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Carrot (Daucus carota L.), a widely cultivated economically vegetable from the Apiaceae family, is grown globally. However, carrots can be adversely impacted by cadmium (Cd) pollution in the soil due to its propensity to accumulate in the fleshy root, thus impeding carrot growth and posing health hazards to consumers. Given the potential of hydrogen sulfide (H 2 S) to improve plant resistance against Cd stress, we treated germinating carrot seeds with varying concentrations of sodium hydrosulfide (NaHS), aiming to alleviate the toxic impacts of Cd stress on carrot seed germination. The results revealed that carrot seeds treated with a concentration of 0.25 mM NaHS displayed better seed germination-associated characteristics compared to seeds treated with NaHS concentrations of 0.1 mM and 0.5 mM. Further investigation revealed a rise in the expression levels of L-cysteine desulfhydrase and D-cysteine desulfhydrase, along with enhanced activity of L-cysteine desulfhydrase and D-cysteine desulfhydrase among the NaHS treatment group, thereby leading to H 2 S accumulation. Moreover, NaHS treatment triggered the expression of pyrroline-5-carboxylate synthase and pyrroline-5-carboxylate reductase and promoted the accumulation of endogenous proline, while the contents of soluble sugar and soluble protein increased correspondingly. Interestingly, since the application of exogenous proline did not influence the accumulation of endogenous H 2 S, suggesting that H 2 S served as the upstream regulator of proline. Histochemical staining and biochemical indices revealed that NaHS treatment led to elevated antioxidant enzyme activity, alongside a suppression of superoxide anion and hydrogen peroxide generation. Furthermore, high performance liquid chromatography analysis revealed that NaHS treatment reduced Cd 2+ uptake, thereby promoting germination rate, seed vitality, and hypocotyl length of carrot seeds under Cd stress. Overall, our findings shed light on the application of NaHS to enhance carrot resistance against Cd stress and lay a foundation for exploring the regulatory role of H 2 S in plants responding to Cd stress., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

Early season carrot (Daucus carota) production is being practiced in Punjab, Pakistan to meet the market demand but high temperature hampers the seed germination and seedling establishment which cause marked yield reduction. Seed priming with potassium nitrate breaks the seed dormancy and improves the seed germination and seedling growth potential but effects vary among the species and ecological conditions. The mechanism of KNO 3 priming in high temperature stress tolerance is poorly understood yet. Thus, present study aimed to evaluate high temperature stress tolerance potential of carrot seeds primed with potassium nitrate and impacts on growth, physiological, and antioxidant defense systems. Carrot seeds of a local cultivar (T-29) were primed with various concentration of KNO 3 (T 0 : unprimed (negative control), T 1 : hydroprimed (positive control), T 2 : 50 mM, T 3 :100mM, T 4 : 150 mM, T 5 : 200 mM, T 6 : 250 mM and T 7 : 300 mM) for 12 h each in darkness at 20 ± 2℃. Seed priming with 50 mM of KNO 3 significantly enhanced the seed germination (36%), seedling growth (28%) with maximum seedling vigor (55%) and also exhibited 16.75% more carrot root biomass under high temperature stress as compared to respective control. Moreover, enzymatic activities including peroxidase, catalase, superoxidase dismutase, total phenolic contents, total antioxidants contents and physiological responses of plants were also improved in response to seed priming under high temperature stress. By increasing the level of KNO 3 , seed germination, growth and root biomass were reduced. These findings suggest that seed priming with 50 mM of KNO 3 can be an effective strategy to improve germination, growth and yield of carrot cultivar (T-29) under high temperature stress in early cropping. This study also proposes that KNO 3 may induces the stress memory by heritable modulations in chromosomal structure and methylation and acetylation of histones that may upregulate the hormonal and antioxidant activities to enhance the stress tolerance in plants., (© 2024. The Author(s).)

The rapid activation of phosphatidylinositol-specific phospholipase C (PI-PLC) occurs early after the stimulation of biotic and abiotic stress in plants, which directly associated with the calcium channel-induced calcium ion (Ca 2+ ) influx. Exogenous calcium chloride (CaCl 2 ) mediates the calcium signaling transduction to promote the γ-aminobutyric acid accumulation and nutritional quality in shredded carrots whereas the generation mechanism remains uncertain. Therefore, the involvement of PI-PLC-associated phospholipid metabolism was investigated in present study. Our result revealed that CaCl 2 treatment promoted the expression and activity of PI-PLC and increased the inositol 1,4,5-trisphosphate and hexakisphosphate content in shredded carrots. The transcripts of multi-glutamate receptor-like channels (DcGLRs), the glutamate and γ-aminobutyric acid (GABA) content, and Ca 2+ influx were induced by CaCl 2 treatment in shredded carrots during storage. However, PI-PLC inhibitor (U73122) treatment inhibited the activation of PI-PLC, the increase of many DcGLRs family genes expression levels, and Ca 2+ influx. Moreover, the identification of DcPI-PLC4/6 and DcGLRs proteins, along with the analysis of characteristic domains such as PLCXc, PLCYc, C2 domain, transmembranous regions, and ligand binding domain, suggests their involvement in phospholipid catalysis and calcium transport in carrots. Furthermore, DcPI-PLC4/6 overexpression in tobacco leaves induced the Ca 2+ influx by activating the expressions of NtGLRs and the accumulation of glutamate and GABA. These findings collectively indicate that CaCl 2 treatment-induced PI-PLC activation influences DcGLRs expression levels to mediate cytosolic Ca 2+ influx, thus, highlighting the "PI-PLC-GLRs-Ca 2+ " pathway in calcium signaling generation and GABA biosynthesis in shredded carrots., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Fruit and vegetable consumption has increased due to their tremendous health benefits. However, recent studies have shown that contaminated products may serve as vehicles for foodborne pathogens and harmful chemicals. Therefore, fresh vegetables must be decontaminated before consumption to ensure food safety., Results: In this study, the combined decontamination treatment of lactic acid (2.5 mL L -1 ) and ozone (9 mg L -1 ) for 10 min showed better efficacy in the removal of contaminants from fresh vegetables as compared to individual treatments. The combined treatment resulted in a reduction of 1.5-3.5 log CFU of native mesophilic bacteria per gram and 1.6-2.9 log CFU of artificially inoculated Escherichia coli per gram from tomato, cucumber, carrot and lettuce. The combined treatment also removed spiked pesticides, which represent artificial chemical contamination (28-97% chlorpyrifos and 62-100% λ-cyhalothrin residues), from fresh vegetables. No significant difference (P > 0.05) in various sensory attributes of vegetables was observed between untreated and treated (lactic acid and ozone) vegetables., Conclusions: The combination treatment provides a novel approach to target two groups of contaminants using a single procedure. The combination treatment can be used as an alternative to currently used decontamination techniques for the supply of safe vegetables to consumers. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

The study was aimed to investigate the combined effect of acid blanching (AB) and high-voltage electric field cold plasma (HVCP) on carrot juice quality. Before juice extraction, carrots were separated into three parts: control, blanched (100 °C for 5 min) with non-acidified water, and blanched with acidified water (35 g/L citric acid at pH 1.34). Carrot juice was then subjected to dielectric barrier discharge at 80 kV for 4 min. Results indicated that AB treatment significantly influenced the efficiency of HVCP. AB-HVCP resulted in antimicrobial synergism, which is an outcome of acidified NO 2 - , H 2 O 2 , O - , and peroxynitrites (ONOO - ) or its precursor OH/NO 2 , along with other species. In addition, plasma treatment also promotes the accumulation of coloring compounds, chlorogenic acid, and sugar contents by surface erosion of the epidermal layer, cis isomerization, rupturing of phenol-sugar and phenolic-cell matrix bonds, and depolymerized long-chain polysaccharides by cleavage of the glycoside bond. Therefore, AB-HVCP is a potential emerging hurdle strategy for fresh produce.

The exposure to combinations of heavy metals can affect the genes of vegetables and heavy metals would accumulate in vegetables and thereby indirectly affecting human health. Exploring the links between genetic changes and phenotypic changes of carrot under the combined pollution of Cd and Cu is of great significance for studying the mechanism of heavy metal pollution. Therefore, this study examined the effects of mixtures of cadmium (Cd) and copper (Cu) on physiological measures (malondialdehyde (MDA), proline, and antioxidant enzyme) and expression of growth-related genes (gibberellin gene, carotene gene, and glycogene) in carrot under greenhouse cultivation. The results showed in the additions with mixtures of Cd and Cu at higher concentration, the MDA content increased significantly (p < 0.05), whereas the proline content was not significantly different from those in the control. In the mixed treatments with high Cd concentrations, the activity of superoxide dismutase (SOD) was significantly lower than that in the control (p < 0.05); whereas the activity of peroxidase (POD) increased to different degrees compared to the control. In the additions with mixtures of Cd and Cu, compared with the control, the expression of the gibberellin gene was downregulated from 1.97 to 20.35 times (not including the 0.2 mg kg -1 Cd and 20 mg kg -1 Cu mixture, the expression of gibberellin gene in this treatment was upregulated 1.29 times), which lead to decreases in the length and dry weight of carrots. The expression of the carotene gene in mixed treatments downregulated more than that in single treatments, which could reduce the ability of carrots to resist oxidative damage, as suggested by the significant increase in the MDA content. In the addition with mixtures of Cd and Cu, compared with the control, the expression of the glycogene was downregulated by 1.42-59.40 times, which can cause a significant reduction in the sugar content in carrots and possibly further reduce their ability to resist heavy metal damage. A cluster analysis showed that in the additions with mixtures of Cd and Cu, the plant phenotype was affected first, and then with increases in the added concentration, the expression of genes was also affected. In summary, in the additions with mixtures of Cd and Cu, plants were damaged as Cd and Cu concentrations increased., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: During the production of fresh-cut products, crops are exposed to wounding stress, and as a stress response, phenolic antioxidants are synthesized. This stress response is elicited by extracellular adenosine triphosphate, released from wounded cells and recognized by receptors of unwounded cells. The phenolic antioxidants produced as a stress response are beneficial for human health. However, a common practice in the fresh-cut industry is the application of washing/sanitizing procedures after cutting. These procedures could be highly detrimental, since they partially remove the wound signal that elicits the biosynthesis of phenolics in plants. In this study, the impact of different washing/sanitizing treatments post-shredding on the wound-induced accumulation of chlorogenic acid (CHA) in carrot was evaluated. Peeled carrots were shredded and dipped in aqueous solutions containing chlorine (100 ppm, 2 min), hydrogen peroxide (1.5%, 2 min) or water (2 min). The content of CHA in treated carrots was evaluated before and after 48 h of storage (19 ± 2 °C)., Results: The control carrots sanitized only before peeling and shredding showed 4000% higher content of CHA as compared with time 0 h samples. However, carrots treated with washing/sanitizing procedures post-shredding including water, chlorine and hydrogen peroxide showed a decrease in the accumulation of CHA by 46.9%, 53.6% and 89.9%, respectively., Conclusions: The results demonstrated that washing/sanitizing procedures applied after fresh-cutting are potentially detrimental to the wound-induced accumulation of health-promoting compounds during storage of fresh produce. Thus, the fresh-cut industry could consider avoiding washing procedures after cutting and implement alternative sanitizing procedures that avoid the partial removal of the wound signal, such as sanitizing only before cutting. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Carrot is an annual or biennial herbaceous plant of the Apiaceae family. Carrot is an important vegetable, and its fresh taproot, which contains rich nutrients, is the main edible part. In the life cycle of carrot, NAC family transcription factors (TFs) are involved in almost all physiological processes. The function of NAC TFs in carrot remains unclear. In this study, 73 NAC family TF members in carrot were identified and characterized using transcriptome and genome databases. These members were divided into 14 subfamilies. Multiple sequence alignment was performed, and the conserved domains, common motifs, phylogenetic tree, and interaction network of DcNAC proteins were predicted and analyzed. Results showed that the same group of NAC proteins of carrot had high similarity. Eight DcNAC genes were selected to detect their expression profiles under abiotic stress treatments. The expression levels of the selected DcNAC genes significantly increased under treatments with low temperature, high temperature, drought, and salt stress. Results provide potentially useful information for further analysis of the roles of DcNAC transcription factors in carrot.

Application of municipal biosolids in agriculture present a concern with potential uptake and bioaccumulation of pharmaceutical compounds from biosolids into agronomic plants. We evaluated the efficacy of biochar as a soil amendment to minimize uptake of antimicrobial agents (ciprofloxacin, triclocarban, and triclosan) in lettuce ( Lactuca sativa ) and carrot ( Daucus carota ) plants. Biochar reduced the concentration of ciprofloxacin and triclocarban in lettuce leaves and resulted in a 67% reduction of triclosan in carrot roots. There was no substantial difference in pharmaceutical concentrations in carrot and lettuce plant matter at low (2.0 g kg -1 soil) and high (20.4 g kg -1 soil) rates of applied biochar. The co-amendment of biochar and biosolids increased soil pH and nutrient content which were positively correlated with an increase in lettuce shoot biomass. Our results demonstrate the potential efficacy of using walnut shell biochar as a sorbent for pharmaceutical contaminants in soil without negatively affecting plant growth.

The aim of a three-year study was to assess the effect of combined biofortification with I and Se in carrot. Four cultivars ('Askona' F 1 , 'Samba' F 1 , 'Kazan' F 1 and 'White Satin') were grown in soil fertilized with KI (4 kg I ha -1 ) and Na 2 SeO 4 (0.25 kg Se ha -1 ). The I + Se fertilization did not affect yield but the plants of all cultivars accumulated both elements in leaves and roots. On average, the I and Se contents in roots increased 7.7-times for I and 4.9-times for Se as well as the average I:Se molar ratio was 0.28:1. The contents of both elements in roots remained well below the hazard threshold thus the intake of 100 g of biofortified carrot would substantially cover the RDA for I and Se. The changes in chemical composition of roots (nitrates, phenolic compounds, sugars, carotenoids, macro-, microelements and cadmium) were rather year-dependent than affected by the applied I + Se fertilization., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The present study aims at optimisation of enhanced solute impregnation through ultrasonication (US) at 37 kHz using central composite design of response surface methodology (RSM). Black carrots were blanched at 98 °C/3 min followed by immersion in salt-spices based solutions for 8 h, treated with bath sonication at specific intervals (2, 4 & 6 h) for 5, 10 and 15 min. Responses for optimum water loss (WL), solute gain (SG), anthocyanins retention, texture and highest antioxidant activity were optimized in effect of NaCl%, spices mix% and US time. The suggested models were of good fit while lack of fit as non-significant (p < 0.0500). Best combination was compared for anthocyanins retention on pressure-cooking (120 °C/15 psi) with fresh, blanched, salt-spiced without US subjected to hot-air convective drying (55 °C/5 h). Ultrasonication-assisted salt-spiced convective dried (USPCD) carrots retained highest anthocyanin content (41%) post cooking which correlated with increased glass transition (T g ) and antimicrobial efficacy (as zone of inhibition) in comparison to fresh, blanched convective dried (BCD) and salt-spiced convective dried (SPCD) carrots. FTIR spectra suggested the correlation of textural effects with pectin degradation and solute impregnation. XRD spectra revealed changes in pectin (2θ = 22.06°) and NaCl crystallinity peaks at 2θ = 27.49, 31.86, 45.62, 56.68, 66.45 and 75.43°, owing to the processing of black carrots. The spectra also confirmed higher solute impregnation in USPCD than SPCD carrots. Scanning electron micrographs of USPCD also indicated higher impregnation through smallest pores visible in longitudinal section of carrot tissue structure, which also appeared to be polymeric network filled with higher solutes than SPCD. Taken together, US assisted solute impregnation might have provided protection to the anthocyanins degradation as well as retention of other quality parameters on processing., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Reutilization of putrescible municipal solid wastes (MSW) in agriculture can provide valuable plant nutrients. However, it may pose serious noncarcinogenic health risks for a human when contaminants, especially the heavy metals in MSW, end up in plants through the waste-soil-plant continuum. This study examined the effects of composting methods viz. aerobically (AC), anaerobically (ANC), and aerobic-anaerobically (AANC) composted MSW material on (i) fertilizer value: vegetable yield, nitrogen (N) mineralization, and apparent N recovery (ANR); and (ii) associated health risks: selected heavy metal concentration, daily intake of metals (DIM), health risk index (HRI), hazard index (HI), and target hazard quotient (THQ) when applied to a loamy soil. All the aforementioned compost materials were incorporated into the sandy loam soil filled in pots and carrot and spinach were cultivated for 85 and 90 days, respectively. After soil application, between 51 and 56% of the applied organic N was mineralized from ANC material, while the values in case of AC and AANC were 26-31% and 34-40%, respectively. Consequently, dry matter yield and vegetable N uptake from composts were in the order ANC > AANC > AC (P < 0.05). Further, vegetable ANR was the highest from ANC (56 and 56%) than AANC (42 and 45%), and AC (30 and 33%) for spinach and carrot, respectively (P < 0.05). Interestingly, plant uptake of lead and cadmium was lowest from ANC as compared to AC or AANC (P < 0.05), irrespective of the vegetable type. Consequently, DIM, HRI, and THQ for these metals were substantially lower in the former as compared to the latter compost materials. Further, HI from ANC material was 50% lower over the unfertilized control indicating the absence of noncarcinogenic human health risks via vegetable intake. This all indicates that from viewpoint of sustainable waste recycling in agriculture, anaerobic composting is superior to the other composting methods.

Main Conclusion: Diflufenican increased 493-fold the level of phytoene. Diflufenican-induced inhibition of phytoene desaturase gene expression in carrot cells resulted in an increased production of phytoene. This work analyzes the effect of diflufenican, an inhibitor of phytoene desaturase, on the gene expression profiles of the biosynthetic pathway of carotenoids related with the production of these compounds in carrot cell cultures. The results showed that the presence of 10 µM diflufenican in the culture medium increased phytoene levels, which was 493-fold higher than in control cells after 7 days of treatment but did not alter cell growth in carrot cell cultures. The maximal production of phytoene was reached with 10 µM diflufenican after 7 days of incubation in the presence of light and with 30 g/L sucrose in the culture medium. Moreover, diflufenican decreased the expression of phytoene synthase and phytoene desaturase genes at all the times studied. This diflufenican-induced inhibition of phytoene desaturase gene expression in carrot cell cultures resulted in an increased production of phytoene. Our results provide new insights into the action of diflufenican in carrot cell cultures, which could represent an alternative more sustainable and environmentally friendly system to produce phytoene than those currently used.

Brassinosteroid (BR) is a predominant plant hormone in regulating cell elongation and cell size. BR-deficient mutants display reduced plant growth and dwarfism in Arabidopsis and rice. In carrot, BRs promote petiole elongation, but its underlying mechanism involving exogenous BR remains unknown. Here, weighted gene co-expression network analysis and promoter region analysis were adopted to identify the potential genes that interacted with DcBZR1/BES1. Bioactive gibberellin (GA) level and cellulose deposition were also determined in the control and treated plants. Quantitative real-time PCR was performed to detect the expression profiles of GA biosynthesis-related genes, GA signaling genes, and cellulose synthase genes. Bioactive GA level and cellulose deposition were upregulated after the petioles were treated with 24-epibrassinolide (24-EBL). The most putative DcBZR1/BES1 genes were clustered in yellow module. The expression level of DCAR&#95;009411 (a GA5-like gene) was significantly induced after 3 h of treatment. The expression levels of DCAR&#95;019754 and DCAR&#95;013973 (CESA-like genes) were also significantly induced after 3 h of 24-EBL treatment. Our results suggested that the effect of BR on carrot petiole growth was quick. These results also provided potential insights into the mechanism by which BRs modulate GA and cellulose synthesis to promote cell elongation in carrot petioles., (Copyright © 2018. Published by Elsevier B.V.)

A pot experiment was undertaken to investigate the effects of Cd and Cu mixtures to growth and nutrients (sugar, carotene or vitamin C) of carrot and pakchoi under greenhouse cultivation condition. The study included: (a) physical-chemical properties of soil and soil animals in response to Cd and Cu stress; (b) bioaccumulation of heavy metals, length, biomass, contents of sugar and carotene (vitamin C) of carrot and pakchoi; (c) estimation the effects of Cd and Cu mixtures by multivariate regression analysis. The results implied that heavy metals impacted negative influence on soil animals' abundance. The metals contents in plants increased obviously with Cd and Cu contamination in soil. The biomass production and nutrients declined with Cd and Cu contents increasing. Cd (20 mg kg -1 ) treatment caused maximum reduction of sugar content (45.29%) in carrot root; maximum reduction in carotene content (75.73%) in carrot, 75.1% sugar content reduction and 70.58% vitamin C content reduction in pakchoi shoots were observed with addition of Cd (20 mg kg -1 ) and Cu (400 mg kg -1 ) mixture. The results of multivariate regression analysis indicated that combination of Cd and Cu exerts negative effects to both carrot and pakchoi, and both growth and nutrients were negatively correlated with metals concentrations. It is concluded that the Cd and Cu mixtures caused toxic damage to vegetable plants as Cd and Cu gradient concentrations increased., (Copyright © 2018. Published by Elsevier Inc.)

Background: The effect of hydrogen sulfide (H 2 S) on the inhibition of the surface whitening of fresh-cut carrots was investigated. Whitening index; H 2 O 2 and malondialdehyde (MDA) contents; activities of catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR), phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO) and peroxidase (POD); and antioxidant capacities were evaluated., Results: H 2 S treatment significantly inhibited the surface whitening of fresh-cut carrots (P < 0.05), reduced the accumulation of H 2 O 2 and MDA, and enhanced antioxidant enzymes (CAT, APX and GR) activities and antioxidant capacities (P < 0.05). The PAL, PPO and POD activities of fresh-cut carrots were inhibited by treatment with H 2 S (P < 0.05). Furthermore, correlation analysis revealed that whitening index was significantly positively correlated with H 2 O 2 content, lipid peroxidation, PPO and POD activities., Conclusion: H 2 S inhibited the surface whitening of fresh-cut carrots by reducing H 2 O 2 accumulation and lipid peroxidation, and also inhibited PPO and POD activities. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

Key Message: Terbinafine induced a significant increase of squalene production. Terbinafine increased the expression levels of squalene synthase. Cyclodextrins did not work as elicitors due to the gene expression levels obtained. Plant sterols are essential components of membrane lipids, which contributing to their fluidity and permeability. Besides their cholesterol-lowering properties, they also have anti-inflammatory, antidiabetic and anticancer activities. Squalene, which is phytosterol precursor, is widely used in medicine, foods and cosmetics due to its anti-tumor, antioxidant and anti-aging activities. Nowadays, vegetable oils constitute the main sources of phytosterols and squalene, but their isolation and purification involve complex extraction protocols and high costs. In this work, Daucus carota cell cultures were used to evaluate the effect of cyclodextrins and terbinafine on the production and accumulation of squalene and phytosterols as well as the expression levels of squalene synthase and cycloartenol synthase genes. D. carota cell cultures were able to produce high levels of extracellular being phytosterols in the presence of cyclodextrins (12 mg/L), these compounds able to increase both the secretion and accumulation of phytosterols in the culture medium. Moreover, terbinafine induced a significant increase in intracellular squalene production, as seen after 168 h of treatment (497.0 ± 23.5 µg g dry weight -1 ) while its extracellular production only increased in the presence of cyclodextrins.The analysis of sqs and cas gene expression revealed that cyclodextrins did not induce genes encoding enzymes involved in the phytosterol biosynthetic pathway since the expression levels of sqs and cas genes in cyclodextrin-treated cells were lower than in control cells. The results, therefore, suggest that cyclodextrins were only able to release phytosterols from the cells to the extracellular medium, thus contributing to their acumulation. To sum up, D. carota cell cultures treated with cyclodextrins or terbinafine were able to produce high levels of phytosterols and squalene, respectively, and, therefore, these suspension-cultured cells of carrot constitute an alternative biotechnological system, which is at the same time more sustainable, economic and ecological for the production of these bioactive compounds.

The present study aimed to optimize ozone (O 3 ) treatments, as gas and dissolved in water, to remove difenoconazole and linuron in carrots. We employed a central composite design to study three variables governing the efficacy of treatments: O 3 concentration, temperature and treatment time. The temperature did not influence the efficacy of treatments. The removal percentage of pesticides increases with increases in ozone concentration and the time of treatment. O 3 application promoted the removal of more than 80% of pesticides when the roots were exposed for approximately 120min at 5 and 10mgL -1 , respectively, in treatments with O 3 as gas and dissolved in water. After storage, pesticide removal was higher than 98% for difenoconazole and 95% for linuron. The degradation products from the pesticides resulting from treatment were monitored, but none were found. This is the first report demonstrating the removal of difenoconazole and linuron from carrots by ozone., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background: Black carrots (Daucus carota ssp. sativus var. atrorubens Alef.) constitute a valuable source of anthocyanins, which are used as natural red, blue and purple food colourants. Anthocyanins and phenolic compounds are specialised metabolites, accumulation of which often requires elicitors, which act as molecular signals in plant stress responses. In the present study, ethephon, an ethylene-generating compound was explored as enhancer of anthocyanin and phenolic contents during growth of 'Deep Purple' black carrots. The effects of ethephon on several parameters were investigated, and the expression of biosynthetic anthocyanin genes was studied during growth and anthocyanin accumulation., Results: Roots of ethephon-treated carrot plants exhibited an increase in anthocyanin content of approximately 25%, with values ranging from 2.25 to 3.10 mg g -1 fresh weight, compared with values ranging from 1.50 to 1.90 mg g -1 fresh weight in untreated roots. The most rapid accumulation rate for anthocyanins, phenolic compounds, soluble solids and dry matter was observed between 10 and 13 weeks after sowing in both untreated and ethephon-treated carrots. The differences in anthocyanin contents between untreated and treated carrots increased for several weeks after the ethephon treatment was terminated. Five cyanidin-based anthocyanin forms were identified, with variable relative abundance values detected during root growth. Overall, the expression of the anthocyanin biosynthetic genes analysed (PAL1, PAL3, F3H1, DFR1, LDOX2) increased in response to ethephon treatment, as did the expression of the MYB1 transcription factor, which is associated with activation of the phenylpropanoid pathway under stress conditions. In addition, a correlation was proposed between ethylene and sugar contents and the induction of anthocyanin synthesis., Conclusions: This study presents a novel method for enhancing anthocyanin content in black carrots. This finding is of economic importance as increased pigment concentration per unit of biomass implies improved profitability parameters in food colour production. We provide new insight into the accumulation patterns of the different cyanidin-based anthocyanins and phenolic compounds during root growth. Moreover, we show that enhanced anthocyanin content in ethephon-treated carrots is accompanied by increased expression of anthocyanin biosynthetic genes.

Gibberellins (GAs) are important growth regulators involved in plant development processes. However, limited information is known about the relationship between GA and xylogenesis in carrots. In this study, carrot roots were treated with GA 3 . The effects of applied GA 3 on root growth, xylem development, and lignin accumulation were then investigated. Results indicated that GA treatment dose-dependently inhibited carrot root growth. The cell wall significantly thickened in the xylem parenchyma. Autofluorescence analysis with ultraviolet (UV) excitation indicated that these cells became lignified because of long-term GA 3 treatment. Moreover, lignin content increased in the roots, and the transcripts of lignin biosynthesis genes were altered in response to applied GA 3 . Our data indicate that GA may play important roles in xylem growth and lignification in carrot roots. Further studies shall focus on regulating plant lignification, which may be achieved by modifying GA levels within plant tissues.

Toxic cyanobacterial blooms are often observed in freshwaters and may reflect the increased eutrophication of these environments and alterations in climate. Cyanotoxins, such as microcystins (MCs), are an effective threat to many life forms, ranging from plants to humans. Despite the research conducted to date on cyanotoxins, the risks associated to the use of contaminated water in agriculture require further elucidation. To tackle this aim, a research was conducted with the root-vegetable Daucus carota. The specific aims of this work were the following: (i) to evaluate the effects of MC-LR on the plant growth and photosynthesis; (ii) to evaluate the nutritional quality of carrot roots; and (iii) to measure bioaccumulation. To this purpose, young carrots were grown in soil during 1 month in natural conditions and exposed to Mycrocystis aeruginosa aqueous extracts containing environmentally realistic concentrations of MC-LR (10 and 50 MC-LR μg/L). The results showed that MC-LR may decrease root growth after 28 days of exposure to 50 μg/L and increase photosynthetic efficiency. We also observed changes in mineral and vitamin content in carrots as a result of the exposure to contaminated water. Moreover, MC-LR was detected in carrot roots by ELISA at very low concentration 5.23 ± 0.47 ng MC eq./g FW. The soil retained 52.7 % of the toxin potentially available for plants. This result could be attributed to MC-LR adsorption by soil particles or due to microbial degradation of the toxin. We conclude that the prolonged use of MC-LR-contaminated water may affect crop growth, alter the nutritional value of vegetable products, and potentiate contamination.

Exogenous reactive oxygen species (ROS) produced by nanobubble (NB) water offer a reasonable explanation for NBs' physiological promotion and oxidation effects. To develop and exploit the NB technology, we have performed further research to identify the specific ROS produced by NBs. Using a fluorescent reagent APF, a Fenton reaction, a dismutation reaction of superoxide dismutase and DMSO, we distinguished four types of ROS (superoxide anion radical (O 2 · - ), hydrogen peroxide (H 2 O 2 ), hydroxyl radical (·OH), and singlet oxygen ( 1 O 2 )). ·OH was confirmed to be the specific ROS produced by NB water. The role of ·OH produced by NB water in physiological processes depends on its concentration. The amount of exogenous ·OH has a positive correlation with the NB number density in the water. Here, spinach and carrot seed germination tests were repeatedly performed with three seed groups submerged in distilled water, high-number density NB water, and low-number density NB water under similar dissolved oxygen concentrations. The final germination rates of spinach seeds in distilled water, low-number density NB water, and high-number density NB water were 54%, 65%, and 69%, respectively. NBs can also promote sprout growth. The sprout lengths of spinach seeds dipped in NB water were longer than those in the distilled water. For carrot seeds, the amount of exogenous ·OH in high-number density NB water was beyond their toxic threshold, and negative effects were shown on hypocotyl elongation and chlorophyll formation. The presented results allow us to obtain a deeper understanding of the physiological promotion effects of NBs.

Consumption of vegetables is often the predominant route whereby humans are exposed to the toxic metal Cd. Health impacts arising from Cd consumption may be influenced by changes in the mineral nutrient content of vegetables, which may occur when plants are exposed to Cd. Here, we subjected model root (carrot) and leaf (lettuce) vegetables to soil Cd concentrations of 0.3, 1.5, 3.3, and 9.6 μg g(-1) for 10 weeks to investigate the effect of Cd exposure on Cd accumulation, growth performance, and mineral nutrient homeostasis. The findings demonstrated that Cd accumulation in lettuce (20.1-71.5 μg g(-1)) was higher than that in carrot (3.2-27.5 μg g(-1)), and accumulation exceeded the maximum permissible Cd concentration in vegetables when soil contained more than 3.3 μg g(-1) of Cd. There was a marked hormetic effect on carrot growth at a soil Cd concentration of 3.3 μg g(-1), but increasing the Cd concentration to 9.6 μg g(-1) caused decreased growth in both crops. Additionally, in most cases, there was a positive correlation between Cd and the mineral nutrient content of vegetables, which was due to physiological changes in the plants causing increased uptake and/or translocation. This may suggest a general mechanism whereby the plant compensated for disrupted mineral nutrient metabolism by increasing nutrient supply to its tissues. Increased nutrient levels could potentially offset some risks posed to humans by increased Cd levels in crops, and we therefore suggest that changes in mineral nutrient levels should be included more widely in the risk assessment of potentially toxic metal contamination. Graphical abstract The Cd concentration (μg g-1 in dry matter) in the root, shoot and translocation factor (TF) of Cd from root to shoot in the carrot and lettuce, and the percentage of root Cd to the gross Cd contents (%) in carrot (C) and lettuce (D) exposed to soil Cd (0 (control), 1, 3, and 9 μg g-1) for 70 days. Values are means ± SD (n = 5).

Carrot (Daucus carota L.) is a vegetable crop that is grown throughout the year across various regions of Brazil in rotation or in succession to other cultures. Herbicide residual effect has emerged as a concern, because of the possibility of carryover. Thus, the objective of this study was to evaluate the effect of tembotrione and atrazine residues - in mixture and isolated - on carrot planted in succession to corn. The experiment was designed in randomized blocks with five replications. Treatments consisted of tembotrione (50.4 g ha(-1)), tembotrione (100.8 g ha(-1)), tembotrione + atrazine (50.4 g ha(-1)+ 2 L ha(-1)), tembotrione + atrazine (100.8 g ha(-1)+ 2 L ha(-1)), and atrazine (2.00 L ha(-1)) applied eight months before carrot seeding, plus a control treatment with no herbicide application. Investigated variables were shoot dry mass, productivity, and classification of carrot roots. The presence of atrazine and tembotrione decreased dry mass in the area, and only tembotrione reduced total root productivity. Thus, there is a carryover effect to tembotrione application that reduces the dry matter accumulation of shoot and total productivity, and an atrazine + tembotrione (100.8 g ha(-1)) mixture reduces the total productivity after application of these herbicides to soil.

Large quantities of veterinary antibiotics (VAs) are being used worldwide in agricultural fields through wastewater irrigation and manure application. They cause damages to the ecosystem when discharged into the environment, but there is a lack of information on their toxicity to plants and animals. This study evaluated the phytotoxic effects of five major VAs, namely tetracycline (TC), sulfamethazine (SMZ), norfloxacin (NOR), erythromycin (ERY) and chloramphenicol (CAP), on seed germination and root elongation in lettuce, tomato, carrot and cucumber, and investigated the relationship between their physicochemical properties and phytotoxicities. Results show that these compounds significantly inhibited root elongation (p<0.05), the most sensitive endpoint for the phytotoxicity test. TC was associated with the highest level of toxicity, followed by NOR, ERY, SMZ and CAP. Regarding crop species, lettuce was found to be sensitive to most of the VAs. The median effect concentration (EC50) of TC, SMZ, NOR, ERY and CAP to lettuce was 14.4, 157, 49.4, 68.8 and 204 mg/L, respectively. A quantitative structure-activity relationship (QSAR) model has been established based on the measured data. It is evident that hydrophobicity was the most important factor governing the phytotoxicity of these compounds to seeds, which could be explained by the polar narcosis mechanism. Lettuce is considered a good biomarker for VAs in the environment. According to the derived equation, phytotoxicities of selected VA compounds on different crops can be calculated, which could be applicable to other VAs. Environmental risks of VAs were summarized based on the phytotoxicity results and other persistent factors., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Arbuscular mycorrhizal fungi (AMF), ubiquitous soil fungi that form symbiotic relationships with the majority of terrestrial plants, are known to play an important role in plant tolerance to chromium (Cr) contamination. However, the underlying mechanisms, especially the direct influences of AMF on the translocation and transformation of Cr in the soil-plant continuum, are still unresolved. In a two-compartment root-organ cultivation system, the extraradical mycelium (ERM) of mycorrhizal roots was treated with 0.05 mmol L(-1) Cr(VI) for 12 days to investigate the uptake, translocation, and transformation of Cr(VI) by AMF using inductively coupled plasma mass spectrometry (ICP-MS), scanning electron microscopy equipped with energy-dispersive spectroscopy (SEM-EDS), transmission electron microscopy equipped with energy-dispersive spectroscopy (TEM-EDS), and X-ray-absorption fine structure (XAFS) technologies. The results indicated that AMF can immobilize quantities of Cr via reduction of Cr(VI) to Cr(III), forming Cr(III)-phosphate analogues, likely on the fungal surface. Besides this, we also confirmed that the extraradical mycelium (ERM) can actively take up Cr [either in the form of Cr(VI) or Cr(III)] and transport Cr [potentially in the form of Cr(III)-histidine analogues] to mycorrhizal roots but immobilize most of the Cr(III) in the fungal structures. Based on an X-ray absorption near-edge spectroscopy analysis of Cr(VI)-treated roots, we proposed that the intraradical fungal structures can also immobilize Cr within mycorrhizal roots. Our findings confirmed the immobilization of Cr by AMF, which plays an essential role in the Cr(VI) tolerance of AM symbioses.

Background: Espresso coffee grounds constitute a residue which is produced daily in considerable amounts, and is often pointed out as being potentially interesting for plant nutrition. Two experiments (incubations and field experiments) were carried out to evaluate the potential nitrogen (N) and phosphorus (P) supply for carrot (Daucus carota L.), spinach (Spinacea oleracea L.) and lettuce (Lactuca sativa L.) nutrition., Results: Immobilisation of nitrogen and phosphorus was detected in all the incubations and, in the field experiments, germination and yield growth were decreased by the presence of espresso coffee grounds, in general for all the species studied., Conclusion: The study showed an inhibition of N and P mineralisation and a reduction of plant germination and growth. Further research is required to determine whether this is related to the immobilising capacity of the residue or possibly due to the presence of caffeine., (© 2015 Society of Chemical Industry.)

Reliable empirical models describing lead (Pb) transfer in soil-plant systems are needed to improve soil environmental quality standards. A greenhouse experiment was conducted to develop soil-plant transfer models to predict Pb concentrations in carrot (Daucus carota L.). Soil thresholds for food safety were then derived inversely using the prediction model in view of the maximum allowable limit for Pb in food. The 2 most important soil properties that influenced carrot Pb uptake factor (ratio of Pb concentration in carrot to that in soil) were soil pH and cation exchange capacity (CEC), as revealed by path analysis. Stepwise multiple linear regression models were based on soil properties and the pseudo total (aqua regia) or extractable (0.01 M CaCl2 and 0.005 M diethylenetriamine pentaacetic acid) soil Pb concentrations. Carrot Pb contents were best explained by the pseudo total soil Pb concentrations in combination with soil pH and CEC, with the percentage of variation explained being up to 93%. The derived soil thresholds based on added Pb (total soil Pb with the geogenic background part subtracted) have the advantage of better applicability to soils with high natural background Pb levels. Validation of the thresholds against data from field trials and literature studies indicated that the proposed thresholds are reasonable and reliable., (© 2015 SETAC.)

Phthalate esters (PAEs) are large-volume chemicals and are found ubiquitously in soil as a result of widespread plasticulture and waste disposal. Food plants such as vegetables may take up and accumulate PAEs from soil, potentially imposing human health risks through dietary intake. In this study, we carried out a cultivation study using lettuce, strawberry, and carrot plants to determine the potential of plant uptake, translocation, and metabolism of di-n-butyl phthalate (DnBP) and di(2-ethylhexyl) phthalate (DEHP) and their primary metabolites mono-n-butyl phthalate (MnBP) and mono(2-ethylhexyl) phthalate (MEHP). All four compounds were detected in the plant tissues, with the bioconcentration factors (BCFs) ranging from 0.16 ± 0.01 to 4.78 ± 0.59. However, the test compounds were poorly translocated from roots to leaves, with a translocation factor below 1. Further, PAEs were readily transformed to their monoesters following uptake. Incubation of PAEs and monoalkyl phthalate esters (MPEs) in carrot cell culture showed that DnBP was hydrolyzed more rapidly than DEHP, while the monoesters were transformed more quickly than their parent precursors. Given the extensive metabolism of PAEs to monoesters in both whole plants and plant cells, metabolism intermediates such as MPEs should be considered when assessing human exposure via dietary intake of food produced from PAE-contaminated soils.

Carrot, a biennial herb of the Apiaceae family, is among the most important vegetable crops in the world. In this study, nine candidate reference genes (GAPDH, ACTIN, eIF-4α, PP2A, SAND, TIP41, UBQ, EF-1α, and TUB) were cloned from carrot. Carrot plants were subjected to abiotic stresses (heat, cold, salt, and drought) and hormone stimuli (gibberellin, salicylic acid, methyl jasmonate, and abscisic acid). The expression profiles of the candidate reference genes were evaluated in three technical and biological replicates. Real-time qPCR data analyses were performed using three commonly used Excel-based applets namely, BestKeeper, geNorm, and NormFinder. ACTIN and TUB were the most stable genes identified among all sample groups, but individual analysis revealed changes in their expression profiles. GAPDH displayed the maximum stability for most of single stresses. To further validate the suitability of the reference genes identified in this study, the expression profile of DcDREB-A1 gene (homolog of AtDREB-A1 gene of Arabidophsis) was studied in carrot. The appropriate reference genes were selected that showed stable expression under the different experimental conditions.

Soil environmental quality standards in respect of heavy metals for farmlands should be established considering both their effects on crop yield and their accumulation in the edible part. A greenhouse experiment was conducted to investigate the effects of chromium (Cr) on biomass production and Cr accumulation in carrot plants grown in a wide range of soils. The results revealed that carrot yield significantly decreased in 18 of the total 20 soils with Cr addition being the soil environmental quality standard of China. The Cr content of carrot grown in the five soils with pH>8.0 exceeded the maximum allowable level (0.5mgkg(-1)) according to the Chinese General Standard for Contaminants in Foods. The relationship between carrot Cr concentration and soil pH could be well fitted (R(2)=0.70, P<0.0001) by a linear-linear segmented regression model. The addition of Cr to soil influenced carrot yield firstly rather than the food quality. The major soil factors controlling Cr phytotoxicity and the prediction models were further identified and developed using path analysis and stepwise multiple linear regression analysis. Soil Cr thresholds for phytotoxicity meanwhile ensuring food safety were then derived on the condition of 10 percent yield reduction., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Our previous studies of action spectra for UV-B-induced anthocyanin accumulation in cultured carrot cells indicated that a reduced form of pterin, possibly tetrahydrobiopterin, contributes to UV-B photoreception. In this report, we provide additional evidence for the involvement of pterin in UV-B light sensing. UV-B-induced phenylalanine ammonia-lyase (PAL) activity was considerably suppressed by N-acetylserotonin (an inhibitor of tetrahydrobiopterin biosynthesis), and this suppression was partially recovered by adding biopterin or tetrahydrobiobiopterin. In addition, protein(s) specifically bound to biopterin were detected by radiolabeling experiments in N-acetylserotonin-treated cells. Furthermore, diphenyleneiodonium, a potent inhibitor of electron transfer, completely suppressed UV-B-induced PAL activity. These results suggest the occurrence of an unidentified UV-B photoreceptor (other than UVR8, the tryptophan-based UV-B sensor originally identified in Arabidopsis) with reduced pterin in carrot cells. After reexamining published action spectra, we suggest that anthocyanin synthesis is coordinately regulated by these two UV-B sensors., (© 2014 The American Society of Photobiology.)

Suspension cultured-cells (SCC) of Daucus carota were used to evaluate the effect of methyl jasmonate and cyclodextrins, separately or in combination, on the induction of defense responses, particularly the accumulation of pathogenesis-related proteins. A comparative study of the extracellular proteome (secretome) between control and elicited carrot SCC pointed to the presence of amino acid sequences homologous to glycoproteins which have inhibitory activity against the cell-wall-degrading enzymes secreted by pathogens and/or are induced when carrot cells are exposed to a pathogen elicitor. Other amino acid sequences were homologous to Leucine-Rich Repeat domain-containing proteins, which play an essential role in defense against pathogens, as well as in the recognition of microorganisms, making them important players in the innate immunity of this plant. Also, some tryptic peptides were shown to be homologous to a thaumatin-like protein, showing high specificity to abiotic stress and to different reticuline oxidase-like proteins that displayed high levels of antifungal activity, suggesting that methyl jasmonate and cyclodextrins could play a role in mediating defense-related gene product expression in SCC of D. carota. Apart from these elicitor-inducible proteins, we observed the presence of PR-proteins in both control and elicited carrot SCC, suggesting that their expression is mainly constitutive. These PR-proteins are putative class IV chitinases, which also have inhibitory activity against pathogen growth and the class III peroxidases that participate in response to environmental stress (e.g. pathogen attack and oxidative), meaning that they are involved in defense responses triggered by both biotic and abiotic factors., (Copyright © 2014 Elsevier Masson SAS. All rights reserved.)

The use of plants to produce chemical compounds with pharmaceutical and nutraceutical applications has intensified in recent years. In this regard, genetic engineering is the most commonly used tool to generate crop lines with enhanced concentrations of desirable chemicals. However, growing genetically modified plants is still limited because they are perceived as potential biological hazards that can create an ecological imbalance. The application of postharvest abiotic stresses on plants induces the accumulation of secondary metabolites and thus can be used as an alternative to genetic modification. The present project evaluated the feasibility of producing shikimic acid (SA) and phenolic compounds (PC) in wounded carrots ( Daucus carota ) treated with glyphosate. The spray application of a concentrated glyphosate solution on wounded carrot tissue increased the concentrations of SA and chlorogenic acid by ∼1735 and ∼5700%, respectively. The results presented herein demonstrate the potential of stressed carrot tissue as a biofactory of SA and PC.

Persistent environmental contaminants may enter agricultural fields via the application of sewage sludge, by irrigation with treated municipal wastewater or by manuring. It has been shown that such contaminants can be incorporated into crop plants. The metabolism of the bacteriostatic agents triclocarban, triclosan, and its transformation product methyl triclosan was investigated after their uptake into carrot cell cultures. A fast metabolization of triclosan was observed and eight so far unknown phase II metabolites, conjugates with saccharides, disaccharides, malonic acid, and sulfate, were identified by liquid chromatography-mass spectrometry. Triclocarban and methyl triclosan lack a phenolic group and remained unaltered in the cell cultures. Phase I metabolization was not observed for any of the compounds. All eight triclosan conjugates identified in the cell cultures were also detected in extracts of intact carrot plants cultivated on triclosan contaminated soils. Their total amount in the plants was assessed to exceed the amount of the triclosan itself by a factor of 5. This study shows that a disregard of conjugates in studies on plant uptake of environmental contaminants may severely underestimates the extent of uptake into plants and, eventually, the potential human exposure to contaminants via food of plant origin.

Polyunsaturated fatty acids (PUFAs) are powerful modulators of several animal ion channels. It is shown here that PUFAs strongly affect the activity of the Slow Vacuolar (SV) channel encoded by the plant TPC1 gene. The patch-clamp technique was applied to isolated vacuoles from carrot taproots and Arabidopsis thaliana mesophyll cells and arachidonic acid (AA) was chosen as a model molecule for PUFAs. Our study was extended to different PUFAs including the endogenous alpha-linolenic acid (ALA). The addition of micromolar concentrations of AA reversibly inhibited the SV channel decreasing the maximum open probability and shifting the half activation voltage to positive values. Comparing the effects of different PUFAs, it was found that the length of the lipophilic acyl chain, the number of double bonds and the polar head were critical for channel modulation.The experimental data can be reproduced by a simple three-state model, in which PUFAs do not interact directly with the voltage sensors but affect the voltage-independent transition that leads the channel from the open state to the closed configuration. The results indicate that lipids play an important role in co-ordinating ion channel activities similar to what is known from animal cells.

Very few studies have provided information about the effects of cadmium (Cd) at histoanatomical and ultrastructural levels, along with potential localization of the metal in planta. In particular, from this standpoint, almost nothing is known in Daucus carota L. (carrot), a particularly important species for in vitro and in vivo functional investigations. In this work we hypothesized that 36 μM Cd, supplied for 1, 2, 3, 4, 7 and 14 days to 30-day-old in vitro-cultured plants, might induce an early acclimation, but a final collapse of roots and leaves. In fact, as a general feature, a biphasic root response to Cd stress actually took place: in the first phase (1-4 days of Cd exposure), the cytological and functional events observed - by light microscopy, TEM, epifluorescence, as well as by the time-course of thiol-peptide compounds - can be interpreted as acclimatory responses aimed at diminishing the movement of Cd across the root. The second phase (from 4 to 14 days of Cd exposure) was instead characterized by cell hypertrophy, cell-to-cell separation events, increase in α-β-γ-tocopherol levels and, not least, endocytogenic processes, coupled with a dramatic drop in the amount of thiol-peptide compounds. These events led to a progressive root collapse, even if they did not ingenerate macro/microscopic injury symptoms in leaf blades and petioles., (Copyright © 2012 Elsevier Masson SAS. All rights reserved.)

Methyl-jasmonate (MJ)-treated hairy roots of Daucus carota L. were used to study the influence of alternative oxidase (AOX) in phenylpropanoid metabolism. Phenolic acid accumulation, as well as total flavonoids and lignin content of the MJ-treated hairy roots were decreased by treatment with salicylhydroxamic acid (SHAM), a known inhibitor of AOX. The inhibitory effect of SHAM was concentration dependent. Treatment with propyl gallate (PG), another inhibitor of AOX, also had a similar inhibitory effect on accumulation of phenolic acid, total flavonoids and lignin. The transcript levels of two DcAOX genes (DcAOX2a and DcAOX1a) were monitored at selected post-elicitation time points. A notable rise in the transcript levels of both DcAOX genes was observed preceding the MJ-induced enhanced accumulation of phenolics, flavonoids and lignin. An appreciable increase in phenylalanine ammonia-lyase (PAL) transcript level was also observed prior to enhanced phenolics accumulation. Both DcAOX genes showed differential transcript accumulation patterns after the onset of elicitation. The transcript levels of DcAOX1a and DcAOX2a attained peak at 6hours post elicitation (hpe) and 12hpe, respectively. An increase in the transcript levels of both DcAOX genes preceding the accumulation of phenylpropanoid-derivatives and lignin showed a positive correlation between AOX activity and phenylpropanoid biosynthesis. The results provide important new insight about the influence of AOX in phenylpropanoid biosynthesis., (Copyright © 2012 Elsevier GmbH. All rights reserved.)

Turnip (Brassica rapa var. rapa L.), beetroot (Beta vulgaris L.) and carrot (Daucus carota L.) are common vegetables in South Africa. The allelopathic potential of aqueous leaf and root extracts of Aloe ferox Mill.- a highly valued medicinal plant- was evaluated against seed germination and seedling growth of the three vegetables in Petri dish experiments. The extracts were tested at concentrations of 2, 4, 6, 8, and 10 mg/mL. Leaf extract concentrations above 4 mg/mL inhibited the germination of all the crops, while the root extract had no significant effect on germination irrespective of concentration. Interestingly, the lowest concentration of leaf extract stimulated root length elongation of beetroot by 31.71%. Other concentrations significantly inhibited both root and shoot growth of the vegetable crops except the turnip shoot. The most sensitive crop was carrot, with percentage inhibition ranging from 29.15 to 100% for root and shoot lengths. Lower percentage inhibition was observed for the root extract than the leaf extract against shoot growth of beetroot and carrot. The results from this study suggested the presence of allelochemicals mostly in the leaves of A. ferox that could inhibit the growth of the turnip, beetroot and carrot.

Pectobacterium carotovorum (formerly Erwinia carotovora ssp. carotovora) is a phytopathogenic bacterium that causes soft rot disease, characterized by water-soaked soft decay, resulting from the action of cell wall-degrading exoenzymes secreted by the pathogen. Virulence in soft rot bacteria is regulated by environmental factors, host and bacterial chemical signals, and a network of global and gene-specific bacterial regulators. We isolated a mini-Tn5 mutant of P. carotovorum that is reduced in the production of extracellular pectate lyase, protease, polygalacturonase and cellulase. The mutant is also decreased in virulence as it macerates less host tissues than its parent and is severely impaired in multiplication in planta. The inactivated gene responsible for the reduced virulent phenotype was identified as corA. CorA, a magnesium/nickel/cobalt membrane transporter, is the primary magnesium transporter for many bacteria. Compared with the parent, the CorA(-) mutant is cobalt resistant. The mutant phenotype was confirmed in parental strain P. carotovorum by marker exchange inactivation of corA. A functional corA(+) DNA from P. carotovorum restored exoenzyme production and pathogenicity to the mutants. The P. carotovorum corA(+) clone also restored motility and cobalt sensitivity to a CorA(-) mutant of Salmonella enterica. These data indicate that CorA is required for exoenzyme production and virulence in P. carotovorum., (© 2011 THE AUTHORS. MOLECULAR PLANT PATHOLOGY © 2011 BSPP AND BLACKWELL PUBLISHING LTD.)

In the present study, inhibitory effects of the hydrosols of thyme, black cumin, sage, rosemary and bay leaf were investigated against Salmonella Typhimurium and Escherichia coli O157:H7 inoculated to apple and carrots (at the ratio of 5.81 and 5.81 log cfu/g for S. Typhimurium, and 5.90 and 5.70 log cfu/g for E. coli O157:H7 on to apple and carrot, respectively). After the inoculation of S. Typhimurium or E. coli O157:H7, shredded apple and carrot samples were washed with the hydrosols and sterile tap water (as control) for 0, 20, 40 and 60 min. While the sterile tap water was ineffective in reducing (P>0.05) S. Typhimurium and E. coli O157:H7, 20 min hydrosol treatment caused a significant (P<0.05) reduction compared to the control group. On the other hand, thyme and rosemary hydrosol treatments for 20 min produced a reduction of 1.42 and 1.33 log cfu/g respectively in the E. coli O157:H7 population on apples. Additional reductions were not always observed with increasing treatment time. Moreover, thyme hydrosol showed the highest antibacterial effect on both S. Typhimurium and E. coli O157:H7 counts. Inhibitory effect of thyme hydrosol on S. Typhimurium was higher than that for E. coli O157:H7. Bay leaf hydrosol treatments for 60 min reduced significantly (P<0.05) E. coli O157:H7 population on apple and carrot samples. In conclusion, it was shown that plant hydrosols, especially thyme hydrosol, could be used as a convenient sanitizing agent during the washing of fresh-cut fruits and vegetables., (Copyright © 2011 Elsevier B.V. All rights reserved.)

We applied a screening-level phytotoxicity assay to evaluate the effects of 10 antibiotics (at concentrations ranging from 1 to 10,000 μg/L) on germination and early plant growth using three plant species: lettuce (Lactuca sativa), alfalfa (Medicago sativa), and carrot (Daucus carota). The range of phytotoxicity of the antibiotics was large, with EC₂₅s ranging from 3.9 μg/L to >10,000 μg/L. Chlortetracycline, levofloxacin, and sulfamethoxazole were the most phytotoxic antibiotics. D. carota was the most sensitive plant species, often by an order of magnitude or more, followed by L. sativa and then M. sativa. Plant germination was insensitive to the antibiotics, with no significant decreases up to the highest treatment concentration of 10,000 μg/L. Compared with shoot and total length measurements, root elongation was consistently the most sensitive end point. Overall, there were few instances where measured soil concentrations, if available in the publicly accessible literature, would be expected to exceed the effect concentrations of the antibiotics evaluated in this study. The use of screening assays as part of a tiered approach for evaluating environmental impacts of antibiotics can provide insight into relative species sensitivity and serve as a basis by which to screen the potential for toxic effects of novel compounds to plants.

Bioballistic transformation of carrot Daucus carota L. callus cultures with a plasmid containing the aadA (aminoglycoside 3'-adenyltransferase) gene and subsequent selection oftransformants on a selective medium containing spectinomycin (100-500 mg/l) yielded ten callus lines resistant to this antibiotic. PCR analysis did not detect exogenous DNA in the genomes of spectinomycin-resistant calluses. Resistance proved to be due to spontaneous mutations that occurred in two different regions of the chloroplast rrn16 gene, which codes for the 16S rRNA. Six lines displayed the G > T or G > C transverions in position 1012 of the rrn16 gene, and three lines had the A > G transition in position 1138 of the gene. Chloroplast mutations arising during passages of callus cultures in the presence of spectinomycin were described in D. carota for the first time. The cause of spectinomycin resistance was not identified in one line. The mutations observed in the D. carota plastid genome occurred in the region that is involved in the formation of a double-stranded region at the 3' end of the 16S rRNA and coincided in positions with the nucleotide substitutions found in spectinomycin-resistant plants of tobacco Nicotiana tabacum L. and bladderpod Lesquerella fendleri L.

Ethylenediurea (EDU) is suggested for use to evaluate plant response under ambient ozone (O(3)) concentrations. Four EDU treatments, viz. 0 (non-EDU), 150, 300 and 450 mg L(-1), applied as soil drench at 10 days interval to carrot (Daucus carota L. var. Pusa Kesar), grown at a tropical suburban site of Varanasi experiencing mean O(3) concentration of 36.1 ppb during the experimental period. EDU treated plants showed significantly higher antioxidative defense, assimilation capability and reduced membrane lipid peroxidation, which led to better growth and significant yield increments compared to non-EDU treated ones. The magnitude of positive responses was highest at 150 mg L(-1) EDU treatment at 60 DAG, representing the metabolically most active phase of root filling in carrot. This study suggests that the lowest EDU concentration was sufficient to provide protection against negative effects of O(3)., (Copyright (c) 2010 Elsevier Inc. All rights reserved.)

Biosynthesis of hydroxybenzoates even at enzymatic level is poorly understood. In this report, effect of feeding of putative biosynthetic precursors and pathway-specific enzyme inhibitors of early phenylpropanoid pathway on p-hydroxybenzoic acid accumulation in chitosan-elicited hairy roots of Daucus carota was studied. Three selective metabolic inhibitors of plant phenylpropanoid pathway, namely, aminooxyacetic acid (AOAA), piperonylic acid (PIP) and 3,4-methylenedioxycinnamic acid (MDCA), which are known to inhibit phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H) and 4-coumarate-CoA ligase (4CL) respectively, the three early enzymes of phenylpropanoid metabolism, were chosen with the anticipation that selective inhibition of these enzymes in vivo may provide information on the metabolic route to p-hydroxybenzoic acid formation. Supplementation of AOAA (0.2-1.0 mM) and PIP (0.2-1.0 mM) resulted in the reduced accumulation of p-hydroxybenzoic acid in the wall-bound fraction. However, addition of MDCA (0.2-1.25 mM), did not suppress p-hydroxybenzoic acid accumulation but suppressed lignin and total flavonoid accumulation, suggesting that 4CL enzyme activity is not required for p-hydroxybenzoic acid formation. Feeding of elicited hairy roots with phenylalanine, coumaric acid and p-hydroxybenzaldehyde had a stimulatory effect on p-hydroxybenzoic acid accumulation; however, maximum stimulatory effect was shown by p-hydroxybenzaldehyde. This suggests that p-hydroxybenzaldehyde might be the immediate precursor in p-hydroxybenzoic acid biosynthesis. Finally, in vitro conversion of p-coumaric acid to p-hydroxybenzoic acid with p-hydroxybenzaldehyde as intermediate using cell-free extract provided an unequivocal support for CoA-independent and non-beta-oxidative route of p-hydroxybenzoic acid biosynthesis in Daucus carota.

A pot culture study was carried out at Avinashilingam Deemed University, Coimbatore to assess the effect of few organic manures (herbal microfertilizer, humic acid, farm yard manure (FYM), biofertilizer) and NPK on the growth attributes of carrot. Among the parameters observed, the germination percentage was highest in FYM treated pots on 7 DAS (Days After Sowing) and in NPK treated pots on 21, 28 and 30 DAS. The parameters, such as root length, crown length and vigour index, were studied on 30, 60, 90 and 120 DAS. The root length on 60 and 120 DAS was increased by FYM, whereas on 30 and 90 DAS, increase was observed in NPK treated plants. The crown length was increased by NPK on 30 and 120 DAS, micro herbal fertilizer showed an increase of crown length on 60 DAS and by FYM on 90 DAS. Highest vigour was in FYM treated plants on 30 DAS, in herbal micro fertilizer on 60 DAS in NPK treatment on 90 and 120 DAS.

In this study, methyl jasmonate (MJ)-elicited hairy root cultures of Daucus carota were explored to study the enzymatic route to p-hydroxybenzoic acid (p-HBA) biosynthesis. Treatment with 100microM MJ caused an enhanced accumulation of p-HBA as well as total phenolic content in elicited root lines as compared to untreated (controls) lines. Using cell-free extract as the source of crude enzymes, attempt was made to reveal the enzymatic route to p-HBA formation. The accumulation of p-HBA was preceded by a substantial upliftment of p-hydroxybenzaldehyde dehydrogenase (HBD) activity in elicited lines as compared to controls. A rapid 6-fold enhancement of phenylalanine ammonia-lyase (PAL) activity, the first enzyme of the phenylpropanoid pathway was also observed. Finally, we demonstrated here for the first time, in D. carota, the evidence of a quite unusual p-hydroxybenzaldehyde synthase (HBS)-type enzyme, which catalyzes the penultimate step of p-HBA biosynthesis by making phenylpropanoid side-chain cleavage of p-coumaric acid without involvement of any cofactor(s), but uplifted by supplementation of a thiol reagent such as DTT in the reaction buffer. This enzyme showed activity in a relatively broad pH range (7-8.4) and the temperature optimum was found to be at 34 degrees C. The MJ-treated roots showed highest HBS activity at 24h (52nkat/mg protein), which was nearly 5-fold higher than that in the control lines.

KDC1 is a voltage-dependent Shaker-like potassium channel subunit cloned from Daucus carota which produces conductive channels in Xenopus oocytes only when coexpressed with other plant Shaker potassium subunits, such as KAT1 from Arabidopsis thaliana. External Zn(2+) determines a potentiation of the current mediated by the dimeric construct KDC1-KAT1, which has been ascribed to zinc binding at a site comprising three histidines located at the S3-S4 (H161, H162) and S5-S6 (H224) linkers of KDC1. Here we demonstrate that also glutamate 164, located in close proximity of the KDC1 S4 segment, is an essential component of the zinc-binding site. On the contrary, glutamate 159, located in symmetrical position with respect to E164 in the sequence E(159)XHHXE(164) but more distant from the voltage sensor, does not play any role in zinc binding. The effects of Zn(2+) can be expressed as a "shift" of the gating parameters along the voltage axis. Kinetic modeling shows that Zn(2+) slows the closing kinetics of KDC1-KAT1 without affecting the opening kinetics. Possibly, zinc affects the movement of the voltage sensor in and out of the membrane phase through electrostatic modification of a site close to the voltage sensor.

The application of triazole fungicides is a common practice in the cultivation of carrot (Daucus carota L.) plants. It is there for seems important to test the changes that are occurring in this food crop under triazoles, the non-traditional plant growth regulators, treatments in order to identify the extent to which it tolerate the fungicide application and thereby make it an economical food crop. A field experiment was conducted to find out the effects of two triazole fungicides (hexaconazole (HEX) and paclobutrazol (PBZ) at 20mg l(-1) plant(-1)) on the biomass, yield, electrolyte leakage, lipid peroxidation and antioxidant potential of carrot. The treatments were given to plants on 15, 30 and 45 days after sowing (DAS). The plants were uprooted for analyses of growth and biochemical parameters on 60 DAS. It was found that both HEX and PBZ have significant effects on the growth and biochemical parameters of this plant. Among the triazoles used, PBZ performed best in terms of anthocyanin, protein, amino acid, proline, starch and sugar, contents whereas HEX enhanced carotenoids, fresh weight, dry weight and biomass. There was no significant variation in chlorophyll ('a' and 'b') contents between the two triazole treated plants, but HEX and PBZ proved best when compared to untreated control plants. HEX and PBZ increased alpha- and beta-amylases enzymes activities to a significant level. Out of these two triazoles, PBZ performed best in increasing the starch hydrolyzing enzymes activities. The non-enzymatic antioxidant, reduced glutathione (GSH) and antioxidant enzyme ascorbate peroxidase (APX) were increased under fungicide applications. The data suggests that, the application of triazole fungicides may be a useful tool to increase the tuber quality as well as quantity in carrot plants, apart from their fungicidal properties.

Carrot callus was centrifuged at 10 g and compared to callus growing at 1 g on agar in the presence of increasing sodium chloride concentrations. Growth after 14 days was enhanced in the centrifuged samples versus samples kept at 1 g. This effect was not found when the samples were grown on potassium chloride. At 50 mM NaCl, the calcium ionophore ionomycin was applied to centrifuged and noncentrifuged callus samples. In both experiments, the growth of callus increased with increasing ionomycin concentrations but under 10 g this increase was more enhanced. As inhibitors of calcium influx, lanthanum and gadolinium chloride were chosen in the presence of 50 mM NaCl. Both inhibitors inhibited growth at 1 g at low concentrations of around 2 microM, whereas the centrifuged samples were not or much less so inhibited. We tested an involvement of actin by application of cytochalasin D to callus grown in the presence of 50 mM NaCl. In both types of samples, growth at 1 g and growth at 10 g, cytochalasin D enhanced growth but the effect was clearly stronger at 10 g than at 1 g. As increased halotolerance was only observed in the presence of increased sodium ions, not potassium ions, and as halotolerance is known to be induced by an influx of calcium, the data suggest that a calcium influx induced by hypergravity and possibly modulated by actin caused the observed increase in halotolerance at 10 g.