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Starch serves as a crucial energy source for both plants and humans, predominantly synthesized and stored in endosperms, tubers, rhizomes, and cotyledons. Given the significant role of amylose in determining the quality of starchy crops, optimizing its content has become a key objective in current crop breeding efforts. Tartary buckwheat, a dicotyledonous plant, notably accumulates high levels of amylose in its endosperm, surpassing common cereals like rice and maize. However, the mechanisms underlying amylose accumulation, distribution, and regulation in Tartary buckwheat remain unclear. Here, amylose content was determined across various tissues and organs of Tartary buckwheat, identifying with the endosperm as the primary site for its biosynthesis and accumulation. RNA sequencing analysis of endosperms from different developmental stages identified 35 genes potentially involved in starch biosynthesis, with 13 genes showing high endosperm-specific expression, suggesting crucial roles in starch biosynthesis. Additionally, the transcription factor FtNF-YB2, which was specifically highly expressed in the endosperm, was discovered to enhance amylose synthesis. Moreover, promoters with potential endosperm-specific activity were identified, advancing our understanding of amylose regulation. Additionally, this study also demonstrates that brassinosteroids (BR) positively influence amylose biosynthesis in Tartary buckwheat endosperm. These findings provide essential insights into the mechanisms of understanding amylose biosynthesis, accumulation and regulation in Tartary buckwheat, offering significant implications for future breeding strategies., Competing Interests: Declaration of competing interest The authors declare no competing financial interest that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The ubiquity of microplastic is widely recognized as pollution. Microplastic can affect the growth performances of plants. Buckwheat is a potential model crop to investigate plant responses to hazardous materials. Still, little is known about the response of buckwheat to microplastics. Thus, this study investigated the effect and uptake of polyethylene (PE) in buckwheat plant growth by monitoring the morphological and photosynthetic merits, antioxidant systems and transcriptome analysis of gene expression. Results confirmed that the impacts of PE on buckwheat growth were dose-dependent, while the highest concentration (80 mg/L) exposure elicited significantly negative responses of buckwheat. PE can invade buckwheat roots and locate in the vascular tissues. PE exposure disturbed the processes of carbon fixation and the synthesis of ATP from ADP + Pi in buckwheat leaves. The promotion of photosynthesis under PE exposure could generate extra energy for buckwheat leaves to activate antioxidant systems by increasing the antioxidant enzyme activities at an expense of morphological merits under microplastic stresses. Further in-depth study is warranted about figuring out the interactions between microplastics and biochemical responses (i.e., photosynthesis and antioxidant systems), which have great implications for deciphering the defense mechanism of buckwheat to microplastic stresses., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Tartary buckwheat flavonoids (TBF) are active components extracted from Tartary buckwheat, which have abundant biological effects. According to this study, we investigated the effect of TBF on high-fat diet (HFD)-induced kidney fibrosis and its related mechanisms. In vivo, we established an HFD-induced kidney fibrosis model in mice and administered TBF. The results showed that TBF was able to alleviate kidney injury and inflammatory response. Subsequently, the mRNA levels between the HFD group and the TBF + HFD group were detected using RNA-seq assay. According to the gene set enrichment analysis (GSEA) and Kyoto Encyclopedia of Genes and Genomes (KEGG) results, the differential genes were enriched in lipid metabolism and mitogen-activated protein kinases(MAPK) signaling pathways. We examined the protein expression of lipid metabolism-related pathways and the level of lipid metabolism. The results showed that TBF significantly activated the adenosine monophosphate activated protein kinase/acetyl-CoA carboxylase (AMPK/ACC) pathway and effectively reduced kidney total cholesterol (TC), triglyceride (TG) and low-density lipoproteinc cholesterol (LDL-C) levels and increased high-density lipoprotein cholesterol (HDL-C) levels in mice. TBF also inhibited transforming growth factor-β1/Smad (TGF-β1/Smad) and MAPK signaling pathways, thus slowing down the kidney fibrosis process. In vitro, using palmitic acid (PA) to stimulate TCMK-1 cells, the in vivo results similarly demonstrated that TBF could alleviate kidney fibrosis in HFD mice by inhibiting TGF1/Smad signaling pathway and MAPK signaling pathway., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Tartary buckwheat (Fagopyrum tataricum L. Gaertn., TB) is an ancient minor crop and an important food source for humans to supplement nutrients such as flavonoids and essential amino acids. Amino acid transporters (AATs) play critical roles in plant growth and development through the transport of amino acids. In this study, 104 AATs were identified in TB genome and divided into 11 subfamilies by phylogenetic relationships. Tandem and segmental duplications promoted the expansion of FtAAT gene family, and the variations of gene sequence, protein structure and expression pattern were the main reasons for the functional differentiation of FtAATs. Based on RNA-seq and qRT-PCR, the expression patterns of FtAATs in different tissues and under different abiotic stresses were analyzed, and several candidate FtAATs that might affect grain development and response to abiotic stresses were identified, such as FtAAP12 and FtCAT7. Finally, combined with the previous studies, the expression patterns and phylogenetic relationships of AATs in multiple species, the functions of multiple high-confidence FtAAT genes were predicted, and the schematic diagram of FtAATs in TB was initially drawn. Overall, this work provided a framework for further functional analysis of FtAAT genes and important clues for the improvement of TB quality and stress resistance., (Copyright © 2022. Published by Elsevier B.V.)

Drought stress has been the main abiotic factor affecting the growth, development and production of common buckwheat (Fagopyrum esculentum). To explore the response mechanisms of regulating buckwheat drought stress on the post-transcriptional and translational levels, a comparative proteomic analysis was applied to monitor the short-term proteomic variations under the drought stress in the seedling stage. From which 593 differentially abundant proteins (DAPs) were identified using the TMT-based proteomics analysis. A number of DAPs were found to be intimately correlated with the styrene degradation, phenylpropanoid biosynthesis and stimulus response, within which. The acyl-CoA oxidase 4 (ACX4), a key regulator in plant abiotic stress response, was selected for further elucidation. Overexpression of the FeACX4 not only conferred drought and salt tolerance in the Arabidopsis, but also significantly increased the root length and fresh weight in the overexpression lines plant relative to the wild type (WT) plant, accompanied by the elevated activities of catalase (CAT) and lowered malonaldehyde (MDA) and H 2 O 2 contents, therefore allowing plants to better adapt to adverse environments. Our results provided information in the exploring of the molecular regulation mechanism responding to drought tolerance in common buckwheat., (Copyright © 2022. Published by Elsevier B.V.)

Tartary buckwheat (Fagopyrum tataricum Gaertn.) is an economical crop with excellent edible, nutritional, and medicinal values. However, the production of Tartary buckwheat is very low and it is urgent to breed high-yield varieties for satisfying the increasing market demand. Heterotrimeric G-protein signaling involves in the regulation of agronomical traits and fruit development in several plant species. In this study, fifteen genes involved in G-protein signaling were characterized in Tartary buckwheat and their potential roles in fruit development were revealed by expression analysis. The exon-intron organization and conserved motif of Tartary buckwheat G-protein signaling genes were similar to those in other dicot plants. All these genes were ubiquitously and differently expressed in five tissues. The expression patterns of Tartary buckwheat G-protein signaling genes in fruit suggested they may play important roles in the fruit at early development stage, which was supported by meta-analysis of G-protein signaling genes' expression in the fruits from different species. Furthermore, we found the expression of G-protein signaling genes in fruit showed high correlation with 178 transcription factors, which indicated a transcriptional regulatory loop moderating G-protein signaling genes' expression during fruit development. This paper provides new insights into the physiological functions of G-protein signaling in fruit., Competing Interests: Declaration of competing interest The authors confirm that there is no conflict of interest associated with this publication., (Copyright © 2021. Published by Elsevier B.V.)

Flavonoids can not only help plants resist ultraviolet and pathogen attacks, but also show a wide range of therapeutic prospects for human health, including antioxidant, anti-inflammatory and anti-hypertension. Tartary buckwheat, as medicinal and food homologous crop, is rich in flavonoids, among which rutin may prevent liver damage. The one of the major objectives of Tartary buckwheat breeding is to cultivate varieties that have large fruits, high flavonoids and nutrient contents. Members of the cytochrome P450 monooxygenase (CYP) superfamily play a vital role in the synthesis of flavonoids, plant growth and development. Whole-genome analyses of the CYP family have been performed in several plants, but the CYP family has not been characterized in Tartary buckwheat. In this study, 285 FtCYPs were identified from the genome to improve the rutin content and quality of Tartary buckwheat. By exploring the structure, motif composition, tandem and segmental duplication events of FtCYPs, as well as evolutionary relationships with CYPs in other plants, we preliminarily screened potential FtCYPs regulating rutin synthesis, growth and development. The expression levels of the FtCYPs in different organs and fruits at various periods were measured. This study provides a solid foundation for verifying the function of FtCYPs, cultivating high rutin Tartary buckwheat varieties., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The seed of Tartary buckwheat (Fagopyrum tataricum) is rich in nutrients and functional ingredients and is recommended as a healthy cereal food. The total proteins of Tartary buckwheat seed (TBS) were extracted and digested; then, the phosphopeptides and glycopeptides were separately enriched and identified by nano liquid chromatography/tandem mass spectrometry. A total of 2613 phosphorylation sites from 1670 phosphoproteins and 404 N-glycosylation sites from 285 N-glycoproteins were identified in TBS. Function and pathway analyses showed that TBS phosphoproteins were significantly enriched in transport, energy metabolism, amino acids biosynthesis/metabolism, and signaling and TBS N-glycoproteins were significantly enriched in modification regulation. The present study reports the first profiles of the phosphoproteome and N-glycoproteome of TBS and provides important post-translational modifications information on the proteins in TBS. The results of this study will aid the understanding of the underlying mechanism of the germination of TBS during cultivation and edible quality changes during storage and processing., (Copyright © 2019. Published by Elsevier B.V.)

Nitrogen is an essential nutrient for Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) because this substance affects the yield and quality of Tartary buckwheat. The physicochemical characteristics of starch represent the important parameters of Tartary buckwheat quality. The effects of different levels of nitrogen application at different levels (0, 45, 135 and 225kg/ha in 2015 and 0, 90, 180 and 270kg/ha in 2017) on the physicochemical characteristics of Tartary buckwheat starch were studied. The amylose content, particle size and retrogradation of Tartary buckwheat starch with nitrogen were lower than those of without nitrogen. The structure complexity, pasting temperature, gelatinization enthalpy, relative crystallinity, light transmittance and solubility of the former were higher than those of the latter. Nitrogen application did not change the 'A'-type crystalline pattern of Tartary buckwheat starch. This study indicated that nitrogen level and years and the interactions among nitrogen fertilizer levels and years significantly affected the physicochemical properties of Tartary buckwheat starch. These integrated results also provided information about the management of fertilization conditions to obtain starches with special properties for applications in food or nonfood industries., (Copyright © 2019. Published by Elsevier B.V.)

Glycoproteins play pivotal roles in a series of biological processes and their glycosylation patterns need to be structurally and functionally characterized. However, the lack of versatile methods to release N-glycans as functionalized forms has been undermining glycomics studies. Here a novel method is developed for dissociation of N-linked glycans from glycoproteins for analysis by MS and online LC/MS. This new method employs aqueous ammonia solution containing NaBH 3 CN as the reaction medium to release glycans from glycoproteins as 1-amino-alditol forms. The released glycans are conveniently labeled with 9-fluorenylmethyloxycarbonyl (Fmoc) and analyzed by ESI-MS and online LC/MS. Using the method, the neutral and acidic N-glycans were successfully released without peeling degradation of the core α-1,3-fucosylated structure or detectable de-N-acetylation, revealing its general applicability to various types of N-glycans. The Fmoc-derivatized N-glycans derived from chicken ovalbumin, Fagopyrum esculentum Moench Pollen and FBS were successfully analyzed by online LC/MS to distinguish isomers. The 1-amino-alditols were also permethylated to form quaternary ammonium cations at the reducing end, which enhance the MS sensitivity and are compatible with sequential multi-stage mass spectrometry (MS n ) fragmentation for glycan sequencing. The Fmoc-labeled N-glycans were further permethylated to produce methylated carbamates for determination of branches and linkages by sequential MS n fragmentation., Significance of the Study: N-Glycosylation represents one of the most common post-translational modification forms and plays pivotal roles in the structural and functional regulation of proteins in various biological activities, relating closely to human health and diseases. As a type of informational molecule, the N-glycans of glycoproteins participate directly in the molecular interactions between glycan epitopes and their corresponding protein receptors. Detailed structural and functional characterization of different types of N-glycans is essential for understanding the functional mechanisms of many biological activities and the pathologies of many diseases. Here we describe a simple, versatile method to indistinguishably release all types of N-glycans as functionalized forms without remarkable side reactions, enabling convenient, rapid analysis and preparation of released N-glycans from various complex biological samples. It is very valuable for studies on the complicated structure-function relationship of N-glycans, as well as for the search of N-glycan biomarkers of some major diseases and N-glycan related targets of some drugs., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Glutaredoxins (Grxs) are ubiquitous thioltransferases and members of the thioredoxin (Trx) fold superfamily. They have multiple functions in cells including oxidative stress responses and cell signaling. A novel glutaredoxin from buckwheat (rbGrx) with higher catalytic activity was identified, cloned, and purified. The structures of glutathionylated rbGrx and an rbGrx mutant, in which cysteine 39 was mutated to alanine, were solved by x-ray diffraction at a resolution of 2.05Å and 2.29Å, respectively. In rbGrx, GSH (glutathione) is bound at the conserved GSH-binding site, and its structure shows that it has the potential to function as a scaffold protein for the assembly and delivery of GSH. The crystal structure shows that GSH does not bind to the C39A rbGrx mutant, and the C39A mutant had no catalytic activity, indicating that C39 is a key residue that is involved in both the binding of rbGrx to GSH and the regulation of its catalytic activity. The model showing the binding of GSH with rbGrx provides a basis for understanding its molecular function and its potential future applications in medicinal food science., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Rutin-degrading enzymes (RDEs) specifically hydrolyze the glycosidic linkages of rutin, producing quercetin and rutinose. Here we report a reliable and sensitive polyacrylamide gel electrophoresis and staining method for the detection of RDE isozymes, which is based on the aqueous solubility difference between rutin and quercetin, as well as the ultraviolet absorbance of quercetin. With this novel method, we achieved a detection limit of 12 ng with 107 U of RDE activity, enabling us to detect at least five RDE isozymes in tartary buckwheat seeds., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The present work extends knowledge about possible biosynthesis of fagopyrin in buckwheat plants by providing possible candidate genes for its biosynthesis and the role of type III polyketide synthases (PKSs). Moreover, new information is presented about the possible connection between naphthodianthrones and phenolic biosynthesis. Possible regulation of fagopyrin biosynthesis and production under different growth conditions is also discussed., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Currently hydrocolloids are applied as baking aids in order to enhance the quality of gluten-free bread. Microbial exopolysaccharides (EPS) can also have a positive impact on gluten-free baked goods. Provided that yields are sufficient, in situ produced EPS may enable clean (additive free) labels. Thus, in situ EPS optimization was conducted to achieve high amounts of EPS in gluten-free sourdoughs. The influence of flours, dough yield, cell counts, sucrose concentration and sucrose fed-batch were investigated for Lactobacillus (L.) animalis TMW 1.971, Lactobacillus reuteri TMW 1.106 and Lactobacillus curvatus TMW 1.624. Maximal yields of 17.93 g EPS kg(-1) flour were obtained with L. animalis TMW 1.971 in buckwheat core doughs, 15.69 g EPS kg(-1) with L. reuteri TMW 1.106 in quinoa, and 16.28 g EPS kg(-)(1) flour with L. curvatus TMW 1.624 in buckwheat sourdoughs. This study evidences that EPS amounts can be manifolded to reach effectual levels through optimization and corroborate the application of EPS-forming starter cultures as a promising approach to improve gluten-free baked goods., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The flavonoid rich grain of buckwheat (Fagopyrum esculentum Moench, Fam. Polygonaceae) is of high nutritional value. With the aim to improve its agronomic productivity, cultivars were crossed with the wild species F. homotropicum which, however, differs in its flavonoid content. The intention of this work was to determine the flavonoid composition in developed interspecific hybrids and to elucidate the proanthocyanidin structures. Seven compounds were purified from methanol extracts of buckwheat (Fagopyrum esculentum Moench) grains by Sephadex LH-20 column chromatography. Beside the procyanidin epicatechin-[4-8]-epicatechin-3-O-(3,4)-dimethylgallate the following propelargonidins were identified: epiafzelechin-[4-6]-epicatechin, epiafzelechin-[4-8]-epiafzelechin-[4-8]-epicatechin, epiafzelechin-[4-8]-epicatechin-3-O-(3,4-dimethyl)-gallate, epiafzelechin-[4-8]-epiafzelechin-[4-8]-epicatechin-3-O-(3,4-dimethyl)-gallate, epiafzelechin-[4-8]-epicatechin-3-O-4-methyl-gallate and epiafzelechin-[4-8]-epicatechin-p-OH-benzoate on the basis of HPLC and LC-MS/MS.

The paper describes the amino acid sequence of a 26 kDa basic subunit of 13S globulin of common buckwheat (Fagopyrum esculentum Moench). The protein has 93 and 75% sequence homology with 11S globulin of Coffea arabica and beta subunit of 11S globulin of Cucurbita pepo respectively. The subunit has the "globally conserved" N-terminal sequence consisting of Gly-Ile-Asp-Glu and the cysteine at P7' from the proteolytic processing site. A conserved 7 residue domain of Pro-His-Trp-Asn-Ile-Asn-Ala, characteristic of basic subunits of legumins from non-leguminous angiosperms, is also present in this protein. A distinguishing features of this subunit is the relatively high level of lysine and methionine.

Dextrorotatory 1-amino-3',4'-dichlorobenzylphosphonic acid was found to be a potent inhibitor of the plant enzyme phenylalanine ammonia-lyase both in vitro and in vivo from among the ring-substituted 1-aminobenzylphosphonic acids and other analogues of phenylglycine. A structure activity relationship analysis of the results obtained permits predictions on the geometry of the pocket of the enzyme and is a basis in the strategy of better inhibitor synthesis.