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Attila Ombódi, Hussein Gehad Daood and Lajos Helyes

rainfall decreased β-carotene content of carrot. Vegetables are not the main dietary sources of tocopherols; the richest sources are cereals and plant originated fats and oils ( DellaPenna and Méne-Safranné, 2011 ). However, carotenoids and tocopherols

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Sofia Caretto, Angelo Parente, Francesco Serio and Pietro Santamaria

tocopherols, known as vitamin E, have been shown to carry out essential functions in slowing or preventing degenerative disease processes in humans ( Kaur and Kapoor, 2001 ; Traber and Sies, 1996 ). Of the four known tocopherol forms—α, β, γ, and δ—α-tocopherol

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Carlos Calderón-Vázquez, Mary L. Durbin, Vanessa E.T.M. Ashworth, Livia Tommasini, Kapua K.T. Meyer and Michael T. Clegg

binding free radical intermediates. It acts as an inhibitor of platelet aggregation and protects lipids by preventing the oxidation of polyunsaturated fatty acids ( Traber and Stevens, 2011 ). In this study we are measuring α-tocopherol, the most

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Christine M. Bradish, Gad G. Yousef, Guoying Ma, Penelope Perkins-Veazie and Gina E. Fernandez

locations in central and western North Carolina were evaluated to determine the effects of a warm production climate and high tunnel cultivation on anthocyanin, carotenoid, tocopherol, and ellagitannin content, among a number of other fruit quality factors

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I.L. Goldman

Carrots contribute ≈14% of the total Vitamin A to the human diet in the United States due to the presence of the provitamin A carotenoids α- and β-carotene. We have described a recessive gene (rp) that inhibits carotenoid biosynthesis in carrot by 93%, resulting in whitish-yellow roots. The rp mutation is also associated with relatively high levels of a tocopherol (Vitamin E, 0.61±0.15 mg α-tocopherol/100 g FW). Vitamin E is a powerful antioxidant that must be obtained from the diet. The biosynthesis of a tocopherol in carrot has not been studied in any detail; however, the rp gene may provide clues as to its mechanism. The production of carotenoids and tocopherols is biosynthetically linked by their common precursor, geranylgeranyl diphosphate (GGDP). GGDP is converted into phytoene by phytoene desaturase to produce carotenoids and combined with homogentisic acid to produce tocopherols. Carotenoid and tocopherol profiles for various carrot genotypes are presented alongside a model describing the potential relationship between root carotenoids and tocopherols in carrot. The presence of significant amounts of tocopherols in carrot could significantly raise the nutritional profile of this vegetable.

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Thomas C. Koch and Irwin L. Goldman

Carotenoids and tocopherols are health-functional phytochemicals that occur in a wide range of fruit and vegetable crops. These two classes of compounds are synthesized from a common precursor, geranyl-geranyl pyrophosphate, and are typically analyzed separately via high-performance liquid chromatography (HPLC) techniques. Because carotenoids and tocopherols are present in many edible horticultural crops, it would be advantageous to measure them simultaneously in plant tissues. Herein we report a one-pass reverse-phase HPLC method for extraction and analysis of carotenoids and tocopherols in carrot that can be extended to other high-moisture plant organs. Elution times ranged from 5 minutes for α-tocopherol to 24 minutes for β-carotene. This method improves the efficiency of analyzing these compounds by up to 50%, and should increase the efficiency of assessing carotenoid and tocopherol profiles in horticultural crops.

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Kanta Kobira, Khalid Ibrahim, Elizabeth Jefferey and John Juvik

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.

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Khalid Ibrahim and John Juvik

Vegetables are a rich source of dietary carotenoids and tocopherols, powerful antioxidants that have the capacity to protect cells against oxidative damage caused by free radical reactions. There is evidence for a negative correlation between the incidence of certain types of cancer, age-related macular degeneration, cataract development, and cardiovascular disease with increased carotenoid and tocopherol intake. Development of elite vegetable germplasm with enhanced levels of these phytochemicals will potentially promote health among the consuming public. To assess the feasibility for genetic improvement in phytochemical content, it is necessary to partition the phenotypic variability into its component sources (genotype, environment, and genotype by environment interaction). To provide data for comparison and partition of phenotypic variation, 41 sweet corn and 13 broccoli genotypes were grown and harvested in one location for 3 years and analyzed for phytochemical content by HPLC. The most abundant form of carotenoids and tocopherols were lutein and gamma-tocopherol in sweet corn and beta-carotene and alpha-tocopherol in broccoli. Analysis of variance showed that, in sweet corn, the differences among genotypes described most of the phenotypic variation (76% for lutein, and 78% for gamma-tocopherol). Genotype by year interaction was a second significant factor, while variation affiliated with the year was found to be a minor component. In contrast, in broccoli, the three sources of variability contributed equally to describe the total phenotypic variation for beta-carotene and alpha-tocopherol. These results suggest that elite sweet corn and broccoli germplasm with improved carotenoid and tocopherol levels can be developed using conventional breeding protocols.

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Jennifer L. Baeten, Thomas C. Koch and Irwin L. Goldman

Carrot has been bred for increased levels of pro-vitamin E α-tocopherol. This vitamin is lipid soluble. Carrot root has been shown to have measurable levels of lipid, but it is not certain if the lipid level is correlated to α-tocopherol levels. The HPLC method is needed to quantify levels of α-tocopherol. Measuring lipids may be less time consuming in a breeding program. We developed a method for extracting lipids from carrot tissue based on the Soxhlet extraction method. The Soxhlet extraction uses a non-polar ether solvent to pull lipids out of freeze-dried tissue. A collection of carrot accessions ranging in α-tocopherol concentration 0.04–0.18 ppm and carotenoid concentration 10.63–1673.76 ppm were used in this investigation. Root tissue was freeze-dried and lipid levels were measured in an experiment with two replications. The mean lipid level of root tissue was 0.05 g fat/g tissue. The range was 0–1.1 g fat/g tissue. Phenotypic correlations were performed among lipid, α-tocopherol, and β-carotene concentrations in these samples. Twenty-four samples were tested for lipid levels (12 high and 12 low). From these results, percent lipid of the root was determined. Correlations were made between the lipid data and α-tocopherol data of the given samples.

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Khalid E. Ibrahim, Kanta Kobira and John A. Juvik

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.