Scanning Electron Microscopy of the Infection Process of Cercospora henningsii on Cassava Leaves

Germination, penetration and sporulation of Cercospora henningsii (Allesch.) on cassava leaves were studied by scanning electron microscopy. Conidia started to germinate 9 h postinoculation producing one to two germ tubes. The germ tubes entered the leaf tissue through the abaxial surface by direct penetration of the epidermis without forming appressoria. The cassava leaf is characterized by its papillose epidermis on the abaxial surface. The penetrations occurred at smooth areas of the leaf epidermis between the papillae. The germ tubes did not enter stomata even when they passed over stomatal openings. Leaf spots started to appear 9 days after the inoculation (dpi), and the emergence of conidia occurred 14 dpi. The symptoms appeared first on the abaxial leaf surface, followed 2 days later on the adaxial. Conidia emerged in clusters through ruptured epidermis on both sides of the leaves. Conidia emerged also through the epidermal papillae and the leaf veins. Even though small groups of conidia emerged through stomata also, emergence through stomata appeared to be random rather than a preferred route. Each conidium was born on a short conidiophore with a swollen base.     

Field Studies of the Wheat Stomata Resistance Influenced by Soil Water Content

Concurrent observations of soil water potential and leaf stomata diffusion resistance were made on two, plots of wheat grown at Datun Agro-ecological Experimental Station in Beijing under two different soil water conditions. These data were further complemented by weather and physiological observation. In this paper, we mainly analysed the influence of soil water potential on the status of wheat leaf stomatal resistance. The results indicate that: (1) there is a obvious influence of soil water potential on the status of wheat leaf stomata under normal conditions and (2) there is a different upper and lower epidermis stomata of wheat leaf respond to the soil water potential. The lower epidermis stomata are more sensitive to soil water potential than upper epidermis one. (3) There is a linear relationship between the ratio of lower and upper epidermis stomata resistance and soil water potential in root layer, according to this we can diagnose the degree of wheat water deficit.     

Where does all the calcium go? Evidence of an important regulatory role for trichomes in two calcicoles

In previous studies of the calcicoles Centaurea scabiosa and Leontodon hispidus, the stomata on isolated epidermis closed partially when the concentration of calcium in the medium was above 1 mol m^?3. This is a much smaller concentration than that believed to be delivered into the leaves in xylem sap when the plants are growing in a calcium-rich medium, and hence the mechanism for ‘protecting’ stomata from excessive exposure to free calcium is thought to be of great physiological significance. It is shown here that, in the leaves of both species, a substantial amount of the calcium they contain is located within meso-phyll cells, and virtually all of that which does enter the epidermis is contained within trichomes, probably as calcium oxalate. The amounts of calcium in the vicinity of the stomata thus remain small despite high concentrations elsewhere, ensuring that the essential role of Ca²⁺ in intra-cellular signalling in guard cells can continue to be performed without disturbance.     

Model-derived dose rates per unit concentration of radon in air in a generic plant geometry

A model for the derivation of dose rates per unit radon concentration in plants was developed in line with the activities of a Task Group of the International Commission on Radiological Protection (ICRP), aimed at developing more realistic dosimetry for non-human biota. The model considers interception of the unattached and attached fractions of the airborne radon daughters by plant stomata, diffusion of radon gas through stomata, permeation through the plant’s epidermis and translocation of deposited activity to plant interior. The endpoint of the model is the derivation of dose conversion coefficients relative to radon gas concentration at ground level. The model predicts that the main contributor to dose is deposition of ²¹⁴Po α-activity on the plant surface and that diffusion of radon daughters through the stomata is of relatively minor importance; hence, daily variations have a small effect on total dose.     

Research on the Application and Mechanism of Stomatic Resistance Assessment in Monitoring Air TSP Pollution

The stomatic resistance of 9 woody species in different functional areas of Chengde City, North China, was measured with an automatic porometer and the effect of total suspensible particles (TSP) on the plugging of stomata was observed under SEM. It was found that plants in the heavy traffic area and the residential area had relatively higher stomatic resistance, with a mean value (averaged from the upper and lower epidermis) of 10. 5±5.1 s · cm-1. The stomatic resistance of the heavy traffic area was twice as much as that of the relativey clean area, even 8 times more in some species. The great difference, as much as 25 times more, was also found between the upper and lower epidermis even in the same species. In some species such as Fraxinus americana, the stomatic resistance of the upper epidermis was so high that it was beyond the limit of measurement. There was a very good correlation between the stomatic resistance of the lower epidermis and the air TSP, especially in F. americana and Salix matsudana, with r=0. 9968 (<0. 001) and 0. 9951 (<0. 001) respectively. Such relation was very poor for the upper epidermis. Stomata plugging with TSP was studied under SEM. It was noticed that small particles (<5 μm) could enter the stomta to cause plugging and large particles (>30 μm) could plug the stomata by covering the whole orifices, both of which cause stomatic resistance. The findings indicate the main mechanism of monitoring air TSP pollution by stomatal resistance measurement.     

Studies on Foliar Penetration: 2. THE ROLE OF LIGHT IN DETERMINING THE PENETRATION OF 2,4 DICHLOROPHENOXYACETIC, ACID

A further study has been made of the factors determining thelevel of penetration of 2,4-dichlorophenoxyacetic acid (2,4-D)into leaves. The technique involves the use of leaf disks and2,4-D containing carbon-14 in the carboxyl group. For Phaseolus vulgaris the influence of the pH of the appliedsolution is greater in the light than in the dark. Between 0and 1,000 f.c. at 27° C there is a small increase in therate of penetration into the abaxial surface. Under these conditionsthe rates remain constant up to 56 hours. This linear relationshipholds for concentrations ranging from 100 to 1,000 mg/l, andthe rate of penetration is directly proportional to the concentration.For intensities in excess of 1,000 f.c. the light response ismarkedly different: over the first few hours there is a steadyand relatively slow rate of penetration which is followed bya second phase when the rate is greatly accelerated. This acceleratedrate can be reversed by transferring the disks to darkness,and does not take place at 1° C. Likewise, if excised disksare left for more than one hour in either the light or the darkbefore applying 2,4-D then there is subsequently no phase ofaccelerated penetration. The course of penetration into theadaxial surface exhibits no accelerated rate, and compared tothe abaxial surface the rates are lower. For leaves of Ligustrum ovalifolium, which lack stomata on theadaxial surface, the rates of penetration at 27° C intoboth surfaces remain constant in either light or darkness. Forboth the adaxial and abaxial surfaces, the rate progressivelyincreases from 0 to; 2000 f.c. and there is no phase of acceleratedpenetration. Penetration into the adaxial surface is less. At1° C the rates for both surfaces in either light or darknessare depressed. If disks of Phaseolus are irradiated with ultraviolet light,subsequent penetration is markedly depressed in the light at27° C, but in the dark or at 1° C it is enhanced. On the basis of these findings it is concluded that both physicaland metabolic factors control the rate of penetration of 2,4-DTransport through the cuticle will be dependent on adsorptionand the length of the diffusion paths. Once diffusion gradientshave been established between the surface of the cuticle andthe outer surface of the cytoplasm in the epidermal cells, thesteepness will be dependent on the rates at which 2,4-D is eitherconverted into some metabolite or moved away from the surfaceor into other cells. The relative importance of physical andmetabolic process will be dependent on the permeability andthickness of the cuticle and the level of metabolic activity.The possible role of ectodesmata in determining both the lengthand steepness of the diffusion paths is discussed.     

An ethoxylated surfactant enhances the penetration of the sulfated laminarin through leaf cuticle and stomata,leading to increased induced resistance against grapevine downy mildew

Some β‐1,3‐glucans and particularly sulfated laminarin (PS3) are known as resistance inducers (RIs) in grapevine against the downy mildew. However, their efficacy in vineyard is still often too low, which might be caused by a limited penetration through the leaf cuticle following spray application. We used ¹⁴C‐sucrose uptake experiments with grapevine leaves in order to select a surfactant as saccharide penetration enhancer. Our results showed that although sucrose foliar uptake was low, it was strongly enhanced by Dehscofix CO125 (DE), a highly ethoxylated surfactant. Fluorescent saccharides were then produced and laser scanning microscopy was used to analyze their foliar diffusion pattern in Arabidopsis thaliana and grapevine. Interestingly, sucrose and PS3 were seemingly able to penetrate the leaf cuticle only when formulated with DE. Diffusion could preferentially occur via stomata, anticlinal cell walls and trichomes. In grapevine, PS3 penetration rate was much higher on the stomateous abaxial surface of the leaf than on the adaxial surface. Finally, using DE allowed a higher level of downy mildew control by PS3, which corroborated diffusion observations. Our results have practical consequences for the improvement of treatments with saccharidic inducers on grape. That is, formulation of such RIs plays a critical role for their cuticular diffusion and consequently their efficacy. Also, spray application should preferentially target the abaxial surface of the leaves in order to maximize their penetration.     

Studies on the Epidermal Structure of Tingia carbonica

Two leafy specimens of Tingia carbonica were collected as impression from Shanxi formation of Permian, from Inner Mongolia, China. The epidermal structure of its leaves is reported in this paper. Under SEM, well preserved epidermal cells as well as some concaves on the surface of large leaves were clearly recognized. The epidermal cells are approximately rectangular in shape, about 100～150 um long and 20～30 um wide. They are arranged longitudinally parallel to veins. The concaves usually in rows are round or elliptical, about 0.65～0.35 mm long and 0.2～0.35 mm wide. Density of concaves is about 1.8/ mm² and no stomata occur inside the concaves. In all probability, this is the upper epidermis. On the other side of the epidermis, anomocytic stomata are scattered irregularly, each with 5～6 epidermal cells around. The stomatal apertures are about 35.8 µm long, and 18.7 um wide, which is organised parallel to the common epidermal cells. As far as shape and size is concerned, it is similar to that described on the upper epidermis. Density of the stomata is about 60/mm². In all Probability, this is the lower epidermis. The ecological preference and classification of Tingia are discussed according to these new characters of the epidermis andsubordinate struture of the leaf.     

The Epidermal Cells of Tea Leaves

The upper epidrmis of tea leaf consists of cells about 30 40 μ in diameter, with slightly sinuous cell surface and devoid of stomata or hairs. The lower epidermis consists of cells about 50–70 μ in diameter, with more sinuous walls. Stomata confined to the lower surface, surrounded by 3–4 round, subsidiary cells. The upper andlower epidermis of the wild tea of southwest China show the difference in surface texture. Luxuriant wax of the epidermis is in knob or club shape. There are two types of stomata (namely general stomata and stomata of sunken crypts (gland scale)) on the same leaf. The numbers of stomata are distributed 70–100/mm2. Hairs are short and rare, or none. Intercellular flanges between epidermis is steep and thick in wild tea. The protruded parts of the torus are in the form of “foot”. The flanges of the cuticle are rather deep.     

Morphological and Anatomical Observations of Clasping Leaves of Tamarix L.

This paper deals with some morphological and anatomical adaptabilities of clasping leaves of 13 species of Tamarix L. in China. These xeromorphic leaves posses some conspicuous characteristics as follows: 1 ) The ratio of the leaf surface area to its volume is low; 2) The numerous, small stomata are deeply sinked in the surface of the epidermis; 3) The palisade tissue is only found in the abaxial part and the spongy tissue in the adaxial part, which are quite different from most of the dorsivcntral leaves; 4) The upper epidermis of these leaves is extended a different degree into the epidermis of stem. These characteristics may be considered as an expression of xeric habits resulting from desert adaption.     

Evidence for the Uptake of Large Anions through Stomatal Pores

Experiments were conducted with leek (Allium porrum L.) leaves to investigate whether aqueous solutions are able to penetrate stomata. Epidermal strips were used for the determination of transport rates. Stomata were opened by fusicoccin or closed by darkness or abscisic acid. A droplet containing the anionic fluorescent dye, uranine, was placed on the physiologically outer side of the epidermis and allowed to dry. With open stomata 30 times more uranine penetrated through the epidermal strips than with closed stomata (comparison of medians). In another experiment droplets of uranine solution were placed on leaf segments and epidermal strips were removed after drying of the droplets. Penetration of uranine through stomata was detectable under the microscope both with epidermal strips from the transport experiments and with strips obtained after application on leaf segments. As maximum uptake rates occurred during the drying process, it is concluded that penetration took place via water films. These results show that the physical restrictions preventing stomatal penetration of static droplets are not decisive for drying droplets and that stomatal uptake of dissolved ionic substances occurs under natural conditions, i.e. without surfactants or applied pressure.     

麦冬、土麦冬和阔叶土麦冬叶表皮形态结构的观察

用光镜和扫描电镜观察了麦冬（Ophiopogon japonicus（L．f）Ker—Gawl.]、土麦冬（Liriope spicata Lour．）和阔叶土麦冬（L．platyphylla Wanget Tang）叶表皮显微结构、亚显微结构和角质层内表面的形态结构。结果表明,气孔主要分布于麦冬、土麦冬和阔叶土麦冬叶片的下表皮,气孔密度分别为76．4、114．3和99．8个·mm^-2;仅阔叶土麦冬叶片上表皮有少量气孔分布。3种植物的气孔器均不具有副卫细胞,并在叶脉间形成纵向气孔带。表皮细胞长方形,气孔带与非气孔带处表皮细胞的形态和大小差异较明显。麦冬气孔周围的表皮细胞平周壁具明显瘤状突起,导致气孔下陷;土麦冬气孔周围的表皮细胞平周壁呈波浪状突起,使气孔相对下陷;阔叶土麦冬气孔周围的表皮细胞平周壁基本无突起,气孔不下陷。3种植物的叶表皮均有发达的角质层和丰富的蜡质,且蜡质主要分布于下表皮气孔带处。这些结构特征可能与它们所具有的喜阳、耐阴和耐旱等特性有一定的相关性。     

Stomatal responses of Argenteum — a mutant of Pisum sativum L. with readily detachable leaf epidermis

Epidermis is easily detached from both adaxial and abaxial surfaces of leaf four of the Argenteum mutant of Pisum sativum L. The isolated epidermis has stomata with large, easily-measured pores. Hairs and glands are absent. The density of stomata is high and contamination by mesophyll cells is low. In the light and in CO₂-free air, stomata in isolated adaxial epidermis of Argenteum mutant opened maximally after 4 h incubation at 25°C. The response of stomata to light was dependent on the concentration of KCl in the incubation medium and was maximal at 50 mol m^-3 KCl. Stomata did not respond to exogenous kinetin, but apertures were reduced by incubation of epidermis on solutions containing between 10^-5 and 10^-1 mol m^-3 abscisic acid (ABA). The responses of stomata of Argenteum mutant to light, exogenous KCl, ABA and kinetin were comparable with those described previously for stomata in isolated epidermis of Commelina communis. A method for preparing viable protoplasts of guard cells from isolated epidermis of Argenteum mutant is described. The response of guard cell protoplasts to light, exogenous KCl, ABA and kinetin were similar to those of stomata in isolated epidermis except that the increase in volume of the protoplasts in response to light was maximal at a lower concentration of KCl (10 mol m^-3) and that protoplasts responded more rapidly to light than stomata in isolated epidermis. The protoplasts did not respond to exogenous kinetin, but when incubated for 1 h in the light and in CO₂-free air on a solution containing 10^-3 mol m^-3 ABA, they decreased in volume by 30%. The advantages of using epidermis from Argenteum mutant for experiments on stomatal movements are discussed.Abbreviations ABA abscisic acid - MES 2-(N-morpholino)ethanesulfonic acid     

青藏高原不同海拔高度下全缘叶绿绒蒿叶表皮特征研究

采用胶带粘取叶表皮法,利用光学显微镜观测不同海拔高度下全缘叶绿绒蒿叶片的表皮毛、气孔及表皮细胞结构特征,探讨全缘叶绿绒蒿叶表皮特征与海拔高度的关系。结果表明,随着海拔高度增加,全缘叶绿绒蒿叶片上、下表皮毛密度、气孔密度和表皮细胞密度逐渐增加;气孔器及表皮细胞的长度、宽度和面积逐渐减小;表皮细胞的形态随着海拔升高由无规则形向多边形变化,垂周壁由波状向弓形或平直变化。全缘叶绿绒蒿叶表皮结构在不同海拔高度下表现出的差异,可能是植物长期在高原生态环境下的综合反应,以结构上的变化来适应对高海拔地区的环境特点,为进一步研究高海拔地区植物的适应性提供一定依据。     

Environmental scanning electron microscopy of the surface of normal and vitrified leaves of Gypsophila paniculata (Babies Breath) cultured in vitro

Leaf surfaces of non-tissue-cultured, vitrified and non-vitrified plantlets of Gypsophila paniculata (Babies Breath) were examined using an environmental scanning electron microscope. Non-tissue-cultured plants had a complete epidermal surface, recessed stomata and wax present on the leaf surface. The surface of tissue-cultured plantlets appeared similar to non-tissue-cultured plants excepting stomata were slightly protruding and less wax appeared to be present. In both non-tissue-cultured and tissue-cultured plants stomata were found both opened and closed and were observed closing. In contrast vitrified plantlets had abnormal, malformed stomata which appeared non-functional. The ventral surfaces of leaves seemed more normal than the dorsal, this may be due to the former receiving more light. Additionally, discontinuities were found in the epidermis. Often epidermal holes were found in association with stomatal apertures. It is suggested that the main cause of desiccation of vitrified G. paniculata plantlets ex vitro is due to loss of water from the discontinuity in epidermis and not because of non-functional stomata. Liquid water could be seen through the epidermal holes indicating that at least some of the extra water in vitrified plantlets is contained in the intercellular spaces.Abbreviations ESEM Environmental scanning election microscope - IAA Indole acetic acid - NTC Non-tissue-cultured - TC Tissue-cultured - V vitrified     

A Possible Role for Calcium in Oxidative Plant Stress

Disruption of calcium homoeostasis may be crucially important in damaging animal cells under oxidising conditions. In hepatocytes, oxidative stress causes an increase in cytosolic calcium, a process which is thought to be largely responsible for subsequent cell death. The oxidation of glutathione precedes a flux of calcium into the cytoplasm from internal and external sources. The possibility that similar events occur in plant cells has been tested using strips of epidermis taken from leaves of Commelina communis. The closure of stomata in response to many chemical or physical stimuli is thought to involve a transient rise in cytosolic calcium. Paraquat (10^-4 M) and hydrogen peroxide (10^-3 M) caused a marked reduction in stomatal aperture when included in the incubation medium of illuminated epidermal strips. The calcium channel blocker verapamil (10^-5 M) protected stomata from hydrogen peroxide-induced closure while the ion chelator EGTA (2 × 10^-3 M) protected stomata in both oxidising treatments. These results are discussed in so far as they support the hypothesis that a rise in cytosolic calcium may play a role in the pathology of oxidative stress in plant cells through a mechanism analogous to the liver cell system.     

Lipids in the skin of a cat-fish Heteropneustes fossilis (Bloch) (Heteropneustidae,Pisces)

Summary Localization and characterization of different lipids in the cellular constituents of the skin of Heteropneustes fossilis has been made using several histochemical techniques.High contents of cholesterol, its esters and phospholipids have been correlated with the metabolically active state of the basal cells undergoing cell proliferation and differentiation.The polygonal cells in the outermost layer of the epidermis, though rich in phospholipid contain small amounts of cholesterol and its esters. This has been correlated with the metabolically less active state of these cells.Neutral lipids and phospholipids in the polygonal cells of the outermost layer may contribute to the contents of surface slime and act as an efficient barrier for the penetration of water through the skin.The deposits of neutral lipids in the subcutis may provide energy during the period of fasting, act as a barrier for water diffusion through the skin and serve as shock absorbing pads protecting the fish from mechanical injury.Supported by a post doctoral fellowship sponsored by the Council of Scientific and Industrial Research, Government of IndiaSupported by a research scholarship sponsored by Banaras Hindu University     

Uptake of Hydrophilic Solutes Through Plant Leaves: Current State of Knowledge and Perspectives of Foliar Fertilization

Foliar fertilization is an agricultural practice of increasing importance in practical terms. Since nutrient sprays are mostly applied as water solutions, the focus of the article was placed on the penetration of ionic, polar solutes through the leaf surface, although the mechanisms of cuticular penetration of lipophilics are also taken into consideration. In theory, application of foliar nutrient sprays may indeed be a more target-oriented and environmentally friendly fertilization method since nutrients are directly delivered to the target organism in limited amounts, thereby helping to reduce the environmental impact associated with soil fertilization. However, response to foliar sprays is often variable and not reproducible due to the existing lack of knowledge of many factors related to the penetration of the leaf-applied solution.

To overcome the current “spray and pray” situation, there is a need to critically analyze the major factors involved and the existing experimental approaches to safely assess the penetration mechanisms, which is the final aim of this review. Beginning with the significance of foliar fertilization in agriculture, a historical overview regarding foliar uptake related studies is given, with especial emphasis on the penetration of solutes through the cuticle and stomata. The existing models of cuticular and stomatal uptake are analyzed separately considering among other factors the physico-chemistry of the solution including the role of adjuvants and the effect of the environment. Methods employed to estimate the process of cuticular and stomatal penetration of solutes are critically assessed. Finally, conclusions related to multidisciplinary research perspectives for improving the efficiency of foliar sprays are drawn.     

DEVELOPMENT,STRUCTURE AND FUNCTION OF SECRETORY TRICHOMES IN PSYCHOTRIA BACTERIOPHILA (RUBIACEAE)

Mucilage-secreting dendroid trichomes develop from the adaxial epidermis of young stipules surrounding the shoot apex. Each trichome consists of a multicellular stalk from which radiate many branch cells. The trichome has no cuticle and the branch cell walls distally are loose cellulosic frameworks. Dictyosomes produce vesicles whose products are secreted through the plasma-lemma and cell wall. Enlarged portions of the ER are frequently associated with dictyosomes and may be part of the system for synthesis and transport of secretion products. Bacteria, which later occur in leaf nodules, are present in the mucilage surrounding trichomes and young leaves. The latter develop stomata through which the bacteria enter. As stipules and leaves grow out of the apical region, the secretory trichomes degenerate and are replaced by non-secretory ones.     

The influence of black bean aphid, Aphis fabae Scop., and its honeydew on the photosynthesis of sugar beet

The effect of black bean aphids on the photosynthesis of sugar beet plants was studied under glasshouse and field conditions. The presence of up to several hundred aphids per leaf had no significant effect on CO₂ exchange rates over a range of light intensities between complete darkness and light saturation. Artificially prepared honeydew, sprayed onto leaves in the same amounts and composition as was found on severely aphid-infested plants, covered 30% of the stomata on the upper epidermis but did not significantly alter the rate of photosynthesis of these leaves in the light or the rate of respiration in the dark. The stomata on the lower epidermis were uncovered and functional. High pressure liquid chromatography of aphid-produced honeydew detected 20 different amino-acids. Three amino-acids, aspartic acid, glutamic acid and gluta-mine, made up the bulk of the amino-acid weight in the honeydew produced on young plants, up till the 8 leaf-stage. In the 10 to 12 leaf-stage, several different amino-acids occurred in substantial amounts. The amino-acids to sugars ratio of the honeydew produced by the aphids decreased strongly as the sugar beet plants aged: from 1:6 in plants with 3 or 4 leaves to 1:25 in plants having 10 to 12 leaves.