Progressive drought alters the root exudate metabolome and differentially activates metabolic pathways in cotton (Gossypium hirsutum).

Root exudates comprise various primary and secondary metabolites that are responsive to plant stressors, including drought. As increasing drought episodes are predicted with climate change, identifying shifts in the metabolome profile of drought-induced root exudation is necessary to understand the molecular interactions that govern the relationships between plants, microbiomes, and the environment, which will ultimately aid in developing strategies for sustainable agriculture management. This study utilized an aeroponic system to simulate progressive drought and recovery while non-destructively collecting cotton (Gossypium hirsutum) root exudates. The molecular composition of the collected root exudates was characterized by untargeted metabolomics using Fourier-Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR MS) and mapped to the Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Over 700 unique drought-induced metabolites were identified throughout the water-deficit phase. Potential KEGG pathways and KEGG modules associated with the biosynthesis of flavonoid compounds, plant hormones (abscisic acid and jasmonic acid), and other secondary metabolites were highly induced under severe drought, but not at the wilting point. Additionally, the associated precursors of these metabolites, such as amino acids (phenylalanine and tyrosine), phenylpropanoids, and carotenoids, were also mapped. The potential biochemical transformations were further calculated using the data generated by FT-ICR MS. Under severe drought stress, the highest number of potential biochemical transformations, including methylation, ethyl addition, and oxidation/hydroxylation, were identified, many of which are known reactions in some of the mapped pathways. With the application of FT-ICR MS, we revealed the dynamics of drought-induced secondary metabolites in root exudates in response to drought, providing valuable information for drought-tolerance strategies in cotton.

Duration of Retinol Isotope Dilution Studies with Compartmental Modeling Affects Model Complexity, Kinetic Parameters, and Calculated Vitamin A Stores in US Women.

Background: Retinol isotope dilution (RID) indirectly estimates vitamin A (VA) status. Multicompartment modeling of RID data is used to refine study designs and equations to calculate VA stores. Previous studies suggest that VA in slowly turning over pools is not traced if follow-up is not long enough; however, shorter RID studies are being investigated. Few long-term models have been published. Objective: We determined the effect of time on mathematical models of VA kinetics, model parameters, and outcomes. Methods: In this longitudinal study, women (mean ± SD age: 22 ± 3 y; n = 7) were given 2.0 µmol [14,15]-13C2-retinyl acetate. Blood samples were staggered from 4 h to 152 d; the fraction of dose in serum was modeled with compartmental models. Four model-time categories were created: full models that used all data (median: 137 d; range 97-152 d) and truncated shorter studies of 14, 27, and 52 d (range: 42-62 d). Outcomes included number of compartments to adequately model serum data, kinetic parameters, total traced VA mass, and time-to-dose equilibration. To gain insight into longer follow-up, an additional participant was given 17.5 µmol 13C4-VA, and data were modeled as long as enrichment was above baseline (5 y). Results: Longer follow-up times affected kinetic parameters and outcomes. Compared with the 14-d models, long-term full models required an additional compartment for adequate fit (14.3% compared with 100%; P = 0.0056) and had longer [median (quartile 1, quartile 3)] whole-body half-life [15.0 d (10.5, 72.6 d) compared with 135 d (115, 199 d); P = 0.0006], time-to-dose equilibration [3.40 d (3.14, 6.75 d) compared with 18.9 d (11.2, 25.7 d); P < 0.0001], and total traced mass [166 µmol VA (162, 252 µmol VA) compared with 476 µmol VA (290, 752 µmol VA); P = 0.0031]. Conclusions: Extended RID sampling alters numerous mathematically modeled, time-dependent outcomes in women. Length of study should be considered when using mathematical models for calculating total-body VA stores or kinetic parameters related to VA turnover. This study is registered at www.clinicaltrials.gov as NCT03248700.

Retention of Carotenoids in Biofortified Maize Flour and ß-Cryptoxanthin-Enhanced Eggs after Household Cooking.

Biofortification of crops to enhance provitamin A carotenoids is a strategy to increase the intake where vitamin A deficiency presents a widespread problem. Heat, light, and oxygen cause isomerization and oxidation of carotenoids, reducing provitamin A activity. Understanding provitamin A retention is important for assessing efficacy of biofortified foods. Retention of carotenoids in high-xanthophyll and high-ß-carotene maize was assessed after a long-term storage at three temperatures. Carotenoid retention in high-ß-cryptoxanthin maize was determined in muffins, non-nixtamalized tortillas, porridge, and fried puffs made from whole-grain and sifted flour. Retention in eggs from hens fed high-ß-cryptoxanthin maize was assessed after frying, scrambling, boiling, and microwaving. Loss during storage in maize was accelerated with increasing temperature and affected by genotype. Boiling whole-grain maize into porridge resulted in the highest retention of all cooking and sifting methods (112%). Deep-fried maize and scrambled eggs had the lowest carotenoid retention rates of 67-78 and 84-86%, respectively.

Methiozolin sorption and mobility in sand-based root zones.

BACKGROUND: Methiozolin is a herbicide currently used for annual bluegrass control in golf course putting greens. Previous research indicates that maximum weed control efficacy requires root exposure; however, soil sorption and mobility of methiozolin have not been established. Research was conducted to investigate soil sorption and subsequent desorption by dilution of methiozolin, as well as soil mobility using batch equilibrium experiments and thin-layer chromatography in nine root zones. Evaluations focused on sand-based systems typical of many golf course putting greens. RESULTS: Sorption coefficients (Kd values) ranged from 0.4 to 29.4 mL g(-1) and averaged 13.8 mL g(-1) . Sorption was most influenced by organic matter content; conversely, soil pH had a negligible effect. Methiozolin desorption did not occur with a 0.01 M CaCl2 dilution. Methiozolin mobility was low; retardation factors (Rf values) were <0.05 for all media with ≥0.3% organic matter. Sand (0.1% organic matter) resulted in an Rf value of 0.46. CONCLUSION: Approximately 24% of applied methiozolin is available for root uptake, and mobility is limited, suggesting resistance to loss through leaching displacement.

Carotenoid profiles in provitamin A-containing fruits and vegetables affect the bioefficacy in Mongolian gerbils.

Fruits and vegetables are rich sources of provitamin A carotenoids. We evaluated the vitamin A (VA) bioefficacy of a whole foods supplement (WFS) and its constituent green vegetables (Study 1) and a variety of fruits with varying ratios of provitamin A carotenoids (Study 2) in VA-depleted Mongolian gerbils (n = 77/study). After feeding a VA-deficient diet for 4 and 6 weeks in Studies 1 and 2, respectively, customized diets, equalized for VA, were fed for 4 and 3 weeks, respectively. Both studies utilized negative and VA-positive control groups. In Study 1, liver VA was highest in the VA group (0.82 +/- 0.16 micromol/liver, P < 0.05), followed by brussels sprouts (0.50 +/- 0.15 micromol/liver), Betanat (beta-carotene from Blakeslea trispora) (0.50 +/- 0.12 micromol/liver) and spinach (0.47 +/- 0.09 micromol/liver) groups, which did not differ from baseline. The WFS (0.44 +/- 0.06 micromol/liver) and kale (0.43 +/- 0.14 micromol/liver) groups had lower liver VA than the baseline group (P < 0.05), but did not differ from the brussels sprouts, Betanat and spinach groups. In Study 2, liver VA was highest in the orange (0.67 +/- 0.18 micromol/liver), papaya (0.67 +/- 0.15 micromol/liver) and VA (0.66 +/- 0.14 micromol/liver) groups, followed by the mango (0.58 +/- 0.09 micromol/liver) and tangerine (0.55 +/- 0.15 micromol/liver) groups. These groups did not differ from baseline. The banana group (0.47 +/- 0.15 micromol/liver) was unable to maintain baseline stores of VA and did not differ from the control (0.46 +/- 0.13 mumol/liver). These fruits (except banana), vegetables and the WFS were able to prevent VA deficiency in Mongolian gerbils and could be an effective part of food-based interventions to support VA nutrition in developing countries and worldwide.

Mathematical modeling of serum 13C-retinol in captive rhesus monkeys provides new insights on hypervitaminosis A.

Hypervitaminosis A is increasingly a public health concern, and thus noninvasive quantitative methods merit exploration. In this study, we applied the (13)C-retinol isotope dilution test to a nonhuman primate model with excessive liver stores. After baseline serum chemistries, rhesus macaques (Macaca mulatta; n = 16) were administered 3.5 mumol (13)C(2)-retinyl acetate. Blood was drawn at baseline, 5 h, and 2, 4, 7, 14, 21, and 28 d following the dose. Liver biopsies were collected 7 d before and 2 d after dosing (n = 4) and at 7, 14, and 28 d (n = 4/time) after dosing. Serum and liver were analyzed by HPLC and GC-combustion-isotope ratio MS for retinol and its enrichment, respectively. Model-based compartmental analysis was applied to serum data. Lactate dehydrogenase was elevated in 50% of the monkeys. Total body reserves (TBR) of vitamin A (VA) were calculated at 28 d. Predicted TBR (3.52 +/- 2.01 mmol VA) represented measured liver stores (4.56 +/- 1.38 mmol VA; P = 0.124). Predicted liver VA concentrations (13.3 +/- 9.7 micromol/g) were similar to measured liver VA concentrations (16.4 +/- 5.3 micromol/g). The kinetic models predict that 27-52% of extravascular VA is exchanging with serum in hypervitaminotic A monkeys. The test correctly diagnosed hypervitaminosis A in all monkeys, i.e. 100% sensitivity. Stable isotope techniques have important public health potential for the classification of VA status, including hypervitaminosis, because no other technique besides invasive liver biopsies, correctly identifies excessive liver VA stores.

13C natural abundance in serum retinol acts as a biomarker for increases in dietary provitamin A.

The natural isotopic composition of 13C and 12C in tissues is largely determined by the diet. Sources of provitamin A carotenoids (e.g., vegetables) typically have a lower 13C to 12C ratio (13C:12C) than preformed vitamin A sources (i.e., dairy and meat) from corn-fed animals, which are prevalent in the US. The 13C:12C of serum retinol (13C:12C-retinol) was evaluated as a biomarker for vegetable intake in a 3-mo dietary intervention designed to promote weight-loss by increased vegetable consumption or reduced calorie and fat intake. Subjects were 21-50 y of age with a BMI between 30-40 kg/m2 and were enrolled from one geographic area in the US. The high vegetable group (n=20) was encouraged to increase daily vegetable and fruit consumption to 0.95 liter vegetables and 0.24-0.35 liter fruits. The caloric reduction group (n=17) was encouraged to lower caloric intake by 500 kcal and consume

Cassava with enhanced beta-carotene maintains adequate vitamin A status in Mongolian gerbils (Meriones unguiculatus) despite substantial cis-isomer content.

Efforts to increase beta-carotene in cassava have been successful, but the ability of high-beta-carotene cassava to prevent vitamin A deficiency has not been determined. Two studies investigated the bioefficacy of provitamin A in cassava and compared the effects of carotenoid content and variety on vitamin A status in vitamin A-depleted Mongolian gerbils (Meriones unguiculatus). Gerbils were fed a vitamin A-free diet 4 weeks prior to treatment. In Expt 1, treatments (ten gerbils per group) included 45 % high-beta-carotene cassava, beta-carotene and vitamin A supplements (intake matched to high-beta-carotene cassava group), and oil control. In Expt 2, gerbils were fed cassava feeds with 1.8 or 4.3 nmol provitamin A/g prepared with two varieties. Gerbils were killed after 4 weeks. For Expt 1, liver vitamin A was higher (P < 0.05) in the vitamin A (1.45 (sd 0.23) micromol/liver), lower in the control (0.43 (sd 0.10) micromol/liver), but did not differ from the beta-carotene group (0.77 (sd 0.12) micromol/liver) when compared with the high-beta-carotene cassava group (0.69 (sd 0.20) micromol/liver). The bioconversion factor was 3.7 microg beta-carotene to 1 microg retinol (2 mol:1 mol), despite 48 % cis-beta-carotene [(Z)-beta-carotene] composition in cassava. In Expt 2, cassava feed with 4.3 nmol provitamin A/g maintained vitamin A status. No effect of cassava variety was observed. Serum retinol concentrations did not differ. Beta-carotene was detected in livers of gerbils receiving cassava and supplements, but the cis-to-trans ratio in liver differed from intake. Biofortified cassava adequately maintained vitamin A status and was as efficacious as beta-carotene supplementation in the gerbil model.

The xanthophyll composition of biofortified maize (Zea mays Sp.) does not influence the bioefficacy of provitamin a carotenoids in Mongolian gerbils (Meriones unguiculatus).

Maize has been targeted for biofortification with provitamin A carotenoids through traditional breeding. Two studies were conducted in gerbils to evaluate factors that may affect provitamin A activity. Maize diets had equal theoretical concentrations of vitamin A (VA) assuming 100% bioefficacy. Study 1 ( n = 57) varied the ratio of beta-cryptoxanthin and beta-carotene but maintained the same theoretical VA. Study 2 ( n = 67) varied lutein and zeaxanthin. Other treatments were oil, VA, or beta-carotene doses. Serum and livers were analyzed for VA and carotenoids. In study 1, total liver VA did not differ among the maize groups. In study 2, total liver VA of the VA and maize groups were higher than controls ( P < 0.05). Conversion factors were 2.1-3.3 mug beta-carotene equivalents to 1 mug retinol. Twice the molar amount of beta-cryptoxanthin was as efficacious as beta-carotene and the proportion of beta-cryptoxanthin or xanthophylls did not appreciably change the VA value of biofortified maize.

beta-Cryptoxanthin from supplements or carotenoid-enhanced maize maintains liver vitamin A in Mongolian gerbils ( Meriones unguiculatus) better than or equal to beta-carotene supplements.

Maize with enhanced provitamin A carotenoids (biofortified), accomplished through conventional plant breeding, maintains vitamin A (VA) status in Mongolian gerbils (Meriones unguiculatus). Two studies in gerbils compared the VA value of beta-cryptoxanthin with beta-carotene. Study 1 (n 47)examined oil supplements and study 2 (n 46) used maize with enhanced beta-cryptoxanthin and beta-carotene. After 4 weeks' depletion, seven or six gerbils were killed; remaining gerbils were placed into weight-matched groups of 10. In study 1, daily supplements were cottonseed oil, and 35, 35 or 17.5 nmol VA (retinyl acetate), beta-cryptoxanthin or beta-carotene, respectively, for 3 weeks. In study 2, one group of gerbils was fed a 50% biofortified maize diet which contained 2.9 nmol beta-cryptoxanthin and 3.2 nmol beta-carotene/g feed. Other groups were given equivalent b-carotene or VA supplements based on prior-day intake from the biofortified maize or oil only for 4 weeks. In study 1, liver retinol was higher in the VA (0.74 (SD 0.11) micromol) and beta-cryptoxanthin (0.5 (SD 0.10) micromol) groups than in the beta-carotene (0.49 (SD 0.13) micromol) and control (0.41 (SD 0.16) micromol)groups (P<0.05). In study 2, the VA (1.17 (SD 0.19) micromol) and maize (0.71 (SD 0.18) micromol) groups had higher liver retinol than the control (0.42 (SD 0.16) micromol) group (P<0.05), whereas the beta-carotene (0.57 (SD 0.21) micromol) group did not. Bioconversion factors (i.e. 2.74 microg beta-cryptoxanthin and 2.4 microg beta-carotene equivalents in maize to 1 microg retinol) were lower than the Institute of Medicine values.

Evaluation of analytical methods for carotenoid extraction from biofortified maize (Zea mays sp.).

Biofortification of maize with beta-carotene has the potential to improve vitamin A status in vitamin A deficient populations where maize is a staple crop. Accurate assessment of provitamin A carotenoids in maize must be performed to direct breeding efforts. The objective was to evaluate carotenoid extraction methods and determine essential steps for use in countries growing biofortified maize. The most reproducible method based on coefficient of variation and extraction efficiency was a modification of Kurilich and Juvik (1999). Heat and saponification are required to release carotenoids from biofortified maize and remove oils interfering with chromatographic analysis. For maize samples with high oil content, additional base may be added to ensure complete saponification without compromising results. Degradation of internal standard before carotenoids were released from the maize matrix required the addition of internal standard after heating to prevent overestimation of carotenoids. This modified method works well for lutein, zeaxanthin, beta-cryptoxanthin, alpha-carotene, and beta-carotene.

Carotenoid-biofortified maize maintains adequate vitamin a status in Mongolian gerbils.

Efforts to biofortify maize with provitamin A carotenoids have been successful, but the impact on vitamin A (VA) status has not been determined. We conducted two studies that investigated the bioefficacy of provitamin A carotenoids from maize and compared maize percentage and carotenoid concentrations on VA status in VA-depleted Mongolian gerbils (Meriones unguiculatus). Gerbils (n = 40/study) were fed a white maize diet 4 wk prior to treatment. In study 1, treatments (n = 10/group) included oil control, 60% high-beta-carotene maize, and beta-carotene or VA supplements (matched to high-beta-carotene maize). In study 2, gerbils were fed 30 or 60% orange or yellow maize diets. Gerbils were killed after 4 wk. In study 1, liver VA concentrations, compared with the high-beta-carotene maize group (0.25 +/- 0.15 micromol/g), were higher in the VA group (0.56 +/- 0.15 micromol/g, P < 0.05), lower in the control (0.10 +/- 0.04 micromol/g, P < 0.05), and did not differ in the beta-carotene group (0.25 +/- 0.08 micromol/g). Bioconversion was approximately 3 microg beta-carotene to 1 mug retinol (1.5 mol beta-carotene to 1 mol retinol). The liver beta-carotene content was greater in the high-beta-carotene maize group (26.4 +/- 6.0 nmol) than in the beta-carotene supplement group (14.1 +/- 6.0 nmol; P < 0.05). In study 2, the gerbils' VA status improved with increasing dietary beta-carotene. Liver VA in gerbils fed orange maize was greater than in those fed yellow maize, regardless of maize percentage (P < 0.05). Biofortified maize adequately maintained VA status in Mongolian gerbils and was as efficacious as beta-carotene supplementation. In populations consuming maize as a staple food, using orange instead of white maize could dramatically affect VA status.

Twice the amount of alpha-carotene isolated from carrots is as effective as beta-carotene in maintaining the vitamin A status of Mongolian gerbils.

The vitamin A (VA) value of carotenoids from fruits and vegetables is affected by many factors. This study determined the VA value of alpha-carotene isolated from carrots compared with beta-carotene and retinyl acetate supplements fed to Mongolian gerbils (Meriones unguiculatus). Gerbils (n = 38) were fed a VA-free diet for 4 wk. At baseline, 6 gerbils were killed to determine liver VA. Gerbils were divided into 3 treatment groups (n = 9/group) and given 35, 35, or 17.5 nmol retinyl acetate, alpha-carotene or beta-carotene, respectively, in 2 divided doses 5 h apart each day. The remaining 5 gerbils received oil vehicle. Gerbils were killed after 3 wk of supplementation. Serum samples and livers were collected and analyzed for VA. Liver extracts were subsequently saponified to quantify alpha-retinol. Serum retinol concentrations did not differ among the groups. Liver retinyl palmitate concentrations were significantly higher in the retinyl acetate treatment group (0.198 +/- 0.051 micromol/g; P < 0.05) than in all other groups. The alpha- and beta-carotene treatments resulted in similar retinyl palmitate concentrations, i.e., 0.110 +/- 0.026 and 0.109 +/- 0.051 micromol/g, respectively, which did not differ from the concentrations in gerbils killed at baseline (0.123 +/- 0.024 micromol/g). The oil group had significantly less retinyl palmitate (0.061 +/- 0.029 micromol/g; P < 0.05) than all other groups. alpha-Retinol was detected in livers of the alpha-carotene group (0.062 +/- 0.013 micromol/g). Thus, twice the amount of purified alpha-carotene maintained VA status as well as beta-carotene in VA-depleted gerbils. Conversion factors were approximately 5.5 microg alpha-carotene or approximately 2.8 mug beta-carotene to 1 microg retinol.

Localization and speciation of chromium in subterranean clover using XRF, XANES, and EPR spectroscopy.

Optimization of phytoremediation and assessment of potential health hazards from metals in the environment requires an understanding of absorption, localization, and transport of the target metal by plants. The objectives of this study were to localize Cr and determine the oxidation state and possible complexation mode of Cr in intact plant tissue by means of XANES, synchrotron XRF microprobe spectroscopy, and EPR spectroscopy. Subterranean clover (Trifolium brachycalycinum) was grown hydroponically with Cr(VI) (0.04-2.0 mmol L(-1)) and compared with plants grown without Cr and with inorganic Cr(III) and various Cr(III)-organic sources. The uptake, translocation, and form of Cr in the plant were dependent on the form and concentration of supplied Cr. Chromium was found predominately in the +3 oxidation state, regardless of the Cr source supplied to the plant, though at high Cr(VI) treatment concentrations, Cr(VI) and Cr(V) were also observed. At low Cr(VI) concentrations, the plant effectively reduced the toxic Cr(VI) to less toxic Cr(III), which was observed both as a Cr(III) hydroxide phase at the roots and as a Cr(III)-organic complex in the roots and shoots. At low Cr(VI) treatment concentrations, Cr in the leaves was observed predominately around the leaf margins, while at higher concentrations Cr was accumulated at leaf veins.