본 연구의 목적은 토종다래의 용매별 추출물에 따른 약리활성에 대한 검증 및 효능 평가로서 토종다래의 항산화, 항염증에 대한 효과를 확인하였다. 염증 반응은 자극이 가해지면 histamine, serotonin, prostaglandin과 같은 혈관 활성물질에 의해 혈관 투과성이 증대되어 염증을 유발하고 cytokine, free radical, lysosomal enzyme 등 다양한 매개 인자가 관여한다. 자극에 의한 macrophage cell의 염증반응은 tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), interleukin-1β(IL-1β)와 같은 pro-inflammatory cytokine의 발현이 유도되고, inducible nitric oxide synthase(i-NOS)와cyclooxygenase-2(COX-2)에 영향을 받는 유전자의 발현을 자극하게 되어 nitric oxide(NO) 등의 염증 인자가 생성된다. 이에 따라 토종다래 추출물의 항염증에 대한 연구를 위해 이에 영향을 주는 인자인 i-NOS, COX-2의 단백질 발현억제 작용을 확인하였다. 그 결과, HKE > HKA > HKW 순서로 높은 효능을 확인 할 수 있었다. 가장 효과가 좋은 HKE 처리군에서 다양한 염증성 인자의 mRNA 발현량을 확인하였다. 측정 결과, HKE(2,000 μg/mL)는 i–NOS, COX-2, IL-1β, IL-6, TNF-α mRNA 발현이 각각 93.2%, 27.9%, 96.4%, 89.4%, 73.9% 억제되는 효과를 확인할 수 있었다. 또한, HKE의 nuclear factor kappa B(NF-κB) 단백질 발현에 농도의존적으로 유의미한 결과를 확인하였으며, 이에 토종다래의 항염증효과는 LPS에 의한 TLR4의 자극에서 NF-κB 경로의 완화로 나타는 것임을 검증하였다. 결론적으로 토종다래는 70% ethanol 추출물(HKE)의 항염증 효과가 가장 높았으며, HKE는 대식세포에서 NF-κB 염증관련 경로의 억제로 세포 내 mRNA 및 단백질 수준에서의 염증인자들의 생성을 저해하여 항염증 효과가 명백히 확인되었다. 향후 본 연구팀은 토종다래의 항염증과 관련된 유효성분의 분리정제 및 구조분석을 진행할 예정이다.

In this study, soy-powder yogurt (SPY) with enhanced levels of γ-aminobutyric acid (GABA) and isoflavone aglycone was produced from sprouting high-protein soybeans (HPSs). The fermented steam-HPS sprouts (0 to 4 cm) were fermented (72 h) with Lactobacillus brevis, and the total free amino acids (FAAs) of the formed mixtures were determined to be 79.53, 489.93, 877.55, 780.53, and 979.97 mg/100 mL in the fermented HPS (FHPS), and the fermented steam-HPS with 0 cm (FSHPS-0), 1 cm (FSHPS-1), 2 cm (FSHPS-2), and 4 cm sprouting lengths (FSHPS-4), respectively. The levels of glutamic acid (GA) and GABA were observed to be the highest, 100.31 and 101.60 mg/100 mL, respectively, in the unfermented HPS (UFSHPS-1, 1 cm) and FSHPS-1 sprouts, respectively. Moreover, the total contents of the isoflavone glycoside form decreased proportionally to the increasing total levels of isoflavone aglycones after fermentation in FSHPS-0, FSHPS-1, FSHPS-2, and FSHPS-4. The levels of isoflavone aglycones were detected as 350.34, 289.15, 361.61, 445.05, and 491.25 μg/g in FHPS, FSHPS-0, FSHPS-1, FSHPS-2, and FSHPS-4, respectively. While FSHPS-1 exhibited the highest DPPH (63.28%) and ABTS (73.28%) radical scavenging activities, FSHPS-4 contained the highest isoflavone aglycone ratio (81.63%). All in all, the FSHPS-1 mixture prepared in this study exhibited high GABA content and functional prosperity, thereby making it suitable for potential applications in the soy-dairy industry.

Present study aimed to determine the effect of ‘bitter melon’, a popularly used fruit in Bangladesh and several other Asian countries, on high-fat-diet-induced type 2 diabetes. To investigate the effect, ethanol extract from bitter melon (BME) as a dietary supplement with mouse chow was used. BME was found to significantly attenuate the high-fat diet (HFD) -induced body weight and total fat mass. BME also effectively reduced the insulin resistance induced by the HFD. Furthermore, dietary supplementation of BME was highly effective in increasing insulin sensitivity and reducing hepatic fat and obesity. These results indicate that BME could be effective in attenuating type 2 diabetes and could therefore be a preventive measure against type 2 diabetes.

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.

EP was obtained through 20% ethanol extraction of Pueraria lobata root, and the fermented form of EP, FEP, was prepared from the EP after incubating with Lactobacillus rhamnosus vitaP1. There was no significant toxicity by EP and FEP up to 1000 ㎍/㎖ in NIH-3T3, HaCaT, and B16F10 cells. In addition to antioxidant potentials of EP and FEP determined by DPPH and ABST assays, we confirmed increase of procollagen type I and elastin synthesis by supplementation of the EP and FEP at the concentration of 50 ㎍/㎖ using ELISA kits. The protein expression levels of matrix metalloprotease (MMP)-1, -3, and -9, those are involved in the degradation of collagen or other skin matrix proteins, were remarkably suppressed while their inhibitory protein metallopeptidase inhibitor 1 (TIMP-1) was greatly up-regulated by supplementation of the EP and FEP at a concentration of 50 ㎍/㎖. Taken together, both EP and FEP supplementation could be involved in the suppression of the skin wrinkle formation through inhibiting degradation of collagen and stimulating the synthesis of collagen and elastin. The results showed that the anti-wrinkle potential of the EP and FEP will be a promising candidate for developing cosmeceutical compounds or products.

Background : Kenaf (Hibiscus cannabinus L.) is a member of the malvaceae family and has been prescribed in traditional folk medicine in Africa and India. It showed broad biologicas activities such as hepatoprotective activity, antioxidative activity and haematinic activity. Kenaf leaves contain a lot of vegetable calcium, protein, iron and vitamins, so have high potential as functional food. Thus, the present study purposed to analyze useful substances contained in kenaf leaves and their activities and look for their possibilities as the materials of food and medicine. Methods and Results :　We sowed the seeds of three varieties of kenaf, namely, Dowling, Everglade-41 and Tainung-2 at planting space of 20 × 20㎝ on the field and collected plants on the 98th , 127th and 141st days from sowing. The total polyphenol content was measured using a slightly modified Folin-Denis method, which uses the phenomenon that a phenolic substance turns blue by reaction with phosphomolybdate. Conclusion : We analyzed SOD enzyme activity related to the scavenging of superoxide anion radical (․O2-) through NBT reduction. As for the SOD activity of leaf extract by variety, the activity was 92.6%, 91.0% and 92.6%, respectively, in Tainung-2 on day 98, 127 and 141 from sowing, slightly higher than those in the other two varieties but not significantly different. The activity was slightly higher in the middle period of growth than in the harvest period, but not much different according to harvest time. As for the DPPH radical scavenging activity of leaf extract by variety, the activity was 80.87% and 80.71%, respectively, in Tainung-2 on day 98 (30th of October)and day127 (28th of September), slightly higher than the other two varieties.

Background : Sorghum is a major cereal food crop used in many parts of the world. It has been grown on a subsistence level by farmers, under various conditions of environmental stresses in the semi-arid tropics of Africa and Asia. This crop has received significant attention because of its ability to lower cholesterol in the blood, and its anti-dementia, antioxidant, and antimicrobial properties. It is possible to develop a functional and commercially viable sorghum variety by using superior cultivars of sorghum. The objective of this study was to construct a database of superior sorghum accession. Methods and Results : We used 250 sorghum accessions collected from different geographical bioregion in Korea. We determined various growth characters including germination rate and ear length of these accessions. To determine the antioxidant capacity, we measured the DPPH radical scavenging activity, ABTS scavenging activity, total phenolic contents, and total flavonoid contents. 189 accession showed higher germination (> 80%) than other accessions. Higher DPPH radical scavenging activity was observed in 11-SB-078 (RC50: 1.89 ± 2.88 ㎍/ ㎖), and higher ABTS scavenging activity was recorded in 11-SB-116 (RC50: 35.48 ± 2.42 ㎍/㎖) than other accessions. The ear length ranged from 15 ㎝ to 48 ㎝, the total phenolic contents ranged from 3 ㎎·GAE/g to 77 ㎎·GAE/g, and total flavonoid contents ranged from 0.09 ㎎·QE/g to 1.07 ㎎·QE/g. Conclusion : Among 250 sorghum accessions, we selected 10 with both superior agronomic characters and higher functional food quality.

Background : Gangwon-do (Province) is chief producing district of six-year-old raw material of red ginseng and transplanting culture is more than 90%. The 6-year-old red ginseng is easy to differentiate in quality and can secure a stable market, but it has high disadvantages such as high seedling cost, effort cost, and long term cultivation. Therefore, it is necessary to develop environment-friendly products that enhance price competitiveness based on 3 to 4-year-old ginseng by using direct seedling cultivation techniques. When ginseng seed sown directly, it is highly productive and has high disease resistance, which is advantageous for environment-friendly cultivation. Thus, it is advantage to save work force in seed production stage. The purpose of this study is to find the planting density for 4-year-old ginseng. Methods and Results : In order to determine proper planting density, ginseng growth characteristics such as root length, root diameter, raw weight, and yields were investigated at 4-year-old ginseng after planting 34, 67, and 100 roots per ㎡. As a result, the root length was the longest at 33.9 ± 5.45 ㎝ in 67 roots/㎡, and there was no difference in root diameter. The raw weight of 4-year-old ginseng was the highest at 65.1g in 34 roots/㎡. However, yields were 4.3 ± 0.78 and 4.8 ± 0.50 ㎏ in the 67 roots/㎡ and 100 roots/㎡, respectively, which were higher than the 34 roots/㎡. Conclusion : The proper planting density for direct seedling cultivation of yunpung cultivar was 67 - 100 roots/㎡, and these results could be applied to increase productivity in direct seeding cultivation of ginseng.

Background : Angelica dahurica Bentham et Hooker, Angelica gigas Nakai, Ostericum koreanum Maximowicz and Peucedanum japonicum Thumberg　are a major medicinal plant in north Geungbuk province. Using medicinal plants are impotant it`s ingredient. Dry condition and stroage method are not standard manual. The ingredient variation of dry condition and stroage method were not researched. Methods and Results : Using plant material were cutivated on Gyongsangbukdo Bonghwa area. It were studied ingredient variation after dry and storage condition by HPLC methods. Major ingredient of Angelica gigas Nakai are decurusin, decurusinangelate. Heated air bulk dry get more decursin than natuarl dry and decurusinangelate of natural bulk dry was higher than heated air bulk dry. Major ingredient of Ostericum koreanum Maximowicz are imperatorin and isoimperatorin.. Imperatorin of Ostericum koreanum was highest peak on 50℃ heated-air dry after plastic bag sorage and isoimperatorin was highest peak on 40℃ heated-air dry after mountain cultivation. Imperatorin is a major ingredient Angelica dahurica Bentham et Hooker. Heated air bulk dry get more decursin and decursinangelate than natuarl dry and small heated-air dry. Peucedanol-7o_glucoside is a major ingredient Peucedanum japonicum Thumberg. Natural bulk dry get more peucedanol-7o_glucoside than heated-air bulk dry. Conclusion : Ingredient of Angelica dahurica Bentham et Hooker, Angelica gigas Nakai, Ostericum koreanum Maximowicz are different under various cutivation, drying method, storage. Diffent Ingedients of Angelica gigas Nakai, Ostericum koreanum Maximowicz were not accord it’s optical conditon.

Background : Schizandra chinensis Baillon have five tastes and lately it is using a beverage broadly. Schizandra chinensis is one of the top producing medicinal plant in Korea. Mungyeong of Gyongbuk province produce almost of Schizandra chinensis. Maturity of Schizandra chinensis get 3 years and proliferation of Schizandra chinensis was not a manual. It is needed that a new cultivar has a big fruit and high quality chracteristics using processed food and beverage. Methods and Results : 105 lines of Schizandra chinensis were collected on Mungyeong, Yeongwol, Jinan. It were studied it’ characteristics especially it’s fruit trait. Fruit traits of Schizandra chinensis were researched on fruit length, fruit weight, maturity, number of fruit, male and female ratio, powdery mildew. Fruit length of Schizandra chinensis is relation of fruit weight. It were founded 15 lines of long fruit length. 5 lines were studied high fruit weight and it’s weight were 32 to 41g. Number of fruit has relation with fruit weight and high fruit weight gets many fruits. it’s numer of fruits were 3 to 41. Male and female ratio were very impotant characteristic. High level of female ratio has quantity of fruit. High level of female ratio were founded 2 lines. Finally It was selected 3 good breed lines of Schizandra chinensis. Conclusion : 105 lines of Schizandra chinensis Baillons were collected on Mungyeong, Yeongwol, Jinan. It were founded 15 lines of long fruit length and 5 lines were studied high fruit weight. High level of female ratio were founded 2 lines. 3 good breed lines of Schizandra chinensis were selected.

Background : Temperature is major factor for growth plant. Recently, because of global warming, abnormal temperature included drought, deluge, sudden temperature change and heavy snow damaged crops in the world. In Korea, crops have been sensitive to low temperature on early growth stage, e.g. fruit tree and ginseng, were damaged owing to sudden heavy snow and cold on Spring. Therefore, recently interest in cold resistance crops were increased in demand rapidly. This study was performed to establish transgenic Nicotiana benthamiana by transforming cold resistant gene related to cold tolerance S-adenosylmethionine synthetase (SAMS) isolated from Miscanthus sinensis. Methods and Results : Total RNA was extracted from leaves of M. sinensis using Trizol assay and isolated MsSAMS. Isolated MsSAMS was insert into SacⅠ- XbaⅠ sites of pMBP1 vector. The vector was transformed to Agrobacterium tumefaciens strain LBA4404 by DH5α. A. tumefaciens with binary plasmid were selected at YEP medium supplemented with kanamycin. Cut leaves of tobacco were co-cultured with selected A. tumefaciens. Co-cultured leaves was grown on regeneration medium for a month at dark condition, and transferred to at light condition. Regeneration shoot from callus were excised and transferred to root-induction medium. Approximately, 58% of leaves explant produced callus. Nearly, 30% of callus had shoot and approximately, 94% of shoots were rooted in root-induction medium. Conclusion : We established an efficient transformation system of N. benthamiana transformed by using MsSAMS gene related to cold tolerance isolated from M. sinensis. We may use the produced transgenic plants to prevent damages carried by cold.

Background : Perilla frutescens L. is a bisexual, annual plant belonging to the Labiatae. Dormancy period of Perilla seed usually ranged from 85-200 days. The germination enhancing effects in a reduced seed vigor during the GA3 treatment has been reported. Objectives of the present study was to evaluate the effect of GA3 on germination rate and the expression of germination-related genes expression by using perilla seeds. Methods and Results : Three different cultivars (Saeyeopsil, Okdong) and accession (line 141) of perilla were used in the experiment and treated with GA3 at different concentration (50, 100, 300, 500 μM) for one day at dark condition. Germination test for perilla seed was carried out by using 100 perilla seeds treated with GA3 at 25℃. Rate of germination were evaluated after 10 days and the experiments were repeated three times in similar condition. Samples were collected from each cultivar and accessions for RNA extraction. After cDNA synthesis quantification, templates were subjected to RT-PCR using actin primers. EST blast performed for sequencing the RT-PCR products. Conclusion : Result showed that 100μM GA3 treated seeds of three perilla cultivars and accessions showed the highest germination rate. However, concentration of GA3 over 100 μM and lower than 100 μM resulted into reduced germination rate. Furthermore, expression of germination-related genes from 100 μM treated GA3 was higher compared to untreated sample by using 100 μM GA3 treated sample by using pC12, pJ14 primers.

Background : Cordyceps militaris has been an wonder drug to anti-aging efficacy and called the three main drugs with ginseng and deer antler from the past. Cordycepin, cordycepic acid (d-mannitol) and adenosine are known as functional ingredients in Cordyceps militaris. Among them, cordycepin, the representative component, has been reported as antimicrobial substance containing immune enhancement, anti-cancer and anti-inflammatory effects. Methods and Results : After Cordyceps militaris produced from different types of medium mixed with 10-fold volume of purified water, the mixture were extracted at 70±5℃ for a hour and that extracts re-extracted using ultrasonics wave for 30 minutes. Qualitative analysis of the index component was determined by using the Q-TOF (A quadrupole time-of-flight mass spectrometer), and quantitative analysis was performed by using HPLC (high-performance liquid chromatography) with Xselect HSS T3 column (2.1 X 100 mm, 2.5㎛, Waters, USA) and ultrapure water and acetonitrile as mobile phase A and B. Detection column temperature, injection volume and the flow rate were 35℃, 2 μL and 0.3 mL / minute respectively. The cordycepin content of Cordyceps militaris produced from medium mixed with vegetable and animal ingredients higher than single ingredient. Moreover, through a variety of analyzes by varying the type and content of the medium additives, the cordycepin in Cordyceps militaris produced from medium mixed with animal ingredients highest. Furthermore, the cordycepin content of a fruit body was higher than those of the a mycelium. Conclusion : These results provide a method for producing an high cordycepin content of Cordyceps militaris as functional food ingredient.

Background : This study aimed to investigate the antioxidant and anti-inflammatory activities of water chestnut (Trapa japonica Flerow) extract. Methods and Results : The antioxidant and anti-inflammatory activities of 100% methanol extract of water chestnut were investigated. The methanol extract was evaluated for its total phenolic and flavonoid content, DPPH•(1,1-diphenyl-2-picrylhydrazyl) free-radical scavenging activity,reducing power, andeffect on nitric oxide (NO) production and cell viability using real-time polymerase chain reaction (qPCR). The total phenolic content was 438.31 ㎍ allic acid equivalent (GAE)/㎎ extract and the total flavonoid content was 61.40 ㎍ quercetin equivalent (QE)/㎎ extract. In addition, results revealed the extract possessed antioxidant activity (DPPH• free-radical scavenging activity) with IC50 value of 5.28 ㎍㎖ The reducing power of the extract was assayed spectro photometrically and showed Abs of 0.71 at 100 ㎍㎖ Furthermore, extracts of water chestnut exhibited no cytotoxicity in RAW 264.7 cells. In addition, the NO assay revealed that LPS-induced NO production was significantly inhibited following treatment with water chestnut extracts. The expression of pro-inflammatory proteins such as inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 decreased in a concentration-dependent manner. The water chestnut extract also decreased tumor necrosis factor α (TNF-α) release. Conclusion : Therefore, the present findings provide scientific evidence for the nutritional potential, chemical composition, and biological activities of Trapa japonica Flerow anddemonstrate its potential use as a functional food forapplication in the pharmaceutical industry

Background : Miscanthus is a diploid hybrid and a temperate, perennial, cross-pollinating grass used as bioenergy plant, biomass production and high quality cellulose and ethanol production. This study was to determine an efficient transformation system for Miscanthus sinensis, and to optimize factors and conditions required for expression of MsCOMT–AS gene. Methods and Results : An efficient transformation of callus from M. sinensis was established using Agrobacterium tumefaciens strain LBA4404 harboring a binary vector pMBP1. In order to establish stable transformation system, we obtained high transformation rate from callus by various transformation factor explant type, strain, co-culture periods, acetosyringone concentration, and selective marker. Finally, in this study, seven putative transgenic plants were obtained. Through various tests including PCR analysis and southern blot were to detect antisense of COMT digested Xba I and Sac I restriction enzymes. The biomass of the control plant was superior than transgenic plants. Conclusion : This study was to develop transgenic Miscanthus sinensis by Agrobacterium tumerfeciens mediated transformation to produce high bioethanols and to reduce the lignin content of transgenic plants. Detailed characterization of the transgenic plants revealed interesting finding about COMT gene expression in the segregates

Background : Ginsenosides, the main ingredient of ginseng roots can be confirmed various physiological activity such as anticancer, antioxidant, a natural ginsenosides is there a structure to be absorbed into the body does not work well absorbed through this process biologically active thus a high conversion ginsenosides. β-glucosidase enzyme is observed in several of the microorganism with an enzyme that serves to convert a ginsenoside prosper that is absorbed into the body. Methods and Results : To view a primary β-glucosidase activity, the bacteria were innoculated in esculin agar medium and the color change of the media were measured by the time and degree of changing color. In the other method, 5 mM of p-nitrophenyl-β-D-glucopyranoside (pNPG) containing 25 mM phosphate buffer solution (pH 7.0) was added to 50 ul enzyme solution. Then the solution was added to 50 ul reaction for 5 min at 30°C. The amount of p-nitrophenol liberated measured at 405 nm absorbance. The experimental results showed higher β-glucosidase activity in Pediococcus pentosaceus, Leuconostoc mesenteroide, Leuconostoc mesenteroides subsp. cremoris, and Paenibacillus polymyxa by using esculin agar medium method. Similarly in second method, β-glucosidase activity was higher in P. pentosaceus 402.32±11.43 unit/l, L. mesenteroide 353.73±14.64 unit/l, Lactobacillus sakei 198.4±15.47 unit/l Lactobacillus plantarum subsp. plantarum 164.1±8.12 unit/l. Conclusion : The result that the β-glucosidase activity was higher in P. pentosaceus, L. mesenteroide, and L. plantarum subsp. plantarum as compared to tested microbes. Therefore selected bacteria can be used in the industry of functioned foods and beverage to improve human healths.

Lepisorus thunbergianus (Kaulf.) Ching has been used in folk medicine in Korea. In this study, a L. thunbergianus methanol extract and its fractions were investigated for their antioxidant properties. The results showed that the ethyl acetate and butanol fractions of L. thunbergianus possess potent DPPH radical scavenging activities. Both fractions also possessed reducing power and inhibited reactive oxygen species formation. In addition, the cytotoxic activity of the L. thunbergianus n-hexane fraction (HF) was investigated. The results suggested that the HF remarkably suppressed proliferation of human breast, liver and colon cancer cells. These results demonstrate, for the first time, that L. thunbergianus extract induces apoptosis in SW620 cells, suggesting that L. thunbergianus may have potential as a therapeutic agent for colon cancer.

Arabidopsis E3 SUMO ligase SIZ1 (AtSIZ1) controls vegetative growth and development including responses to nutrient deficiency and environment stresses. Here, we analyzed the effect of AtSIZ1 on the stability and amount of seed proteins. Proteomic analysis showed that the amount of three major nutrient reservoir proteins, CRUCIFERIN (CRU) 1, 2 and 3, were decreased in siz1-2 mutants. However, quantitative real-time RT-PCR showed that transcript levels of CRU1, 2 and 3 genes were rather significantly higher in siz1-2 mutants than wild-type plants. Yeast two hybrid analysis revealed that AtSIZ1 interacts with CRU1, CRU2 and CRU3, strongly suggesting that CRU1, 2 and 3 proteins are sumoylated by AtSIZ1. In addition, the analysis of amino acid composition by HPLC showed that the contents of amino acids were a bit high in siz1-2 mutants. Our data indicate that AtSIZ1 plays an important function for accumulation of seed storage proteins through its ligase activity.

Gene expression is regulated by DNA and histone methylation by DNA and histone methyltransferases, respectively. In animal system, DNA methyltransferase with CG methylation activity is modified by SUMO conjugation and then its activity was increased, which means that the activity of DNA methyltransferase is modulated by posttranslational modification. so Chromatin remodeling is a new concept for expression of controlling of gene function. We thus analyzed the effect of E3 SUMO ligase AtSIZ1 in CMT3 (chromometnylase 3)-mediated genome methylation by next-generation sequencing (NGS), methyl binding domain MeDIP-sequencing and gene analysis using siz1-2 and cmt3 mutants. we carried out CG-enrich analysis by MeDIP sequencing revealed that the methylation level of the genome including transposons was significantly low in siz1-2 mutants compared to wild-type. Result showed the genes regulated by methylation, that genes related of embryo and root development, cellulose metabolism, and post-translational modifications. All of our data indicate that the methyltransfearse activity of CMT3 may be able to be regulated by AtSIZ1 and thereby CMT3-mediated gene expression and plant development also can be controlled by E3 SUMO ligase activity. Besides, our data also suggest that ammonium (NH4+) can stimulate AtSIZ1- and CMT3- mediated DNA methylation.

The transition from vegetative growth to flowering is a major developmental switch in the plant cycle and the timing of flowering is very critical for reproduction of plant species. In transition to flowering in plants, Flowering locus C (FLC) is one of the crucial factors. Here, we showed How the stability and activity of FLC are regulated by sumoylation mechanism. By pull-down assay, we showed that FLC interact with E3 SUMO ligase in vitro and vivo. And we showed that FLC is sumoylated in vitro condition with AtSUMO1 protein. In transgenic plants with overexpression of FLC and inducible expression of AtSIZ1, sumo E3 ligase led to increase of FLC protein level and delayed the post-translation degradation of FLC indicating that Arabidopsis E3 sumo ligase AtSIZ1 stabilizes FLC. Also, the plants with overexpression of mutant FLC (K154R, a mutation of the sumoylation site on FLC) flowered considerably earlier than plants with overexpression of FLC but comparable with wild type indicating that sumoylation is a important part for function of FLC. Our data indicate that the sumoylation of FLC is critical for its role in the control of flowering time.