Objectives : The process through which mesenchymal cells condense and differentiate into chondrocytes to form new bone is known as endochondral bone formation. Chondrogenic differentiation and hypertrophy are essential steps in bone formation and are influenced by various factors. The stem bark and root bark of Eleutherococcus sessiliflorus (ES) have been widely used to treat growth retardation and arthritis in traditional Korean Medicine. In this study, we aimed to investigate the possible role of the stem bark of ES in the stimulation of chondrogenic differentiation in clonal murine chondrogenic ATDC5 cells. Methods : In ATDC5 cells treated with ES extract, cell viability and extracellular matrix production were determined using CCK-8 assay and Alcian blue staining, respectively, and alkaline phosphatase activity was measured. We also examined mRNA and protein expression levels of genes related to chondrogenic expression in ATDC5 cells using reverse transcription-polymerase chain reaction and western blot analyses. Results : ES extract increased the accumulation of Alcian blue-stained cartilage nodules and alkaline phosphatase activity in ATDC5 cells. It increased the mRNA expressions of chondrogenic markers including bone sialoprotein (BSP), cartilage collagens, Runt-related transcription factor-2 (RUNX-2), osteocalcin (OCN), β-catenin, and bone morphogenetic protein-2 (BMP-2), as well as the protein expressions of β-catenin, RUNX-2, BMP-2, and alkaline phosphatase (ALP). Conclusion : Taken together, these results suggest that ES extract exhibits a chondromodulating activity and therefore may be a possible agent for the treatment of bone growth disorders.
Journal of The Korean Society of Grassland and Forage Science
/
v.23
no.2
/
pp.81-90
/
2003
This pot experiment was conducted to find out the effects of boron application(B$_{0}$; 0.0, B$_1$; 0.2, $B_2$;2.0, $B_3$;6.0, $B_3$;6.0, $B_4$;15.0me B/pot) on the forage performance in the pure and mixed swards of orchardgrass and white clover. This 1st part was concerned with the changes in the growth, flowers and flower buds, and roots/nodules of forages. The results obtained are summarized as follows: 1. At the $B_3$ and $B_4$ treatments, the B toxicity was more seriously in white clover than in orchardgrass in the first half of cutting orders, and reduced in the latter half. In white clover, it was more worsened in a mixture than in a pure culture. It was somewhat reduced at the best growth stage of each forage. 2. In orchardgrass, the B toxicity($B_3$,$ B_4$) showed the chlorosis on leaf tips, shallow leaf, little tillers, and weak stems. Whereas it showed the chlorosis/necrosis on old leaf edge, little and weak stolons in white clover. 3. Comparing with the B deficiency($B_{0}$ , $B_1$) and toxicity($B_3$, $B_4$), the optimum B application($B_2$) influenced markedly good growth of shoot, root, nodule, and flower (flower number, blooming period, early full flower) in white clover. 4. Comparing with orchardgrass, white clover was greatly influenced by the boron application. However, this responses of white clover to boron were reduced in a grass-clover mixture with additional fertilization. It was recognized that the good forage performance in a grass-clover mixture could be regulated by the adequate applications of boron and additional fertilizers.s.
The effect of allelochemicals on growth, root nodule nitrogen fixation activity, and ion patterns of soybeans were investigated. We prepared 50 g/L (T50), 100 g/L (T100), and 200 g/L (T200) extract concentrations by soaking fresh red pine needles in a nutrient solution. Adding needles to the nutrient solution increased the content of total phenolic acids, osmolality, and total ions. The total phenolic content in the T50, T100, and T200 extracts were $206{\pm}12.61$, $335{\pm}24.16$, and $603{\pm}12.30$ mg gallic acid equivalents, respectively. The $K^+$, $Mg^{2+}$, $Ca^{2+}$, and $PO_4^{3-}$ content increased by adding needles to the nutrient solutions, whereas $SO_4^{2-}$ content decreased. The growth inhibition of soybeans was proportional to the needle extract concentrations, and the T100 and T200 concentrations resulted in remarkable growth inhibition. On day 20 after treatment, dry weight and nitrogen fixation activity of the root nodules were reduced by the T100 and T200 treatments, whereas the T50 treatment was not difference from the control. After day 10, total ion content in all treatment groups was not different in comparison with the control. However, total ionic content in all treatment groups decreased significantly compared with that in the control after day 20. The lowest total ion value was found for the T200 concentration. The T200 treatment also resulted in significantly reduced $SO_4^{2-}$ content. The amounts of $Mg^{2+}$, $Ca^{2+}$, and $Mn^{2+}$ were higher than those of the control for the T50 treatment on day 10 and for T100 on day 20 after treatment. A significant increase in osmolality was observed in the T200 treatment on day 10 and in the T100 treatment on day 20. These results suggest that under severe allelochemical stress conditions, a remarkable reduction in nodule formation, nitrogen fixation activity, and ion uptake eventually resulted in a decrease in leaf production. Furthermore, increased $K^+$, $Mg^{2+}$, $Ca^{2+}$, $Mn^{2+}$, and osmolality in soybeans exposed to lower concentrations of allelochemicals than the critical stress level helped overcome the stress.
This study was conducted to identify the effect of planting date and cultivation method on soybean growth and yield in paddy field. The plant height of soybeans in single cropping(SC) was higher than those in cultivating after barley culture (CB). Considering planting date and cultivation method, plant height tend to be higher in SC with level row cultivation(LR) and in CB with high ridge cultivation(HR). In this SC method, nodule formation in LR was better than in HR, but number of nodules of cv. Hwangkeumkong was highest, regardless of planting date and cultivation method. In the progress of growth stage, leaf areas of all cultivars were distinctively increased in CB than in SC. However, dry weight of top plants and roots in SC was comparably higher than that in CB due to growing periods of the soybeans. In cultivation methods, general type of dry weight of top plants was higher in LR than in HR, however, root dry weight was via verses. Growth responses varied depending on cultivars, cultivation methods, and planting date and these factors affected to shoot root (T/R) ratio. The T/R ratios in LR and SC were higher than those in HR and CB. In R8 stage, number of pods and ripened seed varied depending on cultivars. cv. Hwaeomputkong, which showed early maturing trait, was lowest. However, both yield factors tended to be higher in HR and CB than in LR and SC. The ratios of ripened seeds percentage of cv. Hwangkeumkong and cv. Eunhakong were higher in CB than in SC. However, yields of cv. Daewonkon and cv. Taekwangkong were higher in CB than in SC.
The nitrogen fixation activity and environmental factors of Lespedeza bicli!oy Turcz, forming annual root nodules by symbiotic Rhizobium sp. were analyzed in the field conditions during the growing period. Seasunal changes of $N_2-fixation$ activity showed the maximum value of $120\;\mu\textrm{M}\;C_2_H4{\cdot}noduie\;g\;fw^{-1}{\cdot}hr^{-1}$ during the active growing period (June) and varied significantly depending on the growth phase and environmental factors. The maximum activities were attained at the conditions of pH 7, $30^{\circ}C$ of temperature, 18 Kpa of oxygen partial pressure and inhibited by water stress and nitrogen sources. The habitat soil was weak acidic and poor in nitrogen, phosphorus and organic matter contents. The leaf area ratios and chlorophyll contents were ranged from 442 to $48;\textrm{cm}^2{\cdot}g\;dw^{-1}$ and from 33 to $38\;\mu\textrm{g}\;chI{\cdot}\textrm{cm}^2$. Nitrogen contents in each organ showed the maximum of 46, 19 and $11\;mg{\cdot}g\;dw^{-1}, respectively for leaf, rool and stem in the early period. The highest phosphorus contents were 4.2, 1.2 and $0.6\;mg{\cdot}g\;dw^{-1}$, respectively for leaf. root and stem in early growing period. The allocation ratios of nitrogen quantity showed 60% for leaves and 73% for roots during the active and late growth period, and 22% [or stems in average. The allocalion ralios o[ phosphorus quantity showed 58% for leaves during the most productive period, 70% for roots in the pre-growth stage and 26% for stems in average.verage.
Journal of The Korean Society of Grassland and Forage Science
/
v.7
no.1
/
pp.25-30
/
1987
There are differences in opinion as to whether nitrogen fertilizer should be used when establishing alfalfa (Medicago sativa L.). Various reports show that under a hot environment, rhizobia (Rhizobium meliloti) are not as effective in fixing atmospheric nitrogen as they are under moderate temperatures. It is also believed that the addition of nitrogen fertilizer inhibits nodulation of alfalfa seedlings. A replicated experiment was conducted under controlled environmental conditions at the University of Nevada-Reno, Reno, Nevada, USA, to determine the effects of nitrogen application on seedling growth and nodulation of alfalfa grown in a hot environment. Sterile sand was used as the growing media to which a complete nutrient solution minus nitrogen was applied volumetrically to each pot daily. In addition, half of the pots received NH4-$NO_3$, at the rate of 11.2 kg per ha at seeding and at two and four weeks after planting giving a total nitrogen application rate of 33.6 kg per ha during the seven-week experimental period. Rhizobia inoculant (R-12) consisted of a mixture of strains 171-15a, 1682c and 80 PI 265 of (Rhizobium meliloti). Inoculant was applied to the seeds prior to planting and to the sand media at two and four weeks after seeding. Twenty seeds were planted in pots 14.0 cm in diameter and 11.5 cm deep. Plants were thinned to ten plants per pot after emergence and were grown in a controlled environment chamber with a 16-hour light period. Soil temperature at 6 cm depth ranged from 17.4^{\circ}C.$ to 31.1^{\circ}C.$ and had a daily mean of 26.5^{\circ}C.$. Plants were harvested at weekly intervals for seven weeks. Root, shoot and total length, dry weight, volume and number of nodules per plant were determined. Root, shoot, and total length were greater in seedlings grown in soil where nigrogen was applied than that grown in soil to which no nitrogen was applied. The average size of the seedlings as determined by volume and weight was more than two times greater where plants were fertilized with nitrogen. Nodule number per seedling was also greater when nitrogen was applied compared to those which received no nitrogen. The differences were greater as the plants became older. The rhizobia did not fix enough nitrogen for adequate growth of seedlings. This is probably due to high temperature growing conditions that caused the rhizobia to become relatively ineffective as compared to cooler growing conditions. Data suggests it would be desirable to apply nitrogen at seeding when alfalfa is established under hot conditions that occur in mid- or late summer.
These experiment were conducted to investigate the seasonal changes of nodule formation and $N_2$-fixing activitues of soybean root nodules which grown in pot on five cultivars in different soils. The results obtained were as follows; 1. It was found that the development of nodules starts from 20 days after planting and continues to increase up to 50-60 days after planting. 2. $N_2$-fixing activities were different with cultivars of soybean. The trend was high in order H-25 > Suweon 63 > Hill > Yugwoo 3# > Gwanggyo 3. Rhizobium japonicum isolated from different soybean cultivars showed different acetylene reducing activities in vitro ; high in order A62-1 > Suweon 63 > Hill > Gwanggyo > Yugwoo 3#.
To obtain the basic information necessary for the development of soybean varieties well adaptable to upland-paddy field rotational croppings, the difference of growth characteristics between upland and paddy-field including yield potentials of current recommending soybean varieties were evaluated. The growth characteristics, both above and under-ground, which were measured at flowering stage were generally greater in paddy-field but the number of root nodules was much greater in upland, thus the artificial inoculation was practically recommended for soybean growing in paddy-fields. Mean seed yield was generally higher in paddy-fields than in upland. All soybean varieties showed higher seed yield in the early planting date, April 20, were somewhat susceptible to soybean mosaic virus(SMV), thus they could be escaped from the disasterous endemic necrotic soybean mosaic virus(SMV-N). Soybean varieties showed over 4.0 tons/ha seed yield in the paddy-field were Williams 79, Union, SS77053, and Namhaekong. At the same time, Jangyeobkong and Danyeobkong were the most stable soybean varieties among the tested soybean varieties with less than 10% of coefficient variation values in all planting dates in paddy fields. Compared with Hwangkeumkong which is most widely being cultivated on farmer's fields, soybean varieties showed high yields in paddy-field were higher in plant height, less in the number of branches, and more in the number of nodes on main stem. At the same time, they had medium seed size which would bring the good germination and stands. Disease resistance especially for necrotic soybean mosaic virus was also one of the most decisive factors in seed yields for the early planted soybeans.
The field experiment was conducted to find out the effect of inoculation of Rhizobium and interaction between amount of nitrogen and inoculating in soybean with 4 replicated split plot design. Main plot was 4 level of nitrogen 0, 2, 6, 12kg/10a and subplot was inoculation and uninoculation. The results are summarized as follows. 1. The yield of soybean did not show any significant difference among nitrogen level or between inoculating but yield of inoculated plot was a little increased than that of uninoculation. 2. At four leaves stage and flowering stage, the weight of nodules in root was decreased with incresing amount of nitrogen significantly and uninoculation caused a reduction compared with modulation significantly and those tendency was same in plant height, and those difference among treatment was more distinct in four leaves stage than in flowering stage. 3. After harvest, total nitrogen content in soil was not only increased with increasing nitrogen level but also was more increased in inoculated plot than in uninoculated plot except nitrogen 0kg/10a level. Inoculation of Rhizobium raised 15kg of nitrogen per l0a at 6kg nitrogen level compared with uninoculation.
A total of 1-0 colostrum-deprived pigs (1 or 2-day-old) and 6 pigs (35-day-old), which had been raised by natural maternal nursing, were used to study the pathogenicity of the porcine enteroviruses by the intracerebral and intramuscular routes of inoculation, which the enterovirus were isolated from the diseased pigs in Korea. The porcine enteroviruses produced an identical polioencephalomyelitis in colostrum-deprived pigs and 35-day-old pigs, which manifested clinical signs and histopathological changes. Clinically it was characterized by incoordination, rise in rectal temperature, ataxia, flaccid paralysis in all the experimental groups. Histopathologically, the lesions were present in both grey and white matter at all levels of central nervous system, though usually more severe in the grey matter. These changes were characterized by meningeal infiltration, degeneration of nerve cells, neuronophagia, diffuse and focal gliosis, glial nodules and perivascular lymphocytic infiltrations. Ganglionitis of the dorsal root ganglia was frequently observed. On the basis of the clinical and histopathological changes mentioned above, it was concluded that porcine enteroviruses isolated in Korea were pathogenic strains which could produce polioencephalomyelitis in pigs. The most severe Jisease was prcduced by the inoculation of both enterovirus and hog cholera vaccine in the 35-day-old pigs at a time when colostral immunity presumably was low. The porcine enterovirus infections seemed to be associated with certain stress factor such as hog cholera vaccine in or immediately following the weanling period.
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