Antioxidant activity of tartary (Fagopyrum tataricum (L.) Gaertn.) and common (Fagopyrum esculentum moench) buckwheat sprouts

This study compared the differences of two types of buckwheat sprouts, namely, common buckwheat ( Fagopyrum esculentum Moench) and tartary buckwheat ( Fagopyrum tataricum (L.) Gaertn.), in general composition, functional components, and antioxidant capacity. The ethanol extracts of tartary buckwheat sprouts (TBS) had higher reducing power, free radical scavenging activity, and superoxide anion scavenging activity than those of common buckwheat sprouts (CBS). As for chelating effects on ferrous ions, CBS had higher values than TBS. Rutin was the major flavonoid found in these two types of buckwheat sprouts, and TBS was 5 fold higher in rutin than CBS. The antioxidant effects of buckwheat sprouts on human hepatoma HepG2 cells revealed that both of TBS and CBS could decrease the production of intracellular peroxide and remove the intracellular superoxide anions in HepG2 cells, but TBS reduced the cellular oxidative stress more effectively than CBS, possibly because of its higher rutin (and quercetin) content.     

Origin of cultivated Tatary buckwheat (Fagopyrum tataricum Gaertn.) revealed by RAPD analyses

The phylogenetic relationships among cultivated landraces and natural populations of wild subspecies of Tatary buckwheat were investigated at the individual level by constructing a phylogenetic tree based on RAPD markers. As the PCR templates, DNA of individuals rather than bulked samples, was used. Ten individuals from 10 cultivated landraces, 71 individuals from 21 natural populations of wild subspecies, and 7 individuals from 3 weedy Tatary buckwheat were provided for RAPD analyses. Three groups were recognized: (1) all cultivated landraces and wild subspecies from central Tibet and northern Pakistan; (2) 10 individuals of wild subspecies from northwestern Yunnan; (3) the remaining individuals of wild subspecies from northwestern Yunnan and all individuals of wild subspecies from Sichuan. Group (2) was phylogenetically closely related to group (1). The origin of cultivated Tatary buckwheat, the hybrid origin of weedy Tatary buckwheat and of the wild populations from central Tibet and northern Pakistan are discussed. We arrive at the conclusion that cultivated Tatary buckwheat probably originated in northwestern Yunnan in China.     

Variation of major minerals and trace elements in seeds of tartary buckwheat (Fagopyrum tataricum Gaertn.)

Tartary buckwheat (Fagopyrum tataricum Gaertn.) contains protein of high nutritional value, polyphenols, vitamins and minerals. It is one of the most important minor crops in China and has a great potential as a health and functional food. However, information on the elemental mineral composition of its seeds remains limited. The concentrations of Cu, Zn, Fe, K, and Mg in seeds of 123 tartary buckwheat accessions from the same cultivation were studied by means of flame atomic absorption spectrometry. The results revealed that the average concentrations of Cu (x₁), Zn (x₂), Fe (x₃), K (x₄), and Mg (x₅) elements in the accessions are 19.49 (with a range of 5.74–36.01 mg/kg), 27.41 (8.44–66.63 mg/kg), 656.24 (21.8–3,990 mg/kg), 3,639.23 (1,737–5,831 mg/kg), and 1,523.89 mg/kg (729–3,104 mg/kg) respectively. Among them, Fe concentration has the highest coefficient of variation (114.7 %). The results also revealed five significant positive correlations among Cu, Zn, Fe, K, and Mg concentrations. Therefore, distinct genotypes with high concentration of mineral elements should be effective for the development of special buckwheat varieties and improvement of its food nutritional quality.     

Purification and characterization of the antitumor protein from Chinese tartary buckwheat (Fagopyrum tataricum Gaertn.) water-soluble extracts

A novel antitumor protein, coded as TBWSP31, was isolated from tartary buckwheat water-soluble extracts and purified by DEAE-Sepharose Fast Flow anion exchange, Sephadex G-100 gel filtration, and Sephacryl S-200 gel filtration column chromatography. TBWSP31 was identified to a homogeneous fraction by native PAGE. The antitumor effect of TBWSP31 against human mammary cancer cell Bcap37 was measured by an MTT assay. TBWSP31 showed higher antitumor activity, and time- and concentration-dependent effects were observed. SDS-PAGE analysis showed that TBWSP31 is composed of a single polypeptide with an approximate molecular weight of 57 kDa. TBWSP31 was rich in Glx (Gln+Glu), Arg, and Asx (Asp+Asn) according to amino acid analysis. Secondary structural analysis by CD spectroscopy revealed that TBWSP31 has the following: alpha-helix (33.9%), beta-sheet (22.8%), beta-turn (11.3%), and random coil (32.0%).     

Genetic diversity and relationships among cultivated and wild accessions of tartary buckwheat (Fagopyrum tataricum Gaertn.) as revealed by RAPD markers

The methodology of sampling and the selection of a proper marker systemfor the analysis of accessions are major concerns in the evaluation of gene bank material. In our study the RAPD analysis of bulked DNA samples and single seedsDNA was successfully employed to evaluate intra- and inter-population geneticvariability of cultivated and wild tartary buckwheat accessions. The bulkingapproach enabled the distinction of all 40 analysed accessions and theirseparation into geographically well defined clusters. Three wild populations,two from Sichuan and one from Qinghai, formed a group that was geneticallyrelatively distant from wild populations from Tibet and all cultivatedlandraces which, on the other hand, exhibited very close relationships. Thesingle seed study that was used after bulked DNA analysis provided detailedinformation of the genetic variation present within some accessions of specialinterest. A moderate level of genetic variability was detected betweenaccessions and the variability was partitioned into between- andwithin-population components. On average, most of the detected variation ispresent between F. tataricumpopulations. The genetic and geographic distribution of variability is furtherdiscussed. We demonstrated the usefulness of combining bulking and single seedstudy approaches for the effective evaluation of genetic variability inF. tataricum accessions that couldalso have wider applicability in the management of plant genetic resources andphylogenetic studies.     

Phylogenetic position of east Tibetan natural populations in Tartary buckwheat (Fagopyrum tataricum Gaert.) revealed by RAPD analyses

Phylogenetic relationships among cultivated landraces and natural populations of wild subspecies of Tartary buckwheat were investigated by constructing an NJ tree based on RAPD markers focussing on east Tibetan natural populations. Ten plants from three cultivated landraces and 29 plants from five natural populations of wild subspecies in eastern Tibet were used for RAPD analyses. The wild subspecies from eastern Tibet was classified into three types; (1) same type as cultivated landraces; (2) closely related to natural populations of northwestern Yunnan; and (3) an exceptional population, Zhuka, which was closely related to Sichuan populations. Since the type (2) is considered as the wild ancestral type of cultivated Tartary buckwheat, we conclude that eastern Tibet too may be one of the center of origin of this crop.     

Extraction of rutin from buckwheat (Fagopyrum esculentumMoench) seeds and determination by capillary electrophoresis.

The content of the flavonoid rutin was determined in different milling fractions of buckwheat seeds and in buckwheat stems, leaves, and flowers. The extraction was performed by using a solvent containing 60% of ethanol and 5% of ammonia in water. The extracts were analyzed by capillary electrophoresis (running buffer of 50 mM borate (pH 9.3), 100 mM sodium dodecyl sulfate; determination at 380 nm). In bran fractions the concentration of rutin was 131-476 ppm, and in flour fractions 19-168 ppm. On average, about 300, 1000, and 46000 ppm of rutin were found in leaves, stems, and flowers, respectively. The results indicate that buckwheat could be an important nutritional source of flavonoids, especially in countries with a low mean daily flavonoid intake.     

Distribution of Vitamin E, squalene, epicatechin, and rutin in common buckwheat plants (Fagopyrum esculentum Moench)

Buckwheat leaves and young parts of the plant are consumed in some countries as a vegetable. Green flour, obtained by milling of the dried plants, is used as a natural food colorant. The distribution of vitamin E, squalene, epicatechin, and rutin (as the most important antioxidants) within buckwheat plants, as well as changes of their content within leaves during the growing season, were determined by GC-MS and HPLC analyses. alpha-Tocopherol was found as the main component of vitamin E in all parts of the plant; epicatechin and squalene were also detected. For the use of buckwheat as an antioxidant source in the human diet, the most suitable part of the plants seems to be the leaves and the flowers at the stage of full flowering due to the considerable amounts of rutin and epicatechin. alpha-Tocopherol content correlates positively with temperature, drought, and duration of solar radiation. Certain differences appear among varieties of buckwheat, especially in their squalene and rutin contents.     

Guava fruit (Psidium guajava L.) as a new source of antioxidant dietary fiber.

Guava (Psidium guajava L.) is a tropical fruit, widely consumed fresh and also processed (beverages, syrup, ice cream, and jams). Pulp and peel fractions were tested, and both showed high content of dietary fiber (48.55-49.42%) and extractable polyphenols (2.62-7.79%). The antioxidant activity of polyphenol compounds was studied, using three complementary methods: (i) free radical DPPH* scavenging, (ii) ferric reducing antioxidant power assay (FRAP), and (iii) inhibition of copper-catalyzed in vitro human low-density lipoprotein (LDL) oxidation. All fractions tested showed a remarkable antioxidant capacity, and this activity was correlated with the corresponding total phenolic content. A 1-g (dry matter) portion of peel contained DPPH* activity, FRAP activity, and inhibition of copper-induced in vitro LDL oxidation, equivalent to 43 mg, 116 mg, and 176 mg of Trolox, respectively. These results indicate that guava could be a suitable source of natural antioxidants. Peel and pulp could also be used to obtain antioxidant dietary fiber (AODF), a new item which combines in a single natural product the properties of dietary fiber and antioxidant compounds.     

Metabolomic analysis of phenolic compounds in buckwheat (Fagopyrum esculentum M.) sprouts treated with methyl jasmonate

The effects of exogenous methyl jasmonate (MeJA) on phytochemical production in buckwheat sprouts cultivated under dark conditions (0, 1, 3, 5, and 7 d) were investigated by metabolomic analysis, using ultra performance liquid chromatography-quadrupole-time-of-flight (UPLC-Q-TOF) mass spectroscopy (MS) and partial least-squares-discriminant analysis (PLS-DA). MeJA-treated and control groups showed no differences in growth but were clearly discriminated from each other on PLS-DA score plots. The metabolites contributing to the discrimination were assigned as chlorogenic acid, catechin, isoorientin, orientin, rutin, vitexin, and quercitrin, which have various health effects. Moreover, isoorientin, orientin, rutin, and vitexin were assigned as the main phytochemicals of sprouts cultivated under dark conditions. The accumulation of these metabolites caused the phenolic compound content and antioxidant activity of the sprouts to increase. Further, this study revealed that their accumulation resulted from the stimulation of the phenylpropanoid pathway by MeJA treatment. Therefore, these metabolites may be useful for better understanding the effects of MeJA on buckwheat sprout phytochemicals and contribute to improving the functional quality of the sprouts.     

Social and environmental influences on tartary buckwheat (<Emphasis Type="Italic">Fagopyrum tataricum</Emphasis> Gaertn.) varietal diversity in Yunnan,China

Effective conservation strategies aimed to protect crop genetic resources require multiple sources of information. We used a combination of AFLP genotyping and farmer surveys to understand the extent, distribution and management of tartary buckwheat (Fagopyrum tataricum Gaertn.) diversity in its center of origin in Yunnan Province, China. We found genetic evidence of gene flow in tartary buckwheat throughout the study area, with small but statistically significant regional and village-level components. We also found genetic differentiation by seed color. Although most farmers reported exchanging seed in localized kinship networks, our results imply homogenizing gene flow is occurring. Yi ethnic farmers tend to plant more buckwheat than non-Yi farmers, and we found that in some communities, Yi farmers serve as seed sources for farmers of other ethnicities. Different tartary buckwheat varieties did not have different end uses; rather farmers maintained varietal diversity in order to protect crop yield and quality. Individual farmers’ seed exchange practices reflect their ideas about components of seed quality, as well as priorities in protecting buckwheat yield. From the standpoint of genetic resources conservation, the presence of a culturally rich farmer exchange network and hierarchical structuring of tartary buckwheat genetic diversity demonstrates the importance of maintaining an interlinked community of tartary buckwheat farmers in Yunnan.     

Exudation of allelopathic substances in buckwheat (Fagopyrum esculentum Moench)

Root exudates of the common buckwheat, especially phenolic compounds, were studied. Their contents, both in the soil during the growing season and in agar medium during germination, were determined by HPLC and GC-MS. The allelopathic activity of the soil from a buckwheat stand was evaluated, as well. Palmitic acid, squalene, epicatechin, vitexin, a gallic acid derivative, and a quercetin derivative were the main compounds of the agar medium. In the soil, palmitic acid methyl ester, vanillic acid, rutin, a gallic acid derivative, and a 4-hydroxyacetophenone derivative were identified. The effects of vitexin, squalene, epicatechin, 4-hydroxyacetophenone, and vanillic and gallic acids were tested on eight plant species. Inhibitive effects were observed in the cases of 4-hydroxyacetophenone and vanillic and gallic acids. Comparisons of the identified compounds and inhibitive effects of soil extracts indicated that palmitic acid and the gallic acid derivative probably have an important function in the allelopathic root response of buckwheat.     

Determination of rutin, catechin, epicatechin, and epicatechin gallate in buckwheat Fagopyrum esculentum Moench by micro-high-performance liquid chromatography with electrochemical detection

A simple and sensitive method has been developed for determining rutin, catechin, epicatechin, and epicatechin gallate in buckwheat (Fagopyrum esculentum Moench) flour and seeds by micro-high-performance liquid chromatography with electrochemical detection. Chromatography was performed using an octadecylsilica column, acetonitrile-water-formic acid (13:87:1, v/v/v) as a mobile phase, and an applied potential at +0.5 V vs Ag/AgCl. We found that Japanese buckwheat flour contains rutin (12.7 mg/100 g), catechin (3.30 mg/100 g), epicatechin (20.5 mg/100 g), and epicatechin gallate (1.27 mg/100 g). The relative standard deviations for rutin, catechin, epicatechin, and epicatechin gallate peak heights were less than 0.86% (n = 5). The detection limit of rutin was 0.86 ng/mL. Moreover, the present method was applied to the distribution analysis of these compounds in buckwheat seed. The embryo proper and cotyledons of a mature buckwheat seed contained rutin with the highest concentration as compared to other parts. This method is useful in determining rutin, catechin, epicatechin, and epicatechin gallate in buckwheat with a small amount of sample for quality control in the food industry.     

Performance of common buckwheat (Fagopyrum esculentum M.) in response to row spacing under ridge and furrow cropping systems in a semiarid region of China

Ridge and furrow systems as well as the wide-narrow row spacing are recognized as good management practices in crop production. To obtain some available information concerning the suitable agricultural practices for buckwheat cultivation in arid and semi-arid area, a two-year field study was conducted to investigate the performance of common buckwheat grown under six cropping patterns: conventional flat single/double row planting (NS/ND); furrow single/double row planting (FS/FD); ridge single/double row planting (RS/RD); single row planting with equal row spacing and double row planting with wide-narrow row spacing practice. Plants grown under the FD system exhibited higher water use efficiency (WUE) and rainfall use efficiency (RUE) than plants cultivated with the NS system, increases with 13.7% and 21.9%, respectively, in 2014, while 9.8% and 14.0%, respectively, in 2015. Plants of the FD system also displayed the greatest leaf area index and canopy openness at growth stage of 40, 60, and 80 days after sowing, maximum biomass production (13.96 t ha^?1) and grain yield (3486.2 kg ha^?1) in 2014, and the least pronounced soil nitrate consumption as compared to plants from other tillage systems. We conclude that FD is the optimum planting pattern for common buckwheat cultivation in semi-arid and arid environments.     

Genetic diversity of Fagopyrum homotropicum, a wild species related to common buckwheat

Nineteen natural populations of Fagopyrum homotropicum, a self-fertilizing close relative of common buckwheat, from the Yunnan and Sichuan provinces of China were investigated for their chromosome number and allozyme variation at 16 loci of 11 enzymes. Three populations, 'Deqin', 'Zhongdian', and 'Xiancheng', were revealed to be allotetraploid. Judging from allozyme constitution of the tetraploid and their possible progenitors, diploid progenitors are probably the diploid population of F. homotropicum from Lijiang and a natural population of F. esculentum ssp. ancestralis. Diploid populations of F. homotropicum are fixed for a given allele at almost all isozyme loci. Allozyme variation has been maintained in natural populations mainly by fixing different alleles in different populations, as they are highly differentiated among the populations (G_st = 0.969). The position of populations in the phylogenetic tree constructed from genetic distance nearly corresponded with the geographical position of the populations.     

Characterization of an aspartic proteinase activity in buckwheat (Fagopyrum esculentum Moench) seeds

The pepstatin A sensitive acidic proteolytic activity of total protein extracts of buckwheat seeds has been analyzed in developing, mature, and germinating seeds by activity measurements as well as by electrophoretic and immunochemical techniques. Immunoblot analysis using cross-reactive antibodies raised against barley phytepsin suggested that specific proteolytic activity could be attributed to a 47 kDa heterodimeric polypeptide, composed of two subunits: 31 and 16 kDa polypeptides. The analysis of time course expression revealed that the 47 kDa heterodimer accumulated during seed maturation starting from 12 days after pollination and was also present at the beginning of germination. Milk-clotting activity of this proteinase was also indicated.     

Effect of buckwheat (Fagopyrum esculentum) on soil‐phosphorus availability and organic acids

As a cover crop, buckwheat (Fagopyrum esculentum) may increase soil‐P availability. Buckwheat was grown in low‐P and P‐fertilized field plots, and organic anions were measured in rhizosphere soil. Soil‐P availability was not affected by buckwheat, but the concentration of rhizosphere tartrate^2– was significantly higher (p < 0.005) in low‐P vs. P‐fertilized plots. This suggests that organic‐anion root exudation may have a role in buckwheat‐rhizosphere P dynamics.     

Trace element water improves the antioxidant activity of buckwheat (Fagopyrum esculentum Moench) sprouts

Buckwheat (Fagopyrum esculentum Moench) was grown in trace element water (TEW) (100, 200, 300, 400, and 500 ppm) and deionized water (DIW) to evaluate whether the beneficial effects of trace elements on the antioxidant activity could be accomplished with the supplement of TEW. At 300 ppm, TEW significantly increased the Cu, Zn, Mn, and Fe contents in buckwheat sprout but not the Se content. However, the levels of rutin, isoorientin, vitexin, and isovitexin did not differ between buckwheat sprouts grown in TEW and DIW. The ethanolic extract from buckwheat sprout grown in 300 ppm of TEW showed higher ferrous ion chelating activity and inhibitory activity toward lipid peroxidation than that grown in DIW. The extract in the TEW group also enhanced intracellular superoxide dismutase activity and lowered reactive oxygen species and superoxide anion in the human Hep G2 cell. It was concluded that TEW could increase the antioxidant activities of buckwheat sprouts.     

Chemical and productive properties of two Sardinian milk thistle (Silybum marianum (L.) Gaertn.) populations as sources of nutrients and antioxidants

Silybum marianum (L.) Gaertn. (milk thistle), grown as a medicinal plant in several countries, is considered as a weed in pastures and cereal crops but also as an interesting plant for biomass production. As an additional contribution to the full exploitation of a such promising species, two Sardinian populations of S. marianum were investigated for chemical composition, bioactive compounds and antioxidant properties at vegetative and reproductive stages. Dry matter yield was affected by the phenogical stage and differed between populations, ranging from 148 to 246 g plant^?1. Chemical composition did not differ between populations. Antioxidant capacity detected by means of ABTS [(2,2′-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt)] and by DPPH (1,1-diphenyl-2-picrylhydrazyl) methods ranged from 3.45 to 5.42 and 3.83 to 6.32 mmol/100 g dry weight of Trolox equivalent antioxidant capacity, respectively. Differences in antioxidant capacity and bioactive compound contents in the different plant organs were found and also a significant linear correlation between antioxidant capacity and total phenolics and flavonoids, at flowering compared to vegetative stage. Research highlights antioxidant capacity in different organs of milk thistle and encourages the exploitation of biomass also as functional food, source of natural antioxidants and as a complementary fodder.     

Expression, cloning, and immunological analysis of buckwheat (Fagopyrum esculentum Moench) seed storage proteins

cDNA of buckwheat (Fagopyrum esculentum Moench) was isolated from immature seeds harvested 14 days after pollination. Two genes, designated FA02 and FA18, were found to encode legumin-like proteins and were expressed during seed development. The deduced amino acid sequence of FA02 was identical to the N-terminal amino acid domain of BW24KD, which was believed to be a major buckwheat allergen (Urisu, A.; Kondo, Y.; Morita, Y.; Yagi, E.; Tsuruta, M.; Yasaki, T.; Yamada, K.; Kuzuya, H.; Suzuki, M.; Titani, K.; Kurosawa, K. Isolation and characterization of a major allergen in buckwheat seeds. In Current Advances in Buckwheat Research; Shinshu University Press: Matsumoto, Japan, 1995; pp 965--974). It was predicted that FA02 would be cleaved to generate two separate components, a 41.3 kDa alpha-subunit and a 21 kDa beta-subunit. Antiserum was raised against the deduced FA02 beta-subunit, and immunoblotting of total protein from buckwheat seeds (F. esculentum M. and Fagopyrum tartaricum Gaertn.) revealed that several groups of proteins reacted with the antiserum. Polypeptides in the 23--25 kDa range displayed the greatest reactivity.