본 연구는 한국산 재래종 메밀가루를 40%, 70% 에탄올(발효주정)로 각각 처리하여 메밀 추출물을 제조하였고 이를 동결건조한 다음 고지방식이에 혼합하여 흰쥐에게 섭취시킨 후 나타나는 식이성 고지혈증의 개선효과를 확인하고자 하였다. 실험군은 정상대조군(NC), 고지방대조군(HFC), 고지방-40%에 탄올메밀추출물군(HF-40), 고지방-70%에탄올메밀추출물군(HF- 70) 4군으로 나누어 4주간 실험식이로 사육하였으며 실험 결과는 다음과 같다. 1. 실험동물의 식이섭취량과 체중증가량 및 사료효율은 실험군들 간에 유의적 차이를 보이지 않았다. 2. 실험동물의 장기 중간무게를 비교한 결과 정상대조군에 비해 고지방대조군에서 유의적으로 높았고 고지방대조군에 비해 HF-70군은 유의적으로 낮았고 (p<0.05) 정상대조군도 비슷한 수준이었다. 3. 혈중 GOT, GPT 수치는 정상대조군이 가장 낮았고 고지방대조군이 가장 높았으며 HF-40군과 HF-70군은 고지방대조군보다 낮은 경향을 보였다. 혈당 농도는 전체 실험군 중에서 HF-70군이 가장 낮은 경향을 보였다. 4. 혈중 중성지질 농도는 정상대조군에 비해 고지방대조군이 가장 높았고 정상대조군과 고지방대조군에 비해 HF-40군 이 유의적으로 낮았으며(p<0.05) 혈중 유리지방산 농도는 실험군 간에 유의적인 차이를 보이지 않았다. 5. 혈중 콜레스테롤 농도는 정상대조군에 비해 고지방대조군에서 유의적으로 높았고(p<0.05) 고지방대조군과 비교했을 때 HF-70군에서 낮은 경향을 보였으며, 혈중 유리콜레스테롤 농도는 실험군 간에 유의적인 차이가 없었다. 혈중 콜레스테롤에스터 수치는 정상대조군이 71.6mg/dl로 유의적으로 가장 낮았고 고지방대조군의 125.0mg/dl와 비교하여 HF-40군, HF- 70군은 각각 116.9mg/dl, 88.5mg/dl로 유의적으로 낮았다 (p<0.05). 특히 HF-70군은 HF-40군에 비해서도 유의하게 낮은 것으로 나타났다. 6. 혈중 LDL-콜레스테롤은 정상대조군에 비해 고지방대조군이 유의적으로 높았으며 고지방대조군(130.6mg/dl)에 비해 HF-70군(85.1mg/dl)에서 유의적으로 낮았다(p<0.05). 7. 혈중 HDL-콜레스테롤은 정상대조군에 비해 고지방식이군들이 유의적으로 낮았으며(p<0.05) 고지방실험군들 간에는 유의적인 차이가 없었다. 그러나 HDL-콜레스테롤/총콜레스테 롤의 비율은 정상대조군에 비해 고지방대조군이 유의적으로 낮았고 고지방대조군에 비해 HF-70군이 유의적으로 높았다 (p<0.05). 8. 동맥경화지수는 정상대조군이 가장 낮았으며 고지방대조군이 정상대조군에 비해 유의적으로 높았고 고지방대조군에 비해 HF-70군은 유의적으로 낮았다 (p<0.05). 본 실험에서 고지방식이를 섭취한 쥐의 이상지질혈증은 메밀 에탄올추출물에 의해 개선되었으며 40% 에탄올 메밀추출 물보다 70% 에탄올 메밀추출물에서 더욱 많이 개선되는 것으로 확인되었다.

Fagopyrum esculentum (Buckwheat) is a globally used alternative crop that contains several useful substances with various effects; however, many of these substances (rutin, quercetin, etc.) are water insoluble. To extract these substances, alcohols is required, which is inconvenient because these solvents cause diverse problems. Many studies are underway to achieve effective extraction of these substances with water. Among of these studies, microwave assisted water extraction (MAE) has been performed extensively. In this study, we performed the extraction in various solvents and/or microwave from Fagopyrum esculentum. The analysis of the content of useful substances and the bioactivity were performed and shown to increase in MAE. Liquid chromatography-mass was performed in order to identify of the useful water-insoluble substances. Catechin, quercetin, and rutin, which are all insoluble in water, were hardly extracted with water even on heating (4.4 ppb, 3.9 ppb and 60.3 ppb, respectively). However, MAE was found to extract much more of these substances than water (1204 ppb, 110.8 ppb and 2946 ppb, respectively). Although less efficient than alcohols, MAE showed much higher efficiency than simple water extraction. These results indicate that water extraction using microwave technology is effective in cases where it is difficult to extract useful substances using water.

We analyzed the nuclear ribosomal internal transcribed spacer (ITS) sequence of common buckwheat, Fagopyrum esculentum and tartary buckwheat, F. tataricum. The diversity of the nucleotides and haplotypes, Tajima’s D, and Fu’s Fs was analyzed and compared among the varieties of common buckwheat and tartary buckwheat. The diversity of nucleotides and haplotypes indicated that the buckwheat populations had undergone rapid population expansion but D and Fs did not support their expansion statistically. The phylogenetic analysis of ITS sequences did not clearly establish the phylogenetic relationships between the varieties of common buckwheat. The In/Del sequence of ITS-1 region could, therefore, be used as a DNA marker to distinguish raw or manufactured products derived from common buckwheat and tartary buckwheat.

Breeding and cultivation techniques are being treated very severely regarding ecological and physiological development in buckwheat. This study was conducted to focus on the diversity occurring in the cultivated and tartary buckwheat and provide an overview of the characteristics and genetic resources activities. Morphological results showed that the height of common buckwheat ranges from 82-90cm, common buckwheat induced by 200Gy ranges from 52-75cm, common buckwheat induced by 300Gy ranges from 43-56cm, common buckwheat induced by 400Gy ranges from 33-60cm whereas the tartary buckwheat hight ranges from 65-87cm, and while it exposed to various radiation (200Gy, 300Gy and 400Gy), the obtained height ranges from 73-92cm, 55-80cm and 60-75cm respectively. However, the stems from the both cultivar are hollow and that’s why, the plant is very prone to lodging. The leaf color of common buckwheat was green, 200Gray, 300Gy 400Gy common buckwheat light green and green, whereas the tartary buckwheat green and bottle-green, 200Gray 300Gy 400Gy tatary buckwheat bottle-green, common buckwheat (control, 200Gy, 300Gy, 400Gy) stem color is light green and pink, flower color is white, tartary buckwheat (control, 200Gy, 300Gy, 400Gy) flower color is light green. The stem color from tartary buckwheat showed (200Gy, 300Gy, 400Gy) light green and light red color. The results revealed that the two buckwheat cultivars showed diversified characteristics.

Bitter buckwheat, also called tartari buckwheat (F. tartaricum), contains large amount of rutin and it has antioxidant activity compared to common buckwheat. But after harvesting and processing, the discrimination of two species through visual inspection was almost impossible. Therefore we developed InDel markers to identify common and tartari buckwheat content based on the chloroplast genome sequence. We conducted complete full chloroplast genome sequence of tartari buckwheat and compared with common buckwheat chloroplast genome sequence (NC010776). Based on the mVISTA alignment, we found eight big InDel (above 50bp) regions. Among the InDel, 6 regions are intergenic region and two are genic region in ycf1. We designed InDel specific primers and applied to PCR with buckwheat genomic DNA to check the discrimination of two species. These InDel specific primers also applied to buckwheat germplasm, 75 tartari and 21 common buckwheat. Among the primers, 5 markers could be successfully amplified in all germplasm species specific amplicon. And we can detect 10pg/ul of DNA and processed food such as tea and noodle. These results could improve the QC (Quality control) of tartari buckwheat food

본 연구에서는 메밀 속성장의 저장, 유통 과정 중 품질특성을 조사하고자 30일간 발효, 숙성 시킨 메밀 속성장을 다양한 용기(항아리, 플라스틱, 유리)에 담아 5℃, 15℃, 25℃에 저장시킨 후 기간에 따른 pH, 산도, 아미노태 질소,총 균수, 색도의 변화를 관찰하였다. 그 결과 항아리(15℃,25℃)에 저장되었던 실험군을 제외한 나머지 실험군은 저장기간 동안 초기의 pH 4.5를 유지하는 경향을 보였으며 항아리(15℃, 25℃)시료는 pH가 시간이 지남에 따라 증가하여 저장 98일에는 각각 pH 5.6, pH 7.4로 증가 하였다. 또한 항아리(15℃, 25℃)에 저장되었던 실험군의 산도는 저장기간에 따란 점차 증가하는 경향을 보였는데 항아리(15℃)시료는 저장 98일(1.85%), 항아리(25℃) 시료는 저장 91일(0.71%)에 낮은 산도를 나타내었다. 플라스틱(25℃) 실험군의 아미노태 질소 함량은 저장 98일에 0.751%이었으며, 항아리(25℃)에 저장된 실험군은 0.92%로 다른 실험군에 비하여 그 함량이 크게 증가하였다. 또한 저장기간 동안 샘플의 총균수는 모든 실험군에서 전체적으로 뚜렷한 증가나 감소의 경향을 보이지는 않았다. 색도의 경우, 항아리(25℃)에 저장된 시료의 경우, L, a, b의 모든 값에서 가장 큰 변화를 보였으며 저장 온도가 높을 수록 L 값이 낮아지는 현상을 확인할 수 있었다. 따라서, 본 연구결과 메밀 속성장의 저장, 유통과정에서 25℃보다 5~15℃를 유지하는 것이 품질 변화를 줄일 수 있는 것으로 확인 되었으며, 항아리와 플라스틱에 비하여 유리병에 보관하는 것이 제품을 보존하는데 있어 탁월한 것으로 확인되었다. 이러한 실험 결과는 메밀 속성장의 저장성 및 유통기한 설정에 기초자료로 제시하고자 한다.

Buckwheat sprout is used as vegetable, and also flour for making noodles, and so on. Currently, information about tissue culture in buckwheat is limited and restricted to micro-propagation. We carried out somatic embryogenesis and plant regeneration using hypocotyl segments as explant of the cultivated buckwheat species, Fagopyrum esculentum which differs from existing studies in the growth regulator combinations used. Maximum callus regeneration was induced on MS medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) 2.0 mg · L-1, benzyladenine (BA) 1.0 mg · L-1 and 3% sucrose. Friable callus was transferred to solidified MS media containing BA (1.0 mg · L-1) with various concentrations of 2,4-dichlorophenoxyacetic acid for the induction of embryogenesis. The optimum concentrations of growth regulators (for regeneration of plantlet) were indole-3-acetic acid (2.0 mg · L-1), Kinetin (1.0 mg · L-1), BA (1.0 mg · L-1). Only 2,4-D did not show any significant effect on callus induction or embryogenesis. Regeneration of embryonic callus varied from 5% to 20%. Whole plants were obtained at high frequencies when the embryogenic calli with somatic embryos and organized shoot primordia were transferred to MS media with 3% sucrose. The main objective of this research was to develop an efficient protocol for plant regeneration for common buckwheat, and to apply in future for genetic transformation.

Buckwheat (Fagopyrum esculentum Moench), one of the minor crops grown in Korea belonging to the Polygonaceae family, is an annual crop widely cultivated in Asia, Europe, and America and has a character of outcrossing and self-incompatibility. The objective of this study was to analyze the genetic variability, phylogenetic relationships and population structure of buckwheat landraces of Korea using SSR markers. Ten microsatellite markers have been detected from a total of 79 alleles among the 179 buckwheat accessions were collected from Korea. The number of allele per marker locus (NA) ranged from 2 (GB-FE-001, GB-FE-043 and GB-FE-055) to 31 (GB-FE-035) with an average of 7.9 alleles. GB-FE-035 was the most polymorphic with the highest PIC value 0.93. Major allele frequencies (MAF) for the 10 polymorphic loci varied from 0.12 to 0.97 with a mean allele frequency of 0.57. The expected heterozygosity (HE) values ranged from 0.05 to 0.94 with an average of 0.53. The observed heterozygosity (HO) ranged from 0.06 to 0.92 with an average of 0.42. The overall polymorphic information contents (PIC) values ranged from 0.05 to 0.93 with an average of 0.48. The landrace accessions of buckwheat used in the present study were not distinctly grouped according to geographic distribution. The study concludes that the results revealed genetic differentiation was low according to the geographic region because of outcrossing and self-incompatibility. We reported that our analyses on the genetic diversity of common buckwheat cultivars of Korea were performed by using of microsatellite markers.

Buckwheat sprout is used as vegetable, and also flour for making noodles, and so on. Currently, information about tissue culture in buckwheat is limited and restricted to micropropagation. We carried out somatic embryogenesis and plant regeneration using hypocotyl segments as explant of the cultivated buckwheat species Fagopyrum esculentum, differs from existing studies in the growth regulator combinations used. Maximum callus regeneration was induced on MS medium containing 2,4-D(2.0 mg/L) and benzylaminopurine BAP (1.0 mg/L) and 3% sucrose. Friable callus was transferred to solidified MS media containing BAP (1.0 mg/L) and at various concentrations for the induction of embryogensis. The optimum concentrations of additives were IAA (2 mg/L), KIN(1.0 mg/L), BAP (1.0 mg/L), and 3% (w/v) sucrose. Only 2,4-D did not show any significant effect on callus induction or embryogenesis. Regeneration of embryonic callus varied from 5 % to 20%. Whole plants were obtained at high frequencies when the embryogenic calluses with somatic embryos and organized shoot primordia were transferred to MS media with 3% sucrose. Regenerated plants after acclimation will transfer to green house. The main objective of this research was to develop a efficient protocol for plant regeneration for common buckwheat, and to apply in future for genetic transformation.

Comparison of the distribution and utilization of free amino acids in buckwheat is required of understanding the amino acid metabolism. The buckwheat sprouts contained the most abundant amino acid Val (40%) followed by Tyr (28%) in common buckwheat (CB), whereas Val was the most predominant amino acid in tatary buckwheat (TB), accounting for 62%. The buckwheat stem and root contained commonly Gln (40~42% in stem; 30~37% in root). Thus, soluble amino nitrogen source is used for Gln in buckwheat. The main difference of amino acid distribution in three tissues between CB and TB was Tyr in sprouts. Low level of Tyr in TB is presumably resulted from the conversion to other phenolic metabolites. The content of essential free amino acids in TB sprout was 53% higher than that in CB. Thus, the TB sprouts are benefit to the human nutrition. The basic study of amino acids gives a clear evidence for different interconversion and metabolism of amino acids in two buckwheat species.

Buckwheat sprouts are a vegetable which provides health benefit with their nutritionally important substances. Buckwheat has been considered as preventive medicine in the last decade. The present study was focused on the reference maps common (Fagopyrum esculentum Möench.) and tatary(Fagopyrum tataricum Gaertn.) buckwheat leaf and stem cultured in light and dark condition. Proteins were extracted from 7-day germinated buckwheat sprout sand separated by two-dimensional electrophoresis(2-DE) with isoelectro focusing gel over pH3 to 10. A total of more than 1520 protein spots were revealed on 2-DE gel, in which 165 proteins were identified in the basis of peptide mass fingerprinting. Functional category analysis indicated that these differentially expressed proteins mainly involved in cellular process, defense responsive, energy production, metabolism, photosynthesis, DNA recombination, DNA replication, seed storage, signal transduction, stress responsive, transcription, translation, and energy transport proteins. The pattern at protein level suggested the important roles for energy and protein metabolism-related proteins in growing sprouts under dark and light condition, accompanied by the activated of the stress responsive and growth condition. The proteomic profiling of common and tatary buckwheat will give insight for understanding buckwheat physiology and application to buckwheat industry.

Buckwheat (Fagopyrum esculentum Moench.) is an alternative crop belonging to the Polygonaceae family. Buckwheat has been well known as a plant source of rutin, quercetin, kaempferol-3-rutinoside and other phenolic compounds. Buckwheat contains more rutin than most of the other plants, which exhibits antioxidative, antihemorrhagic and blood vessel protecting properties. Phenylalanin ammonia-lyase (PAL), chalcone synthase (CHS), flavone 3-hydroxylase (F3H), and anthocyanidin synthase (ANS) were important enzymes involved in the flavonoid biosynthesis in buckwheat. In this study, we have purified full-length PAL, CHS, F3H and ANS from buckwheat using RACE PCR with degenerated primers targeted to conserved regions of orthologous sequences PAL, CHS, F3H and ANS available. The open reading frame of FePAL was 2112 nucleotides long, encodes for 704 amino acids. FePAL gene shares 80-81% identity and 88-90% similarity with Arabidopsis thaliana PAL1, Capsicum annuum PAL, Lactuca sativa PAL, and Populus trichocarpa PAL2. The cDNA of FeCHS has 1179 nucleotides open reading frame, encoding for 393 amino acids. FeCHS has identities and similarities ranging 84-86% and 92-95%, respectively, with other CHSs from P. trichocarpa, Citrus sinensis, Gerbera hybrid, and Vitis rotundifolia, whereas this gene shares 93% identity and 98% similarity with Polygonum cuspidatum. The cDNA of FeF3H has 1101 nucleotides open reading frame, encoding for 367 amino acids. FeF3H has identities and similarities ranging 76-81% and 92-97%, respectively, with other F3Hs from Gossypium hirsutum, Dimocarpus longan, Garcinia mangostana, Camellia sinensis, Vitis vinifera, Rubus coreanus, Pyrus communis, Glycine max, and Actinidia chinensis. The open reading frame of FeANS was 1074 nucleotides long, encodes for 358 amino acids. FeANS gene shares 74-76% identity and 94-96% similarity with Gypsophila elegans, C. sinensis and G. hirsutum. These result provieded the theoretical basis for the metabolic engineering on buckwheat.

본 실험은 메밀새싹의 추출조건에 따른 수율, 총 페놀, -glucosidase 저해활성능, 및 ACE 저해활성능에 대해 반응표면 분석법을 이용하여 추출조건을 최적화하였다. 중심합성계획에 따라 추출온도(), 추출시간(0~12 hr) 및 에탄올 농도(0~100%)를 달리하였을 때 반응 표면 회귀식의 R2는 수율, 총 페놀, -glucosidase 저해활성능 및 ACE 저해활성능에서 각각 0.9461(p-glucosidase 저해활성능 및 ACE 저해

Single seeds of common buckwheat cultivar Suwon No. 1 when subjected to SDS-PAGE revealed very high polymorphism. High variation existed for protein or protein subunits with molecular weight 54-47kDa, 45-25kDa and 16-11kDa. The electrophoregram showed variation for globulin as well as other protein fractions. About 300 proteins were separated by two-dimensional electrophoresis in common buckwheat (Fagopyrum esculentum Moench.) seed. Seed maturation is a dynamic and temporally regulated phase of seed development that determines the composition of storage proteins reserves in mature seeds. Buckwheat seeds from 5, 10, 15, 20, and 25 days after pollination and matured stage were used for the analysis. This led to the establishment of high-resolution proteome reference maps, expression profiles of 48 spots. It was identified 48 proteins from MALDI-TOF/MS analysis of wild buckwheat seed storage proteins. The 48 proteins were found identical or similar to those of proteins reported in buckwheat and other plants; it is belonging to 9 major functional categories including seed storage proteins, stress/defense response, protein synthesis, photosynthesis, allergy proteins, amino acid, enzyme, metabolism, and miscellaneous. It appears that the major allergenic storage protein separated played the important role in buckwheat breeding and biochemical characterization.

To facilitate the introgression of F. esculentum into the traits of F. homotropicum, several accessions of the hybrids between these two species were pollinated with F. esculentum as the recurrent parent. The embryo in vitro rescue was performed to increase the recovery of backcross progenies. The F2 generation was more amenable than F1 hybrids to produce backcross progenies. The F1 hybrids were backcrossed twice with common buckwheat (pin-type F. esculentum) (recurrent backcrossing). Also, alternate backcrosses with common buckwheat and F. homotropicum (congruity backcrossing) were carried out. Pollen tube growth of BCF1 × F. esculentum (thrum) and F. homotropicum × BCF1 was the disturbed penetration exceeded for all initial interspecific hybrids, and its requirement was proportionally lower when the common buckwheat was used as the recurrent parent and as the last parent of congruity hybrids. Effects of both common buckwheat and F. homotropicum on seed success rate for hybridization were observed. Growth of hybrid embryos before rescue, regeneration of mature hybrids all increased recurrent and congruity backcrosses and inter-crosses between F1 plants and selected fertile plants of the second congruity backcrosses.

To facilitate the introgression of F. esculentum into the traits of F. homotropicum, several accessions of the hybrids between these two species were pollinated with F. esculentum as the recurrent parent. In vitro embryo rescue was performed to increase the recovery of backcross progenies. The F2 generation was more amenable than F1 hybrids to produce backcross progenies. The F1 hybrids were backcrossed twice with common buckwheat (pin-type F. esculentum, recurrent backcrossing). Also, alternate backcrosses with common buckwheat and F. homotropicum (congruity backcrossing) were carried out. Pollen tube growth of BCF1 x F. esculentum (thrum) and F. homotropicum x BCF1 was disturbed penetration exceeded for all initial interspecific hybrids, and its requirement was proportionally lower when the common buckwheat used as the recurrent parent and as the last parent of congruity hybrids. Effects of both common buckwheat and F. homotropicum on seed success rate for hybridization were observed. Growth of hybrid embryos before rescue, regeneration of mature hybrids all increased recurrent and congruity backcrosses, inter-crosses between F1 plants and selected fertile plants of the second congruity backcrosses.

The present study reports isolation and characterization of ten polymorphic SSR markers developed from common buckwheat (Fagopyrum esculentum Moench). These SSR markers produced a total of 59 alleles across 41 common buckwheat accessions with an average of 5.9 alleles per locus. Values for observed (HO) and expected (HE) heterozygosities ranged from 0.071 to 0.924 (mean=0.53) and from 0.073 to 0.902 (mean=0.412), respectively. At significance threshold (P<0.05), seven loci deviated from Hardy-Weinberg equilibrium (HWE), whereas significant linkage disequilibrium (LD) values were observed between 5 pairs of loci. These markers are currently being used for programming of the genetic conservation and classification of common buckwheat germplasm collection.