) and ORAC-H were approximately 8- and 4-fold greater than DCFH-L and ORAC-L, respectively. Whether ORAC-H has more biological relevance than ORAC-L because of its magnitude or whether ORAC-L bears more biological relevance because it relates to cellular estimates of antioxidant activity remains to be determined. Chemical estimates of antioxidant capacity within the plant may not accurately reflect the complex nature of the full antioxidant activity of broccoli extracts within cells.

Genotype-by-environment interaction (G×E) is a fundamental concern in plant breeding since it hinders developing genotypes with wide geographical usefulness. Analysis of variance (ANOVA) has been widely used to interpret G×E, but it does not elucidate the nature and causes of the interaction. Stability analysis provides a summary of the response patterns of genotypes to different growing environments. Two classes of phytochemicals with putative health promoting activity are carotenoids and tocopherols that are relatively abundant in broccoli. Growing clinical and epidemiological evidence suggests that vegetables with enhanced levels of these phytochemicals can reduce the risk of cancer, cardiovascular, and eye diseases. The objective of this study is to have better understanding of the genetic, environmental and G×E interaction effects of these phytochemicals in broccoli to determine the feasibility of the genetic enhancement. The ANOVA and Shukla's stability test were applied to a set of data generated by the HPLC analysis of different carotenoid and tocopherol forms for six broccoli accessions grown over three environments. The ANOVA results show a significant G×E for both phytochemicals that ranged from 22.6% of the total phenotypic variation for beta-carotene to 54.0% for delta-tocopherol while the environmental effects were nonsignificant. The genotypic effects ranged from as low as 1% for alpha-tocopherol to 31.5% and 36.0% for beta-carotene and gamma-tocopherol, respectively. Stability analysis illustrated that the most stable genotype for all phytochemicals is Brigadier. The results suggest that feasibility of the genetic enhancement for major carotenoids and tocopherols. A second experiment that includes a larger set of genotypes and environments was conducted to confirm the results of this study.

Considerable epidemiological evidence exists on the association between consumption of antioxidant-rich vegetables and incidence of chronic diseases, including cancer and cardiovascular disease. Broccoli (Brassica oleracea L. sp. italica) florets are relatively abundant sources of antioxidants, and potentially amenable to genetic manipulation to enhance this vegetable's health-promoting properties. This investigation focuses on the identification of chromosomal segments in the nuclear genome of broccoli associated with antioxidant carotenoid and tocopherol variability. A broccoli F2:3 population consisting of 163 families derived from a cross between two parents (VI-158 and BNC) and previously mapped with 62 polymorphic SSR and SRAP marker loci was evaluated for carotenoid and tocopherol concentration in floret tissue over two growing seasons. Significant differences were observed among F2:3 family means for concentrations of lutein (10-fold difference between the lowest and highest family), beta-carotene 17-fold), alpha-tocopherol (8-fold) and gamma-tocopherol (6-fold). On a concentration basis, beta-carotene, lutein, alpha-tocopherol, and gamma-tocopherol were the most abundant antioxidant forms in broccoli. Heritability estimates of primary phytochemicals ranged from 0.35 to 0.38, 0.40, and 0.44 for beta-carotene, alpha-tocopherol, gamma-tocopherol, and lutein, respectively. Composite interval mapping (CIM) identified two quantitative trait loci (QTL) associated with carotenoid variability on two linkage groups and five QTL associated with tocopherol variability on four linkage groups. The QTL identified in this study have potential for use in marker-assisted crop improvement programs to develop elite germplasm designed to promote health among the consuming public.