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1.
Methods Mol Biol ; 2288: 113-126, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270008

RESUMO

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.


Assuntos
Daucus carota/crescimento & desenvolvimento , Daucus carota/genética , Melhoramento Vegetal/métodos , Aclimatação/genética , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Produtos Agrícolas/fisiologia , Meios de Cultura/química , Daucus carota/fisiologia , Flores/genética , Flores/crescimento & desenvolvimento , Heterozigoto , Homozigoto , Isoenzimas/análise , Biologia Molecular/métodos , Regeneração/genética , Técnicas de Cultura de Tecidos
2.
Int J Mol Sci ; 22(12)2021 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-34204559

RESUMO

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.


Assuntos
Vias Biossintéticas/genética , Sistemas CRISPR-Cas , Carotenoides/metabolismo , Parede Celular/metabolismo , Daucus carota/fisiologia , Edição de Genes , Sequência de Bases , Parede Celular/ultraestrutura , Daucus carota/ultraestrutura , Marcação de Genes , Genes de Plantas , Vetores Genéticos/genética , Mutação , Fenótipo , Plastídeos/genética , Plastídeos/ultraestrutura
3.
Plant Sci ; 302: 110699, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33288012

RESUMO

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.


Assuntos
Ácido Abscísico/metabolismo , Daucus carota/genética , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/fisiologia , Estômatos de Plantas/fisiologia , Fatores de Transcrição/fisiologia , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/fisiologia , Clonagem Molecular , Daucus carota/metabolismo , Daucus carota/fisiologia , Desidratação , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Transcrição/genética , Transcriptoma
4.
Int J Mol Sci ; 21(21)2020 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-33143222

RESUMO

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.


Assuntos
Biomarcadores/análise , Parede Celular/química , Reprogramação Celular , Daucus carota/fisiologia , Hipocótilo/fisiologia , Mucoproteínas/metabolismo , Técnicas de Embriogênese Somática de Plantas , Daucus carota/citologia , Epitopos/imunologia , Hipocótilo/citologia , Mucoproteínas/imunologia , Pectinas/química , Pectinas/metabolismo , Proteínas de Plantas/imunologia , Proteínas de Plantas/metabolismo
5.
Ecotoxicol Environ Saf ; 206: 111363, 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-32977082

RESUMO

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.


Assuntos
Cádmio/toxicidade , Cobre/toxicidade , Daucus carota/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Antioxidantes/metabolismo , Cádmio/análise , Cobre/análise , Daucus carota/fisiologia , Malondialdeído/metabolismo , Metais Pesados/análise , Metais Pesados/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Desenvolvimento Vegetal/genética
6.
Sci Rep ; 10(1): 10808, 2020 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-32616789

RESUMO

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.


Assuntos
Alternaria/isolamento & purificação , Daucus carota/microbiologia , Fungicidas Industriais/farmacologia , Sementes/microbiologia , Alternaria/efeitos dos fármacos , Daucus carota/fisiologia , Relação Dose-Resposta a Droga , Germinação/efeitos dos fármacos , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Fatores de Tempo
7.
DNA Cell Biol ; 39(5): 816-827, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32175765

RESUMO

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.


Assuntos
Daucus carota/genética , Daucus carota/fisiologia , Perfilação da Expressão Gênica , Genômica , Proteínas de Plantas/genética , Estresse Fisiológico/genética , Fatores de Transcrição/genética , Motivos de Aminoácidos , Sequência de Aminoácidos , Temperatura Baixa/efeitos adversos , Sequência Conservada , Daucus carota/efeitos dos fármacos , Daucus carota/crescimento & desenvolvimento , Evolução Molecular , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Sais/farmacologia , Fatores de Transcrição/química , Fatores de Transcrição/metabolismo
8.
Sci Rep ; 9(1): 17384, 2019 11 22.
Artigo em Inglês | MEDLINE | ID: mdl-31757985

RESUMO

Petaloid cytoplasmic male sterility (CMS) is a maternally inherited loss of male fertility due to the complete conversion of stamens into petal-like organs, and CMS lines have been widely utilized in carrot breeding. Petaloid CMS is an ideal model not only for studying the mitochondrial-nuclear interaction but also for discovering genes that are essential for floral organ development. To investigate the comprehensive mechanism of CMS and homeotic organ alternation during carrot flower development, we conducted transcriptome analysis between the petaloid CMS line (P2S) and its maintainer line (P2M) at four flower developmental stages (T1-T4). A total of 2838 genes were found to be differentially expressed, among which 1495 genes were significantly downregulated and 1343 genes were significantly upregulated in the CMS line. Functional analysis showed that most of the differentially expressed genes (DEGs) were involved in protein processing in the endoplasmic reticulum, plant hormone signal transduction, and biosynthesis. A total of 16 MADS-box genes were grouped into class A, B, C, and E, but not class D, genes. Several key genes associated with oxidative phosphorylation showed continuously low expression from stage T2 in P2S, and the expression of DcPI and DcAG-like genes also greatly decreased at stage T2 in P2S. This indicated that energy deficiency might inhibit the expression of B- and C-class MADS-box genes resulting in the conversion of stamens into petals. Stamen petaloidy may act as an intrinsic stress, upregulating the expression of heat shock protein (HSP) genes and MADS-box genes at stages T3 and T4 in P2S, which results in some fertile revertants. This study will provide a better understanding of carrot petaloid CMS and floral development as a basis for further research.


Assuntos
Daucus carota , Flores/crescimento & desenvolvimento , Infertilidade das Plantas/genética , Citoplasma/metabolismo , Daucus carota/genética , Daucus carota/crescimento & desenvolvimento , Daucus carota/fisiologia , Herança Extracromossômica/genética , Flores/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Proteínas de Domínio MADS/genética , Proteínas de Domínio MADS/metabolismo , Herança Materna/genética , Fenótipo , Reguladores de Crescimento de Plantas/genética , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Transcriptoma
9.
Acta Biomater ; 99: 457-468, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31525536

RESUMO

Many physically immobile plants develop passive yet ingenious strategies for active seed dispersal through self-deformation in response to external stimuli, such as humidity. These hygroscopic deformations are usually driven by the internal heterogeneous architecture, which provides valuable, inspiring information for the development of novel actuating systems. The Daucus carota compound umbel is an interesting structure showing a distinct hygroscopic deformation that operates at hierarchical levels among these plants. Here, we investigate the structure of the primary and secondary rays of the umbel associated with their deformation through mechanical analyses. We reveal that through controlling both the cellulose microfibril angle (MFA) and lignification, the multi-level bending behavior of the umbel is achieved, which contributes to efficient seed protection and dispersal. The primary rays generally show more significant bending curvature changes than the secondary rays, and within each level, the outer rays exhibit a larger motion amplitude than the middle and inner rays. Mechanical testing and theoretical analysis support that adjusting the lignin content within the ray structure compensates for the effect of the small differences in cellulose MFA on its bending behavior, which contributes to the overall hygroscopic deformation. Findings also show that the primary outer ray can generate reaction forces that are more than 700 times its weight, which is higher than that for the pine cone scales. The new insights from this work are instructive for bioinspired designs of complex, self-deforming structures and devices. STATEMENT OF SIGNIFICANCE: The carrot (Daucus carota) compound umbels exhibit a unique hierarchical, hygroscopic deformation for seed dispersal among immobile plants. In this work, we elucidate that the multi-level bending behavior of the umbel is achieved through manipulating the cellulose microfibril angle (MFA) and lignification of the primary and secondary rays for the first time. We also discover that adjusting the degree of lignification compensates for the effect of small cellulose MFA differences on the bending behavior theoretically and experimentally. The primary outer rays deform in a highly efficient manner, in which reactions forces about more than 700 times its weight are generated. The findings presented are instructive for bioinspired designs of complex, self-deforming structures and devices.


Assuntos
Daucus carota/fisiologia , Sementes/fisiologia , Estresse Mecânico , Parede Celular , Celulose/química , Módulo de Elasticidade , Lignina/química , Microscopia Eletrônica de Varredura , Pressão , Resistência à Tração , Molhabilidade
10.
Planta ; 250(2): 507-518, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31098709

RESUMO

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.


Assuntos
Daucus carota/genética , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Arabidopsis/anatomia & histologia , Arabidopsis/genética , Arabidopsis/fisiologia , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Quimera , Citoplasma/genética , DNA Mitocondrial/genética , Daucus carota/anatomia & histologia , Daucus carota/fisiologia , Marcadores Genéticos/genética , ATPases Mitocondriais Próton-Translocadoras/genética , ATPases Mitocondriais Próton-Translocadoras/metabolismo , Melhoramento Vegetal , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas
11.
PLoS One ; 14(1): e0210542, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30633764

RESUMO

Plants contain endophytic bacteria, whose communities both influence plant growth and can be an important source of probiotics. Here we used deep sequencing of a 16S rRNA gene fragment and bacterial cultivation to independently characterize the microbiomes of five plant species from divergent taxonomic orders-potato (Solanum tuberosum), carrot (Daucus sativus), beet (Beta vulgaris), neep (Brassica napus spp. napobrassica), and topinambur (Helianthus tuberosus). We found that both species richness and diversity tend to be higher in the peel, where Alphaproteobacteria and Actinobacteria dominate, while Gammaproteobacteria and Firmicutes dominate in the pulp. A statistical analysis revealed that the main characteristic features of the microbiomes of plant species originate from the peel microbiomes. Topinambur pulp displayed an interesting characteristic feature: it contained up to 108 CFUs of lactic acid bacteria, suggesting its use as a source of probiotic bacteria. We also detected Listeria sp., in topinambur pulps, however, the 16S rRNA gene fragment is unable to distinguish between pathogenic versus non-pathogenic species, so the evaluation of this potential health risk is left to a future study.


Assuntos
Bactérias/genética , Ecossistema , Endófitos/genética , Verduras/fisiologia , Bactérias/classificação , Bactérias/crescimento & desenvolvimento , Beta vulgaris/microbiologia , Beta vulgaris/fisiologia , Brassica napus/microbiologia , Brassica napus/fisiologia , DNA Bacteriano/genética , Daucus carota/microbiologia , Daucus carota/fisiologia , Endófitos/classificação , Endófitos/fisiologia , Helianthus/microbiologia , Helianthus/fisiologia , Microbiota/genética , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA/métodos , Solanum tuberosum/microbiologia , Solanum tuberosum/fisiologia , Verduras/classificação , Verduras/microbiologia
12.
Environ Sci Pollut Res Int ; 26(1): 381-391, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30402695

RESUMO

This work examines the strategies adopted by an arbuscular mycorrhizal symbiotic system to ameliorate environmental Pb stress by examining the concentrations of P, Fe, and Pb in the fungal microstructures and the host's root. In vitro cultures of Ri-T DNA-transformed carrot (Daucus carota L.) roots were inoculated with Glomus intraradices and treated with Pb(NO3)2 solution and the extraradical spores and mycelia (S/M) and the root with the vesicles, mycelia, and root cells were subsequently analyzed by polarized energy dispersive x-ray fluorescence (PEDXRF) spectrometry. Upon Pb treatment, within the root, the percentages of mycorrhizal colonization, the vesicles, and mycelia increased as well as the areas of the vesicles and the (extraradical) spores, although the number of spores and arbuscules decreased. The S/M and the mycorrhizal root showed enhanced concentrations of Pb, Fe, and P. These were particularly marked for Fe in the Pb-treated cultures. This indicates a synergistic relationship between the arbuscular mycorrhizal fungus and the host that confers a higher Pb tolerance to the latter by the induction of higher Fe absorption in the host. The intraradical vesicle, mycelia, and arbuscule numbers are interpreted as a "tactic to divert" the intraradical Pb traffic away from the root cells to the higher affinity cell walls of the arbuscular mycorrhizal fungi (AMF) microstructures in the apoplast. The results of this work show that the symbiosis between the AMF G. intraradices and the host plant D. carota distinctly improves the latter's Pb tolerance, and imply that the appropriate metal tolerant host-AMF combinations could be employed in process designs for the phytoremediation of Pb.


Assuntos
Daucus carota/fisiologia , Chumbo/metabolismo , Poluentes do Solo/metabolismo , Simbiose , Transporte Biológico , Daucus carota/microbiologia , Glomeromycota , Micélio , Micorrizas , Raízes de Plantas/microbiologia
13.
Mitochondrial DNA A DNA Mapp Seq Anal ; 29(3): 446-454, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-28335670

RESUMO

The male-sterile carrot is an effective material for carrot breeding. The atp6 gene is involved in carrot fertility. However, the differences in lengths, copies, and expression profiles of the atp6 gene in fertile and male-sterile lines of carrot are unclear. In this study, one copy atp6 gene was found in the mtDNAs of 'Kuroda' (fertility, 954 bp) and 'Wuye-BY' (male sterility, 819 bp) carrot lines, while two copies atp6 genes (Wuye-L and Wuye-D, 954 bp and 819 bp, respectively) were found in the mtDNA of 'Wuye' (fertility). Two putative conserved domains have been detected in the carrot atp6 protein. Evolutionary analysis showed that the atp6 protein sequences of Wuye-L and Kuroda were clustered in the same branch, while Wuye-D and Wuye-BY were clustered in the same branch. The atp6 gene was higher expressed in the flowers of 'Kuroda' and 'Wuye' (Wuye-L), while lower expressed in 'Wuye-BY' and 'Wuye' (Wuye-D).


Assuntos
Daucus carota/fisiologia , Dosagem de Genes , Expressão Gênica , ATPases Mitocondriais Próton-Translocadoras/genética , Daucus carota/classificação , Daucus carota/genética , Fertilidade , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Filogenia , Melhoramento Vegetal , Proteínas de Plantas/genética , Análise de Sequência de DNA
14.
Planta ; 245(4): 737-747, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27999990

RESUMO

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.


Assuntos
Carotenoides/fisiologia , Daucus carota/fisiologia , Regulação da Expressão Gênica de Plantas/fisiologia , Raízes de Plantas/fisiologia , Carotenoides/análise , Carotenoides/metabolismo , Daucus carota/crescimento & desenvolvimento , Immunoblotting , Floema/fisiologia , Raízes de Plantas/química , Xilema/fisiologia
15.
Plant Biol (Stuttg) ; 19(2): 286-297, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27775866

RESUMO

Carrot is generally regarded as a biennial plant with an obligatory vernalization requirement. Early spring cultivation makes plants vulnerable to premature bolting, which results in a loss of commercial value. However, our knowledge of flowering time genes and flowering mechanisms in carrot remain limited. Bolting behavior of D. carota ssp. carota 'Songzi', a wild species sensitive to flower induction by vernalization and photoperiod, and orange cultivar 'Amsterdam forcing', and their offspring were investigated in different growing conditions. We performed RNA-seq to identify the flowering time genes, and digital gene expression (DGE) analysis to examine their expression levels. The circadian patterns of related genes were identified by qPCR. The results showed bolting behavior of carrot was influenced by low temperature, illumination intensity and photoperiod. A total of 45 flowering time-related unigenes were identified, which were classified into five categories including photoperiod, vernalization, autonomous and gibberellin pathway, and floral integrators. Homologs of LATE ELONGATED HYPOCOTYL (LHY) and CONSTANS-LIKE 2 (COL2) were more highly expressed under short day condition than under long day condition. Homologs of COL2, CONSTANS-LIKE 5 (COL5), SUPPRESSION OF OVEREXPRESSION OF CONSTANS 1 (SOC1), FLOWERING LOCUS C (FLC) and GIBBERELLIC ACID INSENSITIVE (GAI) were differentially expressed between 'Songzi' and 'Amsterdam forcing'. The homolog of COL2 (Dct43207) was repressed by light, but that of COL5 (Dct20940) was induced. A preliminary model of genetic network controlling flowering time was constructed by associating the results of DGE analysis with correlation coefficients between genes. This study provides useful information for further investigating the genetic mechanism of flowering in carrot.


Assuntos
Daucus carota/genética , Regulação da Expressão Gênica de Plantas , Redes Reguladoras de Genes , Fotoperíodo , Transcriptoma , Relógios Circadianos , Daucus carota/crescimento & desenvolvimento , Daucus carota/fisiologia , Daucus carota/efeitos da radiação , Flores/genética , Flores/crescimento & desenvolvimento , Flores/fisiologia , Flores/efeitos da radiação , Perfilação da Expressão Gênica , Luz , Filogenia , Fatores de Tempo
16.
Ecotoxicology ; 26(1): 22-31, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27770233

RESUMO

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.


Assuntos
Aphanizomenon , Toxinas Bacterianas/toxicidade , Daucus carota/fisiologia , Minerais/metabolismo , Fotossíntese/efeitos dos fármacos , Uracila/análogos & derivados , Poluentes Químicos da Água/toxicidade , Alcaloides , Poluentes do Solo/toxicidade , Uracila/toxicidade
17.
Planta ; 241(2): 525-38, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25380771

RESUMO

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.


Assuntos
Daucus carota/metabolismo , Daucus carota/fisiologia , Temperatura
18.
Environ Pollut ; 184: 540-6, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24184375

RESUMO

The toxicity of arsenic (As) in the environment is controlled by its concentration, availability and speciation. The aims of the study were to evaluate the accumulation and speciation of As in carrot, lettuce and spinach cultivated in soils with various As concentrations and to estimate the concomitant health risks associated with the consumption of the vegetables. Arsenic concentration and speciation in plant tissues and soils was analysed by HPLC, AAS and XANES spectroscopy. To estimate the plants influence in the rhizosphere, organic acids in lettuce root exudates were analysed by ion chromatography. The results showed that the As accumulation was higher in plants cultivated in soil with higher As extractability. Arsenate predominated in the soils, rhizosphere and root exudates of lettuce. Succinic acid was the major organic acid in lettuce root exudates. Ingestion of the tested vegetables may result in an intake of elevated levels of inorganic As.


Assuntos
Arsênio/análise , Raízes de Plantas/química , Brotos de Planta/química , Rizosfera , Poluentes do Solo/análise , Solo/química , Verduras/química , Daucus carota/química , Daucus carota/fisiologia , Alface/química , Alface/fisiologia , Spinacia oleracea/química , Spinacia oleracea/fisiologia , Verduras/fisiologia , Espectroscopia por Absorção de Raios X
19.
Mol Plant Microbe Interact ; 27(1): 40-55, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24299212

RESUMO

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.


Assuntos
Arabidopsis/microbiologia , Ascomicetos/patogenicidade , Proteínas Fúngicas/metabolismo , Interações Hospedeiro-Patógeno , Doenças das Plantas/microbiologia , Tabaco/microbiologia , Motivos de Aminoácidos , Arabidopsis/fisiologia , Ascomicetos/genética , Ascomicetos/fisiologia , Ascomicetos/ultraestrutura , Morte Celular , Daucus carota/microbiologia , Daucus carota/fisiologia , Carpóforos , Proteínas Fúngicas/genética , Biblioteca Gênica , Inativação Gênica , Modelos Moleculares , Mutagênese Insercional , Ácido Oxálico/metabolismo , Fenótipo , Filogenia , Folhas de Planta/microbiologia , Folhas de Planta/fisiologia , Raízes de Plantas/microbiologia , Raízes de Plantas/fisiologia , Plântula/microbiologia , Plântula/fisiologia , Estresse Fisiológico , Tabaco/fisiologia
20.
Plant Signal Behav ; 8(5): e24030, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23435173

RESUMO

Arbuscular mycorrhizal (AM) symbiosis, established between AM fungi (AMF) and roots of higher plants, occurs in most terrestrial ecosystems. It has been well demonstrated that AM symbiosis can improve plant performance under various environmental stresses, including drought stress. However, the molecular basis for the direct involvement of AMF in plant drought tolerance has not yet been established. Most recently, we cloned two functional aquaporin genes, GintAQPF1 and GintAQPF2, from AM fungus Glomus intraradices. By heterologous gene expression in yeast, aquaporin localization, activities and water permeability were examined. Gene expressions during symbiosis in expose to drought stress were also analyzed. Our data strongly supported potential water transport via AMF to host plants. As a complement, here we adopted the monoxenic culture system for AMF, in which carrot roots transformed by Ri-T DNA were cultured with Glomus intraradices in two-compartment Petri dishes, to verify the aquaporin gene functions in assisting AMF survival under polyethylene glycol (PEG) treatment. Our results showed that 25% PEG significantly upregulated the expression of two aquaporin genes, which was in line with the gene functions examined in yeast. We therefore concluded that the aquaporins function similarly in AMF as in yeast subjected to osmotic stress. The study provided further evidence to the direct involvement of AMF in improving plant water relations under drought stresses.


Assuntos
Adaptação Fisiológica , Aquaporinas/genética , Daucus carota/microbiologia , Daucus carota/fisiologia , Secas , Genes Fúngicos/genética , Glomeromycota/genética , Aquaporinas/metabolismo , Micélio/genética
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