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1.
Theor Appl Genet ; 134(10): 3351-3362, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34282485

RESUMO

KEY MESSAGE: The Or and CH genes are necessary for the accumulation of high amounts of ß-carotene and other carotenoid pigments in carrot roots, in addition to the Y and Y2 genes. Carrot taproot color results from the accumulation of various carotenoid and anthocyanin pigments. Recently, the Or gene was identified as a candidate gene associated with the accumulation of ß-carotene and other provitamin A carotenoids in roots. The specific molecular mechanisms involved with this process, as well as the interactions between Or and the other genes involved in this process are not well understood. In order to better characterize the effect that Or alleles have on conditioning the accumulation of carotenoids in roots, we analyzed an F3 family fixed homozygous recessive for y and y2, derived from a cross between an orange carrot and a white wild carrot, segregating for the two known Or alleles, which we name Orc and Orw. QTL mapping across three different environments revealed that the accumulation of several carotenoids was associated with the Orc allele, with consistent patterns across environments. A second QTL on chromosome 7, harboring a carotene hydroxylase gene homologous to Lut5 in Arabidopsis, was also associated with the accumulation of several carotenoids. Two alleles for this gene, which we name CHc and CHw, were discovered to be segregating in this population. Our study provides further evidence that Or and CH are likely involved with controlling the accumulation of ß-carotene and may be involved with modulating carotenoid flux in carrot, demonstrating that both were important domestication genes in carrot.


Assuntos
Carotenoides/metabolismo , Daucus carota/metabolismo , Regulação da Expressão Gênica de Plantas , Oxigenases de Função Mista/metabolismo , Fenótipo , Proteínas de Plantas/metabolismo , Raízes de Plantas/metabolismo , Daucus carota/genética , Daucus carota/crescimento & desenvolvimento , Oxigenases de Função Mista/genética , Pigmentação , Proteínas de Plantas/genética , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Locos de Características Quantitativas
2.
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
3.
Front Immunol ; 12: 673692, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34305903

RESUMO

In a perspective entitled 'From plant survival under severe stress to anti-viral human defense' we raised and justified the hypothesis that transcript level profiles of justified target genes established from in vitro somatic embryogenesis (SE) induction in plants as a reference compared to virus-induced profiles can identify differential virus signatures that link to harmful reprogramming. A standard profile of selected genes named 'ReprogVirus' was proposed for in vitro-scanning of early virus-induced reprogramming in critical primary infected cells/tissues as target trait. For data collection, the 'ReprogVirus platform' was initiated. This initiative aims to identify in a common effort across scientific boundaries critical virus footprints from diverse virus origins and variants as a basis for anti-viral strategy design. This approach is open for validation and extension. In the present study, we initiated validation by experimental transcriptome data available in public domain combined with advancing plant wet lab research. We compared plant-adapted transcriptomes according to 'RegroVirus' complemented by alternative oxidase (AOX) genes during de novo programming under SE-inducing conditions with in vitro corona virus-induced transcriptome profiles. This approach enabled identifying a major complex trait for early de novo programming during SARS-CoV-2 infection, called 'CoV-MAC-TED'. It consists of unbalanced ROS/RNS levels, which are connected to increased aerobic fermentation that links to alpha-tubulin-based cell restructuration and progression of cell cycle. We conclude that anti-viral/anti-SARS-CoV-2 strategies need to rigorously target 'CoV-MAC-TED' in primary infected nose and mouth cells through prophylactic and very early therapeutic strategies. We also discuss potential strategies in the view of the beneficial role of AOX for resilient behavior in plants. Furthermore, following the general observation that ROS/RNS equilibration/redox homeostasis is of utmost importance at the very beginning of viral infection, we highlight that 'de-stressing' disease and social handling should be seen as essential part of anti-viral/anti-SARS-CoV-2 strategies.


Assuntos
Reprogramação Celular/genética , Herança Multifatorial/genética , SARS-CoV-2/patogenicidade , Acetilserotonina O-Metiltransferasa/genética , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Ciclo Celular/genética , Bases de Dados Genéticas , Daucus carota/genética , Daucus carota/crescimento & desenvolvimento , Fermentação , Perfilação da Expressão Gênica , Humanos , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/metabolismo , Oxirredutases/genética , Oxirredutases/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Espécies Reativas de Nitrogênio/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Tubulina (Proteína)/genética , Vírus/patogenicidade
4.
Genes (Basel) ; 12(5)2021 05 18.
Artigo em Inglês | MEDLINE | ID: mdl-34069875

RESUMO

The AT-hook motif containing nuclear localized (AHL) gene family, controlling various developmental processes, is conserved in land plants. They comprise Plant and Prokaryote Conserved (PPC) domain and one or two AT-hook motifs. DcAHLc1 has been proposed as a candidate gene governing the formation of the carrot storage root. We identified and in-silico characterized carrot AHL proteins, performed phylogenetic analyses, investigated their expression profiles and constructed gene coexpression networks. We found 47 AHL genes in carrot and grouped them into two clades, A and B, comprising 29 and 18 genes, respectively. Within Clade-A, we distinguished three subclades, one of them grouping noncanonical AHLs differing in their structure (two PPC domains) and/or cellular localization (not nucleus). Coexpression network analysis attributed AHLs expressed in carrot roots into four of the 72 clusters, some of them showing a large number of interactions. Determination of expression profiles of AHL genes in various tissues and samples provided basis to hypothesize on their possible roles in the development of the carrot storage root. We identified a group of rapidly evolving noncanonical AHLs, possibly differing functionally from typical AHLs, as suggested by their expression profiles and their predicted cellular localization. We pointed at several AHLs likely involved in the development of the carrot storage root.


Assuntos
Motivos AT-Hook/genética , Daucus carota/crescimento & desenvolvimento , Daucus carota/genética , Desenvolvimento Vegetal/genética , Proteínas de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/genética , Regulação da Expressão Gênica de Plantas/genética , Filogenia , Transcriptoma/genética
5.
PLoS One ; 16(4): e0249613, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33798246

RESUMO

Renewed consumer demand motivates the nutritional and sensory quality improvement of fruits and vegetables. Specialized metabolites being largely involved in nutritional and sensory quality of carrot, a better knowledge of their phenotypic variability is required. A metabolomic approach was used to evaluate phenotypic plasticity level of carrot commercial varieties, over three years and a wide range of cropping environments spread over several geographical areas in France. Seven groups of metabolites have been quantified by HPLC or GC methods: sugars, carotenoids, terpenes, phenolic compounds, phenylpropanoids and polyacetylenes. A large variation in root metabolic profiles was observed, in relation with environment, variety and variety by environment interaction effects in decreasing order of importance. Our results show a clear diversity structuration based on metabolite content. Polyacetylenes, ß-pinene and α-carotene were identified mostly as relatively stable varietal markers, exhibiting static stability. Nevertheless, environment effect was substantial for a large part of carrot metabolic profile and various levels of phenotypic plasticity were observed depending on metabolites and varieties. A strong difference of environmental sensitivity between varieties was observed for several compounds, particularly myristicin, 6MM and D-germacrene, known to be involved in responses to biotic and abiotic stress. This work provides useful information about plasticity in the perspective of carrot breeding and production. A balance between constitutive content and environmental sensitivity for key metabolites should be reached for quality improvement in carrot and other vegetables.


Assuntos
Adaptação Fisiológica , Carotenoides/análise , Daucus carota/metabolismo , Resistência à Doença/imunologia , Metaboloma , Daucus carota/crescimento & desenvolvimento , Daucus carota/imunologia
6.
J Environ Sci Health B ; 55(11): 990-1001, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32877275

RESUMO

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


Assuntos
Carbanilidas/farmacologia , Carvão Vegetal , Ciprofloxacina/farmacologia , Daucus carota/efeitos dos fármacos , Alface/efeitos dos fármacos , Triclosan/farmacocinética , Agricultura/métodos , Anti-Infecciosos/farmacocinética , Biomassa , Biossólidos , Daucus carota/crescimento & desenvolvimento , Daucus carota/metabolismo , Alface/crescimento & desenvolvimento , Alface/metabolismo , Folhas de Planta/química , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Raízes de Plantas/química , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/metabolismo , Brotos de Planta/efeitos dos fármacos , Solo/química , Poluentes do Solo/análise , Poluentes do Solo/farmacocinética
7.
Sci Rep ; 10(1): 13685, 2020 08 13.
Artigo em Inglês | MEDLINE | ID: mdl-32792547

RESUMO

Fungal endophytes can influence production and post-harvest challenges in carrot, though the identity of these microbes as well as factors affecting their composition have not yet been determined, which prevents growers from managing these organisms to improve crop performance. Consequently, we characterized the endophytic mycobiome in the taproots of three carrot genotypes that vary in resistance to two pathogens grown in a trial comparing organic and conventional crop management using Illumina sequencing of the internal transcribed spacer (ITS) gene. A total of 1,480 individual operational taxonomic units (OTUs) were identified. Most were consistent across samples, indicating that they are part of a core mycobiome, though crop management influenced richness and diversity, likely in response to differences in soil properties. There were also differences in individual OTUs among genotypes and the nematode resistant genotype was most responsive to management system indicating that it has greater control over its endophytic mycobiome, which could potentially play a role in resistance. Members of the Ascomycota were most dominant, though the exact function of most taxa remains unclear. Future studies aimed at overcoming difficulties associated with isolating fungal endophytes are needed to identify these microbes at the species level and elucidate their specific functional roles.


Assuntos
Daucus carota/crescimento & desenvolvimento , Fungos/classificação , Análise de Sequência de DNA/métodos , Agricultura , Daucus carota/genética , Daucus carota/microbiologia , Endófitos , Fungos/genética , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala , Micobioma , Filogenia , Raízes de Plantas/microbiologia
8.
Mol Genet Genomics ; 295(6): 1379-1392, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32656704

RESUMO

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.


Assuntos
Vias Biossintéticas/genética , Carotenoides/metabolismo , Biologia Computacional/métodos , Daucus carota/genética , Daucus carota/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Daucus carota/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Pigmentação , Proteínas de Plantas/genética
9.
PLoS One ; 15(6): e0233783, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32497087

RESUMO

Managing pests in carrot production is challenging. Endophytic microbes have been demonstrated to improve the health and productivity of many crops, but factors affecting endophyte dynamics in carrot is still not well understood. The goal of this study was to determine how crop management system and carrot genotype interact to affect the composition and potential of endophytes to mitigate disease caused by Alternaria dauci, an important carrot pathogen. Twenty-eight unique isolates were collected from the taproots of nine diverse genotypes of carrot grown in a long-term trial comparing organic and conventional management. Antagonistic activity was quantified using an in vitro assay, and potential for individual isolates to mitigate disease was evaluated in greenhouse trials using two carrot cultivars. Results confirm that carrot taproots are colonized by an abundant and diverse assortment of bacteria and fungi representing at least distinct 13 genera. Soils in the organic system had greater total organic matter, microbial biomass and activity than the conventional system and endophyte composition in taproots grown in this system were more abundant and diverse, and had greater antagonistic activity. Carrot genotype also affected endophyte abundance as well as potential for individual isolates to affect seed germination, seedling growth and tolerance to A. dauci. The benefits of endophytes on carrot growth were greatest when plants were subject to A. dauci stress, highlighting the importance of environmental conditions in the functional role of endophytes. Results of this study provide evidence that endophytes can play an important role in improving carrot performance and mediating resistance to A. dauci, and it may someday be possible to select for these beneficial plant-microbial relationships in carrot breeding programs. Implementing soil-building practices commonly used in organic farming systems has potential to promote these beneficial relationships and improve the health and productivity of carrot crops.


Assuntos
Alternaria/fisiologia , Produção Agrícola/métodos , Daucus carota/genética , Daucus carota/microbiologia , Endófitos/fisiologia , Genótipo , Doenças das Plantas/microbiologia , Proteção de Cultivos/métodos , Daucus carota/crescimento & desenvolvimento , Endófitos/isolamento & purificação , Germinação , Solo/química , Microbiologia do Solo
10.
Int J Mol Sci ; 21(12)2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32549408

RESUMO

BACKGROUND: Diverse groups of carrot cultivars have been developed to meet consumer demands and industry needs. Varietal groups of the cultivated carrot are defined based on the shape of roots. However, little is known about the genetic basis of root shape determination. METHODS: Here, we used 307 carrot plants from 103 open-pollinated cultivars for a genome wide association study to identify genomic regions associated with the storage root morphology. RESULTS: A 180 kb-long region on carrot chromosome 1 explained 10% of the total observed phenotypic variance in the shoulder diameter. Within that region, DcDCAF1 and DcBTAF1 genes were proposed as candidates controlling secondary growth of the carrot storage root. Their expression profiles differed between the cultivated and the wild carrots, likely indicating that their elevated expression was required for the development of edible roots. They also showed higher expression at the secondary root growth stage in cultivars producing thick roots, as compared to those developing thin roots. CONCLUSIONS: We provided evidence for a likely involvement of DcDCAF1 and/or DcBTAF1 in the development of the carrot storage root and developed a genotyping assay facilitating the identification of variants in the region on carrot chromosome 1 associated with secondary growth of the carrot root.


Assuntos
Biologia Computacional/métodos , Daucus carota/crescimento & desenvolvimento , Proteínas de Plantas/genética , Mapeamento Cromossômico , Mineração de Dados , Bases de Dados Genéticas , Daucus carota/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Polimorfismo de Nucleotídeo Único
11.
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
12.
Nutrients ; 12(2)2020 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-32012793

RESUMO

Carrots' genotype and growing conditions influence their potential properties to fight against cardiovascular and metabolic diseases. The present study evaluated the influence of carrot genotypes contrasted by root color (Bolero, Presto, Karotan, Deep Purple, Kintoki and Blanche des Vosges) growing under standard, water-restricted, biotic stress (Alternaria dauci inoculation), and combined stress conditions (water restriction and A.dauci inoculation). The effect of carrots' polyphenol and carotenoid content was assessed on endothelial and smooth muscle cells, hepatocytes, adipocytes and macrophages functions (oxidative stress, apoptosis, proliferation, lipid accumulation and inflammation). Independently of varieties or growing conditions, all carrot extracts affected vascular cells' oxidative stress and apoptosis, and metabolic cells' oxidative stress and lipid accumulation. Three clusters were revealed and displayed beneficial properties mostly for adipocytes function, smooth muscle cells and hepatocytes, and endothelial cells and hepatocytes, respectively. Karotan and Presto varieties exhibited endothelial tropism while Blanche des Vosges targeted adipocytes. Carrots under biotic stress are more efficient in inducing beneficial effects, with the Bolero variety being the most effective. However, extracts from carrots which grew under combined stress conditions had limited beneficial effects. This report underscores the use of certain carrot extracts as potential effective nutraceutical supplements for metabolic diseases.


Assuntos
Proliferação de Células/efeitos dos fármacos , Daucus carota/genética , Genótipo , Pigmentação , Extratos Vegetais/farmacologia , Raízes de Plantas/química , Animais , Células Cultivadas , Daucus carota/crescimento & desenvolvimento , Daucus carota/metabolismo , Humanos , Camundongos , Extratos Vegetais/química
13.
Plant Sci ; 291: 110327, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31928663

RESUMO

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.


Assuntos
Carotenoides/metabolismo , Daucus carota/genética , Oxirredutases/genética , Proteínas de Plantas/genética , Daucus carota/crescimento & desenvolvimento , Daucus carota/metabolismo , Oxirredutases/metabolismo , Proteínas de Plantas/metabolismo
15.
Sci Rep ; 10(1): 324, 2020 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-31942014

RESUMO

Carotenogenesis has been intensively studied in carrot roots, and transcriptional regulation is thought to be the major factor in carotenoid accumulation in these organs. However, little is known about the transcriptional regulation of carotenoid biosynthetic genes concerning carotenoid accumulation during infestation by the obligate parasite Phelipanche aegyptiaca. HPLC analysis revealed a decrease in carotenoid levels of the different carrot cultivars when parasitized by P. aegyptiaca. Besides, we isolated and analyzed P. aegyptiaca tubercles parasitizing the various carrot root cultivars and show that they accumulate different carotenoids compared to those in non-infested carrot roots. Expression analysis of PHYTOENE SYNTHASE (PSY1) and CAROTENOID ISOMERASE (CRTISO) as well as the strigolactone apocarotenoid biosynthetic genes DWARF27 (D27), CAROTENOID CLEAVAGE DIOXYGENASE 7 (CCD7) and CCD8 revealed that their transcript levels showed significant variation in P. aegyptiaca infested carrot roots. After parasite infestation, the expression of these genes was strongly reduced, as were the carotenoid levels and this was more pronounced in the uncommon non-orange varieties. We also analyzed the parasite genes encoding D27, CCD7 and CCD8 and show that they are expressed in tubercles. This raises important questions of whether the parasite produces its carotenoids and apocarotenoids including strigolactones and whether the latter might have a role in tubercle development.


Assuntos
Carotenoides/metabolismo , Daucus carota/metabolismo , Orobanchaceae/fisiologia , Carotenoides/análise , Cromatografia Líquida de Alta Pressão , Daucus carota/crescimento & desenvolvimento , Dioxigenases/genética , Dioxigenases/metabolismo , Regulação da Expressão Gênica de Plantas , Geranil-Geranildifosfato Geranil-Geraniltransferase/genética , Geranil-Geranildifosfato Geranil-Geraniltransferase/metabolismo , Orobanchaceae/crescimento & desenvolvimento , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/metabolismo , Plantas Daninhas/crescimento & desenvolvimento , Plantas Daninhas/fisiologia , cis-trans-Isomerases/genética , cis-trans-Isomerases/metabolismo
16.
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
17.
Food Chem ; 300: 125202, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31330374

RESUMO

The aim of a three-year study was to assess the effect of combined biofortification with I and Se in carrot. Four cultivars ('Askona' F1, 'Samba' F1, 'Kazan' F1 and 'White Satin') were grown in soil fertilized with KI (4 kg I ha-1) and Na2SeO4 (0.25 kg Se ha-1). The I + Se fertilization did not affect yield but the plants of all cultivars accumulated both elements in leaves and roots. On average, the I and Se contents in roots increased 7.7-times for I and 4.9-times for Se as well as the average I:Se molar ratio was 0.28:1. The contents of both elements in roots remained well below the hazard threshold thus the intake of 100 g of biofortified carrot would substantially cover the RDA for I and Se. The changes in chemical composition of roots (nitrates, phenolic compounds, sugars, carotenoids, macro-, microelements and cadmium) were rather year-dependent than affected by the applied I + Se fertilization.


Assuntos
Biofortificação/métodos , Daucus carota/química , Iodo/farmacologia , Raízes de Plantas/química , Selênio/farmacologia , Cádmio/análise , Carotenoides/análise , Daucus carota/efeitos dos fármacos , Daucus carota/crescimento & desenvolvimento , Fertilizantes , Alimentos Fortificados/análise , Iodo/análise , Iodo/farmacocinética , Polônia , Selênio/análise , Selênio/farmacocinética , Solo/química
18.
Theor Appl Genet ; 132(9): 2485-2507, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31144001

RESUMO

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.


Assuntos
Antocianinas/genética , Daucus carota/genética , Pigmentação/genética , Folhas de Planta/genética , Proteínas de Plantas/genética , Raízes de Plantas/genética , Locos de Características Quantitativas , Antocianinas/metabolismo , Cromossomos de Plantas , Cor , Daucus carota/crescimento & desenvolvimento , Daucus carota/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Filogenia , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Polimorfismo de Nucleotídeo Único , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Transcriptoma
19.
Environ Sci Pollut Res Int ; 26(6): 5463-5474, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30610583

RESUMO

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


Assuntos
Cádmio/toxicidade , Compostagem/métodos , Chumbo/toxicidade , Resíduos Sólidos , Verduras/crescimento & desenvolvimento , Aerobiose , Agricultura/métodos , Anaerobiose , Cádmio/análise , Daucus carota/efeitos dos fármacos , Daucus carota/crescimento & desenvolvimento , Exposição Dietética/prevenção & controle , Fertilizantes , Humanos , Chumbo/análise , Metais Pesados/análise , Metais Pesados/toxicidade , Nitrogênio/metabolismo , Medição de Risco , Solo , Spinacia oleracea/efeitos dos fármacos , Spinacia oleracea/crescimento & desenvolvimento , Verduras/efeitos dos fármacos
20.
Mol Biotechnol ; 61(3): 191-199, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30644027

RESUMO

The clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) system has been successfully used for precise genome editing in many plant species, including in carrot cells, very recently. However, no stable gene-editing carrot plants were obtained with CRISPR/Cas9 system to date. In the present study, four sgRNA expression cassettes, individually driven by four different promoters and assembled in a single CRISPR/Cas9 vector, were transformed into carrots using Agrobacterium-mediated genetic transformation. Four sites of DcPDS and DcMYB113-like genes were chosen as targets. Knockout of DcPDS in orange carrot 'Kurodagosun' resulted in the generation of albino carrot plantlets, with about 35.3% editing efficiency. DcMYB113-like was also successfully edited in purple carrot 'Deep purple', resulting in purple depigmented carrot plants, with about 36.4% rate of mutation. Sequencing analyses showed that insertion, deletion, and substitution occurred in the target sites, generating heterozygous, biallelic, and chimeric mutations. The highest efficiency of mutagenesis was observed in the sites targeted by AtU6-29-driven sgRNAs in both DcPDS- and DcMYB113-like-knockout T0 plants, which always induced double-strand breaks in the target sites. Our results proved that CRISPR/Cas9 system could be for generating stable gene-editing carrot plants.


Assuntos
Citrus sinensis/crescimento & desenvolvimento , Daucus carota/crescimento & desenvolvimento , Mutagênese Sítio-Dirigida/métodos , Proteínas de Plantas/genética , Agrobacterium , Sistemas CRISPR-Cas , Citrus sinensis/genética , Daucus carota/genética , Edição de Genes/métodos , Taxa de Mutação , RNA Guia/genética , Análise de Sequência de DNA , Transformação Bacteriana
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