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1.
BMC Plant Biol ; 24(1): 606, 2024 Jun 26.
Artículo en Inglés | MEDLINE | ID: mdl-38926658

RESUMEN

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 KNO3 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 KNO3 (T0: unprimed (negative control), T1: hydroprimed (positive control), T2: 50 mM, T3:100mM, T4: 150 mM, T5: 200 mM, T6: 250 mM and T7: 300 mM) for 12 h each in darkness at 20 ± 2℃. Seed priming with 50 mM of KNO3 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 KNO3, seed germination, growth and root biomass were reduced. These findings suggest that seed priming with 50 mM of KNO3 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 KNO3 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.


Asunto(s)
Antioxidantes , Daucus carota , Germinación , Nitratos , Compuestos de Potasio , Plantones , Semillas , Antioxidantes/metabolismo , Plantones/crecimiento & desarrollo , Plantones/efectos de los fármacos , Plantones/fisiología , Nitratos/metabolismo , Nitratos/farmacología , Semillas/efectos de los fármacos , Semillas/crecimiento & desarrollo , Semillas/fisiología , Daucus carota/crecimiento & desarrollo , Daucus carota/efectos de los fármacos , Daucus carota/fisiología , Compuestos de Potasio/farmacología , Germinación/efectos de los fármacos , Calor
2.
BMC Plant Biol ; 21(1): 475, 2021 Oct 18.
Artículo en Inglés | MEDLINE | ID: mdl-34663216

RESUMEN

BACKGROUND: Carrot (Daucus carota L.), an important root vegetable, is very popular among consumers as its taproot is rich in various nutrients. Abiotic stresses, such as drought, salt, and low temperature, are the main factors that restrict the growth and development of carrots. Non-heme carotene hydroxylase (BCH) is a key regulatory enzyme in the ß-branch of the carotenoid biosynthesis pathway, upstream of the abscisic acid (ABA) synthesis pathway. RESULTS: In this study, we characterized a carrot BCH encoding gene, DcBCH1. The expression of DcBCH1 was induced by drought treatment. The overexpression of DcBCH1 in Arabidopsis thaliana resulted in enhanced tolerance to drought, as demonstrated by higher antioxidant capacity and lower malondialdehyde content after drought treatment. Under drought stress, the endogenous ABA level in transgenic A. thaliana was higher than that in wild-type (WT) plants. Additionally, the contents of lutein and ß-carotene in transgenic A. thaliana were lower than those in WT, whereas the expression levels of most endogenous carotenogenic genes were significantly increased after drought treatment. CONCLUSIONS: DcBCH1 can increase the antioxidant capacity and promote endogenous ABA levels of plants by regulating the synthesis rate of carotenoids, thereby regulating the drought resistance of plants. These results will help to provide potential candidate genes for plant drought tolerance breeding.


Asunto(s)
Ácido Abscísico/metabolismo , Arabidopsis/genética , Carotenoides/metabolismo , Daucus carota/genética , Oxigenasas de Función Mixta/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Antioxidantes/metabolismo , Arabidopsis/fisiología , Daucus carota/fisiología , Sequías , Expresión Génica , Regulación de la Expresión Génica de las Plantas , Oxigenasas de Función Mixta/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Estrés Fisiológico
3.
Int J Mol Sci ; 22(12)2021 Jun 17.
Artículo en Inglés | MEDLINE | ID: mdl-34204559

RESUMEN

Recent data indicate that modifications to carotenoid biosynthesis pathway in plants alter the expression of genes affecting chemical composition of the cell wall. Phytoene synthase (PSY) is a rate limiting factor of carotenoid biosynthesis and it may exhibit species-specific and organ-specific roles determined by the presence of psy paralogous genes, the importance of which often remains unrevealed. Thus, the aim of this work was to elaborate the roles of two psy paralogs in a model system and to reveal biochemical changes in the cell wall of psy knockout mutants. For this purpose, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR associated (Cas9) proteins (CRISPR/Cas9) vectors were introduced to carotenoid-rich carrot (Daucus carota) callus cells in order to induce mutations in the psy1 and psy2 genes. Gene sequencing, expression analysis, and carotenoid content analysis revealed that the psy2 gene is critical for carotenoid biosynthesis in this model and its knockout blocks carotenogenesis. The psy2 knockout also decreased the expression of the psy1 paralog. Immunohistochemical staining of the psy2 mutant cells showed altered composition of arabinogalactan proteins, pectins, and extensins in the mutant cell walls. In particular, low-methylesterified pectins were abundantly present in the cell walls of carotenoid-rich callus in contrast to the carotenoid-free psy2 mutant. Transmission electron microscopy revealed altered plastid transition to amyloplasts instead of chromoplasts. The results demonstrate for the first time that the inhibited biosynthesis of carotenoids triggers the cell wall remodelling.


Asunto(s)
Vías Biosintéticas/genética , Sistemas CRISPR-Cas , Carotenoides/metabolismo , Pared Celular/metabolismo , Daucus carota/fisiología , Edición Génica , Secuencia de Bases , Pared Celular/ultraestructura , Daucus carota/ultraestructura , Marcación de Gen , Genes de Plantas , Vectores Genéticos/genética , Mutación , Fenotipo , Plastidios/genética , Plastidios/ultraestructura
4.
Int J Mol Sci ; 22(20)2021 Oct 14.
Artículo en Inglés | MEDLINE | ID: mdl-34681749

RESUMEN

Carrot (Daucus carota L.) is widely cultivated as one of the most important root crops, and developing an effective presowing treatment method can promote the development of modern mechanized precision sowing. In the present study, a novel seed priming technology, named hydro-electro hybrid priming (HEHP), was used to promote the germination of carrot seeds. Seed germination experiments showed that HEHP was able to increase the germination index (GI) and vigor index (VI) by 3.1-fold and 6.8-fold, respectively, and the effect was significantly superior to that of hydro-priming (HYD) and electrostatic field treatment (EF). The consumption and utilization rate of seed storage reserves were also greatly improved. Meanwhile, both glyoxysomes and mitochondria were found to appear ahead of time in the endosperm cells of HEHP through observations of the subcellular structure of the endosperm. Activities of isocitrate lyase (ICL), NAD-dependent malate dehydrogenase (MDH), pyruvate kinase (PK), and alcohol dehydrogenase (ADH) were significantly increased by HEHP. From transcriptome results, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways related to the glyoxylate cycle, glycolysis, gluconeogenesis, and the citrate cycle were significantly enriched and real-time quantitative PCR (qRT-PCR) analysis confirmed the expression pattern of 15 critical differentially expressed genes (DEGs) in these pathways. All DEGs encoding MDH, phosphoenolpyruvate carboxykinase (PEPCK), and PK were upregulated in HEHP; thus, it is reasonable to infer that the transformation of malate, oxalacetate, phosphoenolpyruvate, and pyruvate in the cytoplasm may be pivotal for the energy supply during early germination. The results suggest that the optimal effect of HEHP is achieved by initiating stored lipid utilization and respiratory metabolism pathways related to germination.


Asunto(s)
Daucus carota/fisiología , Germinación/fisiología , Metabolismo de los Lípidos , Semillas/metabolismo , Daucus carota/metabolismo , Endospermo/citología , Endospermo/fisiología , Enzimas/metabolismo , Regulación de la Expresión Génica de las Plantas , Glioxilatos/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Semillas/crecimiento & desarrollo , Electricidad Estática , Factores de Transcripción/genética , Factores de Transcripción/metabolismo
5.
Ecotoxicol Environ Saf ; 206: 111363, 2020 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-32977082

RESUMEN

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.


Asunto(s)
Cadmio/toxicidad , Cobre/toxicidad , Daucus carota/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Antioxidantes/metabolismo , Cadmio/análisis , Cobre/análisis , Daucus carota/fisiología , Malondialdehído/metabolismo , Metales Pesados/análisis , Metales Pesados/toxicidad , Estrés Oxidativo/efectos de los fármacos , Desarrollo de la Planta/genética
6.
Int J Mol Sci ; 21(21)2020 Oct 30.
Artículo en Inglés | MEDLINE | ID: mdl-33143222

RESUMEN

Changes in the composition of the cell walls are postulated to accompany changes in the cell's fate. We check whether there is a relationship between the presence of selected pectic, arabinogalactan proteins (AGPs), and extensins epitopes and changes in cell reprogramming in order to answer the question of whether they can be markers accompanying changes of cell fate. Selected antibodies were used for spatio-temporal immunolocalization of wall components during the induction of somatic embryogenesis. Based on the obtained results, it can be concluded that (1) the LM6 (pectic), LM2 (AGPs) epitopes are positive markers, but the LM5, LM19 (pectic), JIM8, JIM13 (AGPs) epitopes are negative markers of cells reprogramming to the meristematic/pluripotent state; (2) the LM8 (pectic), JIM8, JIM13, LM2 (AGPs) and JIM11 (extensin) epitopes are positive markers, but LM6 (pectic) epitope is negative marker of cells undergoing detachment; (3) JIM4 (AGPs) is a positive marker, but LM5 (pectic), JIM8, JIM13, LM2 (AGPs) are negative markers for pericycle cells on the xylem pole; (4) LM19, LM20 (pectic), JIM13, LM2 (AGPs) are constitutive wall components, but LM6, LM8 (pectic), JIM4, JIM8, JIM16 (AGPs), JIM11, JIM12 and JIM20 (extensins) are not constitutive wall components; (5) the extensins do not contribute to the cell reprogramming.


Asunto(s)
Biomarcadores/análisis , Pared Celular/química , Reprogramación Celular , Daucus carota/fisiología , Hipocótilo/fisiología , Mucoproteínas/metabolismo , Técnicas de Embriogénesis Somática de Plantas , Daucus carota/citología , Epítopos/inmunología , Hipocótilo/citología , Mucoproteínas/inmunología , Pectinas/química , Pectinas/metabolismo , Proteínas de Plantas/inmunología , Proteínas de Plantas/metabolismo
7.
Planta ; 250(2): 507-518, 2019 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-31098709

RESUMEN

MAIN CONCLUSION: 'Petaloid' cytoplasmic male sterility is commonly used as a stable genetic mechanism in carrot hybrid breeding. Its introgression in tropical carrot showed morphometric changes and molecular markers were identified for detection at early stage. Cytoplasmic male sterility (CMS) is the only genetic mechanism in carrot for commercial exploitation of heterosis and production of low cost affordable hybrid seeds. The 'petaloid' CMS system is stable and commonly used in hybrid breeding in temperate carrot but there is no information available on existence of natural CMS system in tropical Asiatic carrot. Therefore, the present study was aimed to investigate morphometric traits and organizational features of cytoplasmic atp9 gene sequences in newly converted CMS lines (BC4-7) of tropical carrot. The CMS lines had root traits at par with fertile counterparts while floral traits had variation. Petal colour and length, petaloids colour and shape and style length showed differences among the CMS lines and with their maintainers. Molecular markers are effective to establish male sterility at genetic level, for this, six fixed and stable CMS lines were screened with seven novel primer combinations. Out of which five pairs produced clearly distinguishable bands in CMS lines and their fertile counterparts. The study confirmed that the region between 3' end of atp9-1/atp9-3 gene and 5' end of region of homology to Arabidopsis thaliana mtDNA is ideal for developing the trait specific markers. These new CMS lines have potential to use in hybrid development and molecular markers will be useful to confirm male sterility to rogue out fertile plants.


Asunto(s)
Daucus carota/genética , Infertilidad Vegetal/genética , Proteínas de Plantas/genética , Arabidopsis/anatomía & histología , Arabidopsis/genética , Arabidopsis/fisiología , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Quimera , Citoplasma/genética , ADN Mitocondrial/genética , Daucus carota/anatomía & histología , Daucus carota/fisiología , Marcadores Genéticos/genética , ATPasas de Translocación de Protón Mitocondriales/genética , ATPasas de Translocación de Protón Mitocondriales/metabolismo , Fitomejoramiento , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente
8.
Planta ; 245(4): 737-747, 2017 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-27999990

RESUMEN

Main conclusion Variations in gene expression can partially explain the difference of carotenoid accumulation in secondary phloem and xylem of fleshy carrot roots. The carrot root is well divided into two different tissues separated by vascular cambium: the secondary phloem and xylem. The equilibrium between these two tissues represents an important issue for carrot quality, but the knowledge about the respective carotenoid accumulation is sparse. The aim of this work was (i) to investigate if variation in carotenoid biosynthesis gene expression could explain differences in carotenoid content in phloem and xylem tissues and (ii) to investigate if this regulation is differentially modulated in the respective tissues by water-restricted growing conditions. In this work, five carrot genotypes contrasting by their root color were studied in control and water-restricted conditions. Carotenoid content and the relative expression of 13 genes along the carotenoid biosynthesis pathway were measured in the respective tissues. Results showed that in orange genotypes and the purple one, carotenoid content was higher in phloem compared to xylem. For the red one, no differences were observed. Moreover, in control condition, variations in gene expression explained the different carotenoid accumulations in both tissues, while in water-restricted condition, no clear association between gene expression pattern and variations in carotenoid content could be detected except in orange-rooted genotypes. This work shows that the structural aspect of carrot root is more important for carotenoid accumulation in relation with gene expression levels than the consequences of expression changes upon water restriction.


Asunto(s)
Carotenoides/fisiología , Daucus carota/fisiología , Regulación de la Expresión Génica de las Plantas/fisiología , Raíces de Plantas/fisiología , Carotenoides/análisis , Carotenoides/metabolismo , Daucus carota/crecimiento & desarrollo , Immunoblotting , Floema/fisiología , Raíces de Plantas/química , Xilema/fisiología
9.
Ecotoxicology ; 26(1): 22-31, 2017 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-27770233

RESUMEN

Natural toxins produced by freshwater cyanobacteria, such as cylindrospermopsin, have been regarded as an emergent environmental threat. Despite the risks for food safety, the impact of these water contaminants in agriculture is not yet fully understood. Carrots (Daucus carota) are root vegetables, extensively consumed worldwide with great importance for human nourishment and economy. It is, therefore, important to evaluate the possible effects of using water contaminated with cyanotoxins on carrot cultivation. The aim of this work was to investigate cylindrospermopsin effects on D. carota grown in soil and irrigated for 30 days, with a Chrysosporum ovalisporum extract containing environmentally relevant concentrations of cylindrospermopsin (10 and 50 µg/L). The parameters evaluated were plant growth, photosynthetic capacity, and nutritional value (mineral content) in roots of carrots, as these are the edible parts of this plant crop. The results show that, exposure to cylindrospermopsin did not have a clear negative effect on growth or photosynthesis of D. carota, even leading to an increase of both parameters. However, alterations in mineral contents were detected after exposure to crude extracts of C. ovalisporum containing cylindrospermopsin. A general decline was observed for most minerals (Ca, Mg, Na, Fe, Mn, Zn, Mo, and P), although an increase was shown in the case of K and Cu, pointing to a possible interference of the cyanobacterial extract in mineral uptake. This study is the first to evaluate the effects of C. ovalisporum extracts on a root vegetable, however, more research is necessary to understand the effects of this toxin in environmentally relevant scenarios.


Asunto(s)
Aphanizomenon , Toxinas Bacterianas/toxicidad , Daucus carota/fisiología , Minerales/metabolismo , Fotosíntesis/efectos de los fármacos , Uracilo/análogos & derivados , Contaminantes Químicos del Agua/toxicidad , Alcaloides , Toxinas de Cianobacterias , Contaminantes del Suelo/toxicidad , Uracilo/toxicidad
10.
Planta ; 241(2): 525-38, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25380771

RESUMEN

MAIN CONCLUSION: Calorespirometric measurements proved to be useful for phenotyping temperature response in terms of optimum temperatures for growth and low temperature limits for growth respiration in diverse carrot genotypes. High and low-temperature tolerance is an important trait in many breeding programs, but to date, improvement strategies have had limited success. Developing new, cost efficient and reliable screening tools to identify and select the most tolerant crop plant genotypes is necessary to assist plant breeding on cold and heat tolerance, and calorespirometry is proposed for this. Calorespirometry is a technique to simultaneously measure metabolic heat rates and CO2 emission rates of respiring tissues and can be used as a rapid method to determine how changes in the environment (e.g., temperature) influence plant growth. The main aim of this work was, therefore, to test the usefulness of calorespirometry as a phenotyping tool for carrot taproot growth in response to temperature. Calorespirometric measurements in the carrot taproot meristems of plants from eight carrot inbred lines allowed identification of optimum and minimum temperatures for growth of plants and to distinguish between phenotypes based on those characteristics. The technique proved to be useful for predicting yield-determining temperature responses in diverse carrot genotypes. Preliminary screening of new crop plant genotypes with calorespirometry based on their temperature adaptation and acclimation capability could make the screening process much less laborious by allowing selection of genotypes presenting the best growth performance under particular biotic or abiotic conditions before field tests.


Asunto(s)
Daucus carota/metabolismo , Daucus carota/fisiología , Temperatura
11.
Mol Plant Microbe Interact ; 27(1): 40-55, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-24299212

RESUMEN

To decipher the mechanism of pathogenicity in Sclerotinia sclerotiorum, a pathogenicity-defective mutant, Sunf-MT6, was isolated from a T-DNA insertional library. Sunf-MT6 could not form compound appressorium and failed to induce lesions on leaves of rapeseed though it could produce more oxalic acid than the wild-type strain. However, it could enter into host tissues via wounds and cause typical necrotic lesions. Furthermore, Sunf-MT6 produced fewer but larger sclerotia than the wild-type strain Sunf-M. A gene, named Ss-caf1, was disrupted by T-DNA insertion in Sunf-MT6. Gene complementation and knockdown experiments confirmed that the disruption of Ss-caf1 was responsible for the phenotypic changes of Sunf-MT6. Ss-caf1 encodes a secretory protein with a putative Ca(2+)-binding EF-hand motif. High expression levels of Ss-caf1 were observed at an early stage of compound appressorium formation and in immature sclerotia. Expression of Ss-caf1 without signal peptides in Nicotiana benthamiana via Tobacco rattle virus-based vectors elicited cell death. These results suggest that Ss-caf1 plays an important role in compound appressorium formation and sclerotial development of S. sclerotiorum. In addition, Ss-Caf1 has the potential to interact with certain host proteins or unknown substances in host cells, resulting in subsequent host cell death.


Asunto(s)
Arabidopsis/microbiología , Ascomicetos/patogenicidad , Proteínas Fúngicas/metabolismo , Interacciones Huésped-Patógeno , Nicotiana/microbiología , Enfermedades de las Plantas/microbiología , Secuencias de Aminoácidos , Arabidopsis/fisiología , Ascomicetos/genética , Ascomicetos/fisiología , Ascomicetos/ultraestructura , Muerte Celular , Daucus carota/microbiología , Daucus carota/fisiología , Cuerpos Fructíferos de los Hongos , Proteínas Fúngicas/genética , Biblioteca de Genes , Silenciador del Gen , Modelos Moleculares , Mutagénesis Insercional , Ácido Oxálico/metabolismo , Fenotipo , Filogenia , Hojas de la Planta/microbiología , Hojas de la Planta/fisiología , Raíces de Plantas/microbiología , Raíces de Plantas/fisiología , Plantones/microbiología , Plantones/fisiología , Estrés Fisiológico , Nicotiana/fisiología
12.
Exp Parasitol ; 133(2): 166-78, 2013 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-23201220

RESUMEN

Root lesion nematodes (RLNs, Pratylenchus species) are a group of economically important migratory endoparasitic plant pathogens that attack host roots of major crops such as wheat and sugarcane, and can reduce crop yields by 7-15%. Pratylenchus thornei and Pratylenchus zeae were treated with double stranded RNA (dsRNA) to study gene silencing, (RNA interference, RNAi), as a potential strategy for their control. Mixed stages of nematodes of both species ingested dsRNA when incubated in a basic soaking solution in the presence of the neurostimulant octopamine. Incubation for up to 16 h in soaking solutions containing 10-50 mM octopamine, 0.1-1.0 mg/mL FITC, and 0.5-6 mM spermidine did not affect vitality. Spermidine phosphate salt hexahydrate rather than spermidine or spermidine trihydrochloride increased uptake of FITC by nematodes, and this resulted in more effective gene silencing. Silencing pat-10 and unc-87 genes of P. thornei and P. zeae resulted in paralysis and uncoordinated movements in both species, although to a higher degree in P. thornei. There was also a greater reduction in transcript of both genes in P. thornei indicating that it may be more susceptible to RNAi. For P. thornei treated with dsRNA of pat-10 and unc-87 there was a significant reduction (77-81%) in nematode reproduction on carrot mini discs over a 5 week period. The results show that RLNs are clearly amenable to gene silencing, and that in planta delivery of dsRNA to target genes in these nematodes should confer host resistance. Moreover, for the two genes, dsRNA derived from either nematode species silenced the corresponding gene in both species. This implies cross-species control of nematodes via RNAi is possible.


Asunto(s)
Productos Agrícolas/fisiología , Productos Agrícolas/parasitología , Daucus carota/fisiología , Daucus carota/parasitología , Silenciador del Gen/fisiología , Tylenchoidea/fisiología , Agonistas alfa-Adrenérgicos/farmacología , Animales , Proteínas de Unión al Calcio/genética , Proteínas de Unión al Calcio/fisiología , Fluoresceína-5-Isotiocianato/metabolismo , Colorantes Fluorescentes/metabolismo , Proteínas de Microfilamentos/genética , Proteínas de Microfilamentos/fisiología , Octopamina/farmacología , Enfermedades de las Plantas/parasitología , ARN Bicatenario/metabolismo , Reproducción , Resorcinoles/farmacología , Saccharum/parasitología , Sorghum/parasitología , Espermidina/farmacología , Triticum/parasitología , Troponina C/genética , Troponina C/fisiología , Tylenchoidea/efectos de los fármacos , Tylenchoidea/genética , Australia Occidental , Calponinas
13.
J Plant Res ; 126(2): 243-50, 2013 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-22878456

RESUMEN

Totipotency is the ability of a cell to regenerate the entire organism, even after previous differentiation as a specific cell. When totipotency is coupled with active cell division, it was presumed that cell division is essential for this expression. Here, using the stress-induction system of somatic embryos in carrots, we show that cell division is not essential for the expression of totipotency in somatic/embryonic conversion. Morphological and histochemical analyses showed that the cell did not divide during embryo induction. Inhibitors of cell division did not affect the rate of somatic embryo formation. Our results indicate that the newly acquired trait of differentiation appears without cell division, but does not arise with cell division as a newborn cell.


Asunto(s)
Diferenciación Celular/fisiología , División Celular/efectos de los fármacos , Daucus carota/citología , Daucus carota/fisiología , Técnicas de Embriogénesis Somática de Plantas , Afidicolina/farmacología , Benzamidas/farmacología , Inhibidores Enzimáticos/farmacología , Hibridación in Situ , Estrés Fisiológico , Células Madre Totipotentes/citología
14.
J Exp Bot ; 63(17): 6187-97, 2012 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23105130

RESUMEN

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.


Asunto(s)
Arabidopsis/fisiología , Ácido Araquidónico/farmacología , Daucus carota/fisiología , Ácidos Grasos Insaturados/farmacología , Canales Iónicos/metabolismo , Vacuolas/metabolismo , Arabidopsis/efectos de los fármacos , Proteínas de Arabidopsis/metabolismo , Ácido Araquidónico/química , Calcio/metabolismo , Canales de Calcio/metabolismo , Daucus carota/efectos de los fármacos , Electrofisiología , Ácidos Grasos Insaturados/química , Activación del Canal Iónico , Cinética , Ácido Linoleico/metabolismo , Potenciales de la Membrana , Células del Mesófilo/fisiología , Modelos Biológicos , Ácidos Oléicos/metabolismo , Técnicas de Placa-Clamp , Proteínas de Plantas/metabolismo , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/fisiología , Ácido alfa-Linolénico/metabolismo
15.
Methods Mol Biol ; 2288: 113-126, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34270008

RESUMEN

Carrot is a vegetable of increasing economic importance. New hybrid cultivars are constantly required to meet the changing market needs. The application of anther culture significantly shortens the difficult and long-lasting breeding of carrot. We examined all the stages of the process of generating androgenic plants: induction of embryos in anther cultures, regeneration and acclimatization of produced plants, their evaluation, ploidy and homozygosity, and many other factors affecting their effectiveness. Every factor has been optimized by experimentally selecting the optimal level. As a result, a full protocol of producing homozygous plants using anther cultures was developed, which is presented in this chapter.


Asunto(s)
Daucus carota/crecimiento & desarrollo , Daucus carota/genética , Fitomejoramiento/métodos , Aclimatación/genética , Productos Agrícolas/genética , Productos Agrícolas/crecimiento & desarrollo , Productos Agrícolas/fisiología , Medios de Cultivo/química , Daucus carota/fisiología , Flores/genética , Flores/crecimiento & desarrollo , Heterocigoto , Homocigoto , Isoenzimas/análisis , Biología Molecular/métodos , Regeneración/genética , Técnicas de Cultivo de Tejidos
16.
Plant Sci ; 302: 110699, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33288012

RESUMEN

Abscisic acid-responsive element (ABRE)-binding factors (ABFs) are important transcription factors involved in various physiological processes in plants. Stomata are micro channels for water and gas exchange of plants. Previous researches have demonstrated that ABFs can modulate the stomatal development in some plants. However, little is known about stomata-related functions of ABFs in carrots. In our study, DcABF3, a gene encoding for ABF transcription factor, was isolated from carrot. The open reading frame of DcABF3 was 1329 bp, encoding 442 amino acids. Expression profiles of DcABF3 indicated that DcABF3 can respond to drought, salt or ABA treatment in carrots. Overexpressing DcABF3 in Arabidopsis led to the increase of stomatal density which caused severe water loss. Expression assay indicated that overexpression of DcABF3 caused high expression of stomatal development-related transcription factor genes, SPCH, FAMA, MUTE and SCRMs. Increased antioxidant enzyme activities and higher expression levels of stress-related genes were also found in transgenic lines after water deficit treatment. Changes in expression of ABA synthesis-related genes and AtABIs indicated the potential role of DcABF3 in ABA signaling pathway. Under the treatment of exogenous ABA, DcABF3-overexpression Arabidopsis seedlings exhibited increased root length and germination rate. Our findings demonstrated that heterologous overexpression of DcABF3 positively affected stomatal development and also reduced ABA sensitivity in transgenic Arabidopsis.


Asunto(s)
Ácido Abscísico/metabolismo , Daucus carota/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Proteínas de Plantas/fisiología , Estomas de Plantas/fisiología , Factores de Transcripción/fisiología , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/fisiología , Clonación Molecular , Daucus carota/metabolismo , Daucus carota/fisiología , Deshidratación , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Plantas Modificadas Genéticamente/fisiología , Reacción en Cadena en Tiempo Real de la Polimerasa , Factores de Transcripción/genética , Transcriptoma
17.
J Appl Genet ; 51(2): 141-7, 2010.
Artículo en Inglés | MEDLINE | ID: mdl-20453301

RESUMEN

Microspores were cultured on the modified B5 liquid medium containing 2.4D (0.1 mg L(-1)), NAA (0.1 mg L(-1)), L-glutamine (500 mg L(-1), L-serine (100 mg L(-1)), and sucrose (100 g L(-1)). The developmental stages of microspores and divisions were observed. Initially, the formation of binuclear and multicellular structures was noticed. Plants regenerated in the cultures in which the tetrad stage of microsporogenesis had predominated. Embryoids were still forming 24 weeks after the cultures were set up. Six weeks after the transfer of androgenetic embryos onto the B5 regeneration medium, they were converted into complete plants. Out of 90 androgenetic plants planted in a growth chamber, 42 plants adapted to the new conditions. All of those plants proved to be diploids in cytometric analysis.


Asunto(s)
Daucus carota/fisiología , Polen/fisiología , Técnicas de Cultivo de Célula , División Celular , Cromosomas de las Plantas/genética , Daucus carota/genética , Daucus carota/crecimiento & desarrollo , Gametogénesis en la Planta/fisiología , Haploidia , Polen/citología , Técnicas de Cultivo de Tejidos
18.
Ecotoxicol Environ Saf ; 73(5): 1018-27, 2010 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-20403638

RESUMEN

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).


Asunto(s)
Contaminantes Atmosféricos/toxicidad , Daucus carota/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Ozono/toxicidad , Compuestos de Fenilurea/farmacología , Daucus carota/fisiología , Relación Dosis-Respuesta a Droga , Monitoreo del Ambiente , Peroxidación de Lípido/efectos de los fármacos , Dióxido de Nitrógeno/análisis , Fotosíntesis/efectos de los fármacos , Dióxido de Azufre/análisis
19.
Sci Rep ; 10(1): 10808, 2020 07 02.
Artículo en Inglés | MEDLINE | ID: mdl-32616789

RESUMEN

Carrot (Daucus carota L.), which is one of the 10 most important vegetable crops worldwide, is an edible root vegetable desired for its taste as well as its medicinal uses. However, a fungus isolated from carrot seeds was observed to substantially decrease the germination rate. The isolate was identified as Alternaria alternata based on morphological and molecular characteristics as well as a phylogenetic tree. The maximum seed infection rate of selected carrot cultivars was approximately 60%, with the main infection site just underneath the seed shell. Additionally, the germination rate of infected seeds decreased by 28.7%. However, the seed infection rate varied among the examined carrot cultivars. Regarding the effects of chemical fungicides, the optimal treatment involved immersing seeds in amistar top suspension concentrate (SC) (effective concentration of 0.65 g/L) for 6 h, which effectively killed the fungi inside the carrot seeds. The results of this study provide a theoretical basis for the development of efficient methods for preventing the infection of carrot seeds by specific fungi and increasing the germination rate and vigour index.


Asunto(s)
Alternaria/aislamiento & purificación , Daucus carota/microbiología , Fungicidas Industriales/farmacología , Semillas/microbiología , Alternaria/efectos de los fármacos , Daucus carota/fisiología , Relación Dosis-Respuesta a Droga , Germinación/efectos de los fármacos , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Factores de Tiempo
20.
DNA Cell Biol ; 39(5): 816-827, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32175765

RESUMEN

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.


Asunto(s)
Daucus carota/genética , Daucus carota/fisiología , Perfilación de la Expresión Génica , Genómica , Proteínas de Plantas/genética , Estrés Fisiológico/genética , Factores de Transcripción/genética , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Frío/efectos adversos , Secuencia Conservada , Daucus carota/efectos de los fármacos , Daucus carota/crecimiento & desarrollo , Evolución Molecular , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Raíces de Plantas/crecimiento & desarrollo , Sales (Química)/farmacología , Factores de Transcripción/química , Factores de Transcripción/metabolismo
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