The Structure and Orientation of Guard Cells in Plants Showing Stomatal Responses to Changing Vapour Pressure Difference

Transmission electron micrographs revealed that a substantialpart of the guard cell wall of both Quercus robur L. and Populusnigra ‘italica’ L. was either free of cuticle orcovered with a greatly reduced cuticular layer. In Quercus thestructure of the guard cell was such that the area of limitedcuticular development would be exposed to the evaporating powerof the atmosphere even when the stomata were closed. Lanthanumstaining confirmed that this area might be an important siteof evaporation. A similar evaporation site was identified inthe guard cell wall of Pinus sylvestris L. Light micrographsrevealed that this area could also be exposed on the outsideof the leaf when the stomata were closed. It appears that guardcell orientation with respect to the epidermal plane dependsupon epidermal turgor. Changes in orientation of the guard cellcoupled with the exact location of the cuticle-free area inthe guard cell wall may explain the nature of the stomatal responseof individual species to changing VPD and the effect of othervariables, e.g. water deficit, on this response. Quercus robur L, oak, Populus nigra L, poplar, stomata, guard cells, cuticle, evaporation, vapour pressure difference     

Effect of Phosphorus Nutrition on the Cellular Distribution of Acid Phosphatase in the Leaves of Lycopersicon esculentum L.

A histochemical study using light microscopy has been made ofthe distribution of acid phosphatase (EC 3.1.3.2 [EC] ) activity intransverse sections of fully expanded leaves of Lycopersiconesculentum grown in phosphate-deficient or sufficient media.Leaf tissues were prepared by two methods and were embeddedin paraffin wax. The location of acid phosphatase activity inleaf sections was determined by trapping orthophosphate releasedfrom p-nitrophenyl phosphate with lead acetate and subsequentlyconverting the lead phosphate to optically dense lead sulphide.In leaf sections from control tissue lead sulphide depositswere larpely confined to the spongy mesophyll cells. Whereasthe staining of the palisade cells was limited and of a granularnature, the staining of the spongy mesophyll cells was heavierand coincident with the outline of the individual cells. Moreover,the minor veins were more heavily stained than the surroundingmesophyll cells. Sections of phosphorus-deficient tissues wereheavily stained in both the palisade and spongy mesophyll layersand heavy deposits of lead sulphide were present in the regionsof the minor veins. It is suggested that the enhanced acid phosphataseactivity of the mesophyll cells in fully expanded leaves couldbe involved in the remobilization of phosphate within phosphorus-deficientplants, or be part of a phosphate transporting system, concentratingthe intracellular phosphate from the limiting supply in thesolution bathing the mesophyll cells. Lycopersicon esculentum L., tomato, acid phosphatase, phosphorus nutrition     

Immunological Reactions of Single Pollen Grains, Electrophoresis and Enzymology of Pollen Protein Exudates

Single intact pollen grains of Oenothera organensis, when placedupon a thin layer of agar containing pollen antiserum, producecircular areas of precipitate. Pollen grains from an S₂S₂ plantdo not produce precipitate in S₆ antiserum. Pollen grains froman S₆S₆ plant and an S₂'S₄' self-compatible plant produce precipitatesin S₆ antiserum. Fifty per cent of the pollen grains from anS₂S₆ plant produce precipitate in S₆ antiserum. Protein diffusesinto buffer solutions from intact pollen grains within 212 min.As much as 40 per cent of the total protein diffuses out inan hour. Amylase and invertase were detected in the diffusatefrom pollen grains. Alkaline and acid phosphatases were confinedto the pollen grains and did not diffuse out. The serologicalprecipitates are specific to the incompatibility system.     

野蚕黑卵蜂寄主识别利它素的纯化及氨基酸组成分析

针对利它素特殊的物理及化学性质,采用特定的温度分离方法,成功地从野蚕Theophila mandarina和家蚕Bombyx mori的雌蛾性附腺中分别获得了纯度较高的野蚕黑卵蜂Telenomus theophilae寄主识别利它素。对这两种利它素氨基酸组成的分析表明,来自家蚕和野蚕雌蛾性附腺的利它素在氨基酸的组成和含量方面非常相似,在检测到的15种氨基酸中,甘氨酸、谷氨酸和天冬氨酸的克分子百分数在10%以上。从家蚕得到的分别为28.1%、18.5%和12.6%。从野蚕得到的分别为24.4%、18.1%和10.1%。这3种氨基酸之和在家蚕和野蚕中均超过50%以上。用凝胰乳蛋白酶对这两种利它素进行水解时发现溶液中均产生非水溶性沉淀,电泳表明沉淀的多肽蛋白分子量在10 Kd左右。这显示在野蚕和家蚕利它素结构中,有着与家蚕丝蛋白相类似的结构,存在着结晶区域（沉淀）和无定形区域（上清）。     

Accumulation of Fe, Cr and Ni Metals inside Cells of Acidophilic Bacterium Acidiphilium rubrum that Produces Zn-Containing Bacteriochlorophyll a

The accumulations of metals were studied in cells of aerobicacidophilic proteobacterium Acidiphilium rubrum; this spicesuses Zn, instead of Mg as the central atom of bacteriochlorophyllin its photosynthetic apparatus. Energy dispersive X-ray analysis(EDX) of the cell surfaces gave peaks of P, S, Si and Mg. Transmissionelectron micrograph images showed a high density globule of0.11µm in diameter in each 1.1 x 0.3µm cell. AnEDX analysis of the black precipitate that was collected afterthe centrifuga-tion of disrupted cells showed high amounts ofthe heavy metals Fe, Cr, Ni and traces of K, Ca, P, Si and Alin the order of atomic content. An X-ray powder diffractionanalysis indicated that Fe, Cr and Ni exist as alloys. The metalaccumulations interpret the high tolerance of this bacteriumto the toxic heavy metals abundant in acidic growth medium.Zn seems to specifically exist as Zn-bacteriochlo-rophyll ain this organism, because no significant accumulation of Znwas detected on the cell surface, black precipitate, or cytoplasmicmembranes. (Received December 26, 1997; Accepted April 21, 1998)     

Vapour Loss through Stomatal Pores with the Mesophy II Tissue 1

Measurements of sustained rates of vapour loss via stomatalpores in epidermal strips show that these rates compare withtranspiration rates of intact leaves. The water supply pathwithin the epidermal tissue is thus capable of sustaining ahigh rate of evaporation from subsidiary and guard cells.     

Resistance to Water Loss from the Mesophyll Cell Surface in Plant Leaves

A method is described for the accurate measurement of the apparentresistance to water loss from the mesophyll cell walls of plantleaves (r_w), and for studying the mechanism underlying thisresistance. The method for distinguishing possible mechanismsinvolves a comparison of the calculated values of r_w at differentrates of evaporation. The value of r_w remained below 50 s m^–1at relative water contents greater than 11 ± 3% and 7± 2% for Pelargonium hortorum Bailey and Vicia faba L.respectively. Therefore r_w is relatively insignificant at normalphysiological water contents in these species. When r_w did increaseit was not sensitive to evaporation rate, suggesting that alowering of the vapour pressure at the evaporating sites wasnot involved. This contrasted with the results for cellulosefilter paper, where r_w was more sensitive to evaporative flux.     

毛乌素裸沙丘斑块的实际蒸发量及其对降雨格局的响应

 裸沙表面的蒸发虽然是一个物理问题,但对于沙地植被演替的初始阶段非常重要。目前存在的地表蒸发的机理性模型大多是瞬时或者短时期的 ,而年尺度以上的蒸发量与降水和蒸发驱动下的土壤水分系统的状态变化及其对蒸发过程的反馈密切相关。一些估算毛乌素年蒸发量的实验结 果之间分歧很大且缺乏准确的机理性解释。该文利用生态系统模型中的土壤水分运动和蒸发模块计算了毛乌素裸沙丘从日到年际尺度的实际蒸 发量,发展了一个以单次降雨量和降雨频率为驱动因素的降雨-蒸发模型对年蒸发量进行简单的估算,并研究了年蒸发量对降雨格局的响应。结 果表明毛乌素裸沙丘的多年平均蒸发量为166 mm,占多年平均降雨量的56%。虽然研究区1959～1992年降雨总量无显著变化趋势,但是裸沙丘斑 块的实际蒸发量呈现较明显的增加趋势（1.30 mm&;#8226;a^－1）。小降雨事件对蒸发量贡献的显著增加（0.69 mm&;#8226;a^－1）是导致实际蒸发量增大的重 要原因。大强度降雨事件的频度和雨量对降雨总量的贡献要远高于对蒸发总量的贡献,小于12 mm的降雨事件在年际比较稳定,很大程度上保证 了年蒸发量100 mm左右的基数值。这些因素使得年蒸发量的变异程度小于年降雨量的变异程度。由于降雨格局的年际变化会对蒸发量产生直接 的影响,降雨-蒸发模型可以相对有效地预测年度蒸发量,而用年降雨量预测年蒸发量误差较大。     

Absorption and Secretion of Polyethylene Glycol by Solanaceous Plants

The response of the following species of the Solanaceae to waterstress caused by polyethylene glycol (PEG) as an osmoticum,was examined: Solatium khasianum, S. laciniatum, S. melongena,S. nigrum, S. tuberosum, Capsicum annuum, Lycopersicon esculentumand Hyosciamus boveanus. Secretion of solution from the leaveswas observed in these plants when put into 4.5% solution ofPEG 1500, 4000 and 6000. Secretion of liquid started after 20min in seedlings with mechanically damaged roots, and after24 h in plants with intact roots. After evaporation of the liquid,deposits of white material which remained on the leaves wereidentified as PEG by high pressure liquid chromatography andby PEG induced protoplast fusion. All the species examined bear glandular and non-glandular hairson their leaves. In species with leaves which are highly tomentous,PEG secretion took place mainly through the non-glandular hairs.In species with a small number of hairs, secretion took placethrough ordinary epidermal cells. Under similar conditions mannitolsolution was secreted only through wounds and cracks, and NaClsolution was not secreted at all. Key words: Secretion, PEG, Solanaceous plants     

Localization of ATPase in Sink Tissues of Ricinus

Histochemical localization of ATPase was carried out on phloemtissues from vegetative and reproductive sinks of Ricinus communis,using lead precipitation procedures. Reaction products werelocalized mainly at the plasma membrane of the sieve elements,companion cells and phloem parenchyma cells. Activity was alsopresent in plasmodesmata, the tonoplast of companion cells anddispersed P-protein within the sieve element lumen. The resultsare discussed in relation to the possible involvement of a plasmamembrane ATPase in apoplastic and symplastic unloading fromthe phloem conducting tissues. ATPase, sink tissues, unloading, Ricinus communis     

The Relationship between an Epiphyllous Liverwort and Host Leaves

An investigation was carried out into certain aspects of thestructural and physiological relationships between an epiphyllousliverwort, Radula flaccida Lbg. et G., and a variety of hostleaves. Initial attachment of the epiphylla to the host leafis aided by an adhesive secretion from the rhizoids of the epiphylla.Some rhizoids subsequently penetrate the leaf cuticle and intrudebetween epidermal cells. Penetration results in the death ofsome epidermal cells and rhizoids enter also through the gapsthus created. The presence of well-established epiphyllous colonies was closelyassociated with increased loss of water from leaves by evaporation.Leaves mechanically stripped of their epiphyllae showed a similarlyhigh rate of water loss. It was shown that water and dissolvedradioactive phosphate (salts) can pass from host leaf tissuesinto the epiphylla. It is concluded that, in traditional terminology, epiphyllousR.flaccida is a semi-parasite rather than a simple epiphyte.Some other aspects of the relationship are discussed.     

Stomatal Responses to Sulphur Dioxide and Vapour Pressure Deficit

Stomatal conductances (g_s) of plants of Vicia faba, Raphanussativus, Phaseolus vulgaris, Heilanthus annuus, and Nicotianatabacum were measured in chambers containing either clean airor air containing between 18 and 1000 parts 10^–9 SO₂ atwater vapour pressure deficits (vpd) ranging from 1·0to 1·8 kPa. When vpd was low (<1·3 kPa at 22 °C) and stomatawere open, exposure to SO₂ induced rapid and irreversible increasesin g_s in V. faba. This response persisted throughout the exposure(3 d). The increase in g_s, 20–30% compared with cleanair, was independent of SO₂ concentration up to 350 parts 10^–9Stomatal conductances of polluted plants at night were greaterthan controls. When stomata were closed before exposure to SO₂,there was no effect on g_s. When vpd was varied, g_s of unpolluted plants of P. vulgarisshowed no response, but that of R. sativus increased slightlywith increasing vpd. In both species exposure to SO₂ causedan increase in g_s at all vpd values. g_s of unpolluted plantsof V. faba, H. annuus, and N. tabacum decreased with increasingvpd. At low vpd values exposure to SO₂ in these species causedan increase in g_s, but, above a certain value of vpd, dependingon species, g_s decreased with exposure to SO₂. It is postulated that SO₂, once in the substomatal cavity, entersthe stomatal complex via adjacent epidermal cells and at lowconcentrations leads to a reduction in turgor in these cellsand consequently to stomatal opening. In vpd-sensitive species,increased transpiration from guard cells or epidermal cellsadjacent to the stomata induced by SO₂ may lead to stomatalclosure at large vpd levels. Stomatal sensitivity to vpd insuch cases may be enhanced because adjacent epidermal cell turgoris lowered by SO₂. At high SO₂ concentrations direct disruptionof guard cell structure may lead to a loss of turgor and stomatalclosure.     

The N-glycosylation defect of cwh8Delta yeast cells causes a distinct defect in sphingolipid biosynthesis

CWH8/YGR036c of Saccharomyces cerevisiae has been identifiedas a dolichylpyrophosphate (Dol-PP) phosphatase that removesa phosphate from the Dol-PP generated by the oligosaccharyltransferase(OST), while it adds N-glycans to nascent glycoproteins in theendoplasmic reticulum (ER). Lack of CWH8 was proposed to interruptthe so called dolichol (Dol) cycle by trapping Dol in the formof Dol-PP in the ER lumen. Indeed, cwh8D mutants display a severedeficiency in N-glycosylation. We find that cwh8D mutants havestrongly reduced levels of inositolphosphorylceramide (IPC),whereas its derivative, mannosyl-(inositol-P)₂-ceramide (M(IP)₂C)is not affected. Microsomes of cwh8D contain normal ceramidesynthase and IPC synthesis activities. Within a large panelof mutants affecting Dol dependent pathways such as N- or O-glycosylation,or glycosylphosphatidyl inositol (GPI)-anchoring, only the mutantshaving a deficiency of N-glycan addition show the defect inIPC biosynthesis. By mutating genes required for the additionof N-glycans or by treating cells with tunicamycin (Tm) onecan similarly reduce the steady state level of IPC and exactlyreproduce the phenotype of cwh8D cells. Some potential mechanismsby which the lack of N-glycans could lead to the sphingolipidabnormality were further explored.     

恒河猴皮肤干细胞的体外培养纯化与鉴定

探索恒河猴皮肤干细胞的体外培养及纯化条件,为进一步的研究奠定基础. 通过组织块培养法和消化培养法 在体外培养恒河猴表皮细胞,然后用Ⅳ型胶原吸附法吸附20 min,获得快吸附细胞. 对快吸附细胞进行克隆培养,并进行免疫细胞化学双标染色、RT PCR鉴定 β1 整合素和角蛋白15的表达,用流式细胞仪鉴定纯化前后的细胞中 β1 整合素和角蛋白15的阳性细胞比例,并通过透射电镜观察细胞的超微结构. 组织块培养法和消化培养法均可获得表皮细胞,Ⅳ型胶原纯化后的细胞胞体较小,饱满,核/浆比例大,细胞镶嵌状排列. 细胞克隆分析显示,细胞全克隆生长率高. 细胞免疫荧光显示,分选后的细胞显示 β1 整合素和角蛋白15阳性. RT PCR检查呈现 β1 整合素和角蛋白15的特异性片段. 流式细胞仪检查显示,纯化前的细胞中角蛋白15阳性细胞占总细胞中的比例为8％, β1 整合素阳性细胞的比例为10.7％;纯化后,角蛋白15阳性细胞的比例为89.4％, β1 整合素阳性细胞的比例为88.5％. 通过组织块培养法和消化培养法均可培养获得活性良好的表皮细胞,Ⅳ型胶原吸附法是一种简便、有效的皮肤干细胞分离方法,可以为进一步的眼表上皮替代重建眼表提供足量的高纯度的干细胞建立可靠的物质基础.     

The Water Relations of Young Olive Trees in a Mediterranean Winter: Measurements of Evaporation from Leaves and Water Conduction in Wood

We report measurements of evaporation rate, leaf resistanceto evaporation and water conduction in the stems of young olivetrees (Olea europea L.) growing in Messina, Italy, during thewinter and early spring. We have measured what Zimmermann calls‘leaf specific conductivity’ (LSC) of stem segmentsexcised from olive trees. The LSC is a measure of the specifichydraulic conductivity of stem segments normalized per unitarea of leaves supplied by the stem segment rather than perunit area of sapwood cross-sectional area. We find that theLSC's of primary stems were the largest followed in magnitudeby the LSC's of secondary stems and tertiary stems. Under winterand early spring conditions the maximum evaporative flux fromCoratina and Nocellara varieties of olive trees is about 2.6x 10^–5 kg 8^–1 m^–2. From this and the LSC measurementswe calculate that the pressure gradients needed to maintainthis rate of evaporation in the steady state is 65 kPa m^–1in primary stems, 170 kPa m^–1 in secondary stems and 560kPa m^–1 in tertiary stems. Olive, Olea europea L, evaporation, leaf specific conductivity, hydraulic conductivity, leaf resistance     

Immunochemical Investigations with Highly Purified Glutamate Dehydrogenase from Pea Seeds by Means of the Ouchterlony Test

Monospecific antibodies have been prepared with a homogeneousprotein fraction of the main activity band of pea seed glutamatedehydrogenase. This protein precipitates with its antibodiesin a single band with complete fusion as seen by the Ouchterlonydouble-diffusion test. Identical behaviour is observed withthe protein of the adjacent activity bands of the multiple molecularforms of this enzyme and the antibodies to the former fraction.Organ-specific ‘isoenzymes’ of glutamate dehydrogenasewith preparations of pea roots and epicotyls are not detectedby this procedure. Partially purified glutamate dehydrogenasepreparations from Lemna perpusilla, Zea mays, and Oryza sativaalso precipitate with the antibodies to the pea protein. TheLemna protein is shown to be different from the pea enzyme asjudged from immunological behaviour. The pea antibodies¹ donot cross-react with glutamate dehydrogenases from Candida orbeef liver, nor do the beef liver antibodies react with thepea and Candida enzymes.     

Peroxide-induced membrane blebbing in endothelial cells associated with glutathione oxidation but not apoptosis

Cells underoxidative stress induced by peroxides undergo functional andmorphological changes, which often resemble those observed duringapoptosis. Peroxides, however, also cause the oxidation ofintracellular reduced glutathione (GSH). We investigated the relationbetween these peroxide-induced effects by using human umbilical veinendothelial cells (HUVEC) and two HUVEC-derived cell lines, ECRF24 andECV304. With HUVEC, tert-butylhydroperoxide (tBH) or hydrogenperoxide application in the presence of serum induced, in adose-dependent way, reorganization of the actin cytoskeleton, membraneblebbing, and nuclear condensation. These processes were accompanied bytransient oxidation of GSH. With ECRF24 cells, this treatment resultedin less blebbing and a shorter period of GSH oxidation. However,repeated tBH addition increased thenumber of blebbing cells and prolonged the period of GSH oxidation. ECV304 cells were even more resistant to peroxide-induced bleb formation and GSH oxidation. Inhibition of glutathione reductase activity potentiated the peroxide-induced blebbing response in HUVECand ECRF24 cells, but not in ECV304 cells. Neither membrane blebbingnor nuclear condensation in any of these cell types was due toapoptosis, as evidenced by the absence of surface expression ofphosphatidylserine or fragmentation of DNA, even after prolonged incubations with tBH, although hightBH concentrations lead to nonapoptotic death. We conclude that, in endothelial cells,peroxide-induced cytoskeletal reorganization and bleb formationcorrelate with the degree of GSH oxidation but do not represent anearly stage of the apoptotic process.

     

Effects of Acriflavine on the Formation of the Plasma Membrane of Escherichia coli

When cells of acriflavine-sensitive (acrA) and acriflavine-resistant(acrA⁺) Escherichia coli K-12 strains were treated with a ratherhigh concentration (100 µg ml^-1) of acriflavine in mediumthat had been adjusted to pH 8.1, distinct whirlpool-like structuresderived from the plasma membrane appeared not only in the acrAcells but also in the acrA⁺ cells. Chemical analysis was performedto determine the lipid composition of the cells by thin-layerchromatography on silica gel and gas-liquid chromatography.The amount of total fatty acids was significantly higher inthe acrA cells than in the acrA⁺ cells, when cells were culturedin the presence of acriflavine. This difference seems to becaused by the greater accumulation of unsaturated fatty acids(palmitoleic and cis-vaccenic acid) in the acrA mutant cellsthan in the acrA⁺ cells and by the acceleration of this accumulationas a result of the presence of the dye. A comparison of phospholipidcontents between the acrA and acrA⁺ cells cultured under acriflavine-freeconditions showed that the former cells contained more phosphatidylethanolamine(PE) and, in particular, more cardiolipin (CL) than the lattercells. However, the situation was reversed in the case of phosphatidylglycerol(PG). Addition of acriflavine to the medium led to a markedincrease in levels of PE and CL in both acrA and acrA⁺ cellsbut an increase in levels of PG was found only in the acrA⁺cells. (Received October 13, 1992; Accepted May 31, 1993)     

Differences in Iron Nutrition Strategies of Two Calcifuges,Carex piluliferaL. andVeronica officinalisL.

Veronica officinalisandCarex pilulifera, widespread calcifugeplants in Europe, were cultivated in acid and calcareous soilsto study differences in Fe aquisition strategies indicated inprevious studies. The experiments were performed in a computer-controlledglasshouse at a soil solution moisture content of 50–60%water holding capacity; additional light was supplied at 70W m^-2if ambient light was <100 W m^-2between 0600 and 1800h.Both species developed chlorosis when grown in the calcareoussoil.C. piluliferaproved unable to translocate sufficient amountsof Fe to the leaves when cultivated in calcareous soil, butmuch Fe accumulated in, and especially as a precipitate on thesurface of roots. In contrast,V. officinalistended to increaseFe taken up into the leaves of plants grown on calcareous soil,but a much greater proportion of the leaf tissue Fe was accumulatedas less active forms not extracted by Fe complexing agents,e.g. 1,10-phenanthroline, than was the case in acid-soil grownplants. Considerably less Fe was accumulated in the root biomassofV. officinaliscompared toC. pilulifera.It is concluded thatchlorosis inC. piluliferais related to insufficient Fe uptakein the leaves, whereas leaf immobilization of Fe in physiologicallyless active forms is the problem inV. officinalis. Iron; chlorosis; calcifuge; iron immobilization; leaf tissue; fractionation; Carex pilulifera; Veronica officinalis     

Proximal tubular epithelial cells are generated by division of differentiated cells in the healthy kidney

We searched for evidence for a contribution of stem cells in growth of the proximal S3 segments of healthy rats. According to the stem cell model, stem cells are undifferentiated and slow cycling; the bulk of cycling cells are transit amplifying, rapidly cycling cells. We show the following. 1) By continuous application of a thymidine analog (ThA) for 7 days, S3 proximal epithelial cells in healthy kidneys display a high-cycling rate. 2) Slow-cycling cells, identified by lack of ThA uptake during 14 days of continuous ThA application up to death and by expression of the cell cycle protein Ki67 at death, have the same degree of differentiation as quiescent cells. 3) To detect rapidly cycling cells, rats were killed at various time points after injection of a ThA. Double immunofluorescence for ThA and a cell cycle marker was performed, with colocalization indicating successive divisions. During one week after division, daughter cells display a very low proliferation rate, indicating the absence of rapidly cycling cells. 4) Labeling with cyclin D1 showed that this low proliferation rate is due to cycle arrest. 5) More than 50% of the S3 cells entered the cell cycle 36 h after a potent proliferative stimulus (lead acetate injection). We conclude that generation of new cells in the proximal tubule relies on division of differentiated, normally slow-cycling cells. These may rapidly enter the cycle under an adequate stimulus. immunohistochemistry; cell cycle; proliferation; renal stem cells; proximal tubule; renal epithelial cells