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1.
Molecules ; 29(6)2024 Mar 12.
Artículo en Inglés | MEDLINE | ID: mdl-38542896

RESUMEN

The effects of high-pressure processing (HPP) (450 MPa/600 MPa/3 min) on the carotenoid and vitamin E contents of smoothies made from strawberry, orange juice, banana and apple, and the same smoothies enriched with dietary fiber from discarded carrots were compared. The contents and bioaccessibilities of these compounds were also evaluated over the course of 28 days at 4 °C. The application of HPP in the formulations significantly increased the contents of ß-cryptoxanthin, α-carotene and ß-carotene and retained the contents of lutein, zeaxanthin and vitamin E compared to untreated samples. A decreasing trend in the content of each compound was observed with an increase in storage time. The application of HPP initially led to reductions in the bioaccessibility of individual compounds. However, overall, during storage, there was an increase in bioaccessibility. This suggests that HPP influences cell structure, favoring compound release and micelle formation. HPP is a sustainable method that preserves or enhances carotenoid extractability in ready-to-drink fruit beverages. Furthermore, the incorporation of dietary fiber from carrot processing discards supports circular economy practices and enhances the health potential of the product.


Asunto(s)
Daucus carota , Daucus carota/metabolismo , Vitamina E/análisis , Frutas/química , Carotenoides/análisis , Fibras de la Dieta/análisis
2.
Methods Enzymol ; 671: 273-283, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35878981

RESUMEN

Carrot (Daucus carota) is a useful plant model for the study of carotenoid biosynthesis, specifically in roots which are enriched in carotenoids. Carrot genome and transcriptome sequences, complemented by optimized methods for carrot transformation, contribute to a comprehensive toolbox for exploring pathway regulation. To expand the repertoire of tools available for the study of D. carota, we present protocols for the isolation of protoplasts from D. carota cell suspension cultures and polyethylene glycol (PEG)-mediated transformation. To obtain carrot protoplasts, in vitro somatic embryogenesis from epicotyls is induced. The somatic embryogenic tissue that develops is transferred to liquid medium to obtain a suspension of cells which are homogenized and incubated with cell-wall degrading enzymes to release protoplasts. For transfection, protoplasts are incubated with a plasmid encoding a protein of interest prior to examination of protein localization by light microscopy. As an example, we demonstrate nuclear localization of a carrot transcription factor, DcAREB3.


Asunto(s)
Daucus carota , Carotenoides/metabolismo , Daucus carota/genética , Daucus carota/metabolismo , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Protoplastos/metabolismo
3.
Plant Physiol ; 189(3): 1450-1465, 2022 06 27.
Artículo en Inglés | MEDLINE | ID: mdl-35266544

RESUMEN

Light stimulates carotenoid synthesis in plants during photomorphogenesis through the expression of PHYTOENE SYNTHASE (PSY), a key gene in carotenoid biosynthesis. The orange carrot (Daucus carota) synthesizes and accumulates high amounts of carotenoids in the taproot that grows underground. Contrary to other organs, light impairs carrot taproot development and represses the expression of carotenogenic genes, such as DcPSY1 and DcPSY2, reducing carotenoid accumulation. By means of RNA sequencing, in a previous analysis, we observed that carrot PHYTOCHROME RAPIDLY REGULATED1 (DcPAR1) is more highly expressed in the underground grown taproot compared with those grown in light. PAR1 is a transcriptional cofactor with a negative role in shade avoidance syndrome regulation in Arabidopsis (Arabidopsis thaliana) through the dimerization with PHYTOCHROME-INTERACTING FACTORs (PIFs), allowing a moderate synthesis of carotenoids. Here, we show that overexpressing AtPAR1 in carrot increases carotenoid production in taproots grown underground as well as DcPSY1 expression. The high expression of AtPAR1 and DcPAR1 led us to hypothesize a functional role of DcPAR1 that was verified through in vivo binding to AtPIF7 and overexpression in Arabidopsis, where AtPSY expression and carotenoid accumulation increased together with a photomorphogenic phenotype. Finally, DcPAR1 antisense carrot lines presented a dramatic decrease in carotenoid levels and in relative expression of key carotenogenic genes as well as impaired taproot development. These results suggest that DcPAR1 is a key factor for secondary root development and carotenoid synthesis in carrot taproot grown underground.


Asunto(s)
Arabidopsis , Daucus carota , Fitocromo , Arabidopsis/genética , Arabidopsis/metabolismo , Carotenoides/metabolismo , Daucus carota/genética , Daucus carota/metabolismo , Regulación de la Expresión Génica de las Plantas , Fitocromo/metabolismo
4.
Genes (Basel) ; 12(10)2021 09 22.
Artículo en Inglés | MEDLINE | ID: mdl-34680859

RESUMEN

In purple carrots, anthocyanin pigmentation can be expressed in the entire root, or it can display tissue specific-patterns. Within the phloem, purple pigmentation can be found in the outer phloem (OP) (also called the cortex) and inner phloem (IP), or it can be confined exclusively to the OP. In this work, the genetic control underlying tissue-specific anthocyanin pigmentation in the carrot root OP and IP tissues was investigated by means of linkage mapping and transcriptome (RNA-seq) and phylogenetic analyses; followed by gene expression (RT-qPCR) evaluations in two genetic backgrounds, an F2 population (3242) and the inbred B7262. Genetic mapping of 'root outer phloem anthocyanin pigmentation' (ROPAP) and inner phloem pigmentation (RIPAP) revealed colocalization of ROPAP with the P1 and P3 genomic regions previously known to condition pigmentation in different genetic stocks, whereas RIPAP co-localized with P3 only. Transcriptome analysis of purple OP (POP) vs. non-purple IP (NPIP) tissues, along with linkage and phylogenetic data, allowed an initial identification of 28 candidate genes, 19 of which were further evaluated by RT-qPCR in independent root samples of 3242 and B7262, revealing 15 genes consistently upregulated in the POP in both genetic backgrounds, and two genes upregulated in the POP in specific backgrounds. These include seven transcription factors, seven anthocyanin structural genes, and two genes involved in cellular transport. Altogether, our results point at DcMYB7, DcMYB113, and a MADS-box (DCAR_010757) as the main candidate genes conditioning ROPAP in 3242, whereas DcMYB7 and MADS-box condition RIPAP in this background. In 7262, DcMYB113 conditions ROPAP.


Asunto(s)
Antocianinas/metabolismo , Daucus carota/metabolismo , Perfilación de la Expresión Génica , Floema/metabolismo , Pigmentos Biológicos/metabolismo , Raíces de Plantas/metabolismo , Daucus carota/genética , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Reacción en Cadena en Tiempo Real de la Polimerasa
5.
Sci Rep ; 11(1): 4093, 2021 02 18.
Artículo en Inglés | MEDLINE | ID: mdl-33603038

RESUMEN

Carrot (Daucus carota L.) is one of the most cultivated vegetable in the world and of great importance in the human diet. Its storage organs can accumulate large quantities of anthocyanins, metabolites that confer the purple pigmentation to carrot tissues and whose biosynthesis is well characterized. Long non-coding RNAs (lncRNAs) play critical roles in regulating gene expression of various biological processes in plants. In this study, we used a high throughput stranded RNA-seq to identify and analyze the expression profiles of lncRNAs in phloem and xylem root samples using two genotypes with a strong difference in anthocyanin production. We discovered and annotated 8484 new genes, including 2095 new protein-coding and 6373 non-coding transcripts. Moreover, we identified 639 differentially expressed lncRNAs between the phenotypically contrasted genotypes, including certain only detected in a particular tissue. We then established correlations between lncRNAs and anthocyanin biosynthesis genes in order to identify a molecular framework for the differential expression of the pathway between genotypes. A specific natural antisense transcript linked to the DcMYB7 key anthocyanin biosynthetic transcription factor suggested how the regulation of this pathway may have evolved between genotypes.


Asunto(s)
Antocianinas/metabolismo , Daucus carota/metabolismo , Raíces de Plantas/metabolismo , ARN Largo no Codificante/inmunología , Antocianinas/biosíntesis , Daucus carota/genética , Regulación de la Expresión Génica de las Plantas/genética , Genes de Plantas/genética , Floema/metabolismo , Transcriptoma , Xilema/metabolismo
6.
Mol Genet Genomics ; 295(6): 1379-1392, 2020 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-32656704

RESUMEN

Carotenoids are terpenoid pigments synthesized by all photosynthetic and some non-photosynthetic organisms. In plants, these lipophilic compounds are involved in photosynthesis, photoprotection, and phytohormone synthesis. In plants, carotenoid biosynthesis is induced by several environmental factors such as light including photoreceptors, such as phytochromes (PHYs) and negatively regulated by phytochrome interacting factors (PIFs). Daucus carota (carrot) is one of the few plant species that synthesize and accumulate carotenoids in the storage root that grows in darkness. Contrary to other plants, light inhibits secondary root growth and carotenoid accumulation suggesting the existence of new mechanisms repressed by light that regulate both processes. To identify genes induced by dark and repressed by light that regulate carotenoid synthesis and carrot root development, in this work an RNA-Seq analysis was performed from dark- and light-grown carrot roots. Using this high-throughput sequencing methodology, a de novo transcriptome model with 63,164 contigs was obtained, from which 18,488 were differentially expressed (DEG) between the two experimental conditions. Interestingly, light-regulated genes are preferably expressed in dark-grown roots. Enrichment analysis of GO terms with DEGs genes, validation of the transcriptome model and DEG analysis through qPCR allow us to hypothesize that genes involved in photomorphogenesis and light perception such as PHYA, PHYB, PIF3, PAR1, CRY2, FYH3, FAR1 and COP1 participate in the synthesis of carotenoids and carrot storage root development.


Asunto(s)
Vías Biosintéticas/genética , Carotenoides/metabolismo , Biología Computacional/métodos , Daucus carota/genética , Daucus carota/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/metabolismo , Daucus carota/crecimiento & desarrollo , Perfilación de la Expresión Génica , Pigmentación , Proteínas de Plantas/genética
7.
J Sci Food Agric ; 100(13): 4995-4998, 2020 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-32478414

RESUMEN

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.


Asunto(s)
Antioxidantes/química , Daucus carota/química , Desinfectantes/farmacología , Desinfección/métodos , Manipulación de Alimentos/métodos , Fenoles/química , Antioxidantes/metabolismo , Cloro/farmacología , Ácido Clorogénico/farmacología , Daucus carota/efectos de los fármacos , Daucus carota/metabolismo , Fenoles/metabolismo , Tubérculos de la Planta/química , Tubérculos de la Planta/metabolismo
8.
Plant Sci ; 291: 110327, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31928663

RESUMEN

Daucus carota is a biennale crop that develops an edible storage root. Orange carrots, the most consumed cultivar worldwide, accumulate high levels of ß-carotene and α-carotene in the storage root during secondary growth. Genes involved in ß-carotene synthesis have been identified in carrots and unlike most species, D. carota has two ζ-carotene desaturase genes, named ZDS1 and ZDS2, that share 91.3 % identity in their coding regions. ZDS1 expression falls during leaf, but not root development, while ZDS2 is induced in leaves and storage roots of a mature plant. In this work, by means of post-transcriptional gene silencing, we determined that ZDS1 is essential for initial carrot development. The suppression of the expression of this gene by RNAi triggered a reduction in the transcript levels of ZDS2 and PSY2 genes, with a concomitant decrease in the carotenoid content in both, leaves and storage roots. On the contrary, transgenic lines with reduced ZDS2 transcript abundance maintain the same levels of expression of endogenous ZDS1 and PSY2 and carotenoid profile as wild-type plants. The simultaneous silencing of ZDS1 and ZDS2 resulted in lines with a negligible leaf and root development, as well as significantly lower endogenous PSY2 expression. Further functional analyses, such as a plastidial subcellular localization of ZDS1:GFP and the increment in carotenoid content in transgenic tobacco plants overexpressing the carrot ZDS1, confirmed that ZDS1 codifies for a functional enzyme. Overall, these results lead us to propose that the main ζ-carotene desaturase activity in carrot is encoded by the ZDS1 gene and ZDS2 gene has a complementary and non essential role.


Asunto(s)
Carotenoides/metabolismo , Daucus carota/genética , Oxidorreductasas/genética , Proteínas de Plantas/genética , Daucus carota/crecimiento & desarrollo , Daucus carota/metabolismo , Oxidorreductasas/metabolismo , Proteínas de Plantas/metabolismo
9.
Theor Appl Genet ; 132(9): 2485-2507, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31144001

RESUMEN

KEY MESSAGE: Inheritance, QTL mapping, phylogenetic, and transcriptome (RNA-Seq) analyses provide insight into the genetic control underlying carrot root and leaf tissue-specific anthocyanin pigmentation and identify candidate genes for root phloem pigmentation. Purple carrots can accumulate large quantities of anthocyanins in their root tissues, as well as in other plant parts. This work investigated the genetic control underlying tissue-specific anthocyanin pigmentation in the carrot root phloem and xylem, and in leaf petioles. Inheritance of anthocyanin pigmentation in these three tissues was first studied in segregating F2 and F4 populations, followed by QTL mapping of phloem and xylem anthocyanin pigments (independently) onto two genotyping by sequencing-based linkage maps, to reveal two regions in chromosome 3, namely P1 and P3, controlling pigmentation in these three tissues. Both P1 and P3 condition pigmentation in the phloem, with P3 also conditioning pigmentation in the xylem and petioles. By means of linkage mapping, phylogenetic analysis, and comparative transcriptome (RNA-Seq) analysis among carrot roots with differing purple pigmentation phenotypes, we identified candidate genes conditioning pigmentation in the phloem, the main tissue influencing total anthocyanin levels in the root. Among them, a MYB transcription factor, DcMYB7, and two cytochrome CYP450 genes with putative flavone synthase activity were identified as candidates regulating both the presence/absence of pigmentation and the concentration of anthocyanins in the root phloem. Concomitant expression patterns of DcMYB7 and eight anthocyanin structural genes were found, suggesting that DcMYB7 regulates transcription levels in the latter. Another MYB, DcMYB6, was upregulated in specific purple-rooted samples, suggesting a genotype-specific regulatory activity for this gene. These data contribute to the understanding of anthocyanin regulation in the carrot root at a tissue-specific level and maybe instrumental for improving carrot nutritional value.


Asunto(s)
Antocianinas/genética , Daucus carota/genética , Pigmentación/genética , Hojas de la Planta/genética , Proteínas de Plantas/genética , Raíces de Plantas/genética , Sitios de Carácter Cuantitativo , Antocianinas/metabolismo , Cromosomas de las Plantas , Color , Daucus carota/crecimiento & desarrollo , Daucus carota/metabolismo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Filogenia , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/metabolismo , Proteínas de Plantas/metabolismo , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo , Polimorfismo de Nucleótido Simple , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Transcriptoma
10.
J Exp Bot ; 69(16): 4113-4126, 2018 07 18.
Artículo en Inglés | MEDLINE | ID: mdl-29860511

RESUMEN

Phytoene synthase (PSY) is the first committed enzyme of the carotenoid biosynthesis pathway and the most important point of regulation. Carotenoids are precursors of abscisic acid (ABA), which mediates abiotic stress tolerance responses in plants. ABA activates the synthesis of its own precursors through induction of PSY expression. Carrot, a species that accumulates very high amounts of carotenoids in its reserve root, has two PSY paralog genes that are expressed differentially in the root. Here, we determined that DcPSY2 expression is induced by salt stress and ABA. A DcPSY2 promoter fragment was obtained and characterized. Bioinformatic analysis showed the presence of three ABA responsive elements (ABREs). Through overexpressing pPSY2:GFP in Nicotiana tabacum we determined that all three ABREs are necessary for the ABA response. In the carrot transcriptome, we identified three ABRE binding protein (DcAREB) transcription factor candidates that localized in the nucleus, but only one, DcAREB3, was induced under ABA treatment in carrot roots. We found that AREB transcription factors bind to the carrot DcPSY2 promoter and transactivate the expression of reporter genes. We conclude that DcPSY2 is involved in ABA-mediated salt stress tolerance in carrot through the binding of AREB transcription factors to its promoter.


Asunto(s)
Ácido Abscísico/metabolismo , Daucus carota/metabolismo , Geranilgeranil-Difosfato Geranilgeraniltransferasa/biosíntesis , Estrés Salino , Daucus carota/genética , Inducción Enzimática , Regulación Enzimológica de la Expresión Génica , Regulación de la Expresión Génica de las Plantas
11.
Subcell Biochem ; 79: 199-217, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27485223

RESUMEN

Carrot (Daucus carota) is one of the most important vegetable cultivated worldwide and the main source of dietary provitamin A. Contrary to other plants, almost all carrot varieties accumulate massive amounts of carotenoids in the root, resulting in a wide variety of colors, including those with purple, yellow, white, red and orange roots. During the first weeks of development the root, grown in darkness, is thin and pale and devoid of carotenoids. At the second month, the thickening of the root and the accumulation of carotenoids begins, and it reaches its highest level at 3 months of development. This normal root thickening and carotenoid accumulation can be completely altered when roots are grown in light, in which chromoplasts differentiation is redirected to chloroplasts development in accordance with an altered carotenoid profile. Here we discuss the current evidence on the biosynthesis of carotenoid in carrot roots in response to environmental cues that has contributed to our understanding of the mechanism that regulates the accumulation of carotenoids, as well as the carotenogenic gene expression and root development in D. carota.


Asunto(s)
Carotenoides/biosíntesis , Daucus carota/metabolismo , Pigmentos Biológicos/biosíntesis , beta Caroteno/biosíntesis , Carotenoides/metabolismo , Daucus carota/crecimiento & desarrollo , Regulación de la Expresión Génica de las Plantas , Humanos , Pigmentos Biológicos/metabolismo , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo , Plastidios/genética , Plastidios/metabolismo , Vitamina A/biosíntesis , Vitamina A/metabolismo , beta Caroteno/genética
12.
PLoS One ; 11(6): e0154438, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27253975

RESUMEN

Arbuscular mycorrhizal fungi (AMF) and phosphate solubilizing Pseudomonas bacteria (PSB) could potentially interact synergistically because PSB solubilize phosphate into a form that AMF can absorb and transport to the plant. However, very little is known about the interactions between these two groups of microorganisms and how they influence the growth of each other. We tested whether different strains of bacteria, that have the capacity to solubilize phosphate, are able to grow along AMF hyphae and differentially influence the growth of AMF both outside the roots of carrot in in vitro conditions and inside the roots of potato in the presence of a microbial community. We found strong effects of AMF on the growth of the different bacterial strains. Different bacterial strains also had very strong effects on the growth of AMF extraradical hyphae outside the roots of carrot and on colonization of potato roots by AMF. The differential effects on colonization occurred in the presence of a microbial community. Our results show that these two important groups of rhizosphere microorganisms indeed interact with each other. Such interactions could potentially lead to synergistic effects between the two groups but this could depend on whether the bacteria truly solubilize phosphate in the rhizosphere in the presence of microbial communities.


Asunto(s)
Micorrizas/metabolismo , Fosfatos/metabolismo , Pseudomonas/metabolismo , Rizosfera , Daucus carota/crecimiento & desarrollo , Daucus carota/metabolismo , Daucus carota/microbiología , Micorrizas/crecimiento & desarrollo , Raíces de Plantas/microbiología , Microbiología del Suelo , Simbiosis/genética
13.
Sci Total Environ ; 565: 557-563, 2016 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-27196992

RESUMEN

Consumption of vegetables grown in arsenic (As)-contaminated soils is an important exposure route to the element for humans. The present study is focused on locally-grown, frequently-consumed vegetables, such as carrots (Daucus carota), beets (Beta vulgaris) and quinoa (Chenopodium) from the As-polluted Chiu Chiu area in Northern Chile. The latter region is affected both by As discharge from copper mining activity and natural As contamination, leading to a high As content in local food and water. For the selected vegetables, the following aspects were investigated: i) Their total As, Cu, Pb, Cr, Cd and Mn content; ii) Arsenic speciation in the edible part of the vegetables by liquid chromatography inductively-coupled plasma mass spectrometry (LC-ICPMS) analysis; iii) Arsenic bioaccessibility in the vegetables during in vitro gastrointestinal digestion; iv) Arsenic species present in the extracts obtained from in vitro gastrointestinal digestion; and v) Arsenic dietary exposure estimates for the assessment of the risk posed by the vegetables consumption. A significant degree of As contamination was found in the vegetables under study, their metal content having been compared with that of similar Spanish uncontaminated products. In vitro gastrointestinal digestion of the studied vegetables led to quantitative extraction of As from carrots and beets, whereas efficiency was about 40% for quinoa. For carrots, only As(III) and As(V) species were found, being their concentration levels similar. In the case of quinoa, around 85% of the element was present as As(V). For beets, inorganic As(V) and unknown overlapped As species (probably arsenosugars) were found. No significant transformation of the original As species was observed during in vitro gastrointestinal digestion. Arsenic dietary exposure values obtained for the three vegetables (0.017-0.021µg As person(-1)day(-1)) were much lower than the JFCFA's safety limit of 50µg As person(-1)day(-1). Therefore, no toxicological risk would be expected from the intake of these vegetables.


Asunto(s)
Arsénico/análisis , Contaminación de Alimentos/análisis , Contaminantes del Suelo/análisis , Arsénico/química , Beta vulgaris/metabolismo , Chenopodium quinoa/metabolismo , Chile , Cromatografía Liquida , Daucus carota/metabolismo , Espectrometría de Masas , Contaminantes del Suelo/química
14.
Planta ; 243(3): 675-85, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26613600

RESUMEN

MAIN CONCLUSION: The Taenia solium HP6/TSOL18 antigen was produced in carrot cells, yielding an immunogenic protein that induced significant protection in an experimental murine model against T. crassiceps cysticercosis when orally administered. This result supports the potential of HP6/TSOL18-carrot as a low-cost anti-cysticercosis vaccine candidate. Cysticercosis is a zoonosis caused by Taenia solium that can be prevented by interrupting the parasite life cycle through pig vaccination. Several injectable vaccine candidates have been reported, but the logistic difficulties and costs for its application limited its use in nationwide control programs. Oral plant-based vaccines can deal with this limitation, because of their easy administration and low cost. A stable expression of the HP6/TSOL18 anti-T. solium cysticercosis protective antigen in carrot calli transformed with an optimized transgene is herein reported. An antigen accumulation up to 14 µg g(-1) of dry-weight biomass was achieved in the generated carrot lines. Mouse immunization with one of the transformed calli induced both specific IgG and IgA anti-HP6/TSOL18 antibodies. A statistically significant reduction in the expected number of T. crassiceps cysticerci was observed in mice orally immunized with carrot-made HP6/TSOL18, in a similar extent to that obtained by subcutaneous immunization with recombinant HP6/TSOL18 protein. In this study, a new oral plant-made version of the HP6/TSOL18 anti-cysticercosis vaccine is reported. The vaccine candidate should be further tested against porcine cysticercosis.


Asunto(s)
Antígenos Helmínticos/inmunología , Cisticercosis/veterinaria , Daucus carota/metabolismo , Taenia solium/inmunología , Administración Oral , Animales , Cisticercosis/parasitología , Cisticercosis/prevención & control , Daucus carota/genética , Femenino , Inmunización , Ratones , Ratones Endogámicos BALB C , Proteínas Recombinantes , Porcinos , Transgenes , Vacunas
15.
Genet Mol Res ; 14(4): 13274-88, 2015 Oct 27.
Artículo en Inglés | MEDLINE | ID: mdl-26535641

RESUMEN

The basic-region/leucine-zipper (bZIP) family is one of the major transcription factor (TF) families associated with responses to abiotic stresses. Many members of group A in this family have been extensively examined and are reported to perform significant functions in ABA signaling as well as in responses to abiotic stresses. In this study, 10 bZIP factors in carrot were classified into group A based on their DNA-binding domains. The cis-acting regulatory elements and folding states of these 10 factors were analyzed. Evolutionary analysis of the group A members suggested their importance during the course of evolution in plants. In addition, cis-acting elements and the folding state of proteins were important for DNA binding and could affect gene expression. Quantitative RT-PCR was conducted to investigate the stress response of 10 genes encoding the group A factors. Six genes showed responses to abiotic stresses, while four genes showed other special phenomenon. The current analysis on group A bZIP family TFs in carrot is the first to investigate the TFs of Apiaceae via genome analysis. These results provide new information for future studies on carrot.


Asunto(s)
Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/metabolismo , Daucus carota/genética , Daucus carota/metabolismo , Genómica , Estrés Fisiológico/genética , Secuencia de Aminoácidos , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/química , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/clasificación , Evolución Molecular , Expresión Génica , Genómica/métodos , Datos de Secuencia Molecular , Filogenia , Posición Específica de Matrices de Puntuación , Pliegue de Proteína , Dominios y Motivos de Interacción de Proteínas , Secuencias Reguladoras de Ácidos Nucleicos , Alineación de Secuencia
16.
J Agric Food Chem ; 60(45): 11378-86, 2012 Nov 14.
Artículo en Inglés | MEDLINE | ID: mdl-23101679

RESUMEN

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.


Asunto(s)
Antioxidantes/metabolismo , Daucus carota/efectos de los fármacos , Daucus carota/metabolismo , Glicina/análogos & derivados , Herbicidas/farmacología , Fenoles/metabolismo , Ácido Shikímico/metabolismo , Daucus carota/genética , Glicina/farmacología , Glifosato
17.
J Plant Physiol ; 168(2): 174-80, 2011 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-20655621

RESUMEN

Expression of the protective F1 and V antigens of Yersinia pestis, as a fusion protein, in carrot was pursued in an effort to develop an alternative vaccine production system against the serious plague disease. Transgenic carrot plants carrying the F1-V encoding gene were developed via Agrobacterium-mediated transformation. Presence, integration, and expression of the F1-V encoding gene were confirmed by polymerase chain reaction (PCR), DNA gel blot analysis, and reverse-transcriptase (RT)-PCR analyses, respectively. An ELISA assay confirmed the antigenicity of the plant-derived F1-V fusion protein. Immunogenicity was evaluated subcutaneously in mice using a soluble protein extract of freeze-dried transgenic carrot. Significant antibody levels were detected following immunization. These results demonstrated that the F1-V protein could be expressed in carrot tap roots, and that the carrot F1-V recombinant protein retained its antigenicity and immunogenicity.


Asunto(s)
Antígenos Bacterianos/metabolismo , Proteínas Bacterianas/metabolismo , Daucus carota/metabolismo , Raíces de Plantas/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Proteínas Citotóxicas Formadoras de Poros/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Yersinia pestis/metabolismo , Animales , Antígenos Bacterianos/genética , Proteínas Bacterianas/genética , Daucus carota/genética , Ensayo de Inmunoadsorción Enzimática , Ratones , Ratones Endogámicos BALB C , Peste/inmunología , Vacuna contra la Peste/genética , Vacuna contra la Peste/inmunología , Vacuna contra la Peste/metabolismo , Raíces de Plantas/genética , Plantas Modificadas Genéticamente/genética , Proteínas Citotóxicas Formadoras de Poros/genética , Proteínas Recombinantes de Fusión/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Yersinia pestis/genética
18.
J Environ Radioact ; 100(2): 176-83, 2009 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-19084298

RESUMEN

Vegetables grown with phosphate fertilizer (conventional management), with bovine manure fertilization (organic management) and in a mineral nutrient solution (hydroponic) were analyzed and the concentrations of (238)U, (226)Ra and (228)Ra in lettuce, carrots, and beans were compared. Lettuce from hydroponic farming system showed the lowest concentration of radionuclides 0.51 for (226)Ra, 0.55 for (228)Ra and 0.24 for (238)U (Bq kg(-1) dry). Vegetables from organically and conventionally grown farming systems showed no differences in the concentration of radium and uranium. Relationships between uranium content in plants and exchangeable Ca and Mg in soil were found, whereas Ra in vegetables was inversely correlated to the cation exchange capacity of soil, leading to the assumption that by supplying carbonate and cations to soil, liming may cause an increase of U and a decrease of radium uptake by plants. The soil to plant transfer varied from 10(-4) to 10(-2) for (238)U and from 10(-2) to 10(-1) for (228)Ra.


Asunto(s)
Agricultura , Radio (Elemento)/análisis , Uranio/análisis , Verduras/metabolismo , Daucus carota/metabolismo , Fabaceae/metabolismo , Lactuca/metabolismo , Radio (Elemento)/metabolismo , Uranio/química
19.
Biol Res ; 41(3): 289-301, 2008.
Artículo en Inglés | MEDLINE | ID: mdl-19399342

RESUMEN

Carotenoids are synthesized in prokaryotic and eukaryotic organisms. In plants and algae, these lipophilic molecules possess antioxidant properties acting as reactive oxygen species scavengers and exert functional roles in hormone synthesis, photosynthesis, photomorphogenesis and in photoprotection. During the past decade almost all carotenogenic genes have been identified as a result of molecular, genetic and biochemical approaches utilizing Arabidopsis thaliana as the model system. Studies carried out in leaves and fruits of A. thaliana and tomato determined that light regulates carotenoid biosynthesis preferentially through the modulation of carotenogenic gene transcription. In this work we showed for the first time that light induces accumulation of psy1, pds and zds2 transcripts in leaves of Daucus carota (carrot), a novel plant model. In addition, modified roots of carrots exposed to light accumulate zds1, whereas the pds gene is highly repressed, suggesting that some carotenogenic genes, which are expressed in roots, are regulated by light. Additionally, light negatively regulates the development of the modified carrot root in a reversible manner. Therefore, this suggests that light affects normal growth and carotenogenic gene expression in the modified root of carrot plants. The molecular insight gained into the light-regulated expression of carotenoid genes in this and other model systems will facilitate our understanding of the regulation of carotenoid biosynthesis to improve the prospects for the metabolic engineering of carotenoid production in plants.


Asunto(s)
Carotenoides/genética , Daucus carota/genética , Regulación de la Expresión Génica de las Plantas/genética , Luz , Modelos Genéticos , Carotenoides/biosíntesis , Daucus carota/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de la radiación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
20.
Biol. Res ; 41(3): 289-301, 2008. ilus, tab, graf
Artículo en Inglés | LILACS | ID: lil-511919

RESUMEN

Carotenoids are synthesized in prokaryotic and eukaryotic organisms. In plants and algae, these lipophilic molecules possess antioxidant properties acting as reactive oxygen species scavengers and exert functional roles in hormone synthesis, photosynthesis, photomorphogenesis and in photoprotection. During the past decade almost all carotenogenic genes have been identified as a result of molecular, genetic and biochemical approaches utilizing Arabidopsis thaliana as the model system. Studies carried out in leaves and fruits of A. thaliana and tomato determined that light regulates carotenoid biosynthesis preferentially through the modulation of carotenogenic gene transcription. In this work we showed for the first time that light induces accumulation of psy 1, pds and zds2 transcripts in leaves of Daucus carota (carrot), a novel plant model. In addition, modified roots of carrots exposed to light accumulate zdsl, whereas the pds gene is highly repressed, suggesting that some carotenogenic genes, which are expressed in roots, are regulated by light. Additionally, light negatively regulates the development of the modified carrot root in a reversible manner. Therefore, this suggests that light affects normal growth and carotenogenic gene expression in the modified root of carrot plants. The molecular insight gained into the light-regulated expression of carotenoid genes in this and other model systems will facilitate our understanding of the regulation of carotenoid biosynthesis to improve the prospects for the metabolic engineering of carotenoid production in plants.


Asunto(s)
Carotenoides/genética , Daucus carota/genética , Regulación de la Expresión Génica de las Plantas/genética , Luz , Modelos Genéticos , Carotenoides/biosíntesis , Daucus carota/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de la radiación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
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