Oldenlandia diffusa is a Chinese medicinal herb with antitumor activity capable of suppressing the growth of some cancer cell lines. Oleanolic acid and ursolic acid are triterpenoid compounds that exist in Oldenlandia diffusa. Recently, these have been noted for anti-inflammatory, anti-cancer, and hepato-protective effects. Application of both plant growth regulators, 2,4-D and kinetin, was found to be essential for the initiation of callus and suspension cells. Leaf blades of Oldenlandia diffusa was transformed into callus on Schenk and Hildebrandt medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L kinetin, while optimum initiation condition for suspension cells of Oldenlandia diffusa was determined to be 0.75 mg/L 2,4-D and 0.1 mg/L kinetin. Chromatographic separation of oleanolic acid from its derivatives was achieved using Rexchrom S5-100-ODS column. Analytical conditions for oleanolic acid were determined as follows: flow rate at 1.0 mL/min, UV length at 200 nm and mobile phase of $80\%$ acetonitrile and $20\%$ water. Production of secondary metabolites was found to be increased by the treatment with elicitors or signal transducers. The maximum production of oleanolic acid was 99.6 mg/L in cultures with 0.5 mM salicylic acid. It is 1.74 times higher than that of control.
Korea has plentiful precipitation but rainfall events concentrate on several months of rainy season in her weather condition. Korea, therefore, experiences drought for a given period every year. Moreover the soil has usually low water holding capacity, as it is composed coarse particles originated from the granite. Response of several oaks and the Korean red pine (Pinus densiflora) on water stress showed that water budget was significant factor determining vegetation distribution. In addition, dehydration level due to cold resistance mechanism of several evergreen plants during the winter season was closely related to their distribution in natural condition. Experimental result under water stress showed that the Korean red pine was very tolerant to desiccation but the seedlings showed high mortality during the dry season. The mortality tended to proportionate to soil moisture content of each site. A comparison between soil moisture content during June when it is severe dry season and moisture content of the culture soil when the pine seedlings reached the permanent wilting point due to water withheld proved that high mortality during the dry season was due to water deficit. Water potential of sample plants measured during the exposure experiment to the air pollutant showed a probability that water related factors would dominate the occurrence of visible damage and the tolerance level of sample plants. In both field survey and laboratory experiment, plants exposed to air pollution showed more rapid transpiration than those grown in the unpolluted condition. The result would due to injury of leaf surface by air pollutants. Aluminum (Al/sup 3+/) increased in the acid soil not only inhibits root growth but also leads to abnormal distribution of root system and thereby caused water stress. The water stresses due to air pollution and soil acidification showed a possibility that they play dominating roles in inducing forest decline additionally to the existing water deficit due to weather and soil conditions in Korea. Sludge, which can contribute to improve field capacity, as it is almost composed of organic matter, showed an effect ameliorating the retarded growth of plant in the acidified soil. The effect was not less than that of dolomite known in widely as such a soil ameliorator. Litter extract contributed also to mitigate the water stress due to toxic Al/sup 3+/. We prepared a model showing the potential interaction of multiple stresses, which can cause forest decline in Korea by synthesizing those results. Furthermore, we suggested restoration plans, which can mitigate such forest decline in terms of soil amelioration and vegetation restoration.
To isolate and purify the antimicrobial and antitumor agents in Xanthium strumarium L. hydrothermal extract. The crude extract was extracted in ether or ethylacetate under neutral, acidic, and alkali conditions. The antimicrobial activity of each extract was tested against 16 strains of bacteria, 2 strains of yeast, and 2 strains of fungus. The ether neutral extract (XE-N) exhibited the strongest growth inhibition upon the 8 strains of gram-positive bacteria, 6 strains of gram-negative bacteria and Cryptococcus neoformans. Fluorescein diacetate (FDA) testing of XE-N and XEA-N showed growth inhibition of the 3 strains of E. coli, S. aureus and C. albicans even at 30 ng/mL, with the exception of p. aeruginosa. XE-N-S1 and XE-N-S3 from neutral ether extract (XE-N), XE-N-S3 from the acidic ether extract (XE-A), and XEA-N-S1 from ethylacetate (XEA-N) were purified as antimicrobial and antitumor agents. However all purified compounds decomposed with the exception of XE-N-S1. The results upon the antitumor activities of the crude extract and of its purified compounds, showed that XE-N-S1 had the best antitumor activity against HeLa cells. In terms of antitumor activity against HepG2 cells, XE-N-S1 and XE-N-S3 were superior, and against HT29 cells XE-N and XE-N-Sl were good, against Saos2, NCI H522, NCI H1703, Clone M3 cells XE-N-51 was very good, and against LN CAP cells XE-N-S3 was the best. Comparing of cellular toxicities various extracts and purified compounds with the existing antitumor agents, XE-A, XEA-A and XEA-B had the lowest toxicity, and XE-B had a lower toxicity than etoposide. XE-N-S1 and XE-N-S3 showed higher toxicities than etoposide, and the toxicity of XE-A-S3 was higher than that of etoposide, and lower than that of csplatin.
The objective of this study was to monitor health conditions of genetically identical somatic cells cloned Korean white cattle, endangered indigenous cattle (EIC) and indigenous cattle (IC) by analysis of hematologic characteristics. Naturally ovulated oocytes and donor cells were used for somatic cell nuclear transfer (SCNT). Donor cells and enucleated oocytes were followed by electric fusion, chemical activation and surgical embryo transfer into the oviducts of surrogate females. Two recipients became pregnant; two maintained pregnancy to term, and one live cattle were delivered by caesarean section. The cloned Korean white cattle were genetically identical to the nuclear donor cattle. As a result, the mean values of RBC and platelet of cloned cattle and white cattle were significantly decreased by age (P<0.05). The mean values of RBC, HCT, MCV and MCHC between cloned cattle and IC of the same age (1~2 years) showed the statistical significance (P<0.05). Also, in the WBC of Korean white cattle, the estimated values were decreased according to the age from $12.0{\times}10^3/{\mu}l$ under 1 year to $11.0{\times}10^3/{\mu}l$ over 1 years respectively. Although clone-cattle had lower numbers of RBC than reference range, the most of RBC and WBC related heamatologic results of cloned cattle were not different when compared to reference range. This study suggests that cloned Korean white cattle derived from SCNT did not have remarkable health problems, at least in the growth pattern and hematological parameters. In addition, this study provides a valuable resource for further investigations of the preservation of rare genetic stocks underlying traits of interest in cattle.
It is well known that unidentified factors in sera, hormones and growth factors promote the proliferation of granulosa cells and nuclear maturation of bovine COCs in vitro. Attemps had been developed the simple composition of culture media and similar system to in vivo conditions has been applied. In the present study, we investigated the effect of TGF-${\beta}$ on in vitro maturation and in vitro development of Hanwoo COCs. When the COCs were matured in TCM 199 containing 0.1, 1 or 10 ng/ml TGF-${\beta}$ for 24 hrs, metaphaseⅡ of COCs were obtained 95.8%, 100% of matured COCs, respectively and there were no differences among the concentrations of TGF-${\beta}$. Matured COCs with TGF-${\beta}$ cultured in maturation medium after in vitro fertilization, developmental rate to blastocyst were 0~0.8%. Matured COCs with TGF-${\beta}$ were cultured in TCM 199+10% FBS, 0.8% BSA, 0.1% PVA, blastocyst formation were showed in 12.4%, 12.8%, 8.5% of those and cultured in IVMD or IVD without serum were 38.4%, 34.8%, respectively. There were significant differences among the media (P<0.05). TGF-${\beta}$ is available for i vitro maturation of bovine COCs, but further investigation would be need for finding the synergistic autocrine/paracrine fashion of other growth factors in early bovine development.
In order to elucidate the growth mechanism of sulfide chimney formed as a result of seafloor hydrothermal mineralization, we carried out the mineralogical and fluid inclusion studies on the inactive, sulfide- and silica-rich chimney which has been recovered from a hydrothermal field in the Cleft segment of the Juan de Fuca Ridge. According to previous studies, many active and inactive vents are present in the Cleft segment. The sulfide- and silica-rich chimney is composed of amorphous silica, pyrite, sphalerite and wurtzite with minor amounts of chalcopyrite and marcasite. The interior part of the chimney is highly porous and represents a flow channel. Open spaces within chimneys are typically coated with colloform layers of amorphous silica. The FeS content of Zn-sulfides varies widely from 13.9 to 34.3 mole% with Fe-rich core and Fe-poor rims. This variation possibly reflects the change of physicochemical characteristics of hydrothermal fluids. Chemical and mineralogical compositions of the each growth zone are also varied, possibly due to a thermal gradient. Based on the microthermometric measurements of liquid-rich, two-phase inclusions in amorphous silica that was precipitated in the late stage of mineralization, minimum trapping temperatures are estimated to be about 1140 to 145$^{\circ}$C with the salinities between 3.2 and 4.8 wt.% NaCI equiv. Although the actual fluid temperatures of the vent are not available, this study suggests that the lowtemperature conditions were predominant during the mineralization in the hydrothermal field at Cleft segment. Comparing with the previously reported chimney types, the morphology, colloform texture, bulk chemistry, and a characteristic mineral assemblage (pyrite + marcasite + wurtzite + amorphous silica) of this chimney indicate that the chimney have been formed from a relatively low-temperature (<250$^{\circ}$C) hydrothermal fluid that was changed by sluggish fluid flow and conductive cooling.
This study was conducted to identify the relationship between environmental factors and algal bloom, and provide information for efficient management based on the results of monitoring the environmental parameters and algal diversity in the Jundai reservoir from March 2011 to October 2013. Little change in the weather conditions was observed during the study period except for a slight decrease in rainfall. Concentration of TN and TP in the reservoir exceeded water quality standards for agriculture and significant correlation between algal growth and environmental factors was observed. Phytoplankton in Jundai reservoir included 6 classes, 40 genus, 62 species, and the phytoplankton abundance was in the range of $1.3{\times}10^4{\sim}2.8{\times}10^6$ cells $mL^{-1}$. The annual average of phytoplankton abundance and Chl-a gradually decreased as TN and TP concentrations decreased. Overall Anabaena sp., Oscillatoria sp., and Microcystis sp. were the dominant species in Jundai reservoir. As the water temperature increased, the dominant species were Anabaena sp., Microcystis sp. and Oscillatoria sp., in that order. Anabaena sp. was dominant from spring to early summer with increase in water temperature and pollutant concentrations, and high correlation with environmental factors was observed. Microcystis sp. was dominant depending on changes in the nutrient levels. In the case of Oscillatoria sp., there was no significant correlation between phytoplankton biomess and Chl-a. However, efficient management of water environment and practical control of algal bloom in small scale reservoir polluted by livestock and farm irrigation should be achieved by identification of the relationship between algal growth and environmental factors.
This experiment was conducted to find out the effects of fertilizer-N and co-inoculation of the Rhizobium japonicum and the Azospirillum lipoferum on nodulation, $N_2$-fixation, and growth of soybean under in situ conditions. The results obtained were summarized as follows: 1. The yield of soybean dry matter was significantly greater in the R. japonicum alone, and the mixed inoculation of R. japonicum and A. lipoferum than those of un-inoculation. But inoculum applied by different strains did not significant effect on plant growth. The effects of nitrogen applied on soybean dry matter were higher in the ammonium sulfate than potassium nitrate, and decreased with increasing rates of two forms of nitrogen applied regardless of nitrogen source. 2. Acetylene redution activity was more increased in a single inoculation of R. japonicum than those of the mixed inoculation of the R. japonicum and the A. lipoferum, in cases of Danyeup cultivar, regardless of the form of combined nitrogen used. 3. Nodule mass and total nitrogenase activity per plant showed the positively significant effect in the interrelationship between dry matter of soybean and some factors related to nitrogen fixation efficiency. 4. The highest symbiotic effect in Danyeup cultivar was obtained when a single R. japonicum 84 Dy-1 strain was inoculated and fertilized with 18 mM potassium nitrate.
Kim, Hye-Jin;Lee, Jong-Nam;Kim, Ki-Deog;Im, Ju-Sung;Lim, Hak-Tae;Yeoung, Young-Rok
Journal of Plant Biotechnology
/
v.38
no.3
/
pp.221-227
/
2011
This study was carried out to determine optimal medium conditions for mass propagation by bioreactor culture of ever-bearing strawberry (Fragaria ${\times}$ ananassa D.). Two different type of nitride were that mixed $NH_4NO_3$ and $KNO_3$ or added $KNO_3$ only. And nitride concentrations were at the 4 levels of $1/2{\times}$, $1{\times}$, $2{\times}$ and $3{\times}$ that was included $NH_4NO_3$ and $KNO_3$. Sucrose content ranged at 3 levels of $10g{\cdot}L^{-1}$, $20g{\cdot}L^{-1}$ and $30g{\cdot}L^{-1}$ and medium pH were at the 3 levels of 4.6, 5.6 and 6.6. In bioreactor culture, medium that are included $NH_4NO_3$ and $KNO_3$ together in MS medium was suitable for mass propagation. Medium EC rose rapidly when the nitride concentration was increased. For that reason, plantlet growth was inhibited. Shoots of nitride $1/2{\times}$ concentration was 10.8 ea and $1/2{\times}$ concentration was suitable for shoot propagation. Fresh weight of sucrose $30g{\cdot}L^{-1}$ was 3,101 mg which was heaviest and aerial and ground part were higher than the other concentration. Shoots were increased in proportion to the increasing concentration of sucrose. In the pH condition, from pH 5.6 to 6.8 were appropriate for the optimum growth of aerial and ground part of plant. From the results, in bioreactor culture for mass propagation, MS medium was suitable $1/2{\times}$ concentration that was included $NH_4NO_3$ and $KNO_3$ together, and added $30g{\cdot}L^{-1}$ of sucrose, and then adjusted pH between 5.6 and 6.6.
Lee, Won Jeong;Lee, Ji Hyun;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Heung Tae;Choi, Gyung Ja
Horticultural Science & Technology
/
v.33
no.1
/
pp.70-82
/
2015
This study was conducted to establish an efficient screening system to identify melon resistant to Fusarium oxysporum f. sp. melonis. F. oyxsporum f. sp. melonis GR was isolated from infected melon plants collected at Goryeong and identified as F. oxysporum f. sp. melonis based on morphological characteristics, molecular analyses, and host-specificity tests on cucurbits including melon, oriental melon, cucumber, and watermelon. In addition, the GR isolate was determined as race 1 based on resistance responses of melon differentials to the fungus. To select optimized medium for mass production of inoculum of F. oxysporum f. sp. melonis GR, six media were tested. The fungus produced the most spores (microconidia) in V8-juice broth. Resistance degrees to the GR isolate of 22 commercial melon cultivars and 6 rootstocks for melon plants were investigated. All tested rootstocks showed no symptoms of Fusarium wilt. Among the tested melon cultivars, only three cultivars were susceptible and the other cultivars displayed moderate to high resistance to the GR isolate. For further study, six melon cultivars (Redqueen, Summercool, Superseji, Asiapapaya, Eolukpapaya, and Asiahwanggeum) showing different degrees of resistance to the fungus were selected. The development of Fusarium wilt on the cultivars was tested according to several conditions such as plant growth stage, root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease. On the basis of the test results, we suggest that an efficient screening method for melon plants resistant to F. oxysporum f. sp. melonis is to remove soil from roots of seven-day-old melon seedlings, to dip the seedlings without cutting in s pore s uspension of $3{\times}10^5conidia/mL$ for 30 min, to transplant the inoculated seedlings to plastic pots with horticulture nursery media, and then to cultivate the plants in a growth room at 25 to $28^{\circ}C$ for about 3 weeks with 12-hour light per day.
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