RESUMO
Numerous putative glycosyltransferases (GTs) have been identified using bioinformatic approaches. However, demonstrating the activity of these GTs remains a challenge. Here, we describe the development of a rapid in vitro GT-array screening platform for activity of GTs. GT-arrays are generated by cell-free in vitro protein synthesis and binding using microplates precoated with a N-terminal Halo- or a C-terminal GST-tagged GT-encoding plasmid DNA and a capture antibody. These arrays are then used for screening of transferase activities and the reactions are monitored by a luminescence GLO assay. The products formed by these reactions can be analyzed directly from the microplates by mass spectrometry. Using this platform, a total of 280 assays were performed to screen 22 putative fucosyltransferases (FUTs) from family GT37 (seven from Arabidopsis and 15 from rice) for activity toward five acceptors: non-fucosylated tamarind xyloglucan (TXyG), arabinotriose (Ara3), non-fucosylated rhamnogalacturonan I (RG-I), and RG-II from the mur1-1 Arabidopsis mutant, and the celery RG-II monomer lacking Arap and MeFuc of chain B and l-Gal of chain A. Our screen showed that AtFUT2, AtFUT5, and AtFUT10 have activity toward RG-I, while AtFUT8 was active on RG-II. Five rice OsFUTs have XyG-FUT activity and four rice OsFUTs have activity toward Ara3. None of the putative OsFUTs were active on the RG-I and RG-II. However, promiscuity toward acceptors was observed for several FUTs. These findings extend our knowledge of cell wall polysaccharide fucosylation in plants. We believe that in vitro GT-array platform provides a valuable tool for cell wall biochemistry and other research fields.
Assuntos
Ensaios Enzimáticos , Fucosiltransferases , Glicosiltransferases , Proteínas de Plantas , Apium/enzimologia , Apium/genética , Arabidopsis/enzimologia , Arabidopsis/genética , Arabidopsis/metabolismo , Parede Celular/química , Parede Celular/enzimologia , Parede Celular/metabolismo , Ensaios Enzimáticos/instrumentação , Ensaios Enzimáticos/métodos , Fucosiltransferases/análise , Fucosiltransferases/classificação , Fucosiltransferases/metabolismo , Glicosiltransferases/análise , Glicosiltransferases/metabolismo , Espectrometria de Massas , Oryza/enzimologia , Proteínas de Plantas/análise , Proteínas de Plantas/metabolismo , Polissacarídeos/química , Polissacarídeos/metabolismoRESUMO
The genus Apium, belonging to the family Apiaceae, comprises roughly 20 species. Only two species, Apium graveolens and Apium leptophyllum, are available in China and are both rich in nutrients and have favorable medicinal properties. However, the lack of genomic data has severely constrained the study of genetics and evolution in Apium plants. In this study, Illumina NovaSeq 6000 and Nanopore sequencing platforms were employed to identify the mitochondrial genomes of A. graveolens and A. leptophyllum. The complete lengths of the mitochondrial genomes of A. graveolens and A. leptophyllum were 263,017 bp and 260,164 bp, respectively, and contained 39 and 36 protein-coding genes, five and six rRNA genes, and 19 and 20 tRNA genes. Consistent with most angiosperms, both A. graveolens and A. leptophyllum showed a preference for codons encoding leucine (Leu). In the mitochondrial genome of A. graveolens, 335 SSRs were detected, which is higher than the 196 SSRs found in the mitochondrial genome of A. leptophyllum. Studies have shown that the most common RNA editing type is C-to-U, but, in our study, both A. graveolens and A. leptophyllum exhibited the U-C editing type. Furthermore, the transfer of the mitochondrial genomes of A. graveolens and A. leptophyllum into the chloroplast genomes revealed homologous sequences, accounting for 8.14% and 4.89% of the mitochondrial genome, respectively. Lastly, in comparing the mitochondrial genomes of 29 species, it was found that A. graveolens, A. leptophyllum, and Daucus carota form a sister group with a support rate of 100%. Overall, this investigation furnishes extensive insights into the mitochondrial genomes of A. graveolens and A. leptophyllum, thereby enhancing comprehension of the traits and evolutionary patterns within the Apium genus. Additionally, it offers supplementary data for evolutionary and comparative genomic analyses of other species within the Apiaceae family.
Assuntos
Apiaceae , Apium , Daucus carota , Genoma de Cloroplastos , Genoma Mitocondrial , Magnoliopsida , Filogenia , Apium/genética , Genoma Mitocondrial/genética , Apiaceae/genética , Daucus carota/genética , Magnoliopsida/genéticaRESUMO
Celery (Apium graveolens L.) is an important vegetable crop cultivated worldwide for its medicinal properties and distinctive flavor. Volatile organic compound (VOC) analysis is a valuable tool for the identification and classification of species. Currently, less research has been conducted on aroma compounds in different celery varieties and colors. In this study, five different colored celery were quantitatively analyzed for VOCs using HS-SPME, GC-MS determination, and stoichiometry methods. The result revealed that γ-terpinene, d-limonene, 2-hexenal,-(E)-, and ß-myrcene contributed primarily to the celery aroma. The composition of compounds in celery exhibited a correlation not only with the color of the variety, with green celery displaying a higher concentration compared with other varieties, but also with the specific organ, whereby the content and distribution of volatile compounds were primarily influenced by the leaf rather than the petiole. Seven key genes influencing terpenoid synthesis were screened to detect expression levels. Most of the genes exhibited higher expression in leaves than petioles. In addition, some genes, particularly AgDXS and AgIDI, have higher expression levels in celery than other genes, thereby influencing the regulation of terpenoid synthesis through the MEP and MVA pathways, such as hydrocarbon monoterpenes. This study identified the characteristics of flavor compounds and key aroma components in different colored celery varieties and explored key genes involved in the regulation of terpenoid synthesis, laying a theoretical foundation for understanding flavor chemistry and improving its quality.
Assuntos
Apium , Compostos Orgânicos Voláteis , Apium/genética , Cor , Cromatografia Gasosa-Espectrometria de Massas , Microextração em Fase Sólida , VerdurasRESUMO
Apiose is a unique branched-chain pentose found in plant glycosides and a key component of the cell wall polysaccharide pectin and other specialized metabolites. More than 1,200 plant-specialized metabolites contain apiose residues, represented by apiin, a distinctive flavone glycoside found in celery (Apium graveolens) and parsley (Petroselinum crispum) in the family Apiaceae. The physiological functions of apiin remain obscure, partly due to our lack of knowledge on apiosyltransferase during apiin biosynthesis. Here, we identified UGT94AX1 as an A. graveolens apiosyltransferase (AgApiT) responsible for catalyzing the last sugar modification step in apiin biosynthesis. AgApiT showed strict substrate specificity for the sugar donor, UDP-apiose, and moderate specificity for acceptor substrates, thereby producing various apiose-containing flavone glycosides in celery. Homology modeling of AgApiT with UDP-apiose, followed by site-directed mutagenesis experiments, identified unique Ile139, Phe140, and Leu356 residues in AgApiT, which are seemingly crucial for the recognition of UDP-apiose in the sugar donor pocket. Sequence comparison and molecular phylogenetic analysis of celery glycosyltransferases suggested that AgApiT is the sole apiosyltransferase-encoding gene in the celery genome. Identification of this plant apiosyltransferase gene will enhance our understanding of the physioecological functions of apiose and apiose-containing compounds.
Assuntos
Apium , Flavonas , Apium/genética , Glicosídeos , FilogeniaRESUMO
Single-cell RNA sequencing (scRNA-seq) has revolutionized our understanding of cellular heterogeneity in health and disease. However, the lack of physical relationships among dissociated cells has limited its applications. To address this issue, we present CeLEry (Cell Location recovEry), a supervised deep learning algorithm that leverages gene expression and spatial location relationships learned from spatial transcriptomics to recover the spatial origins of cells in scRNA-seq. CeLEry has an optional data augmentation procedure via a variational autoencoder, which improves the method's robustness and allows it to overcome noise in scRNA-seq data. We show that CeLEry can infer the spatial origins of cells in scRNA-seq at multiple levels, including 2D location and spatial domain of a cell, while also providing uncertainty estimates for the recovered locations. Our comprehensive benchmarking evaluations on multiple datasets generated from brain and cancer tissues using Visium, MERSCOPE, MERFISH, and Xenium demonstrate that CeLEry can reliably recover the spatial location information for cells using scRNA-seq data.
Assuntos
Apium , Transcriptoma , Transcriptoma/genética , Apium/genética , Análise da Expressão Gênica de Célula Única , Análise de Sequência de RNA/métodos , Análise de Célula Única/métodos , Perfilação da Expressão Gênica/métodosRESUMO
BACKGROUND: Water shortage caused by global warming seriously affects the yield and quality of vegetable crops. ß-carotene, the lipid-soluble natural product with important pharmacological value, is abundant in celery. Transcription factor MYB family extensively disperses in plants and plays regulatory roles in carotenoid metabolism and water scarcity response. RESULTS: Here, the AgMYB5 gene encoding 196 amino acids was amplified from celery cv. 'Jinnanshiqin'. In celery, the expression of AgMYB5 exhibited transactivation activity, tissue specificity, and drought-condition responsiveness. Further analysis proved that ectopic expression of AgMYB5 increased ß-carotene content and promoted drought tolerance in transgenic Arabidopsis thaliana. Moreover, AgMYB5 expression promoted ß-carotene biosynthesis by triggering the expression of AtCRTISO and AtLCYB, which in turn increased antioxidant enzyme activities, and led to the decreased contents of H2O2 and MDA, and the inhibition of O2- generation. Meanwhile, ß-carotene accumulation promoted endogenous ABA biosynthesis of transgenic Arabidopsis, which resulted in ABA-induced stomatal closing and delayed water loss. In addition, ectopic expression of AgMYB5 increased expression levels of AtERD1, AtP5CS1, AtRD22, and AtRD29. CONCLUSIONS: The findings indicated that AgMYB5 up-regulated ß-carotene biosynthesis and drought tolerance of Arabidopsis.
Assuntos
Apium , Arabidopsis , Arabidopsis/metabolismo , beta Caroteno , Apium/genética , Apium/metabolismo , Resistência à Seca , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Verduras/genética , Verduras/metabolismo , Peróxido de Hidrogênio/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Estresse Fisiológico/genética , Antioxidantes/metabolismo , Secas , Água/metabolismo , Regulação da Expressão Gênica de Plantas , Ácido Abscísico/metabolismoRESUMO
Male sterility is a common phenomenon in the plant kingdom and based on the organelles harboring the male-sterility genes, it can be classified into the genic male sterility (GMS) and the cytoplasmic male sterility (CMS). In every generation, CMS can generate 100% male-sterile population, which is very important for the breeders to take advantage of the heterosis and for the seed producers to guarantee the seed purity. Celery is a cross-pollinated plant with the compound umbel type of inflorescence which carries hundreds of small flowers. These characteristics make CMS the only option to produce the commercial hybrid celery seeds. In this study, transcriptomic and proteomic analyses were performed to identify genes and proteins that are associated with celery CMS. A total of 1255 differentially expressed genes (DEGs) and 89 differentially expressed proteins (DEPs) were identified between the CMS and its maintainer line, then 25 genes were found to differentially expressed at both the transcript and protein levels. Ten DEGs involved in the fleece layer and outer pollen wall development were identified by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, most of which were down-regulated in the sterile line W99A. These DEGs and DEPs were mainly enriched in the pathways of "phenylpropanoid/sporopollenin synthesis/metabolism", "energy metabolism", "redox enzyme activity" and "redox processes". Results obtained in this study laid a foundation for the future investigation of mechanisms of pollen development as well as the reasons for the CMS in celery.
Assuntos
Apium , Infertilidade Masculina , Masculino , Humanos , Feminino , Transcriptoma , Apium/genética , Proteômica , Perfilação da Expressão Gênica , Verduras/genética , Flores/genética , Infertilidade das Plantas/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genéticaRESUMO
Celery (Apium graveolens L.), a medicinal crop, occupies a significant position in the human diet possessing several essential macro- and microelements. For proper proximate analysis, an experiment was executed by taking 20 celery genotypes. The genotypes were analyzed for macro- and microminerals which include nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), sodium (Na), sulfur (S), zinc (Zn), iron (Fe), copper (Cu), and manganese (Mn). Results from analysis revealed that the amount of N, P, Ca, Na, and S was higher in microgreens, whereas a higher value for K was found in mature leaves. Zn, Cu, and Mn contents were found to be higher in mature leaves, while no significant difference was observed for Fe content in microgreens and mature leaves. The inclusion of celery microgreens in our daily diet would fulfill a significant portion of our daily mineral requirement. This is the first report on mineral comparison between microgreens and mature leaves of celery. It opens a new avenue for further enhancement of minerals via biofortification of this medicinal wonder crop through systematic breeding efforts. On the basis of mineral analysis, three genotypes, namely PAU2, PAU4, and PAU16, were found superior in terms of mineral composition in microgreens and mature leaves of celery. Principal component and cluster analyses divide the genotypes into two different clusters on the basis of variability in mineral composition.
Assuntos
Apium , Humanos , Apium/genética , Melhoramento Vegetal , Minerais , Manganês , Zinco , Cálcio da Dieta , Sódio , Verduras , Folhas de PlantaRESUMO
Celery (Apium graveolens L.) is one of the most popular leafy vegetables worldwide. The main edible parts of celery are the leaf blade and especially the petiole, which typically has a white, green and red color. To date, there are very few reports about the inheritance and gene cloning of celery petiole color. In this study, bulked segregant analysis-sequencing (BSA-Seq) and fine mapping were conducted to delimit the white petiole (wp1) loci into a 668.5-kb region on Chr04. In this region, AgWp1 is a homolog of a DAG protein in Antirrhinum majus and a MORF9 protein in Arabidopsis, and both proteins are involved in chloroplast development. Sequencing alignment shows that there is a 27-bp insertion in the 3'-utr region in AgWp1 in the white petiole. Gene expression analysis indicated that the expression level of AgWp1 in the green petiole was much higher than that in the white petiole. Further cosegregation revealed that the 27-bp insertion was completely cosegregated with the petiole color in 45 observed celery varieties. Therefore, AgWp1 was considered to be the candidate gene controlling the white petiole in celery. Our results could not only improve the efficiency and accuracy of celery breeding but also help in understanding the mechanism of chlorophyll synthesis and chloroplast development in celery.
Assuntos
Apium , Apium/genética , Apium/metabolismo , Verduras/genética , Melhoramento Vegetal , Perfilação da Expressão Gênica , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Celery is an important nutritionally rich crop in the family Apiaceae. It is cultivated worldwide for food as well as for use in pharmaceutics. It is an excellent source of minerals, vitamins, and phytochemicals. Identification of superior genotypes with improved nutritional content is the requirement to develop cultivars for commercial cultivation. For mineral analysis of celery, an experiment was carried out taking 20 diverse genotypes. These genotypes were analysed for macro- and micronutrients which include nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), iron (Fe), zinc (Zn), manganese (Mn), copper (Cu), and sodium (Na). The study revealed high content of K (20.3-26.1 mg/g dry weight (DW)) and Zn (0.09-0.14 mg/g DW) in leaves while the stalks were rich in Ca (41.5-51.3 mg/g DW) and Na (5.2-8.0 mg/g DW). High contents of P (5.2-6.8 mg/g DW), Fe (0.41-0.56 mg/g DW), Cu (0.015-0.026 mg/g DW), and Mn (0.020-0.029 mg/g DW) were observed in seeds. Based on the mineral content, three genotypes, viz., PAU2, PAU4, and PAU7, were found to be superior in terms of mineral composition in leaves, stalks, and seeds. Cluster analysis divided the genotypes into two major groups. These genotypes can be used in crosses as they showed great potential for use in biofortification. This study opens newer avenues for future research, encouraging researchers to enhance the product quality and production efficiency of the leaves, stalks, and seeds valuable for human consumption.
Assuntos
Apium , Humanos , Apium/genética , Apium/química , Minerais/análise , Sódio/análise , Verduras/química , Manganês/análise , Folhas de Planta/genética , Folhas de Planta/química , Sementes/genética , Sementes/química , Cálcio da Dieta/análise , GenótipoRESUMO
MAIN CONCLUSION: Transcriptome and biochemical analyses are applied to individual plant cell types to reveal potential players involved in the molecular machinery of cell wall formation in specialized cells such as collenchyma. Plant collenchyma is a mechanical tissue characterized by an irregular, thickened cell wall and the ability to support cell elongation. The composition of the collenchyma cell wall resembles that of the primary cell wall and includes cellulose, xyloglucan, and pectin; lignin is absent. Thus, the processes associated with the formation of the primary cell wall in the collenchyma can be more pronounced compared to other tissues due to its thickening. Primary cell walls intrinsic to different tissues may differ in structure and composition, which should be reflected at the transcriptomic level. For the first time, we conducted transcriptome profiling of collenchyma strands isolated from young celery petioles and compared them with other tissues, such as parenchyma and vascular bundles. Genes encoding proteins involved in the primary cell wall formation during cell elongation, such as xyloglucan endotransglucosylase/hydrolases, expansins, and leucine-rich repeat proteins, were significantly activated in the collenchyma. As the key players in the transcriptome orchestra of collenchyma, xyloglucan endotransglucosylase/hydrolase transcripts were characterized in more detail, including phylogeny and expression patterns. The comprehensive approach that included transcriptome and biochemical analyses allowed us to reveal peculiarities of collenchyma cell wall formation and modification, matching the abundance of upregulated transcripts and their potential substrates for revealed gene products. As a result, specific isoforms of multigene families were determined for further functional investigation.
Assuntos
Apium , Apium/genética , Celulose/metabolismo , Perfilação da Expressão Gênica , Plantas/genética , Glicosiltransferases/genética , Verduras/genética , Verduras/metabolismo , Parede Celular/metabolismoRESUMO
Celery seed is known to be difficult to germinate due to its morphological dormancy. Light is the key signal to release morphological dormancy and promote seed germination. However, this mechanism has rarely been studied. We performed physiological, transcriptome analyses on celery seed exposed to light and dark to decipher the mechanism by which light promotes germination of celery seed. The results showed that light significantly enhanced the expression of gibberellin synthesis genes and abscisic acid degradation genes and inhibited the expression of abscisic acid synthesis genes and gibberellin degradation genes. Moreover, gibberellin synthesis inhibitor could completely inhibit the germination capacity of celery seed, indicating that gibberellin is indispensable in the process of celery seed germination. Compared with dark, light also increased the activity of α-amylase and ß-amylase and the expression of related coding genes and promoted the degradation of starch and the increase of soluble sugar content, suggesting that light enhanced the sugar metabolism of celery seed. In addition, transcriptome analysis revealed that many genes related to endosperm weakening (cell wall remodeling enzymes, extension proteins) were up-regulated under light. It was also found that light promoted the accumulation of hydrogen peroxide in the radicle, which promoted the endosperm weakening process of celery seed. Our results thus indicated that light signal may promote the release of morphological dormancy through the simultaneous action of multiple factors.
Assuntos
Apium , Reguladores de Crescimento de Plantas , Ácido Abscísico/metabolismo , Ácido Abscísico/farmacologia , Apium/genética , Apium/metabolismo , Endosperma/genética , Endosperma/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Germinação , Giberelinas/metabolismo , Giberelinas/farmacologia , Dormência de Plantas/genética , Reguladores de Crescimento de Plantas/metabolismo , Sementes/metabolismo , Açúcares/metabolismoRESUMO
Celery (Apium graveolens L.), a plant from Apiaceae, is one of the most important vegetables and is grown worldwide. Carotenoids can capture light energy and transfer it to chlorophyll, which plays a central role in photosynthesis. Here, by performing transcriptomics and genomics analysis, we identified and conducted a comprehensive analysis of chlorophyll and carotenoid-related genes in celery and six representative species. Significantly, different contents and gene expression patterns were found among three celery varieties. In total, 237 and 290 chlorophyll and carotenoid-related genes were identified in seven species. No notable gene expansion of chlorophyll biosynthesis was detected in examined species. However, the gene encoding ζ-carotene desaturase (ZDS) enzyme in carotenoid was expanded in celery. Comparative genomics and RNA-seq analyses revealed 16 and 5 key genes, respectively, regulating chlorophyll and carotenoid. An intriguing finding is that chlorophyll and carotenoid-related genes were coordinately regulated by transcriptional factors, which could be distinctively classified into positive- and negative-regulation groups. Six CONSTANS (CO)-like transcription factors co-regulated chlorophyll and carotenoid-related genes were identified in celery. In conclusion, this study provides new insights into the regulation of chlorophyll and carotenoid by transcription factors.
Assuntos
Apium , Apium/genética , Apium/metabolismo , Carotenoides/metabolismo , Clorofila/metabolismo , Regulação da Expressão Gênica de Plantas , Genômica , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Transcriptoma , Verduras/metabolismoRESUMO
Plant metabolites are important for plant development and human health. Plants of celery (Apiumgraveolens L.) with different-colored petioles have been formed in the course of long-term evolution. However, the composition, content distribution, and mechanisms of accumulation of metabolites in different-colored petioles remain elusive. Using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), 1159 metabolites, including 100 lipids, 72 organic acids and derivatives, 83 phenylpropanoids and polyketides, and several alkaloids and terpenoids, were quantified in four celery cultivars, each with a different petiole color. There were significant differences in the types and contents of metabolites in celery with different-colored petioles, with the most striking difference between green celery and purple celery, followed by white celery and green celery. Annotated analysis of metabolic pathways showed that the metabolites of the different-colored petioles were significantly enriched in biosynthetic pathways such as anthocyanin, flavonoid, and chlorophyll pathways, suggesting that these metabolic pathways may play a key role in determining petiole color in celery. The content of chlorophyll in green celery was significantly higher than that in other celery cultivars, yellow celery was rich in carotenoids, and the content of anthocyanin in purple celery was significantly higher than that in the other celery cultivars. The color of the celery petioles was significantly correlated with the content of related metabolites. Among the four celery cultivars, the metabolites of the anthocyanin biosynthesis pathway were enriched in purple celery. The results of quantitative real-time polymerase chain reaction (qRT-PCR) suggested that the differential expression of the chalcone synthase (CHS) gene in the anthocyanin biosynthesis pathway might affect the biosynthesis of anthocyanin in celery. In addition, HPLC analysis revealed that cyanidin is the main pigment in purple celery. This study explored the differences in the types and contents of metabolites in celery cultivars with different-colored petioles and identified key substances for color formation. The results provide a theoretical basis and technical support for genetic improvement of celery petiole color.
Assuntos
Antocianinas , Apium , Apium/genética , Apium/metabolismo , Clorofila/metabolismo , Cor , Regulação da Expressão Gênica de Plantas , Humanos , Metabolômica , Proteínas de Plantas/genética , Espectrometria de Massas em TandemRESUMO
Ascorbic acid (AsA) is an important nutrient in celery, the conversion of D-mannose-1-P to GDP-D-mannose catalyzed by GDP-D-mannose pyrophosphorylase (GMPase) represents the first committed step in the biosynthesis of AsA. To clarify the function of the AgGMP gene of celery, the AgGMP gene was cloned from celery cv. 'Jinnan Shiqin' . It contains an open reading frame (ORF) with the length of 1,086 bp, encoding 361 amino acids. AgGMP protein was highly conserved among different plant species. Phylogenetic analysis demonstrated that the GMP proteins from celery and carrot belonged to the same branch. AgGMP protein was mainly composed of three α-helixes and certain random coils. No signal peptide was found in the AgGMP protein. The subcellular localization indicated that the AgGMP protein was located in the cytoplasm. The relative expression levels of AgGMP in 'Jinnan Shiqin' were significantly up-regulated at 2 h and 4 h under drought stress treatments. AsA contents in transgenic Arabidopsis lines hosting AgGMP gene were higher than that in wild type plants, and the root lengths were also longer in the MS medium containing 300 mM mannitol. The present study provides useful evidence for the functional involvement of AgGMP in regulating AsA accumulation and response to drought stress in celery.
Assuntos
Apium , Arabidopsis , Ácido Ascórbico , Arabidopsis/genética , Apium/genética , Manose/metabolismo , Proteínas de Plantas/química , Secas , Filogenia , Verduras/metabolismoRESUMO
KEY MESSAGE: Overexpression of AgMYB12 in celery improved the accumulation of apigenin by interacting with the AgFNS gene. Celery is a common vegetable, and its essential characteristic is medicine food homology. A natural flavonoid and a major pharmacological component in celery, apigenin plays an important role in human health. In this study, we isolated a novel R2R3-MYB transcription factor that regulates apigenin accumulation from the celery cultivar 'Jinnan Shiqin' through yeast one-hybrid screening and designated it as AgMYB12. The AgMYB12 protein was located in the nucleus. It showed transcriptional activation activity and bound specifically to the promoter of AgFNS, a gene involved in apigenin biosynthesis. Phylogenetic tree analysis demonstrated that AgMYB12 belongs to the flavonoid branch. It contains two flavonoid-related motifs, SG7 and SG7-2, and shared a highly conserved R2R3 domain with flavonoid-related MYBs. The homologous overexpression of AgMYB12 induced the up-regulation of AgFNS gene expression and accumulation of apigenin and luteolin in celery. Additionally, the expression levels of apigenin biosynthesis-related genes, including AgPAL, AgCHI, AgCHS, Ag4CL, and AgC4H, increased in transgenic celery plants. These results indicated that AgMYB12 acted as a positive regulator of apigenin biosynthesis and activated the expression of AgFNS gene. The current study provides new information about the regulation mechanism of apigenin metabolism in celery and offers a strategy for cultivating the plants with high apigenin content.
Assuntos
Apigenina/biossíntese , Apium/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Sequência de Aminoácidos , Apium/metabolismo , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Alinhamento de Sequência , Fatores de Transcrição/metabolismoRESUMO
Celery is a stalky green vegetable that is grown and consumed globally and used in many cuisines for its distinctive taste and flavour. Previous investigations identified the aroma composition of celery and profiled its sensory characteristics using a trained panel; however, evaluation of the sensory characteristics of celery combined with a consumer panel, where consumer preferences and acceptability are determined, is novel. In this study, three parental genotypes (12, 22 and 25) and three new hybrids (12x22, 22x12 and 25x12) were presented to a trained sensory panel (n = 12) for profiling and a consumer panel (n = 118), where liking and preference were assessed. Celery samples were analysed by SPME GC-MS and significant differences in aroma composition between all samples were identified, causing significant differences in the sensory profile. Furthermore, significant differences in attributes assessed for liking (appearance, aroma, texture and overall) were identified. Consumer segmentation identified three groups of consumers exhibiting differences in the hedonic reaction to the samples. Sweet and bitter taste along with overall flavour were identified as drivers of liking. Hybrid 25x12 was found to be the hybrid that exhibited high intensities for most of the attributes assessed.
Assuntos
Apium/genética , Verduras/genética , Comportamento do Consumidor , Preferências Alimentares , Genes de Plantas , Humanos , Odorantes/análise , PaladarRESUMO
Numerous varieties of celery are grown in multiple countries to maintain supply, demand and availability for all seasons; thus, there is an expectation for a consistent product in terms of taste, flavour, and overall quality. Differences in climate, agronomy and soil composition will all contribute to inconsistencies. This study investigated the volatile and sensory profile of eight celery genotypes grown in the UK (2018) and Spain (2019). Headspace analysis determined the volatile composition of eight genotypes, followed by assessment of the sensory profile using a trained panel. Significant differences in the volatile composition and sensory profile were observed; genotype and geographical location both exerted influences. Two genotypes exhibited similar aroma composition and sensory profile in both locations, making them good candidates to drive breeding programmes aimed at producing varieties that consistently display these distinctive sensory properties. Celery samples harvested in the UK exhibited a higher proportion of sesquiterpenes and phthalides, whereas samples harvested in Spain expressed a higher aldehyde and ketone content. Studying the relationship between growing environment and genotype will provide information to guide growers in how to consistently produce a high-quality crop.
Assuntos
Apium/genética , Apium/metabolismo , Odorantes , Sesquiterpenos/análise , Paladar , Compostos Orgânicos Voláteis/análise , Apium/química , Genótipo , Espanha , Reino UnidoRESUMO
ζ-Carotene desaturase (ZDS) is one of the key enzymes regulating carotenoids biosynthesis and accumulation. Celery transgenic efficiency is low and it is difficult to obtain transgenic plants. The study on ZDS was limited in celery. Here, the AgZDS gene was cloned from celery and overexpressed in Arabidopsis thaliana and celery to verify its function. The AgZDS has typical characteristic of ZDS protein and is highly conserved in higher plants. Phylogenetic analysis showed that AgZDS has the closest evolutionary relationship with ZDSs from Solanum lycopersicum, Capsicum annuum and Tagetes erecta. Overexpression of AgZDS gene in A. thaliana and celery resulted in increased accumulations of lutein and ß-carotene and up-regulated the expression levels of the genes involved in carotenoids biosynthesis. The contents of lutein and ß-carotene in two lines, AtL1 and AgL5, were the highest in transgenic A. thaliana and celery, respectively. The relative expression levels of 5 genes (AtPDS, AtZISO, AtZEP, AtNCED3, and AtCCD4) were up-regulated compared to the wild type plants. The relative expression levels of most genes in carotenoids biosynthesis pathway, such as AgPDS, AgCRTISO1, and AgZISO, were up-regulated in transgenic celery plants. The antioxidant capacity of A. thaliana and photosynthetic capacity of celery were also enhanced. This research is the first report on the function of structure gene related to carotenoid biosynthesis in transgenic celery plants. The findings in this study demonstrated the roles of AgZDS in regulating carotenoids metabolism of celery, which laid a potential foundation for quality improvement of celery.
Assuntos
Apium/genética , Apium/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Luteína/biossíntese , Oxirredutases/metabolismo , beta Caroteno/biossíntese , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Luteína/genética , Oxirredutases/genética , Plantas Geneticamente Modificadas , Verduras/genética , beta Caroteno/genéticaRESUMO
Underdeveloped (small) embryos embedded in abundant endosperm tissue, and thus having morphological dormancy (MD) or morphophysiological dormancy (MPD), are considered to be the ancestral state in seed dormancy evolution. This trait is retained in the Apiaceae family, which provides excellent model systems for investigating the underpinning mechanisms. We investigated Apium graveolens (celery) MD by combined innovative imaging and embryo growth assays with the quantification of hormone metabolism, as well as the analysis of hormone and cell-wall related gene expression. The integrated experimental results demonstrated that embryo growth occurred inside imbibed celery fruits in association with endosperm degradation, and that a critical embryo size was required for radicle emergence. The regulation of these processes depends on gene expression leading to gibberellin and indole-3-acetic acid (IAA) production by the embryo and on crosstalk between the fruit compartments. ABA degradation associated with distinct spatiotemporal patterns in ABA sensitivity control embryo growth, endosperm breakdown and radicle emergence. This complex interaction between gibberellins, IAA and ABA metabolism, and changes in the tissue-specific sensitivities to these hormones is distinct from non-MD seeds. We conclude that the embryo growth to reach the critical size and the associated endosperm breakdown inside MD fruits constitute a unique germination programme.