Tartary Buckwheat Cultivars Research Articles

Effect of Auxins on Anthocyanin Accumulation in Hairy Root Cultures of Tartary Buckwheat Cultivar Hokkai T10

This study investigated the influence of auxins on the growth of hairy roots and accumulation of anthocyanins, including cyanidin 3-O-glucoside (C3gl) and cyanidin 3-0-rutinoside (C3r), in the hairy root culture of tartary buckwheat cultivar Hokkai TIO. C3gl and C3r contents were evaluated using high- performance liquid chromatograph (HPLC). Four auxins, 2,4-dichlorophenoxyacetic acid (2,4-D), indoleacetic acid (IAA), indolebutyric acid (IBA) and naphthaleneacetic acid (NAA), were added to the medium of the hairy root cultures at diverse concentrations. IAA, IBA and 2,4-D promoted the growth of hairy roots since the dry weight of the roots was slightly higher than or comparable with that of the control. However, NAA at all concentrations suppressed the growth of hairy roots. Generally, auxin treatments resulted in higher accumulation of C3gl and C3r than that of the control except for 2.85 μM IAA and 2.69 μM NAA. The amount of C3gl and C3r after treatment with 4.92 μM IBA was the highest among all treatments and was 3.24 times more than that of the control. Our results suggested that auxins at appropriate concentrations might facilitate hairy root growth of tartary buckwheat and enhance the production of C3gl and C3r.

Seed Oils of Five Black Tartary Buckwheat Cultivars with Biochemical Characterization and Antioxidant Properties

AbstractBlack tartary buckwheat oils (BTBOs) were extracted from five major industrial tartary buckwheat cultivars grown under similar agronomical activities and environmental conditions. These oils were characterized for the bioactive compounds containing fatty acids, β‐carotene, lutein, α‐, β‐, δ‐ and γ‐tocopherol, and for their antioxidant properties. The total tocopherol contents that were obtained ranged from 704.66 to 1156.19 mg/kg, with γ‐tocopherol (588.98–977.91 mg/kg) as the main component. The concentration of lutein ranged from 253.14 to 429.63 mg/kg, which was almost ten times higher than that of β‐carotenoid (46.71–69.2 mg/kg), indicating that black tartary buckwheat seed oils were a good source of lutein. The predominant fatty acids were unsaturated oleic acid (C18:1) (35.27–40.61 %) and linoleic acid (C18:2) (38.25–42.90 %). Excellent values of 2,2‐diphenyl‐1‐picrylhydrazyl radical (DPPH), diammonium salt (ABTS) radical scavenging activities were obtained and the highest oxygen radical absorbance capacity (ORAC) value of 13.89 mmol Trolox equiv/g oil was detected in the variety of Chuanqiao No. 1, which was clearly separated by principal component analysis (PCA) on the basis of the highest content of tocopherols and carotenoids. Moreover, the correlation analysis showed that tocopherols and carotenoids were the major contributors to the antioxidant activities of the BTBOs. This study demonstrates that lipophilic extraction in the tartary buckwheat seed contains many interesting bioactive compounds, which are beneficial for human health.

ダッタンソバ品種 「満天きらり」 の子実水分が製粉歩留りに及ぼす影響

ダッタンソバ品種 「満天きらり」 の子実水分が製粉歩留りに及ぼす影響

The complete chloroplast genome sequence of Tartary Buckwheat Cultivar Miqiao 1(Fagopyrum tataricum Gaertn.)

Chloroplasts (cp) are indispensable organelles in plant cells that perform photosynthesis amongst other functions, including producing pigments, synthesizing sugars and certain amino acids. The complete chloroplast genome of Tartary Buckwheat Cultivar Miqiao 1 is sequenced in this study. Miqiao 1 is currently the only Tartary Buckwheat variety that can produce the whole nutritive Tartary Buckwheat Rice by hulling. The Miqiao 1 cp genome size is 159,272 bp in length, including 79 protein-coding genes, 30 tRNA genes and 4 rRNA genes. A pair of inverted repeats (IRs) of 30,817 bp were disconnected by a large single copy (LSC) of 84,397 bp and a small single copy (SSC) of 13,241 bp. The cp genome with 37.9% GC content contained 113 annotated genes.

Accumulation of phenylpropanoids and correlated gene expression in hairy roots of tartary buckwheat under light and dark conditions.

Differential expression patterns of flavonoid biosynthetic pathway genes in the hairy roots of tartary buckwheat cultivars "Hokkai T8" and "Hokkai T10" were studied over a time course of the light-dark cycle. The Agrobacterium rhizogenes-mediated transformation system was applied for inducing hairy roots. Further, a total of six phenolic compounds and two anthocyanins were analyzed in the hairy roots which were exposed to both light and dark conditions, and their amounts were estimated by HPLC. The gene expression levels peaked on day 5 of culture during the time course of both dark and light conditions. Notably, FtPAL, Ft4CL, FtC4H, FtCHI, FtF3H, FtF3'H-1, and FtFLS-1 were more highly expressed in Hokkai T10 than in Hokkai T8 under dark conditions, among which FtPAL and FtCHI were found to be significantly upregulated, except on day 20 of culture. Significantly higher levels of phenolic compound, rutin, along with two anthocyanins were detected in the hairy roots of Hokkai T10 under both conditions. Furthermore, among all the phenolic compounds detected, the amount of rutin in Hokkai T10 hairy roots was found to be ∼5-fold (59,01 mg/g dry weight) higher than that in the control (12.45 mg/g dry weight) at the respective time periods under light and dark conditions.

Breeding of 'Manten-Kirari', a non-bitter and trace-rutinosidase variety of Tartary buckwheat (Fagopyrum tataricum Gaertn.).

Here, we developed a new Tartary buckwheat cultivar ‘Manten-Kirari’, whose flour contains only trace amounts of rutinosidase and lacked bitterness. The trace-rutinosidase breeding line ‘f3g-162’ (seed parent), which was obtained from a Nepalese genetic resource, was crossed with ‘Hokkai T8’ (pollen parent), the leading variety in Japan, to improve its agronomic characteristics. The obtained progeny were subjected to performance test. ‘Manten-Kirari’ had no detectable rutinosidase isozymes in an in-gel detection assay and only 1/266 of the rutinosidase activity of ‘Hokkai T8’. Dough prepared from ‘Manten-Kirari’ flour contained almost no hydrolyzed rutin, even 6 h after the addition of water, whereas the rutin in ‘Hokkai T8’ dough was completely hydrolyzed within 10 min. In a sensory evaluation of the flour from the two varieties, nearly all panelists detected strong bitterness in ‘Hokkai T8’, whereas no panelists reported bitterness in ‘Manten-Kirari’. This is the first report to describe the breeding of a Tartary buckwheat cultivar with reduced rutin hydrolysis and no bitterness in the prepared flour. Notably, the agronomic characteristics of ‘Manten-Kirari’ were similar to those of ‘Hokkai T8’, which is the leading variety in Japan. Based on these characteristics, ‘Manten-Kirari’ is a promising for preparing non-bitter, rutin-rich foods.

Transcripts of Anthocyanidin Reductase and Leucoanthocyanidin Reductase and Measurement of Catechin and Epicatechin in Tartary Buckwheat

Anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR) play an important role in the monomeric units biosynthesis of proanthocyanidins (PAs) such as catechin and epicatechin in several plants. The aim of this study was to clone ANR and LAR genes involved in PAs biosynthesis and examine the expression of these two genes in different organs under different growth conditions in two tartary buckwheat cultivars, Hokkai T8 and T10. Gene expression was carried out by quantitative real-time RT-PCR, and catechin and epicatechin content was analyzed by high performance liquid chromatography. The expression pattern of ANR and LAR did not match the accumulation pattern of PAs in different organs of two cultivars. Epicatechin content was the highest in the flowers of both cultivars and it was affected by light in only Hokkai T8 sprouts. ANR and LAR levels in tartary buckwheat might be regulated by different mechanisms for catechin and epicatechin biosynthesis under light and dark conditions.

Composition and Antioxidant Capacity of Flour and Hull Extracts from Different Tartary Buckwheat Cultivars

The content of rutin, quercetin, total phenol, total flavonoid, antioxidant properties in ethanol extract of flour and hull from 13 tartary buckwheat cultivars widely grown in China was investigated. HPLC showed that the content of rutin ranged from 7.33 to 440.43 in flour and from 5.18 to 168.7 mg/g DW in hull. Trace quercetin were detected in both flour and hull extracts of all cultivars and the values were determined as 0.74-275.90 and 0.77-15.65 mg/g DW, respectively. The contents of total phenol/flavonoid in the flour were significantly greater than that in hull, and their content varied between samples. The result showed that JQ, QW, and NP are potential candidates for developing of functional food.

Characterization of cDNA Encoding Resveratrol Synthase and Accumulation of Resveratrol in Tartary Buckwheat

Resveratrol synthase (RS) is the key enzyme for biosynthesis of resveratrol which come from coumaroyl-coenzyme A (CoA) and malonyl-CoA. Here, we report the cloning and characterization of a RS gene and accumulation of resveratrol in tartary buckwheat (Fagopyrum tataricum). FtRS was composed of 1173 bp open reading frame and 390 amino acid residues and had a theoretical molecular weight and isoelectric point value of 43.70 kDa and 6.24, respectively. The FtRS expression levels were examined in sprouts and different organs of two tartary buckwheat cultivars, Hokkai T8 (T8) and Hokkai T10 (T10). FtRS transcript levels and resveratrol contents were higher under the dark condition compared with light condition. The expression levels of different organs of T10 was not observed significant variations compared to different organs of T8. Interestingly, resveratrol was detected in the sprouts developmental stages, but no resveratrol could not detect in any other organs of both T8 and T10. Therefore, we suggest that the resveratrol content in tartary buckwheat sprouts may be attributed mainly to the dark condition. The characterization of FtRS will be helpful for better understanding of the resveratrol biosynthesis in tartary buckwheat.

Correction: Metabolomic Analysis and Phenylpropanoid Biosynthesis in Hairy Root Culture of Tartary Buckwheat Cultivars

Correction: Metabolomic Analysis and Phenylpropanoid Biosynthesis in Hairy Root Culture of Tartary Buckwheat Cultivars

Differential stress-response expression of two flavonol synthase genes and accumulation of flavonols in tartary buckwheat

Flavonoids are ubiquitously present in plants and play important roles in these organisms as well as in the human diet. Flavonol synthase (FLS) is a key enzyme of the flavonoid biosynthetic pathway, acting at the diverging point into the flavonol subclass branch. We isolated and characterized a FLS isoform gene, FtFLS2, from tartary buckwheat (Fagopyrum tataricum). FtFLS2 shares 48% identity and 67% similarity with the previously reported FtFLS1, whereas both genes share 47–65% identity and 65–69% similarity with FLSs from other plant species. Using quantitative real-time PCR and high-performance liquid chromatography (HPLC), the expression of FtFLS1/2 and the production of 3 main flavonols (kaempferol, myricetin and quercetin) was detected in roots, leaves, stems, flowers and different stages of developing seeds. The relationship between the expression of the 2 FLS genes and the accumulation of the 3 basic flavonols was analyzed in 2 tartary buckwheat cultivars. FtFLS1 and FtFLS2 exhibited differential transcriptional levels between the tartary buckwheat cultivars ‘Hokkai T10’ and ‘Hokkai T8’. Generally, higher transcript levels of FtFLS1 and FtFLS2 and a higher amount of flavonols were observed in the ‘Hokkai T10’ cultivar than ‘Hokkai T8’. The content of flavonols showed tissue-specific accumulation between the 2 cultivars. The transcription of FtFLS1 was inhibited by the exogenous application of abscisic acid (ABA), salicylic acid (SA) and sodium chloride (NaCl), while FtFLS2 was not affected by ABA but up-regulated by SA and NaCl. These data indicate that the 2 FtFLS isoforms of buckwheat have different functions in the response of buckwheat to environmental stress.

Antioxidant and antiproteinase effects of buckwheat hull extracts

Buckwheat is known not only due to its appropriate nutritional composition but the content of prophylactic compounds, too. These are responsible for buckwheat beneficial impact on human health. Most of them are concentrated in outer layers of buckwheat grain. The subject of this work was to screen hulls of nine common and one tartary buckwheat cultivar for the content of flavonoids and its antioxidant and antiproteinase effects. The highest content of total flavonoids was determined for tartary buckwheat cultivar Madawaska (0.6% of hulls weight). Among common buckwheat cultivars the best values reached samples Bamby (0.23%) and KASHO-2 (0.11%). Antioxidant activity as detected via binding radical ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)) and monitoring reducing power was the most effective for samples with highest flavonoid content. Buckwheat hulls effectively inhibited pathophysiological proteases thrombin and urokinase, whereas only little effects were seen to trypsin and elastase. In this testing there were again the best samples with highest flavonoid content. Only tartary buckwheat Madawaska effectively inhibited elastase at tested concentrations. No significant correlation was determined between flavonoid content and measured antioxidant or protease inhibitory action. Obtained results allow us to commend tartary buckwheat cultivar Madawaska as well as common buckwheat cultivars Bamby and KASHO-2 for further experiments.

Molecular characterisation and the light–dark regulation of carotenoid biosynthesis in sprouts of tartary buckwheat (Fagopyrum tataricum Gaertn.)

Seven partial-length cDNAs and 1 full-length cDNA that were involved in carotenoid biosynthesis and 2 partial-length cDNAs that encoded carotenoid cleavage dioxygenases were first isolated and characterised in 2 tartary buckwheat cultivars (Fagopyrum tataricum Gaertn.), Hokkai T8 and Hokkai T10. They were constitutively expressed at high levels in the leaves and flowers, where carotenoids are mostly distributed. During the seed development of tartary buckwheat, an inverse correlation between transcription level of carotenoid cleavage dioxygenase and carotenoid content was observed. The light-grown sprouts exhibited higher levels of expression of carotenoid biosynthetic genes in T10 and carotenoid content in both T8 and T10 compared to the dark-grown sprouts. The predominant carotenoids in tartary buckwheat were lutein and β-carotene, and very abundant amounts of these carotenoids were found in light-grown sprouts. This study might broaden our understanding of the molecular mechanisms involved in carotenoid biosynthesis and indicates targets for increasing the production of carotenoids in tartary buckwheat.

Metabolomic Analysis and Phenylpropanoid Biosynthesis in Hairy Root Culture of Tartary Buckwheat Cultivars

Buckwheat, Fagopyrum tataricum Gaertn., is an important medicinal plant, which contains several phenolic compounds, including one of the highest content of rutin, a phenolic compound with anti-inflammatory properties. An experiment was conducted to investigate the level of expression of various genes in the phenylpropanoid biosynthetic pathway to analyze in vitro production of anthocyanin and phenolic compounds from hairy root cultures derived from 2 cultivars of tartary buckwheat (Hokkai T8 and T10). A total of 47 metabolites were identified by gas chromatography–time-of-flight mass spectrometry (GC-TOFMS) and subjected to principal component analysis (PCA) in order to fully distinguish between Hokkai T8 and T10 hairy roots. The expression levels of phenylpropanoid biosynthetic pathway genes, through qRT-PCR, showed higher expression for almost all the genes in T10 than T8 hairy root except for FtF3’H-2 and FtFLS-2. Rutin, quercetin, gallic acid, caffeic acid, ferulic acid, 4-hydroxybenzoic acid, and 2 anthocyanin compounds were identified in Hokkai T8 and T10 hairy roots. The concentration of rutin and anthocyanin in Hokkai T10 hairy roots of tartary buckwheat was several-fold higher compared with that obtained from Hokkai T8 hairy root. This study provides useful information on the molecular and physiological dynamic processes that are correlated with phenylpropanoid biosynthetic gene expression and phenolic compound content in F. tataricum species.

Characterization of Genes for a Putative Hydroxycinnamoyl-coenzyme A Quinate Transferase and p-Coumarate 3′-Hydroxylase and Chlorogenic Acid Accumulation in Tartary Buckwheat

Tartary buckwheat ( Fagopyrum tataricum Gaertn.) contains a high level of flavonoid compounds, which have beneficial and pharmacological effects on health. In this study, we isolated full-length cDNAs encoding hydroxycinnamoyl-coenzyme A quinate hydroxycinnamoyltransferase (HQT) and p-coumarate 3'-hydroxylase (C3H), which are involved in chlorogenic acid (CGA) biosynthesis. We examined the expression levels of HQT and C3H using real-time RT-PCR in different organs and sprouts of two tartary buckwheat cultivars (Hokkai T8 and T10) and analyzed CGA content using high-performance liquid chromatography. Among the organs, the flowers in both cultivars showed the highest levels of CGA. We concluded that the expression pattern of FtHQT and FtC3H did not match the accumulation pattern of CGA in different organs of T8 and T10 cultivars. Gene expression and CGA content varied between the cultivars. We presume that FtHQT and FtC3H levels might be controlled by multiple metabolic pathways in different organs of tartary buckwheat. Probably, FtC3H might have a greater effect on CGA biosynthesis than FtHQT. Our results will be helpful for a greater understanding of CGA biosynthesis in tartary buckwheat.

Cooking, textural, sensorial, and antioxidant properties of common and tartary buckwheat noodles

Noodles made from flour of 3 common and 2 tartary buckwheat cultivars were investigated into cooking, textural, sensorial, antioxidant properties, and polyphenol composition. Common buckwheat noodles possessed greater tensility, lower adhesiveness, and better sensory properties. Buckwheat noodles were found to be good sources of dietary polyphenols and tartary buckwheat noodles contained a higher level of total phenolics (182.65–221.27 mg GA eq./100 g d.w.) and flavonoids (183.33–218.69 mg rutin eq./100 g d.w.) than their counterparts (p<0.05). Tartary buckwheat noodles also showed higher antioxidant capacity measured by reducing power, DPPH radical scavenging, and ABTS•+ scavenging assay. HPLC analysis identified and quantified 12 polyphenol compounds and revealed that rutin and quercetin were the dominant phenolic compounds. Among them, p-hydroxybenzoic acid, protocatechuic acid, rutin, and quercetin were higher in tartary buckwheat noodles, while common buckwheat noodles had greater contents of chlorogenic acid. Overall, buckwheat noodles are good alternative food with high nutritional quality.

Accumulation of flavonoids and expression of flavonoid biosynthetic genes in tartary and rice-tartary buckwheat

Tartary buckwheat seeds are highly nutritious and excellent dietary sources of phenolic compounds. Rice-tartary buckwheat is a special type of tartary buckwheat with a non-adhering hull trait. Here, we investigated the levels of expression of genes involved in the flavonoid biosynthetic pathway in common tartary and rice-tartary buckwheat. The flavonoid biosynthetic genes, from FtPAL to FtANS, showed different expression levels, especially in the reproductive organs of the 2 cultivars. The transcription levels of these genes, except FtC4H and FtANS, were higher in the mature seeds of rice-tartary buckwheat than in those of common buckwheat. We also analyzed the flavonoid compounds found in the different organs of common tartary buckwheat and rice-tartary buckwheat. Even though the content of rutin was similar in both the cultivars, the distribution of the 3 main flavonoids—rutin, quercetin, and kaempferol—was different in the 2 cultivars, both across organs and within the seed. These results indicate that in the 2 tartary buckwheat cultivars, flavonoid biosynthesis is carried out by different mechanisms.

Genetic analyses of agronomic traits in Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.).

The consumption of products made from Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) has increased in recent years in Japan. Increased consumer demand has led to recognition of the need for early varieties of this crop with high and stable yields. In order to accomplish this, more information is needed on the genetic mechanisms affecting earliness and yield. We conducted genetic analysis of 3 agronomic traits (days to flowering, plant height and total seed weight per plant) to segregate F2 and F3 populations derived from a cross between Tartary buckwheat cultivars ‘Hokuriku No. 4’ and ‘Ishisoba’. Broad-sense heritability estimates for days to flowering, plant height and total seed weight were 0.70, 0.62 and 0.75, respectively, in F3 population. Narrow-sense heritability for total seed weight (0.51) was highest, followed by heritability for days to flowering (0.37), with heritability for plant height (0.26) lowest. Later flowering was associated with increased plant height and higher yields. From the F4 generation, we identified twelve candidate plants with earlier maturity and reduced plant height compared to ‘Hokuriku No. 4’, but almost the same total seed weight. These results suggest that hybridization breeding using the single seed descent (SSD) method is an effective approach for improving agronomic characteristics of Tartary buckwheat.

Differential Expression of Anthocyanin Biosynthetic Genes and Anthocyanin Accumulation in Tartary Buckwheat Cultivars ‘Hokkai T8’ and ‘Hokkai T10’

Six genes involved in anthocyanin biosynthesis in tartary buckwheat have been cloned, namely, FtC4H, Ft4CL, FtCHI, FtF3H, FtF3'H, and FtANS, which encode cinnamate 4-hydroxylase (C4H), 4-coumarate:CoA ligase (4CL), chalcone isomerase (CHI), flavones 3-hydroxylase (F3H), flavonoid 3'-hydroxylase (F3'H), and anthocyanidin synthase (ANS), respectively. Then, these cDNAs were used, along with previously isolated clones for phenylalanine ammonia-lyase (PAL) and chalcone synthase (CHS), to compare gene expression in different organs, flowering stages, and maturing seeds of tartary buckwheat cultivars 'Hokkai T8' and 'Hokkai T10'. Quantitative real-time polymerase chain reaction analysis showed that these anthocyanin biosynthetic genes were most highly expressed in the stems and roots of Hokkai T10. The FtANS gene was more highly expressed than other genes during flowering and maturing seeds. In addition, the anthocyanin concentration was higher in 'Hokkai T10' than in 'Hokkai T8'; however, naringenin chalcone, a flavonoid, was absent from 'Hokkai T10' seedlings based on fluorescence microscopy.

Geographical Differences in Agroecological Traits in Tartary Buckwheat Cultivars from Different Regions of the World

Geographical Differences in Agroecological Traits in Tartary Buckwheat Cultivars from Different Regions of the World