ABSTRACT
Anthocyanin is one important nutrition composition in Tartary buckwheat (Fagopyrum tataricum) sprouts, a component missing in its seeds. Although anthocyanin biosynthesis requires light, the mechanism of light-induced anthocyanin accumulation in Tartary buckwheat is unclear. Here, comparative transcriptome analysis of Tartary buckwheat sprouts under light and dark treatments and biochemical approaches were performed to identify the roles of one B-box protein BBX22 and ELONGATED HYPOCOTYL 5 (HY5). The overexpression assay showed that FtHY5 and FtBBX22 could both promote anthocyanin synthesis in red-flower tobacco. Additionally, FtBBX22 associated with FtHY5 to form a complex that activates the transcription of MYB transcription factor genes FtMYB42 and FtDFR, leading to anthocyanin accumulation. These findings revealed the regulation mechanism of light-induced anthocyanin synthesis and provide excellent gene resources for breeding high-quality Tartary buckwheat.
Subject(s)
Anthocyanins , Fagopyrum , Gene Expression Regulation, Plant , Light , Plant Proteins , Transcription Factors , Fagopyrum/genetics , Fagopyrum/metabolism , Fagopyrum/growth & development , Fagopyrum/radiation effects , Anthocyanins/biosynthesis , Anthocyanins/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Transcription Factors/metabolism , Transcription Factors/genetics , Gene Expression Profiling , Nicotiana/genetics , Nicotiana/metabolism , Nicotiana/growth & developmentABSTRACT
To investigate the response and the regulatory mechanism of common buckwheat starch, amylose, and amylopectin biosynthesis to P management strategies, field experiments were conducted in 2021 and 2022 using three phosphorus (P) levels. Results revealed that the application of 75 kg hm-2 phosphate fertilizer significantly enhanced amylopectin and total starch content in common buckwheat, leading to improved grain weight and starch yield, and decreased starch granule size. The number of upregulated differentially expressed proteins induced by phosphate fertilizer increased with the application rate, with 56 proteins identified as shared differential proteins between different P levels, primarily associated with carbohydrate and amino acid metabolism. Phosphate fertilizer inhibited amylose synthesis by downregulating granule-bound starch synthase protein expression and promoted amylopectin accumulation by upregulating 1,4-alpha-glucan branching enzyme and starch synthase proteins expression. Additionally, Phosphate fertilizer primarily promoted the accumulation of hydrophobic and essential amino acids. These findings elucidate the mechanism of P-induced starch accumulation and offer insights into phosphate fertilizer management and high-quality cultivation of common buckwheat.
Subject(s)
Amino Acids , Fagopyrum , Fertilizers , Phosphates , Starch , Fagopyrum/metabolism , Fagopyrum/drug effects , Amino Acids/metabolism , Starch/metabolism , Starch/biosynthesis , Phosphates/metabolism , Plant Proteins/metabolism , Gene Expression Regulation, Plant/drug effects , Amylopectin/metabolism , Amylose/metabolismABSTRACT
In the rosid species Arabidopsis thaliana, the AP2-type AP2 transcription factor (TF) is required for specifying the sepals and petals identities and confers a major A-function to antagonize the C-function in the outer floral whorls. In the asterid species Petunia, the AP2-type ROB TFs are required for perianth and pistil development, as well as repressing the B-function together with TOE-type TF BEN. In Long-homostyle (LH) Fagopyrum esculentum, VIGS-silencing showed that FaesAP2 is mainly involved in controlling filament and style length, but FaesTOE is mainly involved in regulating filament length and pollen grain development. Both FaesAP2 (AP2-type) and FaesTOE (TOE-type) are redundantly involved in style and/or filament length determination instead of perianth development. However, neither FaesAP2 nor FaesTOE could directly repress the B and/or C class genes in common buckwheat. Moreover, the FaesAP1_2 silenced flower showed tepal numbers, and filament length decreased obviously. Interestingly, yeast one-hybrid (Y1H) and dual-luciferase reporter (DR) further suggested that FaesTOE directly up-regulates FaesAP1_2 to be involved in filament length determination in LH common buckwheat. Moreover, the knockdown of FaesTOE expression could result in expression down-regulation of the directly target FaesAP1_2 in the FaesTOE-silenced LH plants. Our findings uncover a stamen development pathway in common buckwheat and offer deeper insight into the functional evolution of AP2 orthologs in the early-diverging core eudicots.
Subject(s)
Fagopyrum , Flowers , Gene Expression Regulation, Plant , Plant Proteins , Fagopyrum/genetics , Fagopyrum/growth & development , Fagopyrum/metabolism , Flowers/genetics , Flowers/growth & development , Flowers/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , Up-Regulation/geneticsABSTRACT
BACKGROUND: The objective of this research was to elucidate the hypocholesterolemic effects of a bioactive compound extracted from buckwheat, and to delineate its influence on the regulatory mechanisms of cholesterol metabolism. The compound under investigation was identified as quercetin. MATERIAL AND RESULTS: In vitro experiments conducted on HepG2 cells treated with quercetin revealed a significant reduction in intracellular cholesterol accumulation. This phenomenon was rigorously quantified by assessing the transcriptional activity of key genes involved in the biosynthesis and metabolism of cholesterol. A statistically significant reduction in the expression of HMG-CoA reductase (HMGCR) was observed, indicating a decrease in endogenous cholesterol synthesis. Conversely, an upregulation in the expression of cholesterol 7 alpha-hydroxylase (CYP7A1) was also observed, suggesting an enhanced catabolism of cholesterol to bile acids. Furthermore, the study explored the combinatory effects of quercetin and simvastatin, a clinically utilized statin, revealing a synergistic action in modulating cholesterol levels at various dosages. CONCLUSIONS: The findings from this research provide a comprehensive insight into the mechanistic pathways through which quercetin, a phytochemical derived from buckwheat, exerts its hypocholesterolemic effects. Additionally, the observed synergistic interaction between quercetin and simvastatin opens up new avenues for the development of combined therapeutic strategies to manage hyperlipidemia.
Subject(s)
Cholesterol 7-alpha-Hydroxylase , Cholesterol , Fagopyrum , Hydroxymethylglutaryl CoA Reductases , Lipid Metabolism , Phytochemicals , Quercetin , Humans , Fagopyrum/chemistry , Fagopyrum/metabolism , Hep G2 Cells , Cholesterol/metabolism , Quercetin/pharmacology , Lipid Metabolism/drug effects , Lipid Metabolism/genetics , Phytochemicals/pharmacology , Hydroxymethylglutaryl CoA Reductases/metabolism , Hydroxymethylglutaryl CoA Reductases/genetics , Cholesterol 7-alpha-Hydroxylase/metabolism , Cholesterol 7-alpha-Hydroxylase/genetics , Anticholesteremic Agents/pharmacology , Simvastatin/pharmacology , Plant Extracts/pharmacology , Gene Expression Profiling/methods , Gene Expression Regulation/drug effectsABSTRACT
This study aimed to investigate the integration of cereal and germinated pseudocereals into set-type yogurt mimic, resulting in a novel and nutritious product. Four groups of yogurts mimic, namely CPY-1, CPY-2, CPY-3, and CPY-4, were prepared using different probiotic cultures, including L. acidophilus 21, L. plantarum 14, and L. rhamnosus 296 along with starter cultures. Notably, CPY-2 cultured with L. plantarum and L. rhamnosus and incubated for 12 h exhibited the most desirable attributes. The resulting yogurt demonstrated an acidity of 0.65%, pH of 4.37 and a probiotic count of 6.38 log CFU/mL. The logistic growth model fit revealed maximum growth rates (k, 1/h) and maximum bacterial counts (Nm log CFU/mL) for each CPY variant. The results revealed that CPY-2 significantly improved protein, dietary fiber, phenols and antioxidant capacities compared to the control. Scanning electron microscopy showed more structured and compact casein network in CPY-2, highlighting its superior textural characteristics. Overall, this study demonstrates the incorporation of cereal and germinated pseudocereals into set-type yogurt mimic offers health benefits through increased dietary fiber and ß-glucan.
Subject(s)
Amaranthus , Antioxidants , Fagopyrum , Germination , Yogurt , Yogurt/analysis , Yogurt/microbiology , Antioxidants/chemistry , Antioxidants/metabolism , Fagopyrum/chemistry , Fagopyrum/growth & development , Fagopyrum/metabolism , Kinetics , Amaranthus/growth & development , Amaranthus/chemistry , Amaranthus/metabolism , Probiotics/analysis , Probiotics/metabolism , Probiotics/chemistry , Fermentation , Lactobacillus/growth & development , Lactobacillus/metabolism , Lactobacillus/chemistry , Food HandlingABSTRACT
BACKGROUND: Proper flower development is essential for plant reproduction, a crucial aspect of the plant life cycle. This process involves precisely coordinating transcription factors, enzymes, and epigenetic modifications. DNA methylation, a ubiquitous and heritable epigenetic mechanism, is pivotal in regulating gene expression and shaping chromatin structure. Fagopyrum esculentum demonstrates anti-hypertensive, anti-diabetic, anti-inflammatory, cardio-protective, hepato-protective, and neuroprotective properties. However, the heteromorphic heterostyly observed in F. esculentum poses a significant challenge in breeding efforts. F. tataricum has better resistance to high altitudes and harsh weather conditions such as drought, frost, UV-B radiation damage, and pests. Moreover, F. tataricum contains significantly higher levels of rutin and other phenolics, more flavonoids, and a balanced amino acid profile compared to common buckwheat, being recognised as functional food, rendering it an excellent candidate for functional food applications. RESULTS: This study aimed to compare the DNA methylation profiles between the Pin and Thrum flower components of F. esculentum, with those of self-fertile species of F. tataricum, to understand the potential role of this epigenetic mechanism in Fagopyrum floral development. Notably, F. tataricum flowers are smaller than those of F. esculentum (Pin and Thrum morphs). The decline in DNA methylation levels in the developed open flower components, such as petals, stigmas and ovules, was consistent across both species, except for the ovule in the Thrum morph. Conversely, Pin and Tartary ovules exhibited a minor decrease in DNA methylation levels. The highest DNA methylation level was observed in Pin stigma from closed flowers, and the most significant decrease was in Pin stigma from open flowers. In opposition, the nectaries of open flowers exhibited higher levels of DNA methylation than those of closed flowers. The decrease in DNA methylation might correspond with the downregulation of genes encoding methyltransferases. CONCLUSIONS: Reduced overall DNA methylation and the expression of genes associated with these epigenetic markers in fully opened flowers of both species may indicate that demethylation is necessary to activate the expression of genes involved in floral development.
Subject(s)
DNA Methylation , Fagopyrum , Flowers , Fagopyrum/genetics , Fagopyrum/growth & development , Fagopyrum/metabolism , Flowers/genetics , Flowers/growth & development , Epigenesis, Genetic , Gene Expression Regulation, PlantABSTRACT
Tartary buckwheat (Fagopyrum tataricum) is an annual coarse cereal from the Polygonaceae family, known for its high content of flavonoid compounds, particularly rutin. But so far, the mechanisms of the flavonoid transport and storage in Tartary buckwheat (TB) remain largely unexplored. This study focuses on ATP-binding cassette transporters subfamily C (ABCC) members, which are crucial for the biosynthesis and transport of flavonoids in plants. The evolutionary and expression pattern analyses of the ABCC genes in TB identified an ABCC protein gene, FtABCC2, that is highly correlated with rutin synthesis. Subcellular localization analysis revealed that FtABCC2 protein is specifically localized to the vacuole membrane. Heterologous expression of FtABCC2 in Saccharomyces cerevisiae confirmed that its transport ability of flavonoid glycosides such as rutin and isoquercetin, but not the aglycones such as quercetin and dihydroquercetin. Overexpression of FtABCC2 in TB hairy root lines resulted in a significant increase in total flavonoid and rutin content (P < 0.01). Analysis of the FtABCC2 promoter revealed potential cis-acting elements responsive to hormones, cold stress, mechanical injury and light stress. Overall, this study demonstrates that FtABCC2 can efficiently facilitate the transport of rutin into vacuoles, thereby enhancing flavonoids accumulation. These findings suggest that FtABCC2 is a promising candidate for molecular-assisted breeding aimed at developing high-flavonoid TB varieties.
Subject(s)
Fagopyrum , Gene Expression Regulation, Plant , Plant Proteins , Rutin , Rutin/metabolism , Fagopyrum/genetics , Fagopyrum/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Promoter Regions, Genetic , Multidrug Resistance-Associated Proteins/metabolism , Multidrug Resistance-Associated Proteins/genetics , Biological Transport , Flavonoids/metabolism , Phylogeny , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae/geneticsABSTRACT
Tartary buckwheat is highly valued for its abundant rutin (quercetin 3-O-rutinoside). As a flavonoid glycoside, rutin is synthesized with the crucial involvement of UDP-dependent glycosyltransferases (UGTs). However, the functions and transcriptional regulation of the UGT-encoded genes remain poorly understood. This study identified a key gene, FtUFGT163, potentially encoding flavonol 3-O-glucoside (1 â 6) rhamnosyltransferase in Tartary buckwheat through omics analysis and molecular docking methods. The recombinant FtUFGT163 expressed in Escherichia coli demonstrated the capacity to glycosylate isoquercetin into rutin. Overexpression of FtUFGT163 significantly enhanced the rutin content in Tartary buckwheat. Further investigation identified a novel bZIP transcription factor, FtGBF1, that enhances FtUFGT163 expression by binding to the G-box element within its promoter, thereby augmenting rutin biosynthesis. Additional molecular biology experiments indicated that the specific positive regulator of rutin, FtMYB5/6, could directly activate the FtGBF1 promoter. Collectively, this study elucidates a novel regulatory module, termed "FtMYB5/6-FtGBF1-FtUFGT163", which effectively coordinates the biosynthesis of rutin in Tartary buckwheat, offering insights into the genetic enhancement of nutraceutical components in crops.
Subject(s)
Fagopyrum , Gene Expression Regulation, Plant , Plant Proteins , Rutin , Fagopyrum/genetics , Fagopyrum/metabolism , Fagopyrum/chemistry , Rutin/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Promoter Regions, Genetic , Molecular Docking SimulationABSTRACT
BACKGROUND: As a newly class of endogenous phytohormones, strigolactones (SLs) regulate crop growth and yield formation by interacting with other hormones. However, the physiological mechanism of SLs affect the yield by regulating the balance of endogenous hormones of Tartary buckwheat is still unclear. RESULTS: In this study, a 2-year field experiment was conducted on Tartary buckwheat (Jinqiao 2) to study the effects of different concentrations (0, 10, and 20 µmol/L) of artificial synthetic analogs of SLs (rac-GR24) and inhibitor of SL synthesis (Tis-108) on the growth, endogenous-hormone content, and yield of Tartary buckwheat. The main-stem branch number, grain number per plant, grain weight per plant, and yield of Tartary buckwheat continuously decreased with increased rac-GR24 concentration, whereas the main-stem diameter and plant height initially increased and then decreased. Rac-GR24 treatment significantly increased the content of SLs and abscisic acid (ABA) in grains, and it decreased the content of Zeatin (Z) + Zeatin nucleoside (ZR). Conversely, Tis-108 treatment decreased the content of SLs and ABA but increased the content of Z + ZR. Results of correlation analysis showed that the content of ABA and SLs, the ratio of SLs/(Z + ZR), SLs/ABA, and ABA/(Z + ZR) were significantly negatively correlated with the yield of Tartary buckwheat, and that Z + ZR content was significantly positively correlated with the yield. Regression analysis further showed that ABA/ (Z + ZR) can explain 58.4% of the variation in yield. CONCLUSIONS: In summary, by adjusting the level of endogenous SLs in Tartary buckwheat, the balance of endogenous hormones in grains can be changed, thereby exerting the effect on yield. The results can provide a new agronomic method for the high-yield cultivation of Tartary buckwheat.
Subject(s)
Fagopyrum , Lactones , Plant Growth Regulators , Fagopyrum/drug effects , Fagopyrum/growth & development , Fagopyrum/metabolism , Plant Growth Regulators/metabolism , Plant Growth Regulators/pharmacology , Lactones/metabolism , Heterocyclic Compounds, 3-Ring/metabolism , Abscisic Acid/metabolismABSTRACT
The MADS-box gene family is a transcription factor family that is widely expressed in plants. It controls secondary metabolic processes in plants and encourages the development of tissues like roots and flowers. However, the phylogenetic analysis and evolutionary model of MADS-box genes in Fagopyrum species has not been reported yet. This study identified the MADS-box genes of three buckwheat species at the whole genome level, and conducted systematic evolution and physicochemical analysis. The results showed that these genes can be divided into four subfamilies, with fragment duplication being the main way for the gene family expansion. During the domestication process from golden buckwheat to tartary buckwheat and the common buckwheat, the Ka/Ks ratio indicated that most members of the family experienced strong purification selection pressure, and with individual gene pairs experiencing positive selection. In addition, we combined the expression profile data of the MADS genes, mGWAS data, and WGCNA data to mine genes FdMADS28/48/50 that may be related to flavonoid metabolism. The results also showed that overexpression of FdMADS28 could increase rutin content by decreasing Kaempferol pathway content in hairy roots, and increase the resistance and growth of hairy roots to PEG and NaCl. This study systematically analyzed the evolutionary relationship of MADS-box genes in the buckwheat species, and elaborated on the expression patterns of MADS genes in different tissues under biotic and abiotic stresses, laying an important theoretical foundation for further elucidating their role in flavonoid metabolism.
Subject(s)
Evolution, Molecular , Fagopyrum , Flavonoids , Gene Expression Regulation, Plant , Genes, Plant , MADS Domain Proteins , Fagopyrum/genetics , Fagopyrum/metabolism , Flavonoids/metabolism , MADS Domain Proteins/genetics , MADS Domain Proteins/metabolism , PhylogenyABSTRACT
This study aimed to clarify the effects of drought during flowering period on the carbon and nitrogen metabolism, growth, and yield of Tartary buckwheat. Tartary buckwheat cultivar Jinqiao 2 was treated with well-watered (CK), slight soil-drought stress (LD), moderate soil-drought stress (MD), and severe soil-drought stress (SD), with the soil water potential maintained at - 0.02 to - 0.03, - 0.04 to - 0.05, - 0.05 to - 0.06, and - 0.06 to - 0.07 MPa, respectively. With prolonged growth period and an increase in drought stress, the antioxidant enzyme activities and the contents of substances and activities of enzymes related to carbon and nitrogen metabolism in Tartary buckwheat leaves initially increased and then decreased. Meanwhile, the contents of malondialdehyde and superoxide anion showed a continuous. LD treatment induced the highest antioxidant enzyme activities and the contents of substances and activities of enzymes related to carbon and nitrogen metabolism but the lowest contents of malondialdehyde and superoxide anion in Tartary buckwheat leaves. Compared with CK, LD treatment increased the grain number, 1000-grain weight (MTS), and yield per plant by 6.52%, 17.37%, and 12.35%, respectively. In summary, LD treatment can increase the antioxidant enzyme activities and the contents of substances and activities of enzymes related to carbon and nitrogen metabolism, thus enhancing the adaptability of Tartary buckwheat to drought stress and increasing the yield per plant.
Subject(s)
Carbon , Droughts , Fagopyrum , Flowers , Nitrogen , Plant Leaves , Fagopyrum/metabolism , Fagopyrum/growth & development , Nitrogen/metabolism , Carbon/metabolism , Flowers/growth & development , Flowers/metabolism , Plant Leaves/metabolism , Plant Leaves/growth & development , Stress, Physiological , Antioxidants/metabolism , Soil/chemistry , Malondialdehyde/metabolism , Water/metabolismABSTRACT
Tartary buckwheat, celebrated as the "king of grains" for its flavonoid and phenolic acid richness, has health-promoting properties. Despite significant morphological and metabolic variations in mature achenes, research on their developmental process is limited. Utilizing Liquid chromatography-mass spectrometry and atmospheric pressure matrix-assisted laser desorption/ionization mass spectrometry imaging, we conducted spatial-temporal metabolomics on two cultivars during achene development. Metabolic profiles including 17 phenolic acids and 83 flavonoids are influenced by both varietal distinctions and developmental intricacies. Notably, flavonols, as major flavonoids, accumulated with achene ripening and showed a tissue-specific distribution. Specifically, flavonol glycosides and aglycones concentrated in the embryo, while methylated flavonols and procyanidins in the hull. Black achenes at the green achene stage have higher bioactive compounds and enhanced antioxidant capacity. These findings provide insights into spatial and temporal characteristics of metabolites in Tartary buckwheat achenes and serve as a theoretical guide for selecting optimal resources for food production.
Subject(s)
Fagopyrum , Metabolomics , Fagopyrum/chemistry , Fagopyrum/growth & development , Fagopyrum/metabolism , Flavonoids/metabolism , Flavonoids/chemistry , Flavonoids/analysis , Liquid Chromatography-Mass Spectrometry , Plant Extracts/metabolism , Plant Extracts/chemistry , Seeds/chemistry , Seeds/growth & development , Seeds/metabolism , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
BACKGROUND: Tartary buckwheat (Fagopyrum tataricum) belongs to Polygonaceae family and has attracted increasing attention owing to its high nutritional value. UDP-glycosyltransferases (UGTs) glycosylate a variety of plant secondary metabolites to control many metabolic processes during plant growth and development. However, there have been no systematic reports of UGT superfamily in F. tataricum. RESULTS: We identified 173 FtUGTs in F. tataricum based on their conserved UDPGT domain. Phylogenetic analysis of FtUGTs with 73 Arabidopsis UGTs clustered them into 21 families. FtUGTs from the same family usually had similar gene structure and motif compositions. Most of FtUGTs did not contain introns or had only one intron. Tandem repeats contributed more to FtUGTs amplification than segmental duplications. Expression analysis indicates that FtUGTs are widely expressed in various tissues and likely play important roles in plant growth and development. The gene expression analysis response to different abiotic stresses showed that some FtUGTs were involved in response to drought and cadmium stress. Our study provides useful information on the UGTs in F. tataricum, and will facilitate their further study to better understand their function. CONCLUSIONS: Our results provide a theoretical basis for further exploration of the functional characteristics of FtUGTs and for understanding the growth, development, and metabolic model in F. tataricum.
Subject(s)
Fagopyrum , Humans , Phylogeny , Fagopyrum/metabolism , Glycosyltransferases/genetics , Glycosyltransferases/metabolism , Plant Proteins/metabolism , Gene Expression Regulation, PlantABSTRACT
The rigid hull encasing Tartary buckwheat seeds necessitates a laborious dehulling process before flour milling, resulting in considerable nutrient loss. Investigation of lignin composition is pivotal in understanding the structural properties of tartary buckwheat seeds hulls, as lignin is key determinant of rigidity in plant cell walls, thus directly impacting the dehulling process. Here, the lignin composition of seed hulls from 274 Tartary buckwheat accessions is analyzed, unveiling a unique lignin chemotype primarily consisting of G lignin, a common feature in gymnosperms. Furthermore, the hardness of the seed hull showed a strong negative correlation with the S lignin content. Genome-wide detection of selective sweeps uncovered that genes governing the biosynthesis of S lignin, specifically two caffeic acid O-methyltransferases (COMTs) and one ferulate 5-hydroxylases, are selected during domestication. This likely contributed to the increased S lignin content and decreased hardness of seed hulls from more domesticated varieties. Genome-wide association studies identified robust associations between FtCOMT1 and the accumulation of S lignin in seed hull. Transgenic Arabidopsis comt1 plants expressing FtCOMT1 successfully reinstated S lignin content, confirming its conserved function across plant species. These findings provide valuable metabolic and genetic insights for the potential redesign of Tartary buckwheat seed hulls.
Subject(s)
Fagopyrum , Lignin , Seeds , Lignin/metabolism , Lignin/genetics , Fagopyrum/genetics , Fagopyrum/metabolism , Seeds/genetics , Seeds/metabolism , MethyltransferasesABSTRACT
Epigenetic programming plays a vital role in regulating pluripotency genes, which become activated or inactivated during the processes of dedifferentiation and differentiation during an organism's development. The analysis of epigenetic modifications has become possible through the technique of immunostaining, where specific antibodies allow the identification of a single target protein. This chapter describes a detailed protocol for the analysis of the epigenetic modifications with the use of confocal microscopy, subsequent image, and statistical analysis on the example of Fagopyrum calli with the use of nine antibodies raised against histone H3 and H4 methylation and acetylation on several lysines as well as DNA methylation.
Subject(s)
Fagopyrum , Fagopyrum/metabolism , Histones/metabolism , Cell Nucleus/metabolism , DNA Methylation , Antibodies/metabolism , Epigenesis, Genetic , AcetylationABSTRACT
Immunostaining is a well-established technique for identifying specific proteins in tissue samples with specific antibodies to identify a single target protein. It is commonly used in research and provides information about cellular localization and protein expression levels. This chapter describes a detailed protocol for immunostaining fixed Fagopyrum calli embedded in Steedman's wax using nine antibodies raised against histone H3 and H4 methylation and acetylation on several lysines and DNA methylation.
Subject(s)
Fagopyrum , Fagopyrum/metabolism , Histones/metabolism , Epigenesis, Genetic , DNA Methylation , Lysine/metabolism , Antibodies/metabolism , AcetylationABSTRACT
Immunohistochemistry is a method that allows the detection of individual components of cell walls in an extremely precise way at the level of a single cell and wall domains. The cell wall antibodies detect specific epitopes of pectins, arabinogalactan proteins (AGP), hemicelluloses, and extensins. The presented method visualization of the selected pectic and AGP epitopes using antibodies directed to wall components is described. The method of the analysis of the chemical composition of the wall is present on the example of the shoot apical meristems of Fagopurum esculentum and Fagopyrum tataricum. Recommended protocols for immunostaining and examination on fluorescence microscopy level are presented.
Subject(s)
Fagopyrum , Fagopyrum/chemistry , Fagopyrum/metabolism , Meristem/metabolism , Pectins/analysis , Immunohistochemistry , Epitopes , Cell Wall/chemistryABSTRACT
Rutin is a significant flavonoid with strong antioxidant property and various therapeutic effects. It plays a crucial role in disease prevention and human health maintenance, especially in anti-inflammatory, antidiabetic, hepatoprotective and cardiovascular effects. While many plants can synthesize and accumulate rutin, tartary buckwheat is the only food crop possessing high levels of rutin. At present, the rutin content (RC) is regarded as the key index for evaluating the nutritional quality of tartary buckwheat. Consequently, rutin has become the focus for tartary buckwheat breeders and has made considerable progress. Here, we summarize research on the rutin in tartary buckwheat in the past two decades, including its accumulation, biosynthesis and breakdown pathways, and regulatory mechanisms. Furthermore, we propose several strategies to increase the RC in tartary buckwheat seeds based on current knowledge. This review aims to provide valuable references for elevating the quality of tartary buckwheat in the future.
Subject(s)
Fagopyrum , Rutin , Humans , Rutin/metabolism , Fagopyrum/metabolism , Biofortification , Flavonoids/metabolism , Metabolic Networks and PathwaysABSTRACT
Tartary buckwheat (Fagopyrum tataricum) is frequently employed as a resource to develop health foods, owing to its abundant flavonoids such as rutin. However, the consumption of Tartary buckwheat (TB) is limited in food products due to the strong bitterness induced by the hydrolysis of rutin into quercetin. This transformation is facilitated by the degrading enzyme (RDE). While multiple RDE isoenzymes exist in TB, the superior coding gene of FtRDEs has not been fully explored, which hinders the breeding of TB varieties with minimal bitterness. Here, we found that FtRDE2 is the most abundant enzyme in RDE crude extracts, and its corresponding gene is specifically expressed in TB seeds. Results showed that FtRDE2 has strong rutin hydrolysis activity. Overexpression of FtRDE2 not only significantly promoted rutin hydrolysis and quercetin accumulation but also dramatically upregulated genes involved in the early phase of flavonoid synthesis (FtPAL1ãFtC4H1ãFt4CL1, FtCHI1) and anthocyanin metabolism (FtDFR1). These findings elucidate the role of FtRDE2, emphasizing it as an endogenous factor contributing to the bitterness in TB and its involvement in the metabolic regulatory network. Moreover, correlation analysis revealed a positive relationship between the catalytic activity of RDE extracts and the expression level of FtRDE2 during seed germination. In summary, our results suggest that FtRDE2 can serve as a promising candidate for the molecular breeding of a TB variety with minimal bitterness.
Subject(s)
Fagopyrum , Quercetin , Quercetin/metabolism , Fagopyrum/genetics , Fagopyrum/metabolism , Plant Breeding , Rutin/metabolism , Seeds/metabolismABSTRACT
To promote the selenium (Se) uptakes in fruit trees under Se-contaminated soil, the effects of water extract of Fagopyrum dibotrys (D. Don) Hara straw on the Se accumulation in peach seedlings under selenium-contaminated soil were studied. The results showed that the root biomass, chlorophyll content, activities of antioxidant enzymes, and soluble protein content of peach seedlings were increased by the F. dibotrys straw extract. The different forms of Se (total Se, inorganic Se, and organic Se) were also increased in peach seedlings following treatment with the F. dibotrys straw extract. The highest total shoot Se content was treated by the 300-fold dilution of F. dibotrys straw, which was 30.87% higher than the control. The F. dibotrys straw extract also increased the activities of adenosine triphosphate sulfurase (ATPS), and adenosine 5'-phosphosulfate reductase (APR) in peach seedlings, but decreased the activity of serine acetyltransferase (SAT). Additionally, correlation and grey relational analyses revealed that chlorophyll a content, APR activity, and root biomass were closely associated with the total shoot Se content. Overall, this study shows that the water extract of F. dibotrys straw can promote Se uptake in peach seedlings, and 300-fold dilution is the most suitable concentration.
The water extract of Fagopyrum dibotrys (D. Don) Hara straw promoted the selenium (Se) uptake in peach seedlings under selenium-contaminated soil. The concentration of F. dibotrys straw extract showed a quadratic polynomial regression relationship with the total root and shoot Se. Furthermore, chlorophyll a content, APR activity, and root biomass were closely associated with the total shoot Se. This study shows that water extract of F. dibotrys straw can promote Se uptake in peach seedlings, and 300-fold dilution is the most suitable concentration.