The effects of light and $CO_2$ on the electrophysiological characteristics of guard cells in the intact leaf and isolated epidermis have been investigated. Fast hyperpolarization of guard cell apoplastic PD in the intact leaf was recorded reaching up to around 7 mV and 20 mV in response to light and $CO_2$. Whenever the experiments were attempted with isolated epidermis, there was no response to light and $CO_2$. In order to determine the influence of the mesophyll cells, the apoplastic PD of guard cells in isolated epidermis was measured in the presence of the mesophyll supernatant or the control medium. The apoplastic PD in isolated epidermis was hyperpolarized to -7mV, changing from -22mV to -29mV at 40 min. But, when isolated epidermis was incubated with the supernatant from mesophyll cells incubated in the light, the apoplastic PD in isolated epidermis was hyperpolarized to -19 mV, changing from -22 mV to -40.5 mV. $CO_2$ also caused a change of 0.1 to 0.3 pH unit in the intact leaf. However, this change was absent in isolated epidermis. A vibrating probe was used to detect the change in electrical currents at the surface of excised intact leaves and isolated epidermis. The reading of excised intact leaves in the dark was $0.5\muA\;cm^{-2},$ remaining steady until illuminated. Light increased the current on the surface of excised leaves to about $0.8\muA\;cm^{-2},$. However, light had no effect in the current on the surface of isolated epidermis. Apoplastic pH changes across the stomatal complex in response to light and dark were measured both in the intact leaves and isolated epidermis over the same time period using pH micro-electrodes. The guard cell wall of intact leaf was acidified to 2.5 pH unit, falling from pH 7.5 to pH 5.0 in the first 10 min. in the light. At the same time the guard cell wall pH of isolated epidermis fell from pH 7.5 to pH 7.0 at 10 min. The guard cell wall pH of isolated epidermis incubated in the mesophyll supernatant fell from pH 7.6 to pH 6.7 at 10 min. Likewise, It could be imagined that an electrical signal, chemicals and hormones propagated from the mesophyll in response to light and $CO_2$ could control a fast stomatal response.
To investigate the influence of the mesophyll cells on stomatal opening in response to white light, the segments of isolated epidermis were transferred on partly exposed mesophyll cells of a leaf and stomatal apertures were measured. Transferring the isolated epidermis on partly exposed mesophyll cells of a leaf caused a marked increase on stomatal apertures while stomata in isolated epidermis incubated in MES buffer hardly opened. Mesophyll infiltration with photosynthetic inhibitors (DCMU, DCCD, $NaN_3$) was performed to elucidate the correlation between stomatal apertures and the degree of photosynthetic activity. It was found that transferring the isolated epidermis on partly exposed mesophyll cells of a leaf caused an increase of stomatal apertures depending on the degree of photosynthetic activities. In $NaN_3$ infiltrated leaf discs, transferring the fresh isolated epidermis on partly exposed mesophyll cells of a leaf showed no significant effect, but a slight increase on stomatal apertures. Isolated epidermis alone did not respond to the light properly, but if it was closely contacted with mesophyil cells, the stomata regained the ability of the light response. Therefore, it could be suggested that stomatai apertures were related with the degree of photosynthetic activity in the mesophyll cells.
Many researchers have been studied with guard cell protoplasts and detached epidermis as they think that properly stabilized protoplasts and detached epidermis retain many of the properties of intact guard cells. However, some studies have shown that stomata in detached epidermis behave differently, both quantitatively and qualitatively, from those in the intact leaf. Stomata in the intact leaf are very sensitive to environmental factors such as light, $CO_2$ and osmotic stress, but stomata in detached epidermis are less sensitive to these factors than those in the intact leaf. The clearest evidence to suggest the different response between detached epidermis and intact leaf obtained from the experiments with heavy metal, cadmium. 3-weeks old Commelina. communis was transferred to and grown in Hoagland solution in the presence or absence of 5 mM $Cd^{2+}$ for 4 days. The application of $Cd^{2+}$ showed about 70% inhibition of stomatal conductance when measured at various light intensity (100-1,000 $\mu$mole $m^{-2}s^{-1}). However, stomata in detached epidermis floated on an incubation medium containing 100 $\mu$M $Cd^{2+}$ opened to a degree of about 8.38 fm, but the stomata treated with no cadmium opened to 3.74 ${\mu}{\textrm}{m}$. These results were unexpected as the intact leaf grown in a Hoagland solution containing cadmium showed very negative physiological responses. These results showed that stomata in detached epidermis and in the intact leaf could respond reversely. Therefore, it is possible that we now misunderstand how stomata open in real natural condition.
The ultrastructural characteristics of epidermis isolated from healthy and rusty ginseng roots(Panax ginseng) were observed by scanning electron microscopy (SEM), and the distribution profiles of inorganic elements were also examined by energy dispersive X-ray analysis (EDX). The epidermis of rusty ginseng was thick and cell walls were also severely disrupted whereas the epidermal image of healthy ginseng showed relatively thin, clear and rectangular structure. A high amount of rod-shaped bacteria was found in rusty ginseng and cells near bacteria were degraded. The bacterial density in epidermis of rusty ginseng was ranged from 2.9$\times$10$^{6}$ to 3.5 $\times$ 10$^{7}$ CFU/g fresh weight, while that of healthy ones was from 4.7$\times$10$^2$ to 1.2$\times$10$^3$CFU/g. Artificial inoculation of bacteria isolated from rusty ginseng induced similar symptom like rusty ginseng. The mineral contents inculding Al, Si and Fe were Higher in the epidermis of rusty ginseng, but K content was lower compared to healthy ones.
The lectins from mucilaginous jelly and green epidermis of Aloe vera were isolated by gel and affinity chromatography. The molecular weights of the lectins were determined by SDS-PAGE. The molecular weights of the lectins from mucilaginous jelly isolated by Sephadex G-100 were 58.7 kD and 33.3 kD, and that isolated by acid-treated Sepharose 4B was 176.4 kD. The molecular weights of the lectins from epidermis isolated by Sephadex G-100 were 221.1, 54.0 and 32.5 kD respectively. And that isolated by acid-treated Sepharose 4B was 222.0 and 158.0 kD. The agglutinating activity of lectin from jelly was inhibited by D-galactose, lactose and D-galactosamine, but that from epidermis was not inhibited by lactose. The activity was stable at the pH range of $7.0{\sim}9.0$ and at the temperature $0{\sim}60^{\circ}C$.
Rusty root of ginseng has been known as one of the limiting factors in ginseng production in Korea. An attempt was, therefore, made to elucidate biological and chemical natures of the rusty root, and the redox Potential of the ginseng cultivated soils were measured and compared with diseased and non-diseased soils. Reddish discoloration was most frequently observed on the epidermis of ginseng root and the pigments were accumulated in all epidermal cells of the diseased lesions. The lower the redox potential of the ginseng cultivated soil was, the more severe the rusty root was observed. Fe content in the diseased epidermis was 3 times higher than that of healthy one. Organic acids such as oxalic, malonic, succinic, and citric acids were also higher in the mss root than in the healthy one. Thin layer chromatogram of phenolic acid fractions obtained from the epidermal cells of the rusty root of ginseng exhibited 3 to 4 unidentified substances not found in the healthy root. Also lignification of the epidermal cells and the activity of phenylalanine ammonia lyase were greater in the rusty root than the healthy root. Colony formation and conidia production of F. solani, And mycelial growth and sclerotium formation of Sclerotinia sp. isolated from ginseng root were suppressed in a nutritionally minimal medium supplemented with water extract of rusty ginseng root epidermis. It is, therefore, suggested that rusty root of ginseng is caused by unfavorable rhizosphere environmental stress or stresses resulting abnormal metabolism in the root as a selfdefence mechanism of non-specific resistance responses.
One of the physiologically important ginseng diseases is red-colored phenomena (RCP) that is caused by accumulation of red-colored substances on the epidermis of ginseng roots. Although RCP severely deteriorates the quality of ginseng products, there has been little information on what red-colored substance is and how RCP occurs. Therefore, the heavy losses of cultivators and ginseng industry are suffering by RCP, For this reason, we have investigated with the morphochernical characteristics of RCP to find out main cause of it. The red-colored substances (RS) on the epidermis of red-colored ginseng (RCG) were examined using inverted light microscope, confocal laser scanning microscope (CLSM)and furier transform infrared (FT/IR) spectrometer. Red brown substances were accumulated in the cell wall of the epidermis from early stage to late stage of RCC. Especially, cell wall of the late stage of RCG was covered with the sub-stances with 80~ 130 fm thick. Therefore, the cell wall of RCG cannot protect the ginseng root cells from the mechanical damages, bacteria and fungi. To analyse red substances of roots, RS were isolated from epidermis of RCG and extracted using various solvents. RS is strongly insoluble but it was bleached by oxidizing agents including 12% (v/v) NaOCl. Therefore, RS was Presumed to make up of high chelation power. The proriles of FT/IR spectra or both healthy ginseng (HEG) and RCG showed a significant difference at two wavelength,2857 cm$\^$-1/(C-H) and 1032 cm$\^$-1/(S=O), respectively. Furthermore, absorption peak of 2857cm$\^$-l/ appears on the only epidermis of RCG. The other peak is shown lower absorption rate on the epidermis of RCG than that of healthy ginseng. Also, FT/IR spectra of the mixture of carboxym-ethylcellulose (CMC) and iron (Fe$\^$3+/) were very similar to RCG spectrum profiles. One of a interesting fact is that the contents of phenolic compounds at the epidermis of healthy ginseng were highest. The results of these experiments sup-port the RCP was closely related with the chemical interaction between inorganic elements (Fe) of rhizosphere and organic matters (cellulose, cellobiose, cell sap, etc.) of ginseng roots.
To elucidate morphologic lesion of porcine exudative epidermitis which is occurred sporadically in Korea, Staphylococcus hyicus subsp. hyicus isolated from the naturally affected pigs was inoculated to suckling pigs. The infected piglets were observed grossly and histopathologically. Although affected piglets were taking acute, subacute, or chronic course, some piglets suffered from chronic disease showed poor prognosis and marked growth depression. Affected peglets had erythematous skin on the face, ear, and abdomen and these localized lesions appear as brownish spots of exudative epidermitis and fromed crust in the early stage. But, after this stage, the skin were covered by viscous greasy exudate and formed blackish brown crust and appeared fissures and hypertrophy. Grossly, there has been hemorrhage with the removal of crust-like materials of epidermis and edematous subcutis. The superficial lymph nodes were edematous and swollen or congested and hemorrhagic. Some piglets had swollen ureters, cysts in the renal cortex, or polyarthritis. A few cases had mild edematous swelling of kidney, intestinal catarrh and congestion of brain. Microscopically, skin lesions had detachment of keralinized layer and parakeratosis of epidermis, hydropic degeneration of epidermal cell, and retrogressive degeneration of hair root sheath. Dermis had edema, and infiltration of neutrophils and mononuclear cells. As the disease was proceeded, there was marked perivasculitis with lots of mononuclear inflammatory cells. More chronic lesions formed granuloma-like bodies(nodules) due to more mononuclear, perivascular inflammatory cell infiltration and proliferation of fibroblast. Lots of plasma cells and eosinophils were also present in dermis. Epidermis was hyperplastic by proliferation of basal cells stratum germinativum and epidermal pegs often extended into the dermis. In secondary infection, lots of neutrophils could be seen in epidermis and derms. Kidney had neutrophilic infiltration, necrotic and cystic glomeruli, and dilation of renal tubules and ureters. Purulent arthritis was sometimes observed in joints. Three days old mice administrated Staphylococcus hyicus subsp hyicus subcutaneously before had focal congestion and hemorrhage, necrosis, and subcutaneous edema of the skin. This observation was also seen in the study of mice administrated exfoliatin toxin of Staphylococcus which evoked human staphylococcal scalded skin syndrome.
Purpose: Collagenoma is an intradermal harmatomatous collagen proliferation among connective tissue nevi of the skin. Although there are some reports of isolated collagenomas that occurred on the sole and the palm, collagenoms at the finger has not been reported in Korea. Methods: An 11-year-old girl presented with a growing mass on the distal interphalangeal joint of the left 5th finger. It was a skin-colored, oval and mild tender mass. There were no associated cutaneous or systemic abnormal findings. Results: The nodule, at the subcutaneous level on the distal phalanx, was completely removed by excision. Grossly it was covered with normal skin, and its surface was smooth having a definite margin with a size of $1.5{\times}1.2cm$. Histopathological examination, the epidermis showed to be normal, increased thickness of collagens arranged with a whirl formation was found in the dermis. No signs of cellular component increase were observed. Conclusion: Isolated collagenoma can be aroused as a solitary nodule in at any place of the body. We report a rare case of a female patient with an isolated finger collagenoma.
In order to elucidate the mechanism of red-colored phenomena(RCP) in ginseng(Panax ginseng C.A. Meyer), distribution of inorganic elements of ginseng root and its surrounding soil, and microflora in the soil were investigated. Red brown colored-substances were accumulated in the cell wall of epidermis at early stage of red-colored ginseng (RCG). Cell wall of the late stage of RCG was disordered and microorganisms were shown in the disordered cell wall. Al, Si and Fe contents among inorpanic elements in the epidermis of RCG were higher at two or three times than that of healthy ginseng. On the other hand, K content was higher at three times in healthy ginseng than that of RCG. Especially, Fe content was higher at three times in lateral roots of RCG than that of healthy ginseng. Total 21 strains of microorganisms were isolated on the 523 medium from surface soil, surrounding soil of both healthy and RCG, and RCG. Six strains of microorganisms among them were resistant to 2 mM Fe. Two species in Bacillus app. and Lactobacillus app. , and one species in Micrococcus sp. and Npisseria sp. respectively were identified. It seemed that RCP was closely related with the distribution and uptake of inorganic elements, was also correlated Fe-independent metabolism of microorganisms.
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