Dioscorea Japonica Research Articles

Molecular cloning, structural analysis and mass spectrometric identification of native dioscorins of various yam species

Dioscorins are the major storage proteins of yam tubers. However, the molecular nature of their heterogeneity in tubers has not been fully elucidated. In this study the authors isolated the dioscorin gene families of Dioscorea japonica and Dioscorea pseudojaponica, performed matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) and elucidated which dioscorin isoforms are the major constituents in tubers. The dioscorin gene families of D. japonica (Dj-dioA1-Dj-dioA4, Dj-dioB1 and Dj-dioB2) and D. pseudojaponica (Dp-dioA1-Dp-dioA5 and Dp-dioB1) were cloned from cDNA libraries of yam tubers. The dioscorins isolated from Dioscorea alata (Da-dioscorins), D. japonica (Dj-dioscorins) and D. pseudojaponica (Dp-dioscorins) were mainly monomers, with a few dimers. The monomers contained one intramolecular disulfide bond (Cys(28)-Cys(187)) and belonged to Class A dioscorins with two cysteine residues. The dimers consisted of Class B dioscorins with one intermolecular disulfide bond (Cys(40)-Cys(40)). Results of MALDI-TOF-MS revealed that the Da-dioscorins were mainly encoded by Da-dioA2, Da-dioA3 and Da-dioA4. The majority of the Dj-dioscorins were encoded by Dj-dioA1, Dj-dioA2, Dj-dioA3 and Dj-dioB2. The Dp-dioscorins mainly comprised proteins encoded by Dp-dioA1, Dp-dioA3, Dp-dioA4, Dp-dioB1 and Dp-dioB2. Determination of the constituents of dioscorin isoforms in yam tubers provides a basis for future studies of their physiological and biomedical functions.

Recombinant dioscorins of the yam storage protein expressed in Escherichia coli exhibit antioxidant and immunomodulatory activities

Dioscorins, the major storage proteins in yam tubers, exhibit biochemical and immunomodulatroy activities. To investigate the potential application of dioscorins in biomedical research, we expressed the dioscorin genes Dj-dioA3 and Dp-dioA2 from Dioscorea japonica and Dioscorea pseudojaponica, respectively, in E. coli and routinely obtained approximately 15mg proteins per liter Escherichia coli culture (mg/L) to 30mg/L of rDj-dioscorinA3 and 4 to 8mg/L of rDp-dioscorinA2. Western blot analyses revealed that both recombinant dioscorins contained epitopes with similar antigenicities to those of the native dioscorins. Results from dithiothreitol (DTT) treatment followed by monobromobimane (mBBr) staining showed that both recombinant dioscorins, like the native dioscorins, contain an intramolecular disulfide bond between Cys28 and Cys187 residues. Circular dichroism spectroscopy findings indicated that the secondary structural contents of the recombinant dioscorins showed high similarity to those of their corresponding native dioscorins. Both recombinant dioscorins, like the native dioscorins, exhibited 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and Toll-like receptor 4 signaling activities, and stimulated the phagocytosis of E. coli by macrophage. Overall, our results indicated that substantial amounts of recombinant dioscorins can be purified easily from E. coli and that these recombinant dioscorins are appropriate for application in future investigations of the biomedical functions of dioscorins.

단마와 장마 영여자의 항산화능 및 항돌연변이 활성 검정

The bulbil of the Dioscorea species is produced, the amount of which is 2,000 tons annually, but it has been discarded without specific use. In this study the antioxidant and antimutagenicity of bulbil of the Dioscorea, which compared to bulbil of Danma(Dioscorea japonica Decaisne) and Jangma(Dioscorea batatas Decaisne), a major domestic cultivation species. The study was done by extracting bulbil of the Dioscorea methanol and the methanol extracts was re-extracted with chloroform, ethyl acetate, n-butanol and water. In methanol extract of Danma and Jangma, polyphenolic compounds contained 2.2 and 3.9 mg/g extract, respectively. The ethyl acetate fraction of Danma and Jangma had higher polyphenolic contents of 33.9 and 39.1 mg/g, whereas water fractions were much lower at 2.4 and 5.8 mg/g. Determination of antioxidant activity showed that the ethylacetate fraction strong DPPH radical scavenging activity and ABTS radical scavenging activity. The inhibitory effects of methanol extracts and chloroform, ethylacetate, n-butanol, water fraction from bulbil of Danma and Jangma on the mutagenicity in 1-NP, AFB1, Trp-P-1 were investigated using S. typhimurium TA98. Danma and Jangma cultivars decreased the reverse mutation induced by 1-NP, AFB1, Trp-P-1 in S. typhimurium TA98. The fraction of chloroform and ethylacetate showed strong inhibitory effects, in a dose dependant manner against the mutagenicities induced by 1-NP, AFB1, Trp-P-1 in S. typhimurium TA98.

Analytical Methods for Enzyme and DPPH Radical Scavenging Activities of Natural Pigments from Some Plants

The enzyme activities in different fractions of Dioscorea japonica Thunb. and 1,1-diphenyl-2-picryl hydrazyl radical scavenging activity in 15 natural plant pigments from black rice, purple sweet potato, yellow bitter melon, yellow paprika, red cabbage, yellow gardenia, blue gardenia, Chinese foxglove, mulberry leave, onion peel, grape peel, mulberry, red beet, gromwell, and cactus were determined. The antioxidant activity in the cosmetic composition of mulberry leaves, grape peel, mulberry, and red cabbage pigments was relatively high in comparison with all other studied plants. Enzyme activities in investigated plants were evaluated as superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX). The cosmetic composition of mulberry leaf pigment had the highest SOD enzyme activity of 67.1% while onion peel pigment showed the lowest SOD enzyme activity of 15.3%. The activity of CAT and APX from cosmetic composition of natural plant pigments has also been investigated. Both CAT and APX showed higher values in the cactus, mulberry, and red cabbage cosmetic compositions in comparison with other plant pigments. The cosmetic composition in EtOAc extract of D. japonica Thunb. had the highest SOD enzyme activity while the BuOH and EtOH extracts were comparatively low. CAT and APX activities showed significantly high values in EtOH and EtOAc extracts. The antioxidant enzyme activities of D. japonica Thunb. differ significantly in different plant pigments during their extraction. In conclusion, we showed that the plant pigments and D. japonica Thunb. had the potent biological activities. Therefore, these plant resources having anti-aging components could be good materials for development of source of natural cosmetics.

除草の風土〔22〕三重県の茶園における雑草防除，特にヤマノイモの抜き取り

除草の風土〔22〕三重県の茶園における雑草防除，特にヤマノイモの抜き取り

마분말 첨가 파스타의 품질특성 및 제조조건 최적화

Abstract This study's objective is to determine the optimum mixing ratio of yam (Dioscorea japonica THUMB) powder and egg for the preparation of pasta. Response surface methodology revealed 10 experimental points, including two replicates for yam powder and egg. Yam pasta formulation was optimized using rheology. Yellowness(p<0.05) and redness displayed a linear model pattern, while lightness was represented by a quadratic model. The texture(p<0.05), including flavor(p<0.05) and overall quality(p<0.05) was measured as a sensory evaluation. In addition, mechanical properties displayed significant values in adhesiveness(p<0.05). These results showed that yam powder affects flavor and appearance, and egg affects adhesiveness and overall quality. The optimum formulations processed by numerical and graphical optimization were determined at 19.50 g of yam powder and 28.07 g of egg. Key words : yam powder, pasta, sensory evaluation, optimization, response surface methodology

Cloning of genes and enzymatic characterizations of novel dioscorin isoforms from Dioscorea japonica

Dioscorin, the major tuber storage protein of yam, has been shown to possess carbonic anhydrase, trypsin inhibitor, dehydroascorbate reductase, and monodehydroascorbate reductase activities. In the present study, dioscorin from Dioscorea japonica was confirmed as a glycoprotein using the enhanced concanavalin A-peroxidase staining method, and the protein was shown to have both N- and O-glycans. Following the gene cloning, four full-length isoforms of dioscorin were expressed in Escherichia coli and purified by affinity purification and anion-exchange chromatography for structural and biochemical experiments. It was clearly observed that the recombinant dioscorins had carbonic anhydrase, trypsin inhibitor, dehydroascorbate reductase, and monodehydroascorbate reductase activities. However, the dehydroascorbate reductase and monodehydroascorbate reductase activities were markedly decreased in recombinant dioscorins compared with native dioscorin. The decreased activities were closely related to the loss of the glycosylation from the protein.

Active Compound Isolated from Dioscorea japonica Roots with Fumigant Activity against House Dust and Stored Food Mites

Active Compound Isolated from Dioscorea japonica Roots with Fumigant Activity against House Dust and Stored Food Mites

Active compound isolated from Dioscorea japonica roots with fumigant activity against house dust and stored food mites

The acaricidal activity of essential oil from Dioscorea japonica roots against house dust and stored food mites was examined using the impregnated fumigant bioassay. The active component of D. japonica roots, isolated by silica gel column chromatography and high performance liquid chromatography, was identified as 2-hydroxy-4-methoxyacetophenone on the basis of spectroscopic analyses. Lethal dose 50 (LD50) values of 2-hydroxy-4-methoxyacetophenone were 1.29 and 1.78 μg/cm3 against Dioscorea farinae and Dioscorea pteronyssinus, respectively. The acaricidal activity of 2-hydroxy-4-methoxyacetophenone against Tyrophagus pteronyssinus was 2.28 μg/cm3. Compared to the commercial acaricide N,N-diethyl-m-toluamide (DEET), 2-hydroxy-4-methoxyacetophenone was found to be 19.52, 10.17, and 5.96 times more toxic against D. farinae, D. pteronyssinus, and T. putescentiae, respectively. These results suggest that 2-hydroxy-4-methoxyacetophenone may be useful as a preventive agent to control the growth of house-dust and stored-food mite populations.

Withanolides from the Rhizomes of Dioscorea japonica and Their Cytotoxicity

Edible yams are tropical crops that serve as important staple foods in many parts of the world. The rhizome of Dioscorea japonica , well-known as "Japanese yam", is a food and medicinal source known as "San Yak" in Korea. Bioassay-guided fractionation and chemical investigation of the extract of this yam resulted in the identification of two new withanolides, named dioscorolide A (1) and dioscorolide B (2). The structures of these new compounds were determined by spectroscopic methods, including 1D and 2D nuclear magnetic resonance (NMR) techniques, high-resolution mass spectrometry (HRMS), and chemical methods. The cytotoxic activities of the isolates (1 and 2) were evaluated by determining their inhibitory effects on four human tumor cell lines (A549, SK-OV-3, SK-MEL-2, and HCT15) and a human normal cell line (HUVEC) using a sulforhodamine B (SRB) bioassay. Compounds 1 and 2 showed cytotoxicity against tumor cell lines (A549, SK-OV-3, SK-MEL-2, and HCT15) with IC(50) values ranging from 6.3 to 26.9 μM and exhibited lower activity against the normal cell line (HUVEC) with IC(50) values ranging from 27.1 to 28.8 μM, suggesting selective toxicity among tumor and normal cells.

Neurotrophic activity of DA-9801, a mixture extract of Dioscorea japonica Thunb. and Dioscorea nipponica Makino, in vitro

Dioscorea japonica Thunb. has been traditionally used to treat polyuria and diabetes in Korea. We previously report the effects of Dioscorea japonica Thunb. extract on glucose control, NGF induction, and neuroprotection in a rodent diabetic model. Since the most potent fraction, DA-9801, was identified from a mixture of Dioscorea japonica Thunb. (DJ) and Dioscorea nipponica Makino (DN) following bioactivity-guided fractionation, here, we investigated the potential mechanism of the extract activity against diabetic peripheral neuropathy (DPN). A 1:3 mixture of DJ and DN was extracted with ethanol (DA-9801) and further fractionated into an ethylacetate-soluble fraction (DA-9801E). Effects of these extracts on neurite outgrowth were measured in PC-12 cells and DRG neurons. Effects on cell viability and TrkA phosphorylation were evaluated in PC-12 cells. NGF induction effect was determined in primary Schwann cells as well as IMS32 cells (immortalized Schwann cells). No cytotoxicity was observed in PC-12 cells at the concentration below 500 μg/ml of either DA-9801 or DA-9801E. DA-9801 and DA-9801E at 100 μg/ml and 10 μg/ml, respectively, showed a significant effect on neurite outgrowth in PC-12 cells and DRG neurons in the presence of or absence a low concentration of NGF (2 ng/ml). The Trk-A phosphorylation effect of DA9801 was confirmed in PC-12 cells. An NGF induction effect of these extracts was not detected in either IMS-32 cells, or primary Schwann cells. The NGF agonistic activity of DA-9801 and DA-9801E was demonstrated, which may contribute to their neuroprotective effect against DPN. Studies of the detailed mechanism of these extracts as well as identification of the active components are warranted for the development of an anti-DPN drug from DJ and DN.

Furostanol saponins from the rhizomes of Dioscorea japonica and their effects on NGF induction

The rhizome of Dioscorea japonica is a food and medicinal source known as 'San Yak' in Korea. Two new furostanol saponins, coreajaponins A (1) and B (2), together with 10 known compounds (3-12) were isolated from the rhizomes of D. japonica. Their structures were determined by spectroscopic methods, including 1D and 2D NMR techniques, HRMS, and chemical methods. Nerve growth factor (NGF), a crucial factor for neuronal survival and differentiation, can potentially improve neurodegenerative diseases and diabetic polyneuropathy. We evaluated the effects of isolates (1-12) on NGF induction in a C6 rat glioma cell line. Coreajaponin B (2) upregulated NGF content without significant cell toxicity, as did 6, 8, 9, and 11.

Fine-scale spatial structure of genets and sexes in the dioecious plant Dioscorea japonica, which disperses by both bulbils and seeds

To understand the evolution of clonal reproduction and the diversity of clonal plants, it is necessary to clarify the characteristics of each clonal habit. There has been little research on whether bulbils alter spatial genetic structure (SGS) because of the lack of connection to maternal ramets. We used simple-sequence-repeat (SSR) markers to determine the fine-scale SGS of the dioecious plant Dioscorea japonica, which disperses both as bulbils and as seeds. We also evaluated the contributions of sexual and clonal reproduction and tested for spatial sex segregation (SSS). We discovered 111 genets from 394 ramets in a 2.8-ha plot. Genotypic richness (R = 0.28) and clonal diversity (Simpson’s D = 0.94, Fager’s E = 0.90) were high. We did not find SSS, suggesting that the population does not suffer from a shortage of mating pairs due to clonal reproduction. The Sp values revealed moderate SGS at the genet level (Sp = 0.013–0.014), and the genets intermingled at a local scale. Significant SGS at the ramet level showed that ramets within the same genet tended to aggregate. We also found a skewed clonal spatial distribution. The spatial extent of genets was positively correlated with the number of ramets within a genet. The contribution of bulbil production to the variance of parent–offspring gene dispersal was about one–fifth the contribution from sexual reproduction. These results suggest that the dispersal via bulbils affects the SGS in D. japonica, although its contribution to gene dispersal is small compared to the contribution of sexual reproduction.

Putative storage root specific promoters from cassava and yam: cloning and evaluation in transgenic carrots as a model system

A prerequisite for biotechnological improvements of storage roots is the availability of tissue-specific promoters enabling high expression of transgenes. In this work, we cloned two genomic fragments, pMe1 and pDJ3S, controlling the expression of a gene with unknown function from cassava (Manihot esculenta) and of the storage protein dioscorin 3 small subunit gene from yam (Dioscorea japonica), respectively. Using beta-glucuronidase as a reporter, the activities of pMe1 and pDJ3S were evaluated in independent transgenic carrot lines and compared to the constitutive CaMV35S and the previously described cassava p15 promoters. Activities of pMe1 and pDJ3S in storage roots were assessed using quantitative GUS assays that showed pDJ3S as the most active one. To determine organ specificities, uidA transcript levels in leaves, stems and roots were measured by real-time RT-PCR analyses showing highest storage root specificity for pDJ3S. Root cross sections revealed that pMe1 was highly active in secondary xylem. In contrast, pDJ3S was active in all root tissues except for the central xylem. The expression patterns caused by the cassava p15 promoter in carrot storage roots were consistent with its previously described activities for the original storage organ. Our data demonstrate that the pDJ3S and, to a lesser extent, the pMe1 regulatory sequences represent feasible candidates to drive high and preferential expression of genes in carrot storage roots.

New Multi-herb Mixture, HT005, Induces Longitudinal Bone Growth in Male Rats

Objectives : It was investigated new herbal prescription HT005 supported by traditional Korean medicine has a activity on the longitudinal bone growth of rats. HT005 were consisted of the root of Angelica sinensis, the fruit of Alpinia oxyphylla, the root of Astragalus membranaceus, the stem of Eleutherococcus senticosus, and the root of Dioscorea japonica, and Poria cocos. Method : To investigate the effect on longitudinal bone growth in adolescent male Sprague-Dawley rats, the longitudinal length of growth plate was measured directly by the tetracycline fluorescent labeling method and chondrocyte staining method. HT005 administered orally for four days, and the tetracycline was injected twice on the growth plate of the animals for fluorescent dying. The rate of longitudinal bone growth was measured the length between the tetracycline bands which fixed on the 3rd day and 5th day after injection. Cresyl violet was also used to stain the chondrocytes in the growth plate. The length of growth plate after administration was compared. Expression of IGF-1 and BMP-2 in the growth plate was investigated by immunohistochemistry. Results : HT005 showed a significant longitudinal bone growth which was at the dose of 100 mg/kg and (p (p (p at the dose of 10 mg/kg compared to the control group by cresyl-violet staining method. Both the number and intensity of BMP-2 and IGF-1 positive cells were increased in the hypertrophic zone of growth plate. There was a significant correlation between BMP-2 and IGF-1 expression and heights of chondrocytes in growth plate. Conclusions : Treatment of HT005 on Sprague-Dawley rats markedly increased the longitudinal bone growth. Therefore, HT005 may be an alternative herbal source to growth hormone as it promotes bone growth in children afflicted by growth retardation.

Quality and Sensory Characteristics of Soybean Dasik by Additions of Dioscorea japonica Flour

Quality and Sensory Characteristics of Soybean Dasik by Additions of Dioscorea japonica Flour

Quality and Sensory Characteristics of Soybean Dasik by Additions of Dioscorea japonica Flour

Quality and Sensory Characteristics of Soybean Dasik by Additions of Dioscorea japonica Flour

Yam Storage Protein Dioscorins from Dioscorea alata and Dioscorea japonica Exhibit Distinct Immunomodulatory Activities in Mice

The aim of this study was to elucidate the effect of the major storage protein dioscorin isolated from two different yam species, Tainong No. 1 (TN1-dioscorins) and Japanese yam (Dj-dioscorins), on the immune activities of mice. Dj-dioscorins, like TN1-dioscorins, could induce expression of the pro-inflammatory cytokines and stimulate phagocytosis of RAW 264.7. Intraperitoneal injection of the TN1-dioscorins into mice stimulated phagocytosis of bone marrow, spleen, and thymic cells. In contrast, the T and B cells in bone marrow, spleen, and thymus isolated from mice injected with Dj-dioscorins had higher proliferative responses to mitogens. Furthermore, Dj-dioscorins enhanced proliferation of CD4(+), CD8(+), and Tim3(+) (Th1) cells in spleen and CD19(+) cells in both spleen and thymus. Supplement of Dj-dioscorins in the lymphoid cells isolated from Dj-dioscorins primed mice induced cell proliferation of both spleen and thymic cells. These findings indicated that TN1-dioscorins have a higher ability to stimulate the phagocytic activity of the lymphoid cells than Dj-dioscorins, whereas Dj-dioscorins possess more abilities than TN1-dioscorins to enhance the proliferation of the lymphoid cells.

A Study on the glucose‐regulating Enzyme Activities of Water Extracts from Korea Medicinal Herbs

This study investigated the anti‐diabetic effects of water extracts from Korea medicinal herbs on hepatic glucose‐regulating enzymes, such as acetyl‐CoA carboxylase (ACC) and glucokinase (GK). The hepatic cytosol faction of a type ? diabetic animal (GK rat, Goto‐kakizaki) was used in acetyl‐CoA carboxylase (ACC) and glucokinase (GK) activity assays. Type 2 diabetes generally characterized by non‐insulin‐dependent hyperglycemia in early stages and the therapeutic approach available is by retarding absorption of glucose through the inhibition of α‐glucosidase enzyme in the intestinal. Thus α‐glucosidase inhibition assay was assayed. The Korea medicinal herbs were Salvia miltiorrhiza (DS), , Panax notoginsengs (JC), Lycium chinense (JGP), Dioscorea japonica (SY), R. glutinosa (SJH), Pyrus pyrifolia fructus (YSB), Cucumis melo var. makuwa stem (CWJ), Coix lachrymajobi var. mayuen (YM), Cornus officinalis (SSY), Paeonia suffruticosa (MDP), Cordyceps sinensis (DCH), Acanthopanax senticosus (GSO). The ACC activity in cytosol fraction of liver was higher in the 0.01mg/ml DCH water extract than that of the control and SSY water extract significantly increased the GK activity. YSB, MDP, GSO water extracts had high α‐glucosidase inhibitory activity and the effect was higher than that of acarbose (reference α‐glucosidase inhibitor).These observations suggest that DCH, SSY, YSB, MDP and GSO water extracts are the effective candidates for preventing type 2 diabetes.

Secondary dispersal of Dioscorea japonica (Dioscoreaceae) bulbils by rodents

We examined rodent dispersal of Dioscorea japonica bulbils. Bulbils dispersed by rodents may travel farther from their parent plants than those dispersed by gravity alone. To confirm the presence of secondary dispersal and the types of vector, we used an automatic camera to photograph animals that approached bulbils. To clarify secondary dispersal distance, we set marked bulbils at eight sites and searched for the bulbils once they were removed from their original locations by a disperser. We took photographs of mice (Apodemus speciosus) approaching the bulbils. In June, 10 of 333 bulbils that were moved germinated. We also measured the primary dispersal distance at four sites by dropping bulbils from heights of 0.5, 3.0, and 12.0 m. We then compared primary and secondary dispersal distances. The mean secondary dispersal distance was significantly greater than the primary dispersal distance for dispersal heights of 0.5 and 3.0 m, but less than that for a 12.0-m dispersal height. This indicates that rodents may disperse bulbils farther than gravity alone when bulbils drop from lower heights but they contribute little to dispersal for bulbils that drop from greater heights (e.g., from the top of the canopy). Thus, we confirmed the occurrence of rodent dispersal of bulbils and rodents might to some extent contribute to the dispersal and establishment of bulbils.