Calcium Inhibits Ion-Stimulated Stomatal Opening in Epidermal Strips of Commelina communis L.

Inoue, H. and Katoh, Y. 1987. Calcium inhibitsion-stimulatedstomatal opening in epidermal strips of Commelina communis L.—J.exp. Bot. 38: 142–149. Ca²⁺ suppressed both the ion-stimulated stomatal opening andH⁺ extrusion of pre-illuminated epidermal strips isolated fromCommelina communis L. In the absence of Ca²⁺, the rate of H⁺release was 18 nmol H⁺ cm^–2 h^–1 per epidermal stripunit area in 150 mol m^–3 KCL at pH 7?4. Half-maximum inhibitionof stomatal opening was observed with 220 mmol m^–3 ofCa²⁺. The hexavalent dye, ruthenium red, showed concentration-dependentprevention of the inhibition by Ca²⁺ of the ion-stimulated stomatalopening. The effect of ruthenium red was non-competitive, andthe K₁ for the calcium inhibition was found to be 3?6 mmol m^–3.The calcium inhibition of H⁺ extrusion was also prevented byruthenium red. These results suggest that Ca²⁺ inhibits theactivity of electrogenic H⁺ translocating ATPase of the guardcell plasma membrane and leads to the suppression of stomatalopening. Key words: Calcium, Commelina communis, ruthenium red, stomata     

Control of Auxin-induced Stem Elongathn by the Epidermis

Segments of the 4th and 5th internodes of light-grown pea seedlings were used for the study of control of stem elongation. With 5th internodes, at low turgor as well as at water saturation auxin primarily appeared to cause a change in cell wall properties of the epidermis but it showed little effect on expansion af the inner tissue. This was confirmed by comparison of expansion between peeled and unpeeled segments, split tests and by measurements of stress-relaxation properties of the epidermal cell wall. Segments with the central part re-moved elongated well in response to auxin, but the isolated epidermis showed neither auxin-induced elongation nor cell wall loosening. A fungal β-1,3-glucanase appeared, at least partly, to have a similar effect as that of auxin on elongation, by changing cell wall properties of the epidermal cell wall. Peeled segments of 4th internodes expanded very little and auxin had little effect on their epidermal cell wall properties.     

Role of the epidermis in auxin-induced elongation of light-grown pea stem segments

1. Effects of auxin on elongation and cell wall properties werestudied using 5th internode segments of light-grown pea epicotyl.The results were:
2. The optimum concentration of 2,4-D for elongationinductionwas about 1 µg/ml, both for unpeeled and peeledsegments.
3. Using stress-relaxation analysis, mechanical propertiesof thecell wall were expressed by the parameters 1/₁, To andT_m. Unpeeledsegments were first treated with 2,4-D, then theepidermis waspeeled off. Parameters of the epidermal cell wallwere conspicuouslychanged by 2,4-D but those of the inner tissuewere not.
4. Actinomycin D and cycloheximide inhibited 2,4-D-inducedchangesin cell wall parameters, as well as in elongation, ofunpeeledsegments apd of the epidermis.
5. 2,4-D did not induceelongation of the isolated epidermis butpromoted that of peeledsegments. This promotion was smalleras compared with unpeeledsegments. 2,4-D did not significantlyinfluence the diffusionpressure deficit of peeled segmentsbut did increase their elasticand plastic extensibilities.
6. We conclude that auxin primarilyinduces cell wall looseningof the epidermis, most likely throughnucleic acid and proteinsynthesis.

¹ Present address: Biological Institute, Department of GeneralEducation, Nagoya City University, Mizuho-ku, Mizuho-cho, Nagoya467, Japan. (Received April 22, 1971; )     

ROLE OF NUCLEUS IN CYTOPLASMIC ACTIVITIES WITH SPECIAL REFERENCE TO THE FORMATION OF SURFACE MEMBRANE IN ELODEA LEAF CELLS

Using leaf cells of Elodea densa, investigations were made onthe role of nucleus in some activities of protoplasm, by comparingthe behaviours of nucleated and enucleated halves of protoplast,which were separated by means of plasmolytic treatment of intactcells.

1. Plasmolytic treatment caused a remarkable chloroplast-systrophe,which was more pronounced in nucleated halves than in enucleatedones, in which the systrophe, if any, was dispersed gradually.
2. In general, protoplasmic streaming was suppressed under plasmolyticconditions, although in nucleated protoplasm a rotation of systrophedchloroplast-bunch was often observed after a longer durationof culture.
3. The membrane which was re-formed on the surfaceof divided protoplasmichalves was found to be more solidifiedand less elastic in thenucleated halves than in the enucleatedhalves.
4. Detection of RNA in the cytoplasm by UNNA-PAPPENHEIM'sstainingmethod showed that the ability of pyronin uptake wasdecreasedin the enucleated halves as compared with that inthe nucleatedhalves.
5. By treatment with ribonuclease, theproperties of nucleatedprotoplasmic halves were rendered tobe similar to those ofthe enucleated halves.
6. Based on theobservations, discussions were made on the r?leof nucleus inthe regulation of the membrane formation by, andthe viscosityand other physico-chemical properties of, thecytoplasm.

¹ This study was supported by a grant-in-aid for DevelopmentalScientific Research from the Ministry of Education, No. 407041(1955), and No. 407084 (1960). (Received November 19, 1960; )     

Nutrient utilization and requirement under photoheterotrophic growth of Marchantia polymorpha: Improvement of the culture medium

Previously we reported on a suspension culture of chlorophyllous cells of a liverwort, Marchantia polymorpha L., under photoheterotrophic conditions. The chemically defined medium was a modified Murashige and Skoog's medium, which contained, besides glucose, inorganic salts, a growth regulator (2,4-D), and twenty-four organic compounds as micro constituents. Because of this complexity, we undertook a simplification of the medium. Having examined the utilization of the major nutrients and the requirements for the micro constituents, we have succeeded in improving the medium. The new medium contains phosphate at 3.13 mM and only eight out of the twenty-four original micro organic constituents. In this new medium, the cells grow under a well-balanced nutritional condition, with richer chlorophyll and at a higher rate during the exponential phase than in the original medium.     

DIFFERENTIATION AND DEDIFFERENTIATION OF RAT LENS EPITHELIAL CELLS IN SHORT- AND LONG-TERM CULTURES

The crystallin synthesis of rat lens cells in cell culture systems was studied in relevance to their terminal differentiation into lens fibers. SDS-gel electrophoresis combined with several immunological techniques showed that γ-crystallin is a fiber-specific lens protein and is not localized in the epithelium of either newborn or adult lenses. When lens epithelial cells of newborn rats were cultured in vitro , α-crystaIlin was detected in many, but not all, of cells cultured for 10 days. Cells with α-crystallin gradually changed their shape into a flattened filmy form and finally differentiated into lentoid bodies. The differentiation of lentoid bodies was also found in cultures of epithelial cells obtained from adult lenses. The molecular constitution of lentoid bodies was the same as that of lens fibers in situ . The differentiation of lentoid bodies occurred successively for 5 months in cultures of lens epithelial cells. Most of the proliferating cells, however, lost α-crystallin during the culture period. Thereafter, they did not show any sign of further differentiation into lens fibers. Four clonal lines were established from these cells. One protein which is specific to the lens epithelium and the neural retina in situ (tentatively named as β_u-crystallin) was maintained in all lines, suggesting that some specific properties of ocular cells remain in the lined cells.     

一种新型网膜对SARS冠状病毒的抑制作用

利用光触媒钛羟基磷灰石网膜（PTAF）具有吸附和酶催化的特点,研究其对SARS病毒的抑制作用.实验结果表明在紫外照射条件下,PTAF膜对SARS冠状病毒的抑制率为100％.在没有紫外照射的条件下,PTAF膜对SARS冠状病毒的抑制率为99.99％, 与对照组相比,PTAF膜抑制病毒的效率是HAF膜的1 000倍以上.研究结果提示,PTAF在预防SARS冠状病毒及其他病毒性疾病流行方面有潜在的应用价值.     

EMBRYO AND ORGAN CULTURES OF THE HORSESHOE CRAB,TACHYPLEUS TRIDENTATUS*

This paper deals with some preliminary technical experiments on cultures of the embryos and organs of Tachypleus tridentatus. In embryo culture, using the embryos at various stages of development, embryonic areas with hardly any yolk remaining in them were cultured in various media and by various methods (Series A); hemispheres of embryos containing the embryonic area (Series B) and whole embryos without egg membranes (Series C) were cultured in sterilized sea water. The suitability of the culture media and methods is described; the medium containing calf serum or 199, and the hanging-drop method were found to be the most appropriate of those used in respective series of experiments. The changes in external and internal structures of the explants from embryos at various developmental stages cultured in various media and by various methods are described; dorsal closure, secretion of inner egg membrane, thickening of the cell layer and the elongation of limb buds were all observed in the Series A experiments. In Series B and C, coelom formation and also normal development of the embryos cultured after the 2nd embryonic moulting were observed. In organ culture, fragments of the embryonic heart were cultured in various media and by various methods. The hanging-drop method resulted in longer survival than the method using agar media.     

Oleic acid synthesis in the fruit of Persea americana MILL

The tissue slices from the mesocarp of avocado could incorporateradioactive acetate into lipids. Oleic, palmitic and stearicacids were the most labeled fatty acids found to accumulatein triglycerides. The conclusions that oleic acid was formedby way of chain elongation of already unsaturated short-chainfatty acids, and that there was no evidence for the desaturationof stearic acid were based on the following observations: 1)Stearic acid-¹⁴C was incorporated into triglycerides by thetissue slices without formation of oleic acid. 2) The oleicacid synthesized from specifically labeled acetate was not randomlylabeled. The specific radioactivity of azelaic acid moiety ofoleic acid was rapidly increased while that of pelargonic acidmoiety was gradually increased. 3) An unexpected rise of stearicacid was observed among commonly occurring fatty acids in thetissue slices. This was accentuated by anaerobiosis which prevailedduring vacuum infiltration of labeled acetate. ¹Present address: Department of Botany, Faculty of Agriculture,Hokkaido University, Sapporo, Japan. (Received January 13, 1969; )