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[Immunopharmacological study of buckwheat hypersensitivity (author's transl)].

Effect of dialysate from buckwheat extract on immediate hypersensitivity reactions. To investigate the hypersensitivity reaction of buckwheat, the aqueous extract of buckwheat was separated into dialysate (BWD) and non-dialysate (BWND). BWD neither decreased a release of anaphylactic mediator from the chopped lung tissue nor inhibited Schultz-dale reaction in the ileum of guinea pig sensitized with BWND. Heterologous PCA mediated by rabbit anti-BWND serum in guinea pig and homologous PCA in rats using anti-BWND IgE serum were inhibited by i.d. or i.v. treatment of BWD. Radioallegosorbent test (RAST) and PK reaction were inhibited in a dose dependent fashion by BWD mixed with human serum sensitive to buckwheat, although BWD required approximately 10,000 times more than BWND to produce a RAST inhibition. RAST, however, was not affected when BWD was added to 125I-labelled anti-human IgE. Anti-buckwheat IgE antibody in sensitized animals and atopic patients was specifically absorbed with an insoluble copolymer of BWD-conjugated BSA. BWD did not form a precipitin line against anti-BWND IgG antibody in immunoelectrophoresis. Rat IgE serum-induced degranulation and histamine release from mast cells were inhibited in a dose dependent fashion by BWD and the inhibition was specific to anti-BWND IgE antibody. BWD may be useful as an hyposensitization agent in buckwheat hypersensitivity, since BWD is a haptenic substance capable of neutralizing specific IgE antibody on mast cells.

Anaphylaxis

[Immunopharmacological study of buckwheat hypersensitivity III. Effect of dialysate-conjugated T cell mitogen on IgE formation (author's transl)].

It has already been reported that dialysate (BWD) separated from the aqueous extract of buckwheat is a haptenic substance capable of neutralizing specific IgE antibody both on mast cells and on B cells in some species. The present work represents a study carried out to determine the effect of BWD conjugated with T cell mitogen such as phyothemagglutinin-P (PHA) and concanavalin A (Con A) on IgE formation in mice. Anti-buckwheat IgE formation was little affected by the pretreatment of these conjugates given i.v. 3 days before the immunization, but was effectively suppressed by the pretreatment of BWD-PHA or BWD-Con A given i.p. together with incomplete Freund's adjuvant 2 weeks before the immunization. BWD-PHA induced a more potent suppression of IgE formation as compared with BWD-Con A, Con A or PHA. The transfer of T cells obtained from spleen cells primed with BWD-PHA, which responded with buckwheat and exerted a slight helper function for adoptive anti-DNP IgE formation to DNP-buckwheat, suppressed anti-buckwheat IgE formation in recipients with no effect on the anti-KLH IgE response. Our findings suggest that suppressor T cells specific to buckwheat are induced in spleen cells of mice treated with BWD-PHA.

Animals

Comparative analysis of DREB gene family in buckwheat: the role of FtDREB02 in the delphinidin biosynthesis and drought stress response.

Dehydration response element binding (DREB) transcription factors play a pivotal role in plant abiotic stress responses, but its evolutionary and functional characterization in buckwheat remains unexplored. Here, we conducted a comprehensive analysis of the DREB gene family across three buckwheat species, revealing segmental duplication as the primary driver of family expansion and potential purifying selection during evolution. A FtDREB02 gene, classified as group A2, was identified through genome-wide association analysis (GWAS) on drought tolerance and delphinidin content. Functional validation in Arabidopsis thaliana and the hairy root of Tartary buckwheat (Fagopyrum tataricum) demonstrated that overexpression of this gene promotes delphinidin biosynthesis and enhances plant resistance to water scarcity. Through the integration of DAP-seq and PEG transcriptome cluster analysis, a FtANS candidate was screened. Functional studies showed that FtDREB02 regulates delphinidin content by binding directly to DRE elements of the FtANS promoter. This research identifies and comprehensively analyzes the DREB family within buckwheat species, elucidating the regulatory mechanisms of FtDREB02 in controlling flavonoid biosynthesis and drought resistance, providing potential genetic resources for breeding buckwheat varieties with excellent agronomic traits.

Anthocyanins

[Immunopharmacological study of buckwheat hypersensitivity (II). Effect of dialysate on antibody formation (author's transl)].

It was reported that dialysate (BWD) separated from the aqueous extract of buckwheat was a haptenic substance capable of neutralizing IgE antibody on mast cells and that the activity was specific to non-dialysate (BWND)-induced hypersensitivity reaction. The effect of BWD on antibody formation was investigated in the present paper. In rats, anti-buckwheat IgE formation was slightly depressed by the administration of BWD, but both anti-DNP IgE and IgG formations in rats immunized with DNP-BWND were unaffected. In mice, anti-buckwheat IgE formation was suppressed by BWD administration. A good correlation was noted between a decrease of surface IgE population on B cells and that of IgE titer in serum. However, the helper function of T cells for adoptive anti-DNP IgE formation was little affected by BWD treatment. Lymphocyte transformation to BWND and other non-specific mitogens using spleen cells obtained from sensitized mice was inhibited in a dose-dependent fashion by the addition of BWD. However, lymphocyte transformation using spleen cells pretreated with BWD was not affected by other non-specific mitogens except for BWND and pokeweed mitogen.

Animals

The Tartary Buckwheat FtMYB46-FtNRAMP3 Module Enhances Plant Lead and Cadmium Tolerance.

The presence of toxic heavy metals lead (Pb) and cadmium (Cd) in polluted soil damage crop production and consequently harms human and livestock health. Tartary buckwheat (Fagopyrum tataricum) is a potential model plant for heavy metal phytoremediation because of its valuable characteristics of high heavy metal tolerance and abundant biomass production. Here, we report that the Tartary buckwheat FtMYB46-FtNRAMP3 module enhances plant Pb and Cd tolerance. RNA sequencing analysis showed that Pb treatment specifically induced expression of FtNRAMP3, a member of the NRAMP (Natural Resistance-Associated Macrophage Protein) transporter gene family. Further cytological and biochemical analysis revealed that FtNRAMP3 was localised to the plasma membrane and significantly contributed to increased tolerance to Pb and Cd in yeast cells. Consistently, transgenic overexpression of FtNRAMP3 in Arabidopsis significantly increased plant tolerance to Pb and Cd applications, reducing Pb concentration but increasing Cd concentration in the overexpression transgenic plants. Subsequent yeast one-hybrid and electrophoretic mobility shift assays showed that the transcription factor FtMYB46 directly binds to the FtNRAMP3 promoter. Further, FtMYB46 promoted FtNRAMP3 expression and increased plant Pb and Cd tolerance. Overall, this study demonstrates the important role of the FtMYB46-FtNRAMP3 module and its potential value in the phytoremediation of Pb and Cd stress.

Cadmium

Interference of L-alpha-aminooxy-beta-phenylpropionic acid with phenylalanine metabolism in buckwheat.

L-alpha-Aminooxy-beta-phenylpropionic acid (AOPP), a potent competitive inhibitor of phenylalanine ammonia-lyase (PAL), blocked light-induced phenylpropanoid synthesis in excised buckwheat hypocotyls and produced an up to 40-fold increase in the endogenous phenylalanine concentration, while the level of all other amino acids was hardly affected. After a 24 h incubation in the light in the presence of 0.3 or 1 mM AOPP phenylalanine alone constituted about 25% of the total soluble amino acids, compared to appr. 1% in the controls. In the presence of AOPP illuminated hypocotyls accumulated nearly 3 times more phenylalanine than hypocotyls kept in the dark, indicating an enhancing effect of light on the flow of carbon through the shikimate pathway. Exogenously added [14C]phenylalanine was extensively metabolized by control tissue, but accumulated in AOPP treated tissue. In the presence of AOPP radioactivity from [14C]shikimate accumulated predominantly in phenylalanine, and the flow of shikimate into tyrosine and phenylalanine was not affected by the inhibitor. Therefore, under these conditions no feedback control of phenylalanine and tyrosine synthesis from shikimate is apparent in buckwheat hypocotyls.

Amino Acids

[Change in the sterol and phospholipid makeup during the hydrothermal processing of buckwheat grain].

The effect of different modes of the hydrothermal treatment of buckwheat and of the grit cooking on a change in the composition of sterols and phospholipids was investigated. While not changing the qualitative composition of sterols and phospholipids, the hydrothermal and culinary treatment was found to produce some changes in the correlation of their individual fractions.

Cooking

Nutritional evaluation of dry-roasted navy bean flour and mixtures with cereal proteins.

A navy bean flour prepared by dry roasting in a salt bed as a medium of heat exchange was found to have a higher PER than beans which had been autoclaved in the conventional manner. This difference was attributed to a small but significant improvement in the digestibility of the protein. The PER of the roasted beans was higher than the autoclaved beans even in the presence of supplemental methionine. When various proportions of roasted beans and corn were fed at a level of 8.3% protein in the diet, a mixture in which 40% to 60% of the protein was provided by either beans or corn had a PER essentially the same as casein. Diets containing roasted beans and various cereal grains (oats, barely, buckwheat, wheat germ, and rice) were formulated in proportions calculated to give the highest chemical scores. In most cases the PER' were not significantly different from that of casein, and, in the case of rice, the PER was higher than that of casein. Supplementation of such diets with their first limiting amino acid failed to produce a further enhancement of the PER.

Amino Acids, Essential

In silico, in vitro, and in vivo characterization of thiamin-binding proteins from plant seeds.

Thiamin, an essential micronutrient, is a cofactor for enzymes involved in the central carbon metabolism and amino acid pathways. Despite efforts to enhance thiamin content in rice by incorporating thiamin biosynthetic genes, increasing thiamin content in the endosperm remains challenging, possibly due to a lack of thiamin stability and/or a local sink. The introduction of storage proteins has been successful in several biofortification strategies, and similar efforts targeting thiamin have been performed, leading to a 3-4-fold increase in white rice. However, only one thiamin-binding protein (TBP) sequence has been described in plants, more specifically from sesame seeds. Therefore, we aimed to identify and characterize TBPs, as well as to evaluate the effect of their expression on thiamin concentration, using a comprehensive approach integrating in silico, in vitro, and in vivo methods. We identified the sequences of putative TBPs from Oryza sativa (Os, rice), Fagopyrum esculentum (Fe, buckwheat), and Zea mays (Zm, maize) and pinpointed the thiamin-binding pockets through molecular docking. FeTBP and OsTBP contained one pocket with binding affinities similar to the Escherichia coli TBP, a well-characterized TBP, supporting their function as TBPs. In vivo expression studies of TBPs in tobacco leaves and rice callus resulted in increased thiamin levels, with FeTBP and OsTBP showing the most pronounced effects. Additionally, thermal shift assays confirmed the thiamin-binding capabilities of FeTBP and OsTBP, as observed by the significant increases in melting temperatures upon thiamin binding, indicating protein stabilization. These findings offer new insights into the diversity and function of plant TBPs and highlight the potential of FeTBP and OsTBP to modulate thiamin levels in crop plants.

Thiamine

Quantitative study on anomeric forms of glucose produced by alpha-glucosidases.

Anomeric forms of glucose produced from phenyl alpha-maltoside, maltose, or phenyl alpha-glucoside have been determined quantitatively by simultaneous measurements of optical rotation and reducing power, for eight kinds of glucose-producing 1,4-alpha-glucosyl hydrolases, including glucose-forming amylase from human urine, and alpha-glucosidases from pig serum, honey bee, buckwheat seed, rice seed, sugar beet seed, flint corn seed, and brewer's yeast. All the eight enzymes studied were found to produce alpha-glucose exclusively.

Animals