목적 : 항산화 기능을 가지는 하이드로겔 콘택트렌즈(Hy)를 제조하기 위해 갈릭산(Gallic acid: GA)을 Hy에 상 분리 없이 도입시키는 방법에 관해 연구하고, 갈릭산이 첨가된 렌즈의 물리적 특성 및 항산화 효과를 살펴보고 자 한다.
방법 : 하이드로겔 콘택트렌즈는 실험실에서 열중합방법으로 제조하였다. 갈릭산과의 결합을 위해 사용된 Chitosan(Cs) 은 100~300 kDa(CL), 600~800 kDa(CH) 분자량을 가진 두 종류를 사용하였으며, Hy에 IPN(Interpenetrating polymer network) 방법으로 첨가하였다. 광 투과율은 300~800 nm 파장 범위에서 측정하였으며, 함수율은 중량 법으로 측정하였다. 키토산과 갈릭산 간의 amide bonding은 FT-IR을 이용하여 확인하였으며, 첨가된 갈릭산의 양 은 298 nm에서 측정한 UV 흡광도 값으로 정량하였다. 렌즈의 항산화 효과는 DPPH radical 소거능 분석 방법을 사용하였다.
결과 : 광 투과율은 Hy에서 95%, Hy-CL-GA와 Hy-CH-GA에서 91%를 보였다. 함수율은 Hy에서 50.5±1.2%, Hy-CL-GA에서 52.4±0.7%, Hy-CH-GA에서 55.6±0.6%로 나타났다. 렌즈에 결합된 갈릭산의 양은 Hy-CLGA와 Hy-CH-GA에서 0.30, 0.37 mmol/cm2로 각각 나타났으며, 항산화 효과는 Hy-CL-GA에서 48.23%, Hy- CH-GA에서는 58.32%로 나타나 갈릭산을 첨가한 두 렌즈에서 매우 높은 항산화 기능을 확인할 수 있었다.
결론 : 키토산과 갈릭산을 활용하여 광 투과율과 함수율이 높고 항산화 효과를 가진 기능성 하이드로겔 콘택트 렌즈를 제조하였다. 렌즈에 결합된 갈릭산의 양이 증가함에 따라 함수율과 항산화 효과가 증가하였다. 콘택트렌즈 에 다당류와 폴리페놀류의 화합물을 첨가하는 방법을 제시함으로써 단백질 억제, 습윤성 향상, 항산화 기능 등을 함유한 다 기능성 렌즈 제조에 기여할 것으로 본다.
Platycodon grandiflorum (Bell flower) is an important plant that has traditionally been used as herbal medicine for the treatment of cough, phlegm, sore throats, lung abscesses, chest pains, dysuria, and dysentery. The present study was initiated to investigate the feasibility of inducing shoot and root organogenesis in cultured explants of P. grandiflorum in a range of culture media and through use of various plant growth regulators (PGRs). The plantlets (Stem containing one node) were isolated and cultured on different concentrations of Murashige and Skoog (MS) medium supplemented with PGRs. We found that proliferation and elongation of shoots and roots could be achieved on ¼ MS for P. grandiflorum with wild and green petals and on ⅛ MS for P. grandiflorum with double petals. The highest levels of development and elongation of adventitious shoots and roots were observed when petal explants were cultured on ¼ MS (pH 3.8) supplemented with 5% sucrose. Increasing the agar concentration reduced shoot growth and rooting potential; nevertheless, the highest number of shoots and roots was observed on 0.6% agar. In the case of growth regulators, ¼ MS supplemented with 1 mg L-1 6-benzylaminopurine (BA) was found to be best for shooting, although higher concentrations of BA tended to reduce shoot and root elongation. The highest number of shoots was achieved on 0.5 mg ․ L-1 thidiazuron (TDZ) from double petal explants grown on ⅛ MS. However, root and shoot elongation were found to decrease when TDZ concentrations were increased. Low concentrations of kinetin, naphthalene acetic acid, indole acetic acid, and 3-indole butyric acid induced shoot and root proliferation and elongation. Taken together, our study showed that low concentrations of PGRs induced the greatest root formation and elongation, showing that the optimal concentration of PGRs for shoot proliferation was species-dependent.
The roots of Platycodon grandiflorum species either dried or fresh, are used as an ingredient in salads and traditional cuisine in Korea. To interpret the root proteins, a systematical and targeting analysis were carried out from diploid and tetraploid roots. Two dimensional gels stained with CBB, a total of 39 differential expressed proteins were identified from the diploid root under in vivo condition using image analysis by Progenesis Same Spot software. Out of total differential expressed spots, 39 differential expressed protein spots (≥ 1.5-fold) were analyzed using LTQ-FTICR mass spectrometry. Except two proteins, the rest of the identified proteins were confirmed as down-regulated such as Isocitrate dehydrogenase, Proteasome subunit alpha type-2-B. However, the most of the identified proteins from the explants were mainly associated with the oxidoreductase activity, nucleic acid binding, transferase activity and catalytic activity. The exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.
Platycodon grandiflorum, commonly known as Doraji in Korea, has a wide range of pharmacologic properties, such as reducing adiposity and hyperlipidemia, and antiatherosclerotic effects. However, the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (≥ 2-fold) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose- 1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). In that way, the exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.
Platycodon grandiflorum is a perennial flowering plant, known as Chinese bell flower, widespread in northeast Asia. The roots of this species are used for centuries to treat diseases, and have extensive pharmacological effects such as reducing adiposity, hyperlipidemia as well as anti-atherosclerotic disorder. In this study, systematical and targeting proteome analysis were executed from the 3, 4 and 5 months aged diploid and tetraploid roots of Platycodon grandiflorum and the proteins were separated by 2-DE and stained by CBB. In diploid roots, a total of 30 protein spots (≥ 2-fold) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Among the 30 differentially expressed proteins, 21 proteins sopts were identified as up-regulated and 9 proteins were identified as down-regulated. In contrary, a total of 40 differentially expressed proteins were confirmed from tetraploid roots whereas 28 protein spots were confirmed as up-regulated and 12 proteins were identified as down-regulated. However, the differentially expressed proteins from diploid and tetraploid roots were classified into 12 and 14 possible functional categories respectively using Protein Information Resources. The results revealed that the identified proteins from diploid and tetraploid roots were mainly involved in oxidoreductase activity, nucleotide binding, transferase activity and catalytic activity in bellflower roots. In conclusion, the exclusive proteins from diploid and tetraploid roots may provide insight clues for better understanding the characteristics and functions of proteins and metabolic activity of Platycodon grandiflorum.
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.
Grain sorghum (Sorghum bicolor) is a major staple for a large portion of the world. The crop ranks fifth among the cereals world-wide with respect to its importance for food and feed applications. To this end, the grain harvested from sorghum, and the millets provides an important source for dietary calories and protein for approximately one billion people in the semi-arid regions of the world. However, grain sorghum products are known to have relatively poor digestibility, only approximately 50%–70%, in comparison with other grains, such as wheat and maize, which tend to have digestibility percentages over 80% and 70%, respectively. Protein with high digestibility is by definition nutritionally superior owing to the increased availability of amino acids. Digestibility can be impacted by both protein–protein and⁄or protein–nonprotein interactions. However, with respect to grain sorghum, it is thought that the major factor influencing digestibility is the former because of high protein cross-linking around the protein body. To understand the mechanism of seed storage proteins in the sorghum, the proteomic analysis was carried out between the wild(BTX623) and mutant(M271207) genotypes of sorghum. Proteins were separated from the mature seed using IEF in the first-dimension and SDS-PAGE in the second dimension along with hybrid LTQ-FTICR mass spectrometry. After image analysis using Progenesis SameSpot software, we identified the 62 differential expressed protein spots out of 293 protein spots. Out of total differential expressed spots, 35 differential expressed protein spots (more than2-fold) were analyzed by mass spectrometry. Out of 35 protein spots, we were identified 20 protein spots as up-regulated and 15 protein spots as downregulated, significantly. In our proteomic investigation, the candidate proteins may provide novel clues for better understanding the characteristics of seed proteins in Sorghum.