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Tartary buckwheat sprouts are highly valued by consumers for their superior nutritional content. Ionic titanium (Ti) has been shown to enhance crop growth and improve nutritional quality. However, there is limited research on the impact of ionic Ti on the nutritional quality of Tartary buckwheat sprouts. This study cultivated Tartary buckwheat sprouts with ionic Ti and found that the high concentration of ionic Ti significantly increased the contents of chlorophyll a, chlorophyll b, and carotenoids (increased by 25.5%, 27.57%, and 15.11%, respectively). The lower concentration of ionic Ti has a higher accumulation of total flavonoids and total polyphenols. Metabolomics analysis by LC-MS revealed 589 differentially expressed metabolites and 54 significantly different metabolites, enriching 82 metabolic pathways, especially including amino acid biosynthesis and flavonoid biosynthesis. This study shows that ionic Ti can promote the growth of Tartary buckwheat sprouts, improve nutritional quality, and have huge development potential in food production., 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 Ltd. All rights reserved.)

The R2R3-MYB transcription factors (TFs) play a crucial role in regulating plant secondary metabolism and abiotic stress responses, yet they are still poorly understood in common buckwheat (Fagopyrum esculentum), a valuable minor grain crop resource. In this study, a candidate gene, FeR2R3-MYB, was cloned from the anthocyanin-rich common buckwheat variety 'QZZTQ'. FeR2R3-MYB was found to contain two MYB DNA-binding domains and be located at the nucleus with transcriptional activation activity. Molecular analysis indicated that FeR2R3-MYB is predominantly expressed in flowering tissue and is highly responsive to environmental factors such as light, drought, and cold. In addition, the promoter of FeR2R3-MYB showed a positive correlation with fragment length. Further functional analysis suggested that FeR2R3-MYB not only participates in the anthocyanin biosynthetic pathway by interacting with leucoanthocyanidin reductase (FeLAR), but also enhances drought tolerance in common buckwheat. To sum up, FeR2R3-MYB exhibits positive effects on both pigment production (e.g., anthocyanin) and abiotic stress resistance, providing valuable insights for future research in buckwheat molecular breeding and resource development., 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 Masson SAS. All rights reserved.)

Tartary buckwheat is known for its ability to adapt to intricate growth conditions and to possess robust stress-resistant properties. Nevertheless, it remains vulnerable to drought stress, which can lead to reduced crop yield. To identify potential genes involved in drought resistance, a genome-wide association study on drought tolerance in Tartary buckwheat germplasm was conducted. A gene encoding pectin methylesterase inhibitors protein (FtPMEI13) was identified, which is not only associated with drought tolerance but also showed induction during drought stress and abscisic acid (ABA) treatment. Further analysis revealed that overexpression of FtPMEI13 leads to improved drought tolerance by altering the activities of antioxidant enzymes and the levels of osmotically active metabolites. Additionally, FtPMEI13 interacts with pectin methylesterase (PME) and inhibits PME activity in response to drought stress. Our results suggest that FtPMEI13 may inhibit the activity of FtPME44/FtPME61, thereby affecting pectin methylesterification in the cell wall and modulating stomatal closure in response to drought stress. Yeast one-hybrid, dual-luciferase assays, and electrophoretic mobility shift assays demonstrated that an ABA-responsive transcription factor FtbZIP46, could bind to the FtPMEI13 promoter, enhancing FtPMEI13 expression. Further analysis indicated that Tartary buckwheat accessions with the genotype resulting in higher FtPMEI13 and FtbZIP46 expression exhibited higher drought tolerance compared to the others. This suggests that this genotype has potential for application in Tartary buckwheat breeding. Furthermore, the natural variation of FtPMEI13 was responsible for decreased drought tolerance during Tartary buckwheat domestication. Taken together, these results provide basic support for Tartary buckwheat breeding for drought tolerance., (© 2024 Society for Experimental Biology and John Wiley & Sons Ltd.)

The auxin response factor (ARF) is a plant-specific transcription factor that regulates the expression of auxin response genes by binding directly to their promoters. They play an important role in the regulation of plant growth and development, as well as in the response to biotic and abiotic stresses. However, the identification and functional analysis of ARFs in Fagopyrum dibotrys are still unclear. In this study, a total of 26 FdARF genes were identified using bioinformatic methods. Their chromosomal location, gene structure, physical and chemical properties of their encoded protein, subcellular location, phylogenetic tree, conserved motifs and cis-acting elements in FdARF promoters were analyzed. The results showed that 26 FdARF genes were unevenly distributed on 8 chromosomes, with the largest distribution on chromosome 4 and the least distribution on chromosome 3. Most FdARF proteins are located in the nucleus, except for the proteins FdARF7 and FdARF21 located to the cytoplasm and nucleus, while FdARF14, FdARF16, and FdARF25 proteins are located outside the chloroplast and nucleus. According to phylogenetic analysis, 26 FdARF genes were divided into 6 subgroups. Duplication analysis indicates that the expansion of the FdARF gene family was derived from segmental duplication rather than tandem duplication. The prediction based on cis-elements of the promoter showed that 26 FdARF genes were rich in multiple stress response elements, suggesting that FdARFs may be involved in the response to abiotic stress. Expression profiling analysis showed that most of the FdARF genes were expressed in the roots, stems, leaves, and tubers of F. dibotrys, but their expression exhibits a certain degree of tissue specificity. qRT-PCR analysis revealed that most members of the FdARF gene were up- or down-regulated in response to abiotic stress. The results of this study expand our understanding of the functional role of FdARFs in response to abiotic stress and lay a theoretical foundation for further exploration of other functions of FdARF genes., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

The diversity of secondary metabolites is an important means for plants to cope with the complex and ever-changing terrestrial environment. Plant biosynthetic gene clusters (BGCs) are crucial for the biosynthesis of secondary metabolites. The domestication and evolution of BGCs and how they affect plant secondary metabolites biosynthesis and environmental adaptation are still not fully understood. Buckwheat exhibits strong resistance and abundant secondary metabolites, especially flavonoids, allowing it to thrive in harsh environments. A non-canonical BGC named UFGT3 cluster is identified, which comprises a phosphorylase kinase (PAK), two transcription factors (MADS1/2), and a glycosyltransferase (UFGT3), forming a complete molecular regulatory module involved in flavonoid biosynthesis. This cluster is selected during Tartary buckwheat domestication and is widely present in species of the Fagopyrum genus. In wild relatives of cultivated buckwheat, a gene encoding anthocyanin glycosyltransferase (AGT), which glycosylates pelargonidin into pelargonidin-3-O-glucoside, is found inserted into this cluster. The pelargonidin-3-O-glucoside can help plants resist UV stress, endowing wild relatives with stronger high-altitude adaptability. This study provides a new research paradigm for the evolutionary dynamics of plant BGCs, and offers new perspectives for exploring the mechanism of plant ecological adaptability driven by environmental stress through the synthesis of secondary metabolites., (© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)

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.)

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 β-glucan., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Competing Interests: Conflict of interest The authors declare no conflicts of interest.

Rutin is a crucial bioactive compound that determines the nutritional value of Tartary buckwheat (TB). However, the potential of utilizing TB as a dietary source of rutin for human consumption remains largely unexplored. This study aims to address these knowledge gaps by conducting a detailed analysis of rutin content distribution in TB tissues. Our findings revealed a significant variation in rutin content across different plant tissues. Notably, higher levels of rutin were found in embryos and cotyledons compared to other tissues, highlighting them as the primary sites of rutin accumulation in TB seeds and sprouts. Additional research on the processing of TB showed that sprouts and seeds retain high rutin levels even after boiling, steaming, deep-frying, stir-frying, and popping. Comparative analysis of different TB-derived products confirmed that cooked seeds and sprouts can serve as significant dietary sources of rutin. This study offers a foundational framework for the development of future dietary recommendations and applications of TB., Competing Interests: Declaration of competing interest All 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 Ltd. All rights reserved.)

The differences in volatile flavor compounds (VFCs) between Monascus-solid-state fermented Tartary buckwheat (MSFTB) and Monascus-liquid-state fermented Tartary buckwheat (MLFTB) were investigated using electronic nose and gas chromatography-ion mobility spectrometry (GC-IMS) analysis. The study revealed several significant differences in the composition and abundance of VFCs between the two states. Compared to MSFTB, MLFTB exhibited notable increases in various elements including protein, crude fat, total flavonoids, total polyphenols, Monacolin K, Monascus pigments. Principal component analysis demonstrated significant increases in the production of specific VFCs in MLFTB compared to MSFTB. A total of 25 VFCs were identified through GC-IMS, including 9 esters, 7 alcohols, 5 ketones, and 4 aldehydes. The content of pleasant VFCs in MLFTB was significantly higher than in MSFTB. These compounds served as both VFCs and key aroma components during fermentation. In conclusion, the Monascus fermentation state played a crucial role in enhancing the flavor quality of Tartary 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. Published by Elsevier Ltd.)

The classic ABC model postulates how three classes of floral homeotic genes (A, B and C) work in a combinational way to confer organ identity to each whorl that make up a perfect flower in core eudicot plants. Fagopyrum esculentum (Polygonaceae) produces dimorphic flowers with single whorl showy tepals, representing a considerable difference with most core eudicots flowers. Here, we explain in detail the function of a duplicated pair of floral homeotic genes involved in the formation of tepals and stamens in the LH F. esculentum. FaesAP1_1 and FaesAP1_2 work together to specify tepal identity. FaesAP3_1/2 or FaesPI_1/2 have redundant function in specifying filament identity, while FaesAP3_2 and FaesPI_2 also retain a conserved role in specifying anther development and gain novel function in style length determination. However, FaesPI_1 gain novel function in floral color formation. In addition, FaesAG can directly regulate stamen and pistil development or binds to the CArG-box of pFaesPI_1 to indirectly regulate stamen and pistil development by a gene regulatory pathway involving FaesAP1_1/2, FaesAP3_1/2 and FaesPI_1/2. Moreover, FaesAP1_1/2 can directly or indirectly regulate B-class gene (FaesAP3_1/2 and FaesPI_1/2) expression to be involved in floral development. Our work has led to detailed insights into the MADS-box gene regulatory networks that control floral developmental process in LH F. esculentum., Competing Interests: Declaration of Competing Interest The authors declare that there are no conflicts of interest in relation to the manuscript titled "Duplicate MADS-box genes with split roles and a genetic regulatory network of floral development in long-homostyle common buckwheat" submitted to Plant Science. We confirm that the results and interpretations reported in the manuscript are original and have not been plagiarized. We certify that we have read and understand the Plant Science conflict of interest policy, and I understand that failure to disclose a conflict of interest may result in the manuscript being rejected or retracted. We also certify that we have disclosed any financial or non-financial relationships that may be interpreted as constituting a conflict of interest in relation to this manuscript. I understand that this information will be subject to peer review, and I am willing to provide further information or clarification if required. We confirm that we have no known conflicts of interest that would influence the results or interpretation of the data presented in this manuscript, and we understand that failure to disclose a conflict of interest is unethical and may result in sanctions being imposed on us., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Flavonoids constitute a class of polyphenolic secondary metabolites synthesised mainly by plants and possessing anticancer, antioxidant, anti-inflammatory, and antiviral properties. Common buckwheat ( F. esculentum Moench) is a dicotyledonous plant rich in different classes of flavonoids (e.g., rutin) and other phenolic compounds. Lipochitooligosaccharides (LCOs), i.e., rhizobial Nod factors and important signalling molecules for the initiation of symbiosis with legumes, are very effective mitogens that stimulate cell division in plant meristems and the production of secondary metabolites. They can also act in this way in non-legume plants. It has been shown that rhizobial Nod factors noticeably improve plant growth. Rhizobial Nod factors influence the production of flavonoids in common buckwheat grown in greenhouse conditions. The amount of rutin and isoorientin in leaves and flowers has been shown to increase in a statistically significant way after application of Nod factors to buckwheat seeds. The presence of rhizobial Nod factors has no influence on the flavonoid content in stems and roots.

The present study investigated the effect of nitrogen fertilization (NF) at the levels of 0, 45, and 90 kg·ha -1 combined with selected sulfur complex fertilization (SCF) levels of 0 and 45 kg·ha -1 on the nutritional and technological characteristics of buckwheat flour from five varieties. The results showed that the genotype was a critical factor affecting the chemical composition and physicochemical properties of buckwheat flour. NF significantly increased protein, total starch, and amylose content as well as mineral composition but decreased particle size, color value, and water hydration properties. However, SCF enhanced the ash content and decreased the protein content but had no significant effect on the pasting temperature. In addition, the combination of NF and SCF significantly reduced granule size, water solubility, viscosity, and rheological properties with increasing fertilization levels. This study can guide the cultivation of buckwheat with the desired physicochemical properties and provide information for buckwheat-based products in the food industry.

Acidic soils have increased due to agricultural practices, climate factors, and the excessive use of nitrogen fertilizers to meet food demand. In these soils, aluminium (Al) is soluble and can be taken up by roots, but it is toxic to most plant species. Fagopyrum esculentum is able to adapt to acidic toxic aluminium conditions. Anatomical studies identifying novel potential cellular structures as sites of Al accumulation are currently lacking. This study provides an anatomical description of the cotyledons, revealing the presence of papillae and glandular trichomes at their margins. In seedlings treated with 100 μM Al, energy-dispersive x-ray spectroscopy (ESEM-EDS) analysis of the cotyledons revealed that the margin has the highest concentration of Al. The margin containing the epidermal papillae was subjected to laser microdissection, and Al was quantified using mass spectrometry with an inductively coupled plasma source ICP-MS and compared with the Al in the remaining leaf blades. The concentration of Al in the microdissected papillae was 3,460 mg Al kg -1 Dry Weight (DW), whereas the blades contained only 1,390 mg Al kg -1 DW. Moreover, histochemical tests for Al and total phenols in the epidermal papillae revealed that Al may be bound to phenolic compounds. Thus, this study demonstrated that the cotyledons of F. esculentum have epidermal papillae that can accumulate Al., (© 2024 The Author(s). Physiologia Plantarum published by John Wiley & Sons Ltd on behalf of Scandinavian Plant Physiology Society.)

The yield of Tartary buckwheat is significantly affected by continuous cropping. Melatonin plays a crucial role in plant defense mechanisms against abiotic stresses. However, the relationship between melatonin and continuous cropping tolerance remains unclear. This study aimed to analyze the physiological mechanism of melatonin in enhancing the continuous cropping tolerance (abiotic stress) of Tartary buckwheat. A field experiment was conducted on Tartary buckwheat cultivar Jinqiao 2 under continuous cropping with five melatonin application rates, 0 (Control), 10, 50, 100, and 200 μmol L -1 , applied during the early budding stage. The chlorophyll content, antioxidant enzyme activity, osmolyte and auxin (IAA) contents, root activity, rhizosphere soil nutrient content, and agronomic traits of Tartary buckwheat initially increased and then decreased with an increase in the concentration of exogenous melatonin application, with the best effects observed at 100 μmol L -1 . Compared with the Control treatment, the 100 μmol L -1 treatment decreased the contents of malondialdehyde, superoxide anion free radical, and abscisic acid (ABA) by an average of 28.79%, 27.08%, and 31.64%, respectively. Exogenous melatonin treatment significantly increased the yield of Tartary buckwheat under continuous cropping. Plants treated with 10, 50, 100, and 200 μM respectively had 1.88, 2.01, 2.20, and 1.78 times higher yield than those of the Control treatment. In summary, melatonin treatment, particularly 100 μmol L -1 , enhanced the continuous cropping tolerance of Tartary buckwheat by increasing antioxidant capacity and osmotica content, coordinating endogenous ABA and IAA content levels, and delaying senescence, ultimately increasing yield., (© 2024 Scandinavian Plant Physiology Society.)

Erosion and leaching of metal(loid)s from contaminated sites can spread pollution to adjacent ecosystems and be a source of toxicity for living organisms. Phytostabilization consists of selecting plant species accumulating little or no metal(loid)s in aerial parts to establish a vegetation cover and thus to stabilize the contaminants in the soil. Seeds of white lupin, common vetch, and buckwheat were sown in greenhouse on soils from former French mines (Pontgibaud and Vaulry) contaminated with metal(loid)s including high concentrations of As and Pb (772 to 1064 and 121 to 12,340 mg kg -1 , respectively). After 3 weeks of exposure, the growth of white lupin was less affected than that of the 2 other species probably because metal(loid) concentrations in roots and aerial parts of lupins were lower (5-20 times less Pb in lupin leaves on Pontgibaud soil and 5-10 times less As in lupin leaves on Vaulry soil than in vetch and buckwheat). To limit oxidation and/or scavenge metal(loid)s, white lupin increased the content of proline and total phenolic compounds (TPC) in leaves and roots by a factor 2 whereas buckwheat stimulated the production of TPC by a factor 1.5-2, and non-protein thiols (NPT) by factors around 1.75 in leaves and 6-12 in roots. Vetch accumulated more proline than white lupin but less NPT than buckwheat and less TPC than the 2 other plant species. The level of oxidation was however higher than in control plants for the 3 species indicating that defense mechanisms were not completely effective. Overall, our results showed that white lupin was the best species for phytostabilization but amendments should be tested to improve its tolerance to metal(loid)s., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Gibberellin (GA 3 ) is an important plant hormone involved in many physiological and developmental processes in plants. However, the physiological mechanism of GA 3 on the regulation yield and grain shell thickness of Tartary buckwheat is still unclear. In this study, the thick-shelled cultivar "Jinqiao 2" and thin-shelled cultivar "Miku 18" were used to study the effects of different concentrations (0, 50, and 100 mg L -1 ) of exogenous GA 3 and chlorocholine chloride (CCC, GA 3 synthesis inhibitor) on the cellulose content, amylase, and sucrose synthase (SS) activity in grain shell and the yield of Tartary buckwheat. The application of exogenous GA 3 can improve the cellulose content and the activity of amylase and SS in the grain shell of the two Tartary buckwheat varieties. It can also increase the main stem node number, main stem branch number, grains per plant, and yield. Compared with the control treatment (CK, 0 mg L -1 ), the 100 mg/L exogenous GA 3 treatment increased the number of grains per plant, grain weight per plant, 1000-grain weight, and yield of Jinqiao 2 by 20.1%, 41.9%, 13%, and 34.7%, respectively. These items of Miku 18 were increased by 26%, 15.2%, 10.2%, and 23.8%. The application of CCC reduced the activity of amylase and SS and cellulose content in grain shell. In addition, it decreased the main stem node number, main stem branch number, grains per plant, and yield of Tartary buckwheat. In summary, exogenous GA 3 treatment not only improved the yield of Tartary buckwheat but also increased the thickness of grain shell by enhancing the activity of amylase and SS and promoting the synthesis and accumulation of cellulose. The results can provide theoretical references for clarifying the physiological mechanism of the difference in shell thickness between Tartary buckwheat varieties., (© 2024. The Author(s).)

Starch is the main component that determines the yield and quality of Tartary buckwheat. As a quantitative trait, using quantitative trait locus (QTL) mapping to excavate genes associated with starch-related traits is crucial for understanding the genetic mechanisms involved in starch synthesis and molecular breeding of Tartary buckwheat varieties with high-quality starch. Employing a recombinant inbred line population as research material, this study used QTL mapping to investigate the amylose, amylopectin, and total starch contents across four distinct environments. The results identified a total of 20 QTLs spanning six chromosomes, which explained 4.07% to 14.41% of the phenotypic variation. One major QTL cluster containing three stable QTLs governing both amylose and amylopectin content, qClu-4-1 , was identified and located in the physical interval of 39.85-43.34 Mbp on chromosome Ft4. Within this cluster, we predicted 239 candidate genes and analyzed their SNP/InDel mutations, expression patterns, and enriched KEGG pathways. Ultimately, five key candidate genes, namely FtPinG0004897100.01, FtPinG0002636200.01, FtPinG0009329200.01, FtPinG0007371600.01, and FtPinG0005109900.01, were highlighted, which are potentially involved in starch synthesis and regulation, paving the way for further investigative studies. This study, for the first time, utilized QTL mapping to detect major QTLs controlling amylose, amylopectin, and total starch contents in Tartary buckwheat. The QTLs and candidate genes would provide valuable insights into the genetic mechanisms underlying starch synthesis and improving starch-related traits of Tartary buckwheat.

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

Buckwheat ( Fagopyrum spp. ) is an underutilized resilient crop of North Western Himalayas belonging to the family Polygonaceae and is a source of essential nutrients and therapeutics. Common Buckwheat and Tatary Buckwheat are the two main cultivated species used as food. It is the only grain crop possessing rutin, an important metabolite with high nutraceutical potential. Due to its inherent tolerance to various biotic and abiotic stresses and a short life cycle, Buckwheat has been proposed as a model crop plant. Nutritional security is one of the major concerns, breeding for a nutrient-dense crop such as Buckwheat will provide a sustainable solution. Efforts toward improving Buckwheat for nutrition and yield are limited due to the lack of available: genetic resources, genomics, transcriptomics and metabolomics. In order to harness the agricultural importance of Buckwheat, an integrated breeding and OMICS platforms needs to be established that can pave the way for a better understanding of crop biology and developing commercial varieties. This, coupled with the availability of the genome sequences of both Buckwheat species in the public domain, should facilitate the identification of alleles/QTLs and candidate genes. There is a need to further our understanding of the molecular basis of the genetic regulation that controls various economically important traits. The present review focuses on: the food and nutritional importance of Buckwheat, its various omics resources, utilization of omics approaches in understanding Buckwheat biology and, finally, how an integrated platform of breeding and omics will help in developing commercially high yielding nutrient rich cultivars in Buckwheat.

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.)

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.

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.

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., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

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 → 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.

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.)

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., (© 2024. The Author(s).)

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., (© 2024 The Authors. Advanced Science published by Wiley‐VCH GmbH.)

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., 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 Masson SAS. All rights reserved.)

Rutin, a flavonoid rich in buckwheat, is important for human health and plant resistance to external stresses. The hydrolysis of rutin to quercetin underlies the bitter taste of Tartary buckwheat. In order to identify rutin hydrolysis genes, a 200 genotypes mini-core Tartary buckwheat germplasm resource was re-sequenced with 30-fold coverage depth. By combining the content of the intermediate metabolites of rutin metabolism with genome resequencing data, metabolite genome-wide association analyses (GWAS) eventually identified a glycosyl hydrolase gene FtGH1, which could hydrolyse rutin to quercetin. This function was validated both in Tartary buckwheat overexpression hairy roots and in vitro enzyme activity assays. Mutation of the two key active sites, which were determined by molecular docking and experimentally verified via overexpression in hairy roots and transient expression in tobacco leaves, exhibited abnormal subcellular localization, suggesting functional changes. Sequence analysis revealed that mutation of the FtGH1 promoter in accessions of two haplotypes might be necessary for enzymatic activity. Co-expression analysis and GWAS revealed that FtbHLH165 not only repressed FtGH1 expression, but also increased seed length. This work reveals a potential mechanism behind rutin metabolism, which should provide both theoretical support in the study of flavonoid metabolism and in the molecular breeding of Tartary buckwheat., (© 2023 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.)

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., (© 2024. The Author(s).)

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., (© 2024. The Author(s).)

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., (© 2024. The Author(s).)

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., 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 Masson SAS. All rights reserved.)

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 (FtPAL1、FtC4H1、Ft4CL1, 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., Competing Interests: Declaration of competing interest The authors declare that there are no competing interests., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

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., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

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.

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., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

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., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Tartary buckwheat is an annual minor cereal crop with a variety of secondary metabolites, endowing it with a high nutritional and medicinal value. Flavonoids constitute the primary compounds of Tartary buckwheat. Recently, metabolomics, as an adjunct breeding method, has been increasingly employed in crop research. This study explores the correlation between the total flavonoid content (TFC) and antioxidant capacity in 167 Tartary buckwheat varieties. Ten Tartary buckwheat varieties with significant differences in flavonoid content and antioxidant capacity were selected by cluster analysis. With the use of liquid chromatography-mass spectrometry, 58 flavonoid compounds were identified, namely, 42 flavonols, 10 flavanols, 3 flavanones, 1 isoflavone, 1 anthocyanidin, and 1 proanthocyanidin. Different samples were clearly separated by employing principal component analysis and partial least-squares discriminant analysis. Eight differential flavonoid compounds were further selected through volcano plots and variable importance in projection. Differential metabolites were highly correlated with TFC and antioxidant capacity. Finally, metabolic markers of kaempferol-3- O -hexoside, kaempferol-7- O -glucoside, and naringenin- O -hexoside were determined by the random forest model. The findings provide a basis for the selection and identification of Tartary buckwheat varieties with high flavonoid content and strong antioxidant activity.

Tartary buckwheat ( Fagopyrum tataricum ) is an important plant, utilized for both medicine and food. It has become a current research hotspot due to its rich content of flavonoids, which are beneficial for human health. Anthocyanins (ATs) and proanthocyanidins (PAs) are the two main kinds of flavonoid compounds in Tartary buckwheat, which participate in the pigmentation of some tissue as well as rendering resistance to many biotic and abiotic stresses. Additionally, Tartary buckwheat anthocyanins and PAs have many health benefits for humans and the plant itself. However, little is known about the regulation mechanism of the biosynthesis of anthocyanin and PA in Tartary buckwheat. In the present study, a bHLH transcription factor (TF) FtTT8 was characterized to be homologous with AtTT8 and phylogenetically close to bHLH proteins from other plant species. Subcellular location and yeast two-hybrid assays suggested that FtTT8 locates in the nucleus and plays a role as a transcription factor. Complementation analysis in Arabidopsis tt8 mutant showed that FtTT8 could not recover anthocyanin deficiency but could promote PAs accumulation. Overexpression of FtTT8 in red-flowering tobacco showed that FtTT8 inhibits anthocyanin biosynthesis and accelerates proanthocyanidin biosynthesis. QRT-PCR and yeast one-hybrid assay revealed that FtTT8 might bind to the promoter of NtUFGT and suppress its expression, while binding to the promoter of NtLAR and upregulating its expression in K326 tobacco. This displayed the bidirectional regulating function of FtTT8 that negatively regulates anthocyanin biosynthesis and positively regulates proanthocyanidin biosynthesis. The results provide new insights on TT8 in Tartary buckwheat, which is inconsistent with TT8 from other plant species, and FtTT8 might be a high-quality gene resource for Tartary buckwheat breeding.

Tartary buckwheat protein-derived peptide (Ala-Phe-Tyr-Arg-Trp, AFYRW) is a natural active peptide that hampers the atherosclerosis process, but the underlying role of AFYRW in angiogenesis remains unknown. Here, we present a system-based study to evaluate the effects of AFYRW on H 2 O 2 -induced vascular injury in human umbilical vein endothelial cells (HUVECs). HUVECs were co-incubated with H 2 O 2 for 2 h in the vascular injury model, and AFYRW was added 24 h in advance to investigate the protective mechanism of vascular injury. We identified that AFYRW inhibits oxidative stress, cell migration, cell invasion, and angiogenesis in H 2 O 2 -treated HUVECs. In addition, we found H 2 O 2 -induced upregulation of phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), phosphorylation of nuclear factor-κB (NF-κB) p65 and nuclear translocation of NF-κB decreased by AFYRW. Taken together, AFYRW attenuated H 2 O 2 -induced vascular injury through the PI3K/AKT/NF-κB pathway. Thereby, AFYRW may serve as a therapeutic option for vascular injuries., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2023 Elsevier Inc. 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.)

Pseudo-cereals such as buckwheat ( Fagopyrum esculentum ) are valid candidates to promote diet biodiversity and nutrition security in an era of global climate change. Buckwheat hulls (BHs) are currently an unexplored source of dietary fibre and bioactive phytochemicals. This study assessed the effects of several bioprocessing treatments (using enzymes, yeast, and combinations of both) on BHs' nutrient and phytochemical content, their digestion and metabolism in vitro (using a gastrointestinal digestion model and mixed microbiota from human faeces). The metabolites were measured using targeted LC-MS/MS and GC analysis and 16S rRNA gene sequencing was used to detect the impact on microbiota composition. BHs are rich in insoluble fibre (31.09 ± 0.22% as non-starch polysaccharides), protocatechuic acid (390.71 ± 31.72 mg/kg), and syringaresinol (125.60 ± 6.76 mg/kg). The bioprocessing treatments significantly increased the extractability of gallic acid, vanillic acid, p-hydroxybenzoic acid, syringic acid, vanillin, syringaldehyde, p-coumaric acid, ferulic acid, caffeic acid, and syringaresinol in the alkaline-labile bound form, suggesting the bioaccessibility of these phytochemicals to the colon. Furthermore, one of the treatments, EC&#95;2 treatment, increased significantly the in vitro upper gastrointestinal release of bioactive phytochemicals, especially for protocatechuic acid ( p < 0.01). The BH fibre was fermentable, promoting the formation mainly of propionate and, to a lesser extent, butyrate formation. The EM&#95;1 and EC&#95;2 treatments effectively increased the content of insoluble fibre but had no effect on dietary fibre fermentation ( p > 0.05). These findings promote the use of buckwheat hulls as a source of dietary fibre and phytochemicals to help meet dietary recommendations and needs.

NAC transcription factors play a significant role in plant stress responses. In this study, an NAC transcription factor, with a CDS of 792 bp encoding 263 amino acids, was cloned from Fagopyrum tataricum (L.) Gaertn. ( F. tataricum ), a minor cereal crop, which is rich in flavonoids and highly stress resistant. The transcription factor was named FtNAC10 (NCBI accession number: MK614506.1) and characterized as a member of the NAP subgroup of NAC transcriptions factors. The gene exhibited a highly conserved N-terminal, encoding about 150 amino acids, and a highly specific C-terminal. The resulting protein was revealed to be hydrophilic, with strong transcriptional activation activity. FtNAC10 expression occurred in various F. tataricum tissues, most noticeably in the root, and was regulated differently under various stress treatments. The over-expression of FtNAC10 in transgenic Arabidopsis thaliana ( A. thaliana ) seeds inhibited germination, and the presence of FtNAC10 enhanced root elongation under saline and drought stress. According to phylogenetic analysis and previous reports, our experiments indicate that FtNAC10 may regulate the stress response or development of F. tataricum through ABA-signaling pathway, although the mechanism is not yet known. This study provides a reference for further analysis of the regulatory function of FtNAC10 and the mechanism that underlies stress responses in Tartary buckwheat.

Objectives: Buckwheat quercetin (QUE) was used as a dietary supplement to investigate the mechanism of QUE on the regulation of lipid metabolism and intestinal flora in hyperlipidemic rats., Methods: Here, using a high-fat diet-induced hyperlipidemia model, the intervention was carried out by gavage of QUE at doses of 50, 100, and 200 mg/kg. Serum lipid levels, liver biochemical parameters, and histopathologic changes in the liver and intestinal microorganisms were measured in rats by enzyme-linked immunosorbent assay, hematoxylin-eosin, and high-throughput sequencing, respectively., Results: Our results found that QUE, at a dose of 200 mg/kg, significantly reduced body weight, liver index, and lipid levels in rats (P < 0.05); improved hepatic oxidative stress; and repaired liver injury. In addition, the upregulation of beneficial bacteria, such as christensenellaceae and Bifidobacterium, in the organism increased the content of short-chain fatty acids, thus interfering with intestinal pH and improving the intestinal environment, while downregulating the relative abundance of Proteobacteria and Eubacterium&#95;coprostanoligenes&#95;group, and regulating the overproduction of butyrate. The real-time fluorescence quantitative polymerase chain reaction results found that QUE inhibited the expression of Toll-like receptor 4 (TLR4) and nuclear factor κB (NF-κB) mRNA content and blocked the activation of the TLR4/NF-κB signaling pathway, thus affecting the downregulation of lipid levels and restoring intestinal homeostasis., Conclusions: A QUE dose of 200 mg/kg may improve lipid levels and the composition of intestinal flora through the TLR4/NF-κB pathway, suggesting that proteobacteria and christensenellaceae abundance changes may be biomarkers of potential diseases., Competing Interests: Declaration of Competing Interest All authors have seen and approved the final version of the manuscript being submitted. We warrant that the article is the authors' original work, hasn't received prior publication and isn't under consideration for publication elsewhere. 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. Published by Elsevier Inc.)

Background: Tartary buckwheat protein peptides have been shown to be able to inhibit angiotensin-converting enzyme (ACE), but the exact protein type has been less studied for ACE activity inhibition, and only a few types of ACE inhibitory peptides have been reported. In this study, we purified and identified ACE inhibitory peptides from albumin hydrolysate (AH)., Results: Albumin, globulin, prolamin and glutelin were extracted from Tartary buckwheat, and their ACE active peptides were obtained by a pepsin-trypsin sequential hydrolysis process. All four hydrolysates exhibited ACE inhibitory activity, and AH displayed the strongest ACE inhibition activity and the highest peptide yield (82.28%). At 0.2 mg mL -1 , the inhibition rate of AH was 79.89%, followed by globulin hydrolysate at 71.84%, while prolamin hydrolysate and glutelin hydrolysate showed lower inhibition rates. The peptides with the highest inhibition rate were then isolated from AH using gel filtration chromatography and reversed-phase high-performance liquid chromatography, and identified using nanoscale high-performance liquid chromatography-tandem mass spectrometry. After isolation and purification, 42 ACE inhibitory peptides were identified in the fraction with the highest inhibition rate, 14 of which were completely novel discoveries in this study. These 14 peptides showed potent ACE inhibitory effects through computer analysis., Conclusion: Tartary buckwheat albumin can be used as a good source of ACE inhibitory peptides and can be further developed and utilized as edible supplements or drugs. © 2023 Society of Chemical Industry., (© 2023 Society of Chemical Industry.)

Satiety hormone cholecystokinin (CCK) plays a vital role in appetite inhibition. Its secretion is regulated by dietary components. The search for bioactive compounds that stimulate CCK secretion is currently an active area of research. The objective of this study was to evaluate the ability of buckwheat ( Fagopyrum esculentum Moench) protein digest (BPD) to stimulate CCK secretion in vitro and in vivo and clarify the structural characteristics of peptides stimulating CCK secretion. BPD was prepared by an in vitro gastrointestinal digestion model. The relative molecular weight of BPD was <10 000 Da, and peptides with <3000 Da accounted for 70%. BPD was rich in essential amino acids Lys, Leu, and Val but lacked sulfur amino acids Met and Cys. It had a stimulatory effect on CCK secretion in vitro and in vivo . Chromatographic separation was performed to isolate peptide fractions involved in CCK secretion, and five novel CCK-releasing peptides including QFDLDD, PAFKEEHL, SFHFPI, IPPLFP, and RVTVQPDS were successfully identified. A sequence length range of 6-8 and marked hydrophobicity (18-28) were observed among the most CCK-releasing peptides. The present study demonstrated for the first time that BPD could stimulate CCK secretion and clarify the structural characteristics of bioactive peptides having CCK secretagogue activity in BPD.

Menopause is a natural aging process characterized by decreased levels of sex hormones in females. Deprivation of estrogen following menopause results in alterations of dendritic arborization of the neuron that leads to neurobehavioral complications. Hormone replacement therapy is in practice to manage postmenopausal conditions but is associated with a lot of adverse effects. In the present study, the efficacy of buckwheat tartary (Fagopyrum tataricum) whole seed extract was investigated against the neurobehavioral complication in middle-aged ovariectomized rats, which mimic the clinical postmenopausal condition. Hydroalcoholic extraction (80% ethanol) was done, and quantification of major marker compounds in the extract was performed using HPLC. Oral treatment of the extract following the critical window period rescued the reconsolidation process of spatial and recognition memory, as well as depression-like behavior. Gene expression analysis disclosed elevated oxidative stress and neuroinflammation that largely disturb the integrity of the blood-brain barrier in ovariectomized rats. Gfap and Pparγ expression also showed reactive astrogliosis in the rats subjected to ovariectomy. The extract treatment reverted the elevated oxidative stress, neuroinflammation and expression of the studied genes. Furthermore, protein expression analysis revealed that Gsk-3β was activated differentially in the brain, as suggested by β-catenin protein expression, which was normalized following the treatment with extract and rescued the altered neurobehavioral process. The results of the current study concluded that Fagopyrum tataricum seed extract is better option to overcome the neurobehavioral complications associated with the menopause., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

This study aims to clarify the effects of different concentrations of sodium chloride on the carbon and nitrogen metabolism and yield of Tartary buckwheat. The salt-sensitive cultivar Yunqiao 2 was pot-grown and treated with four salt concentrations including 0, 2, 4, and 6 g kg -1 . The root morphology index increased from seedling stage to maturate stage. The content of soluble protein in the leaves reached the maximum at the anthesis stage, and the other substances content and the enzymes activity related to carbon and nitrogen metabolism reached the maximum at the grain filling stage. The root morphology index, root activity; invertase, amylase, sucrose synthase, and sucrose phosphate synthase activities; nitrate-nitrogen, ammonium nitrogen, and soluble protein content; and nitrate reductase and glutamate synthase activities increased first and reached the maximum at 2 g kg -1 treatment and then decreased with increasing salt stress concentration. The content of soluble sugars and sucrose and the activity of glutamate dehydrogenase increased continuously with increasing salt concentration, and reached the maximum in the 6 g kg -1 treatment. The grain number per plant, 100-grain weight, and yield per plant increased first and reached the maximum at 2 g kg -1 treatment and then decreased with increasing salt stress concentration. In summary, moderate salt stress (2 g kg -1 ) can promote the root growth, increase the content of carbon and nitrogen metabolism-related substances and enzyme activity, and increase the yield per plant of Tartary buckwheat., (© 2023. The Author(s).)