Vegetable crops can be significant sources of nutritionally important dietary carotenoids and Brassica vegetables are sources that also exhibit antioxidant and anticarcinogenic activity. The family Brassicaceae contains a diverse group of plant species commercially important in many parts of the world. The six economically important Brassica species are closely related genetically. Three diploid species (B. nigra, B. rapa, and B. oleracea) are the natural progenitors of the allotetraploid species (B. juncea, B. napus, and B. carinata). The objective of this study was to characterize the accumulation of important dietary carotenoid pigments among the genetically related Brassica species. The HPLC quantification revealed significant differences in carotenoid and chlorophyll pigment accumulation among the Brassica species. Brassica nigra accumulated the highest concentrations of lutein, 5,6-epoxy lutein, violaxanthin, and neoxanthin. The highest concentrations of beta-carotene and total chlorophyll were found in B. juncea. Brassica rapa accumulated the highest concentrations of zeaxanthin and antheraxanthin. For each of the pigments analyzed, the diploid Brassica species accumulated higher concentrations, on average, than the amphidiploid species. Brassicas convey unique health attributes when consumed in the diet. Identification of genetic relationships among the Brassica species would be beneficial information for improvement programs designed to increase carotenoid values.
Dean A. Kopsell, Scott McElroy, Carl Sams, and David Kopsell
Dean A. Kopsell, David E. Kopsell, and Joanne Curran-Celentano
Therapeutic compounds in herbal crops are gaining recent attention. Sweet basil (Ocimumbasilicum L.) is a popular culinary herbal crop grown for both fresh and dry leaf markets. Recently, basil (unidentified cultivar) was shown to rank highest among spices and herbal crops for xanthophylls carotenoids. This class of carotenoids is associated with decreased risks of certain cancer and age-related eye diseases. The research goal for the current study was to characterize the concentrations of nutritionally important carotenoid pigments among popular varieties of basil. Eight cultivars of sweet basil (`Genovese', `Italian Large Leaf', `Nufar', `Red Rubin', `Osmin Purple', `Spicy Bush', `Cinnamon', and `Sweet Thai') were grown in both field and greenhouse environments and evaluated for plant pigments using HPLC methodology. Environmental and cultivar differences were significant for all of the pigments analyzed. `Sweet Thai' accumulated the highest concentrations of lutein, zeaxanthin, and beta-carotene carotenoids, while `Italian Large Leaf' had the lowest concentrations. Comparing the two environments, cultivar means for carotenoid and chlorophyll pigments were higher in the field environment when expressed on both a fresh and dry weight basis. Exceptions were found only for the purple leaf basils (`Osmin Purple' and `Red Rubin'). Positive and highly significant correlations existed between carotenoid and chlorophyll pigments in both environments. This study demonstrates that sweet basil can accumulate high levels of nutritionally important carotenoids in both field and greenhouse environments.
Lavanya Reddivari and J. Creighton Miller Jr.
Antioxidants have been widely reported to play an important role in disease prevention. In addition to preventing cancer, stroke, heart diseases, and inflammation, they are also involved in immune surveillance. Since the per capita consumption of potatoes in the U.S. is about 137 lb, even moderate levels of antioxidants in this most important vegetable crop probably have an important human health benefit. About 75% to 80% of antioxidant activity in specialty potatoes is due to phenolics and carotenoids. The objectives of this investigation were to evaluate antioxidant activity and total phenolic and carotenoid content of specialty potato selections from the Texas Potato Variety Development Program, and to identify candidate compounds for cancer cell culture investigations. Potato tubers were also used to identify and quantify individual phenolics and carotenoids. Some 320 specialty selections were screened for antioxidant activity (AA), total phenolic content (TP) and carotenoid content (CC) using DPPH (2,2-Diphenyl-1-picrylhydrazyl), FCR (Folin-Ciocalteu Reagent) and colorimetric assays, respectively. After the initial screening, the top 10% were used for analysis of individual phenolics and carotenoids using HPLC. Wide variability for antioxidant activity, phenolic content, and carotenoid content was found among specialty potato selections, providing evidence for genetic control of theses traits. The specialty selection CO112F2-2P/P (purple flesh, purple skin) had the highest AA (832 μg trolox equivalents/g fw), TP (1553 μg chlorogenic acid equivalents/g fw) and CC (590 μg lutein equivalents/100 g fw). Chlorogenic acid (55% to 60%), caffeic acid (≈5%), gallic acid (18% to 20%), and catechin (18% to 20%) were found to be the most prevalent phenolic acids, and lutein and zeaxanthin were the most prominent carotenoids contributing to antioxidant activity. Gallic acid was identified as the candidate compound for use in cancer cell culture investigations.
Guohai Xia and Lailiang Cheng
Four-year-old `Gala'/M.26 trees were grown under low (2.5 mm), medium (12.5 mm), or high (25 mm) N supply with balanced nutrients in sand culture and the cropload was adjusted to 5 fruit/cm2 trunk cross-sectional area at 10 mm king fruit. At about 100 days after bloom, exposed fruit on the south side of the canopy were chosen for monitoring chlorophyll fluorescence and fruit peel samples were taken for measuring xanthophyll cycle pigments, antioxidant enzymes, and metabolites. At noon, the efficiency of excitation transfer (Fv'/Fm') of the sun-exposed peel was higher in the low N treatment than in the medium or high N treatments. Photochemical quenching coefficient did not differ between fruits in different N treatments. The Photosystem II operating efficiency was higher in the peel of low N fruit compared with medium N or high N fruit. However, maximum quantum efficiency (Fv/Fm) of fruit peel after overnight dark adaptation was similar across the N treatments. The xanthophyll cycle pool size expressed on peel area basis was larger in the high N fruit than in the low N fruit. This corresponds well with the thermal dissipation capacity, as indicated by efficiency of excitation transfer. Over 95% of the xanthophyll cycle pool in the sun-exposed side was present in the form of zeaxanthin and antheraxanthin at noon regardless of N treatments. Activities of superoxide dismutase and all the antioxidant enzymes and metabolites in the ascorbate-glutathione cycle were higher in the high N fruit than in low N fruit. The results indicate that apple fruit with a good N status have a higher photoprotective capacity in terms of xanthophyll cycle-dependent thermal dissipation and detoxification of reactive oxygen species compared with low N fruit.
Li-Song Chen and Lailiang Cheng
To determine the cause of a characteristic zonal chlorosis of `Honeycrisp' apple (Malus ×domestica Borkh.) leaves, we compared CO2 assimilation, carbohydrate metabolism, the xanthophyll cycle and the antioxidant system between chlorotic leaves and normal leaves. Chlorotic leaves accumulated higher levels of nonstructural carbohydrates, particularly starch, sorbitol, sucrose, and fructose at both dusk and predawn, and no difference was found in total nonstructural carbohydrates between predawn and dusk. This indicates that carbon export was inhibited in chlorotic leaves. CO2 assimilation and the key enzymes in the Calvin cycle, ribulose 1,5-bisphosphate carboxylase/oxygenase, NADP-glyceraldehyde-3-phosphate dehydrogenase, phosphoribulokinase, stromal fructose-1,6-bisphosphatase, and the key enzymes in starch and sorbitol synthesis, ADP-glucose pyrophosphorylase, cytosolic fructose-1,6-bisphosphatase, and aldose 6-phosphate reductase were significantly lower in chlorotic leaves than in normal leaves. However, sucrose phosphate synthase activity was higher in chlorotic leaves. In response to a reduced demand for photosynthetic electron transport, thermal dissipation of excitation energy (measured as nonphotochemical quenching of chlorophyll fluorescence) was enhanced in chlorotic leaves under full sun, lowering the efficiency of excitation energy transfer to PSII reaction centers. This was accompanied by a corresponding increase in both xanthophyll cycle pool size (on a chlorophyll basis) and conversion of violaxanthin to antheraxanthin and zeaxanthin. The antioxidant system, including superoxide dismutase and ascorbate peroxidase and the ascorbate pool and glutathione pool, was up-regulated in chlorotic leaves in response to the increased generation of reactive oxygen species via photoreduction of oxygen. These findings support the hypothesis that phloem loading and/or transport is partially or completely blocked in chlorotic leaves, and that excessive accumulation of nonstructural carbohydrates may cause feedback suppression of CO2 assimilation via direct interference with chloroplast function and/or indirect repression of photosynthetic enzymes.
Haejeen Bang*, Sungil Kim, Daniel Leskovar, and Stephen King
Carotenoids are plant compounds that serve a variety of essential functions in the plant and have also been found to have several health-promoting activities in humans. Carotenoids found in watermelon (Citrullus lanatus) flesh are responsible for the various colors such as red, yellow and orange. Previous inheritance studies of flesh color revealed that six genes were involved in color determination. The relationship and interaction of these genes suggests that some color-determining genes may be the result of mutations on the structural genes encoding enzymes in the carotenoid biosynthesis pathway. In this study we were able to isolate and sequence six genes encoding enzymes involved in the carotenoid biosynthetic pathway, and determine their expression in different colored watermelon fruit. The cDNA was synthesized from total RNA using RACE (Rapid Amplification of cDNA ends) kit (SMART RACE cDNA Amplification Kit; Clontech, Palo Alto, Calif.). Degenerate primers were designed based on published homologous genes from other species and were used to isolate gene fragments and full-length cDNAs of phytoene synthase, phytoene desaturase, _-carotene desaturase, β-cyclase, β-carotene hydroxylase and zeaxanthin expoxidase. RT-PCR was carried out to examine any differential expression of cloned genes in white, yellow, orange and red-fleshed watermelon. All cloned enzyme-encoding genes were expressed regardless of flesh colors. These results indicate that carotenoid biosynthesis may be regulated at the post-transcriptional level. One interesting feature supports this hypothesis. In case of β-cyclase, a 229-bp leader intron was identified, and an unspliced mRNA with this leader intron existed dominantly in cDNA pool of all samples.
Lailiang Cheng and Fengwang Ma
Xanthophyll cycle conversion and the antioxidant system in the peel of apple fruit (Malus ×domestica Borkh. `Liberty') were monitored in the field over a diurnal course at about 3 months after full bloom. Compared with leaves, sun-exposed peel of apple fruit had much lower photosystem II operating efficiency at any given photon flux density (PFD) and a larger xanthophyll cycle pool size on a chlorophyll basis. Zeaxanthin (Z) level increased with rising PFD in the morning, reached the highest level during midday, and then decreased with falling PFD for the rest of the day. At noon, Z accounted for >90% of the xanthophyll cycle pool in the fruit peel compared with only 53% in leaves. Efficiency of excitation transfer to PSII reaction centers (F v′/F m′) was negatively related to the level of Z in fruit peel and leaves throughout the day. In fruit peel, the antioxidant enzymes in the ascorbate-glutathione cycle, ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR), dehydroascorbate reductase (DHAR) and glutathione reductase (GR) showed a diurnal pattern similar to that of incident PFD. In contrast, the activities of APX and GR in leaves did not change significantly during the day although activities of both MDAR and DHAR were higher in the afternoon than in the morning. In both fruit peel and leaves, superoxide dismutase activity did not change significantly during the day; catalase activity showed a diurnal pattern opposite to that of PFD with much lower activity in fruit peel than in leaves. The total ascorbate pool was much smaller in fruit peel than in leaves on an area basis, but the ratio of reduced ascorbate to oxidized ascorbate reached a maximum in the early afternoon in both fruit peel and leaves. The total glutathione pool, reduced glutathione and the ratio of reduced glutathione to oxidized glutathione in both fruit peel and leaves also peaked in the early afternoon. We conclude that the antioxidant system as well as the xanthophyll cycle responds to changing PFD over the course of a day to protect fruit peel from photooxidative damage.
Brandon R. Smith and Lailiang Cheng
The objective of this study was to quantify how photoprotective mechanisms in the leaves of `Concord' grapevines (Vitis labruscana Bailey) respond to a range of iron (Fe) supply. Own-rooted, 1-year-old container-grown vines were fertigated twice weekly for 11 weeks with a complete nutrient solution containing 1, 10, 20, 50, or 100 μm Fe from ferric ethylenediamine di (o-hydroxyphenylacetic) acid (Fe-EDDHA). Leaf total Fe content did not increase in response to Fe supply; however, “active” Fe (extracted with 2,2′-dipyridyl) and chlorophyll (Chl) increased on a leaf area basis as applied Fe increased. At the lowest active Fe level, leaf absorptance and the efficiency of excitation transfer (Fv′/Fm′) was lower, and nonphotochemical quenching (NPQ) was significantly greater. Photosystem II (PSII) quantum efficiency decreased curvilinearly, and the proportion of PSII reaction centers in the open state (qP) decreased linearly as active Fe content decreased. On a Chl basis, the xanthophyll cycle pool size [violaxanthin (V) + antheraxanthin (A) + zeaxanthin (Z)], lutein, and β-carotene increased curvilinearly as active Fe decreased, and neoxanthin (Neo) increased at the lowest Fe level. On a leaf area basis, as active Fe decreased, V+A+Z and β-carotene decreased curvilinearly, and lutein and Neo decreased linearly. At noon, conversion of V to A and Z increased as active Fe decreased. On a Chl basis, activities of antioxidant enzymes superoxide dismutase (SOD), monodehydroascorbate reductase (MDAR), and dehydroascorbate reductase (DHAR) increased curvilinearly, and glutathione reductase (GR) activity increased linearly as active Fe levels declined. Ascorbate peroxidase (APX) and catalase (CAT), on a Chl basis, were relatively constant. On a leaf area basis, a decrease in active Fe increased SOD and MDAR activity, whereas APX, CAT, DHAR and GR activity decreased. Antioxidant metabolites ascorbate (AsA), dehydroascorbate (DAsA), reduced glutathione (GSH) and oxidized glutathione (GSSG) also increased in response to Fe limitation when expressed on a Chl basis, whereas on a leaf area basis AsA and DAsA decreased and GSH increased curvilinearly. The GSH:GSSG ratio increased as active Fe declined, whereas the AsA:DAsA ratio did not change. In conclusion, both photoprotective mechanisms, xanthophyll cycle-dependent thermal dissipation and the ascorbate-glutathione antioxidant system, are enhanced in response to Fe deficiency to cope with excess absorbed light. In a low soil pH tolerant species such as V. labruscana, the foliar antioxidant system was upregulated in response to excess absorbed light from Fe deficiency-induced chlorosis, and there was no evidence of an increase in oxidative stress from high rates of applied Fe-EDDHA.
Calen McKenzie, Ivette Guzman, Ciro Velasco-Cruz, and Paul W. Bosland
a pump (600 Controller, Waters) and a C 30 carotenoid column (4.6 × 250 mm, 5-μm particle size, C 30 ; YMC America, Allentown, PA) provided separation of lutein, zeaxanthin, β-carotene, Chl a, and Chl b. The solvents used were solvent A [methanol
Neda Keyhaninejad, Richard D. Richins, and Mary A. O’Connell
, capsanthin, capsorubin, β-carotene, β-cryptoxanthin, violaxanthin, and zeaxanthin ( Guzmán et al., 2010) among others. Light plays a key regulatory role for genes and gene products related to photosynthesis including carotenoids ( Pizarro and Stange, 2009