Effect of physiological activities on aluminum uptake in carrot (Daucus carota L.) cells in suspension culture

Responses to aluminum (AI) of carrot (Daucus carota L.) cells in suspension culture (hereafter referred as “carrot suspension cells”) were investigated under different physiological activities. The respiration rate of the cells at the logarithmic growth phase decreased when the external Al concentration increased. A similar tendency was observed for the ATP content of the cells when the external Al concentration of the medium increased. Decrease of about 50% of the respiration rate and of about 35% of the ATP content of the cells was. observed at 300 µm AlCl₃ at 24 h, respectively. The Al content (total and citrate-insoluble AI) in cells increased when the external Al concentration increased. Both the respiration rate and the ATP content in cells were reduced when the culture temperature decreased or ATP synthesis inhibitors, namely CCCP (carbonylcyanide-m-chlorophenylhydrazone) and DNP (2,4-dinitrophenol) were added. The addition of SHAM (salicylhydroxamic acid) did not affect the ATP content in cells. The Al content of the cells treated with 50 µm Al together with SHAM was similar to that of the cells treated with only AI. However, the Al content of the cells increased when the cells were treated with 50 µm Al together with CCCP, DNP, or a low temperature (15°C). The content of citrate Al content in cells increased rapidly when ATP synthesis inhibitors were present or the culture temperature was low. There was a negative linear correlation between the Al content and the ATP content in cells. These results suggest that the cells containing a large amount of ATP and showing normal functions of the plasma membranes may display an apparent tolerance to Al by suppressing Al absorption into their cytoplasm at a cell suspension level.     

Isolation and identification of novel pyranoanthocyanins from black carrot (Daucus carota L.) juice

Six novel pyranoanthocyanins were identified by HPLC-ESI-MSn in black carrot (Daucus carota L. ssp. sativus var. atrorubens Alef.) juice. The two major compounds, namely, the vinylcatechol adducts of cyanidin 3-O-(6-O-feruloyl-beta-D-glucopyranosyl)-(1-->6)-[beta-D-xylopyranosyl-(1-->2)]-beta-D-galactopyranoside and cyanidin 3-O-[beta-D-xylopyranosyl-(1-->2)]-beta-D-galactopyranoside, respectively, were isolated by a combination of high-speed countercurrent chromatography with semipreparative HPLC. Their structures were fully elucidated by means of one- and two-dimensional NMR spectroscopy and high-resolution mass spectrometry. The four remaining pigments were characterized as the vinylphenol and vinylguaiacol adducts of cyanidin 3-O-[beta-D-xylopyranosyl-(1-->2)]-beta-D-galactopyranoside, the vinylguaiacol adduct of cyanidin 3-O-(6-O-feruloyl-beta-D-glucopyranosyl)-(1-->6)-[beta-D-xylopyranosyl-(1-->2)]-beta-D-galactopyranoside, and the vinylcatechol adduct of cyanidin 3-O-(6-O-sinapoyl-beta-d-glucopyranosyl)-(1-->6)-[beta-D-xylopyranosyl-(1-->2)]-beta-D-galactopyranoside. These compounds are formed during storage of the juice through the direct reaction of either caffeic, ferulic, or coumaric acid with the respective genuine anthocyanins.     

Purple carrot (Daucus carota L.) polyacetylenes decrease lipopolysaccharide-induced expression of inflammatory proteins in macrophage and endothelial cells

Carrots ( Daucus carota L.) contain phytochemicals including carotenoids, phenolics, polyacetylenes, isocoumarins, and sesquiterpenes. Purple carrots also contain anthocyanins. The anti-inflammatory activity of extracts and phytochemicals from purple carrots was investigated by determining attenuation of the response to lipopolysaccharide (LPS). A bioactive chromatographic fraction (Sephadex LH-20) reduced LPS inflammatory response. There was a dose-dependent reduction in nitric oxide production and mRNA of pro-inflammatory cytokines (IL-6, IL-1beta, TNF-alpha) and iNOS in macrophage cells. Protein secretions of IL-6 and TNF-alpha were reduced 77 and 66% in porcine aortic endothelial cells treated with 6.6 and 13.3 microg/mL of the LH-20 fraction, respectively. Preparative liquid chromatography resulted in a bioactive subfraction enriched in the polyacetylene compounds falcarindiol, falcarindiol 3-acetate, and falcarinol. The polyacetylenes were isolated and reduced nitric oxide production in macrophage cells by as much as 65% without cytotoxicity. These results suggest that polyacetylenes, not anthocyanins, in purple carrots are responsible for anti-inflammatory bioactivity.     

Aluminum-tolerance of carrot (daucus carota L.) plants regenerated from selected cell cultures

Abstract

Aluminum-tolerant variants from cell cultures of carrot (Daucus carota L. cv. MS Yonsun) were selected by exposing them to excess ionic aluminum. The medium contained A1CI₃ and 0.1 mM phosphate at pH 4.0. Under ionic aluminum stress (0.5-1.0 mM), 282 calli were selected. Twenty-eight fertile plants were regenerated from 7 selected calli. Based on the root elongation tests of the seedlings, the seeds displayed aluminum-tolerance. In the case of the cell line H7-08, the selection occurred at a frequency of ca. 6 × 10^-7 when the materials were plated on agar medium containing 0.5 mM aluminum.     

Impact of egyptian broomrape (Orobanche aegyptiaca (Pers.) parasitism on amino acid composition of carrot (Daucus carota L.)

The relationship between the organic nitrogen status of Egyptian broomrape and one of its hosts, carrot, was studied by comparing amino acid profiles of leaf and root tissues of nonparasitized and broomrape-parasitized carrot plants and by analyzing amino acid profiles of broomrape at different growth stages. Total N concentrations, expressed as a percentage of the dry weight of the tissues, were similar in leaves of nonparasitized and parasitized carrot plants but were lower in parasitized roots than in nonparasitized roots. In both dry and germinated broomrape seeds, N concentrations were lower than or similar to those in broomrape tubercles, shoots, or callus. Individual amino acid concentrations in hydrolysates of leaves of parasitized carrot plants tended to be similar to or greater than those in hydrolysates of nonparasitized carrot plants. Roots of parasitized plants tended to have similar or lower amino acid concentrations than roots of nonparasitized plants. Dry and germinated broomrape seeds had similar amino acid profiles, but individual amino acid concentrations were lower than in the other broomrape tissues examined. The broomrape shoot tended to have lower amino acid concentrations than the tubercle and callus. Free amino acid profiles of leaves and roots of parasitized plants paralleled those of nonparasitized plants, respectively. Individual free amino acids tended to occur at similar or lower levels in dry and germinated broomrape seeds than in the tubercle, shoot, or callus. Free amino acid composition of the broomrape tubercle was similar to that of the parasitized root. Arginine and alanine concentrations in broomrape callus were dramatically higher than those of other amino acids in this or other tissues investigated. These results indicate that changes in the composition of both free and bound amino acids in carrot are associated with broomrape parasitism.     

胡萝卜DcDREB-A6亚族转录因子基因的克隆与非生物胁迫响应分析

干旱响应元件结合蛋白(dehydration responsive element-binding protein,DREB)类转录因子是干旱应答元件的结合蛋白,在植物响应非生物胁迫时发挥重要作用。本研究基于胡萝卜(Daucus carota L.)转录组数据,检索和拼接获得一个胡萝卜DREB类转录因子基因序列,并根据其序列设计引物,采用RTPCR方法从黑田五寸获得该胡萝卜转录因子基因DcDREB-A6(GenBank登录号:KM386646)。序列分析显示,来源于胡萝卜中的DcDREB-A6基因含有993 bp的开放阅读框,编码330个氨基酸。氨基酸组成成分、理化性质、亲水性/疏水性和三级结构分析结果显示,胡萝卜DcDREB-A6转录因子蛋白质分子量为36.68 kD,等电点为6.00,属于亲水性蛋白。进化树分析显示,DcDREB-A6属于APETLA 2/乙烯反应元件结合蛋白(AP2/ERF)家族转录因子中DREB亚族中的A6组。空间结构表明,该转录因子结构域具有典型的植物AP2/ERF家族转录因子的结构特征,有1个α螺旋和3个β折叠。实时定量荧光PCR证实,胡萝卜DcDREB-A6转录因子基因受高温、低温和高盐胁迫诱导,分别在高温、低温处理4和2 h后表达量增加2倍左右;盐胁迫对其影响较大,处理8 h时,基因表达量增加4.6倍;而干旱胁迫下该基因表达变化幅度相对较小。胡萝卜DcDREB-A6转录因子基因能够响应高温、低温、干旱和高盐等非生物胁迫,本研究结果有助于进一步开展胡萝卜DREB类转录因子的逆境调控研究。     

Inheritance of purple colour and its different flavonoids profiling in F2 populations of tropical carrot (Daucus carota L.)

Genetic Resources and Crop Evolution - The results of the inheritance study revealed that the purple root epiderm (Pe) and cortex (Pcx) colour were dominant over the yellow colour of epiderm (ye)...     

Quantitative analysis of aroma compounds in carrot (Daucus carota L.) cultivars by capillary gas chromatography using large-volume injection technique

Dynamic headspace sampling was used to collect aroma compounds from raw samples of four carrot (Daucus carota L.) cultivars (Brasilia, Duke, Fancy, and Cortez). The collected volatiles were analyzed by capillary GC-FID and GC-MS using large-volume cool on-column injection (LVI-COC). Of the 36 compounds identified, 6 had not been previously detected in carrots. Significant differences between the carrot cultivars were found for 31 of the identified volatiles as well as for total monoterpenes, sesquiterpenes, and total volatile content. Mono- and sesquiterpenes accounted for about 98% of the total volatile mass in all cultivars. LVI-COC injection was used to determine the loss of carrot volatiles during concentration of headspace samples under a stream of nitrogen. The loss among major monoterpenes in the concentrated samples varied from 16% for p-cymene to >40% for alpha-pinene as compared to nonconcentrated samples. The loss among high-boiling sesquiterpenes varied from not detectable (beta-caryophyllene, alpha-humulene, and caryophyllene oxide) to approximately 7% for (E)- and (Z)-gamma-bisabolene.     

Genetic analysis of root traits in tropical carrots (Daucus carota L.)

Genetic Resources and Crop Evolution - Carrot root being used as vegetable and salad due to the nutritional value, hence consumers demanding carrot production. Thus, the production can be achieved...     

Structural and sensory characterization of compounds contributing to the bitter off-taste of carrots (Daucus carota L.) and carrot puree

Sequential application of solvent extraction, gel permeation chromatography, and HPLC in combination with taste dilution analyses revealed that not a sole compound but a multiplicity of bitter tastants contribute to the bitter off-taste of cold-stored carrots and commercial carrot puree, respectively. Among these bitter compounds, 3-methyl-6-methoxy-8-hydroxy-3,4-dihydroisocoumarin (6-methoxymellein), 5-hydroxy-7-methoxy-2-methylchromone (eugenin), 2,4,5-trimethoxybenzaldehyde (gazarin), (Z)-heptadeca-1,9-diene-4,6-diin-3,8-diol (falcarindiol), (Z)-heptadeca-1,9-diene-4,6-diin-3-ol (falcarinol), and (Z)-3-acetoxy-heptadeca-1,9-diene-4,6-diin-8-ol (falcarindiol 3-acetate) could be identified on the basis of MS as well as 1D- and 2D-NMR experiments. Due to the low concentrations of <0.1 mg/kg and the high taste thresholds found for eugenin and gazarin, these compounds could be unequivocally excluded as important contributors to the bitter taste of carrots. Calculation of bitter activity values as the ratio of their concentration to their bitter detection threshold clearly demonstrated that neither in fresh and stored carrots nor in commercial carrot puree did 6-methoxymellein contribute to the bitter off-taste. In contrast, the concentrations of falcarindiol in stored carrots and, even more pronounced, in carrot puree were found to be 9- and 13-fold above its low bitter detection concentration of 0.04 mmol/kg, thus demonstrating that this acetylenic diol significantly contributes to the bitter taste of the carrot products investigated.     

Sucrose transporter NtSUT1 confers aluminum tolerance on cultured cells of tobacco (Nicotiana tabacum L.)

The role of plasma membrane-localized sucrose transporter (NtSUT1) was investigated using cultured tobacco cell (Nicotiana tabacum L.) line BY-2. The wild type (WT) cells were first transformed with the NtSUT1 gene or its fragments cloned from tobacco cell line SL to form the over-expression (OX) and suppression (RNAi) cell lines, respectively. Using OX and RNAi transgenics, the role of NtSUT1 in growth capacity of actively growing cells and in aluminum (Al)-treated cells was examined. During the logarithmic phase of growth in nutrient medium containing 2,4-dichlorophenoxyacetic acid (2,4-D), both the rate of sucrose uptake measured with radio-tracer and the content of soluble sugars were higher in OX and lower in RNAi cell lines compared to WT. Overall, the content of soluble sugars negatively correlated with the time necessary for doubling mass (fresh weight). When cells were treated without (control) or with Al in a simple medium containing calcium, sucrose and 2-(N-morpholino)ethanesulfonic acid (MES; pH 5.0) for up to 18 h, the expression of NtSUT1 under its native promoter, or under the control of strong constitutive cauliflower mosaic virus (CaMV) 35S promoter, was strongly dependent on the presence of 2,4-D. Thereafter, the cells were preferentially treated in the presence of 2,4-D. During 6 h after a start of the control treatment, sucrose uptake rates were, compared to WT, slightly higher and lower in OX and RNAi lines respectively. The addition of Al reduced the sucrose uptake rates of OX and WT to the level of RNAi line, indicating that Al inhibits sucrose uptake via NtSUT1. During the post-Al culture of control and Al-treated cells in a nutrient medium, sucrose uptake rates were much higher in OX compared to WT and RNAi lines, which closely and positively correlated with the growth capacity of the cells. Judging from the growth capacity of Al-treated cells relative to that of control cells, OX cells were more tolerant to Al than WT and RNAi. In summary, we conclude that over-expression of NtSUT1 confers higher growth capacity in actively growing cells as well as in Al-treated cells.     

Possible relationships between wild and cultivated carrots (Daucus carota L.) in the Netherlands

Summary A theory about the descent and development of modern carrot cultivars in Europe formulated byBanga (1963) is discussed. According to this author, the cultivated carrot originates from Afghan carrot material. Furthermore, the origin of carrots with fleshy, orange roots is suggested to lay in the Netherlands in the 17th century.However, it is shown that wild populations of the carrot with white coloured roots have occurred in Europe since prehistoric times. During the late Middle Ages and Renaissance these carrots were used as medicinal plants. It is suggested that these indigenous carrots have played a minor role in the development of modern cultivated carrots.

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Verwandtschaftliche Beziehungen zwischen wilden und kultivierten Möhren (Dau-cus carota L.) in den Niederlanden (Daucus carota L.)

Zusammenfassung Die vonBanga (1963) formulierte Abstammungs- und Entwicklungstheorie über die rezenten Möhrensorten wird diskutiert. Nach seiner Meinung sind die kultivierten Möhren aus afghanischen Formen entstanden. Darüber hinaus wird vermutet, daß Möhren mit fleischigen, orangefarbigen Wurzeln in den Niederlanden im 17. Jahrhundert entwickelt worden sind.Es wird jedoch gezeigt, daß es schon seit prähistorischen Zeiten in Europa Wild-populationen von Möhren mit weißfarbigen Wurzeln gegeben hat. Diese Möhren wurden im späten Mittelalter und in der Renaissance als Heilpflanzen verwendet. In vorliegender Arbeit wird darauf hingewiesen, daß diese heimischen Möhrenformen bei der Entstehung unserer heutigen Möhrensorten eine nur geringe Rolle gespielt haben dürften.

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Chelate-enhanced phytoextraction and phytostabilization of lead-contaminated soils by carrot (Daucus carota)

The objective of this study was to study the influence of different ethylenediamine tetraacetate (EDTA), nitrilotriacetic acid (NTA) and oxalic acid (HOx) concentrations on tolerance and lead (Pb) accumulation capacity of carrot (Daucus carota). The results indicated that by increasing Pb, NTA and HOx concentrations in the soil, the shoot, taproot and capillary root dry matters increase effectively. In contrary, EDTA caused to reduce capillary roots biomass. EDTA was more effective than NTA and HOx in solubilizing soil Pb. The highest Pb content in shoots (342.2 ± 13.9 mg kg^?1) and taproots (301 ± 15.5 mg kg^?1) occurred in 10 mM EDTA, while it occurred for capillary roots (1620 ± 24.6 mg kg^?1) in 5 mM HOx, when the soil Pb concentration was 800 mg kg^?1. The obtained high phytoextraction and phytostabilization potentials were 1208 (±25.6) and 11.75 (±0.32) g Pb ha^?1 yr^?1 in 10 mmol EDTA kg^?1 soil and no chelate treatments, respectively. It may be concluded that chelate application increases Pb uptake by carrots. Consequently, this plant can be introduced as a hyperaccumulator to phytoextract and phytostabilize Pb from contaminated soils.     

Effect of storage temperature on the stability of anthocyanins of a fermented black carrot (Daucus carota var. L.) beverage: shalgam

The effect of temperature on the stability of shalgam anthocyanins stored at 4, 25, and 40 degrees C for 90 days was investigated. The effect of pasteurization and sorbate addition on the anthocyanin stability as compared to control was also studied. The monomeric anthocyanin content and color density decreased with increasing time as a function of storage temperature whereas the percent polymeric color and browning increased. The same trends were observed in control, pasteurized, and sorbate-added shalgam samples. Shalgam anthocyanins consisted of two nonacylated and three acylated cyanidin derivatives. Acylated anthocyanins were more stable when compared to nonacylated ones at all storage temperatures. The activation energies, 11.11-11.64 kcal/mol, were calculated from the reaction rate constants evaluated taking first-order reaction kinetics. The highest anthocyanin retention was observed at 4 degrees C storage temperature with a half-life between 231 and 239 days.     

Mechanical weeding effects on soil structure under field carrots (Daucus carota L.) and beans (Vicia faba L.)

The successful production of organic vegetables relies heavily on mechanical weeding, flame weeding and stale seedbeds. These operations involve repeated passes by tractors. Mechanical weeding also involves regular tillage. This combination of repeated tillage and compaction changes soil structure. We studied these structural changes in two fields of organic carrots and one field of beans in eastern Scotland. Structure was described by measuring soil strength with a vane shear tester and a cone penetrometer, by measuring bulk density and by visual assessment. Under beans, vane shear strength below the growing root zone was highly variable and in some areas was high enough to restrict root growth (>50 kPa). The carrots were grown in beds containing crop rows separated by bare soil. The bare soil was regularly weeded mechanically. The structure of this weeded soil in the top 10 cm layer of a loam eventually became disrupted and compacted enough to deter root growth (vane shear strength of 70 kPa). In addition the topsoil and subsoil in the wheel-tracks between the beds became very compact with little distinguishable structure. This compaction extended to the subsoil and persisted into the next cropping season (cone resistance >3 MPa at 35–50 cm depth). Reduced tillage by discing without ploughing was used to incorporate the straw used to protect the carrots overwinter and prepare the soil for the next crop. The resulting topsoil quality was poor leading to anaerobic growing conditions which restricted growth of the following crop and led to losses of the greenhouse gas nitrous oxide. The greatest threat to soil quality posed by mechanical weeding was subsoil compaction by tractor wheeling.     

Changes in volatile compounds of carrots (Daucus carota L.) during refrigerated and frozen storage

Carrots (Daucus carota L.) of cv. Bolero and cv. Carlo were processed into shreds and stored for up to 4 months at -24 degrees C (frozen storage), or the roots were stored for up to 4 months at 1 degrees C (refrigerated storage) followed by processing into shreds. Volatiles from the carrot shreds were collected by dynamic headspace technique and analyzed by GC-FID, GC-MS, GC-MS/MS, and GC-O to determine the volatile composition and aroma active components of carrots stored under different temperature conditions. A total of 52 compounds were quantified, of which mono- and sesquiterpenes accounted for approximately 99% of the total volatile mass. Major volatile compounds were (-)-alpha-pinene, beta-myrcene, (-)-limonene, (+)-limonene, (+)-sabinene, gamma-terpinene, p-cymene, terpinolene, beta-caryophyllene, alpha-humulene, and (E)- and (Z)-gamma-bisabolene. A considerable increase in the concentration of mono- and sesquiterpenes was observed during refrigerated storage, whereas the concentration of terpenoids was around the same level during frozen storage. GC-O revealed that the major volatiles together with (+)-alpha-pinene, (-)-beta-pinene, (+)-beta-pinene, 6-methyl-5-hepten-2-one, (-)-beta-bisabolene, beta-ionone, and myristicin had an odor sensation, which included notes of "carrot top", "terpene-like", "green", "earthy", "fruity", "citrus-like", "spicy", "woody", and "sweet".     

Comparison of aluminum tolerance and phosphate absorption between rape (Brassica napus L.) and Tomato (Lycopersicum esculentum Mill.) in relation to organic acid exudation

Aluminum (Al) tolerance and phosphate absorption in rape and tomato were compared under water culture and field conditions. The relative growth rate in the Al treatment compared with -A1 treatment was similar in the two crops under water culture conditions, while under field conditions, the growth rate was 2- to 3-fold higher in rape than in tomato in spite of the higher Al concentration in the soil solution than in the culture solution. The relative amount of phosphate absorbed in the Al treatment compared with - Al was not appreciably different between rape and tomato under water culture conditions, while under field conditions, it was 3- to 6-fold larger in rape than in tomato. The exudation rate of citric acid by roots was much higher in rape than in tomato. The plant growth, root elongation, and amount of phosphate absorbed in rape were inhibited in the 150 µM Al in the culture solution. However, the inhibition was alleviated by the addition of 200 µM citric acid or 500 µM malic acid. The P concentration in the culture solution decreased by the presence of Al as aluminum phosphate. However, addition of citric and malic acids increased the amount of phosphate released from the precipitated aluminum phosphate. In conclusion, one of the mechanisms for the higher Al tolerance and larger phosphate absorption in rape than in tomato under field conditions was ascribed to the higher concentration of exuded citric acid by Al in the rhizosphere. It was suggested that the exudation of citric acid might contribute to the detoxification of Al and to the increase phosphate availability in the rhizosphere in rape.     

Carotenoid profiles and consumer sensory evaluation of specialty carrots (Daucus carota, L.) of various colors

Five different colored carrots were analyzed for their carotenoid profile and underwent sensory evaluation to determine consumer acceptance (n = 96). Four major carotenoids were identified and quantified by use of HPLC methods. High beta-carotene orange carrots were found to contain the greatest concentration of total carotenoids. Except for the white, all the carrots are a significant source of bioavailable carotenoids. Sensory evaluation showed the high beta-carotene orange and white carrots to be favored over the yellow, red, and purple carrots in both blind and nonblind treatments (P < 0.01). However, all the carrots were well accepted by the consumer panel. With this information, carrot growers should be encouraged to cultivate specialty carrots to provide sources of both vitamin A precursors and phytochemicals.     

Reinvestigation of the bitter compounds in carrots (Daucus carota L.) by using a molecular sensory science approach

In order to reinvestigate the key molecules inducing bitter off-taste of carrots ( Daucus carota L.), a sensory-guided fractionation approach was applied to bitter carrot extracts. Besides the previously reported bitter compounds, 6-methoxymellein (1), falcarindiol (2), falcarinol (3), and falcarindiol-3-acetate (4), the following compounds were identified for the first time as bitter compounds in carrots with low bitter recognition thresholds between 8 and 47 micromol/L: vaginatin (5), isovaginatin (6), 2-epilaserine oxide (7), laserine oxide (8), laserine (14), 2-epilaserine (15), 6,8-O-ditigloyl- (9), 6-O-angeloyl-, 8-O-tigloyl- (10), 6-O-tigloyl-, 8-O-angeloyl- (11), and 6-, 8-O-diangeloyl-6 ss,8alpha,11-trihydroxygermacra-1(10) E,4 E-diene (12), as well as 8-O-angeloyl-tovarol (13) and alpha-angeloyloxy-latifolone (16). Among these bitter molecules, compounds 9, 10, 13, and 16 were not previously identified in carrots and compounds 6, 11, and 12 were yet not reported in the literature.