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.
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.
The different forms of flowers in a species have drawn thoughtful attention of many evolutionary botanists, including Charles Darwin. Common buckwheat (Fagopyrum esculentum Moench.) is regarded as a dimorphic self-incompatible plant which bears either a pin or a thrum flower. It is revealed that the S supergene the key element to govern the self-incompatibility, flower morphology, and pollen size. Already, we have produced self-incompatible buckwheat lines by an interspecific cross between F. esculentum and F. homotropicum by using embryo rescue. We also notice that the self-compatibility allele, Sh, keeps up the heteromorphic incompatibility. In the past decades, two dimensional gel electrophoresis based proteomics approaches have been applied systematically to identify and profile proteins expressed during pollen development of model plant species. Proteome techniques have vastly been applied in the fields of plant genetics, plant development, and plant physiology and ecology to reveal plant genetic diversity, plant development, differentiation of plant tissue and organ, separation and functional identification of novel component of various organelles, mechanisms of plant adapted to abiotic or biotic stresses including high temperature, low temperature, high salt, drought, and pathogens and insects, and interaction of plant with microbe. However, the plethora of studies related to heteromorphic has added remarkably to our knowledge in the field of the multiple aspects of the breeding system and many researches have provided evidence for the connection between these two components. But in spite of its potential biological significance, the high throughput proteomics analysis of this connection has so far been grossly overlooked. So our attempts are to unravel the proteome investigation in common buckwheat.
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
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.
Common buckwheat has the sporophytic self-incompatibility mechanism and that’s why it has the ability to cross pollinate between two plants with different styles (thepin type and thrum type). The S supergene is thought to govern self-incompatibility, flower morphology and pollen size in buckwheat. Already, we have produced self-compatible buckwheat lines by an interspecific hybridization between Fagopyrum esculentum and F. homotropicum by embryo culture. The pollen size of F1 plants produced by a cross between a pin type plant and the self-compatible plant was similar to that of the self-compatible lines and segregated together with flower morphology without exception. The pollen tubes of the self-compatible plants were compatible with styles of the pin plants but incompatible with the styles of thrum plants. But, the pollen tubes of thrum flowers were compatible with the styles of self-compatible plants. Also, the pollen tubes of pin flowers were incompatible with the styles of self-compatible plants. Already, from these results, we have reported a tentative genotype for heterostyle and homostyle flower types. Homomorphism was controlled by a single allele Sh, while the pin/thrum-complex gene was governed by a single genetic locus S, with two alleles, S and s, which control Ss (thrum-type) as well as the ss (pin-type), respectively. Corresponding represents the case of a single locus S with three alleles, Sh, S and s, and the phenotypes, homomorphic, pin and thrum. It can be characterized by relationship of dominance, S>Sh>s. Using the two self-fertile lines, one is considered as the long-homostyle flowers and the other is considered as the short homostyle flowers. If the short-homostyle trait had arisen by recombination in the S supergene, its genotype would be considered to be GIs ip a/GIs ip a. The pollen tubes of the short-homostylous plant should be compatible with the styles of thrum plants. Also, the pollen tubes of short-homostylous plants should be incompatible with the style of long-homostylous plants, and the reciprocal cross also should be incompatible, because the genotype of long homostyle is gis Ip PA/gis Ip PA. Furthermore, the flower morphology of F1plants produced by the cross between cross and short homostyle flowers should be thrum or short homostyle and only short-homostylous plants should be produced by the cross between pin and short homostyle flowers. However, the compatibility or incompatibility of short homostyle flower was not clarified. So, we need to clarify the compatibility or incompatibility of the style of short homostyle flowers for the next step.
Many research results have indicated that many kinds of useful ingredients are rich in buckwheat, which have high nutritional values and medicinal properties, so, buckwheat has been cultivated around Asia and Europe. In this paper, genetic diversity of common and tartary buckwheat germplasms were studied based on morphological and molecular markers in order to provide useful information for conservation and utilization of buckwheat genetic resources. The length, width, thick, value and weight of the seed of the common and tartary buckwheat were measured and analyzed by the statistics methods. The result has shown that there are morphological variation both in common buckwheat and tartary buckwheat seeds collected from different regions. It also has shown that the morphological variation of tartary seeds was significantly correlated to geographical regions. The nuclear ribosomal internal transcribed spacers (ITS) of the tartary buckwheat collected from different countries were cloned, sequenced and statistically analyzed. The data showed that ITS sequences were informative to analyze the phylogeny of the tartary buckwheat and the data also showed that the genetic distances varied among different tartary buckwheat seeds collected from different countries.
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.
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.
Buckwheat is one of the traditional crops and has become a renewed target of interest or a popular crop as a healthy foodstuff, because it is a good source of cereal protein which is rich with essential amino acids. However, what is critical to our health is that buckwheat contains proteins which cause a allergy. Buckwheat allergy resulting from ingestion is caused by the storage proteins in the grain with molecular weights ranging from 15, 22, 35, 39 and 50 kDa proteins of the inner fractions to low, and there were clear differences in the protein compositions between the inner and outer buckwheat flour fractions. A major allergenic protein of buckwheat is Fag e 1 with molecular weight 22 kDa (BW22KD). Buckwheat allergy is an immunoglobulin E (IgE)-mediated hypersensitive response capable of causing anaphylactic shock. Buckwheat seeds were dissected to endosperm and embryo. From each fraction we extracted proteins and analyzed extracts by SDS-PAGE and 2-DE. On electrophoregrams of endosperm proteins, 6 intense bands were detected. The most intense corresponded to molecular weights ranging from 54 to 65 kDa. These proteins have been reported not to be allergenic. We show here that the allergenic buckwheat seed proteins are found only among embryo proteins. No allergenic proteins were found in the buckwheat endosperm. The results presented here lead to the proposal that patients with hypersensitivity to buckwheat flour should use only fine flour from buckwheat endosperm, as this fraction contains no allergenic proteins. At present, specific protein spots will be selected and in-gel digested for MALDI-TOF-TOF/MS analysis.
본 연구는 단메밀과 타타리메밀의 페놀성화합물의 함량을 측정하고 분석, 평가한으로써 생리활성 연구의 기초 자료와 건강기능성 식품소재 개발에도 이용될 수 있는 유용한 기초 자료를 얻고자 수행하였다. 1. 총 페놀의 함량은 타타리메밀 종자가 단메밀 종자보다 약 2배정도 높게 나타났다. 총 페놀 중의 플라보노이드 함량은 단메밀이 약 50~% , 타타리메밀이 약 95~% 를 차지하고 있다. 단메밀과 타타리메밀의 총 페놀 및 플라보노이드 함량은 메밀쌀이 메밀껍질보다 높았으나, 단메밀의 플라보노이드 함량은 메밀껍질 더 높았다. 3. 단메밀의 rutin함량은 메밀껍질(25.2 mg/100g)>껍질을 벗기지 않은 종자(19.8 mg/100 g)>메밀쌀(12.8 mg/100 g)의 순으로 높았으나, 타타리메밀은 메밀쌀(2042.1mg/100g)>껍질을 벗기지 않은 종자(1375.8 mg/100g)>메밀껍질(138.7 mg/100 g)의 순으로 높게 나타났다. 식물체 부위별 rutin의 함량은 단메밀이 메밀싹>잎>줄기>종자의 순이고, 타타리메밀은 잎enli메밀싹>종자>줄기의 순으로 나타났다. 모든 부위에서 타타리메밀이 단메밀보다 높은 rutin함량을 나타냈다. 4. 단메밀과 타타리메밀 종자의 flavanols함량은 큰 차이를 보이지 않았으나, 잎, 줄기, 싹나물부위의 함량은 타타리메밀이 높은 경향을 보였고, 두 종의 flavonols의 함량도 단메밀보다 타타리메밀에서 월등히 높은 경향을 보였다. 5. 식물체 부위별 flavanols의 함량은 단메밀과 타타리메밀의 메밀싹>잎>줄기>종자의 순으로 높게 나타났다. 모든 부위에서 catechin> Epicatechin> Epicatechingallate의 순으로 높게 나타났으나, 메밀싹은 두 종 모두 Epicatechingallate의 함량이 종자의 약 30~~40 배, 잎, 줄기보다 약 15~~20 배가량 높았다.
The isolation of female gametes is precondition for the satisfactory micromanipulation of gametes in common buckwheat (Fagopyrum esculentum Moench.). Embryo sacs were isolated at flowering stage after brief treatment with minimal concentrations of cell-wa