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.
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.
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.
식물에 존재하는 천연물질은 예로부터 건강증진 및 질병치료를 위하여 다양하게 이용되어 왔고, 실제로 morphine, ephedrine 등과 같이 천연물에서 유래한 의약품이 현재에도 질병치료에 널리 응용되고 있다. 메밀 (Fagopyrum esculentum Moench)의 proanghocyanidine, rutin, lignan 등은 항산화, 항균활성 및 항암효과를 나타내는 성분으로 보고 (Cassidy, 1996; Rym of al., 1996)되었다. 따라서 본 실험에서는 메밀의 기능성 물질의 확보와 가공을 통한 원료 고부가가치 창출을 목표로 국내산 메밀을 발이 길이별로 추출물을 제조하여 식품성 성분의 생리활성 인자를 탐색할 목적으로 각 길이별 추출물로 항산화활성 및 항미생물 활성을 측정하였다. 먼저, 항산화 활성에서 DPPH의 50%를 환원 시키는데 필요한 시료의 양(RC50)은 무발아에서 50.41 μg/mL, 발아길이 10 mm에서 80.57 μg/mL, 발아길이 2 mm에서 93.77 μg/mL, 발아길이 5 mm에서 107.09 μg/mL순으로 천연항산화제인 Vit. C 5.98 μg/mL보다는 약하지만, 합성항산화제인 BHT 163.96 μg/mL보다는 월등히 뛰어난 라디칼 소거능이 확인되었다. 발아 길이별 각 추출물의 항미생물 활성은 최고농도 40 mg/mL에서 그람양성균인 S. aureus의 투명저지대의 직경이 4~10 mm 였고, 그람음성균인 P. aeruginosa는 2~9 mm의 범위에서 증식이 억제되어 항균활성은 비교적 높은 것으로 판단되었으며, 그 외의 균주에서는 본 실험에서 사용한 첨가농도로는 완전한 증식억제 효과가 나타나지 않았다. 무발아 시료와 비교할 때 발아가 진행되면서 항균력이 떨어졌다. 암세포 증식 억제효과는 최고농도 800 μg/mL에서 Calu-6 세포의 경우 발아 길이 5 mm 시료에서 95.12%, 무발아 추출물은 87.15%의 높은 암세포 생육억제활성을 나타내었다. 동일 농도에서 발아 길이 5 mm인 시료의 경우 SNU-601에 대하여 85.33%의 억제효과를 보였다. 그러나 유방암세포인 MCF-7과 대장암세포인 Caco-2의 경우 최대농도의 시료를 첨가한 경우에도 세포증식을 억제하지 못하였다. 메밀의 발아 길이별 IC50값을 살펴보면, Calu-6에서 발아 길이 5 mm 추출물에서 301.06 μg/mL, SNU-601에서 2 mm 추출물이 510.20 μg/mL로 탁월한 효과를 보였다. 즉, Calu-6와 SNU-601 세포주에 대한 IC50은 대조군에 비해 발아에 의하여 세포독성 효과를 증가되었지만, MCF-7와 Caco-2에 대한 항암효과는 없음을 알 수 있었다.
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
Korean cultivars of buckwheat(Fagopyrum esculentum Moench.) was crossed with Canadian cultivars in order to improve seed yield as well as leaf production of buckwheat for using as food and medicine. The agronomic characteristics and rutin contents of F2 hybrids are investigated for further selection of superior lines. Dry weight per plant was the highest in a line 1110(6.71g) and leaf weight per plant was the highest in a line 1110(1.91g). Hybrid seeds were 0.55 - 0.70cm long and 0.37 - 0.47cm wide on average. 100 seeds weight ranged from 2.57g to 3.58g. Line 1076 produced the longest seeds(0.70cm) and line 1186 was the longest in seed width. Line 1196 showed the highest 100 seeds weight(3.58g). The highest frequency of the LWR(length/width rate) was 0.66~0.70, indicating that seed shape of the hybrids was mostly oval. Line 1087 showed the highest contents of rutin(77.26ppm). Lines 1090 and 1181 contained respectively rutin of 54.76ppm and 54.35ppm in the seeds. From the yield and rutin point of view, the most superior lines was line 1087 among the lines used for this study.