Protein Phosphorylation in the Sieve Tubes of Rice Plants

Proteins and their phosphorylation were examined in rice phloemsap that had been collected by the insect laser technique. Analysisby SDS-PAGE indicated that rice phloem sap contained over 150proteins. The total protein concentration was 150–200ng µl^-1. Analysis of proteins extracted from leaves, rootsand seeds revealed that several major low-molecular-weight proteinswere confined to the rice phloem sap. Maintenance of rice plantsunder stable environmental conditions was associated with aconstant complement of proteins in the phloem sap. Phosphorylation and dephosphorylation of proteins were detectedin the sieve tubes and occurred in a light dependent mannerwhen [     

Fluid Dynamics Simulating Experiment of Pressure Gradient for Assimilate Translocation in Sieve Tubes

According to the similitude theory of fluid mechanics, a pressure-driven experiment in a sieve tube model has been performed. The relationships between the pressure gradient and the transport velocities at the various diameters of sieve pore have been obtained. The experimental data were compared with the theoretical prediction based on the poiseuille equation. The result shows that the pressure gradient required for translating the assimilate is directly proportional to the translating velocity, which agrees with Poiseuille equation. However, the resistance of the sieve plate to flow is much more than that estimated by the previous theory. Even though the translating velocity is a normal level and the diameter of the sieve tube is a typical size, the pressure gradient required for flow is about 1 bar/m. It will increase sharply with the decrease of the diameter of sieve pore. The ratios of the pressure gradients measured in the test to that predicted by Poiseuille equation are about 2–4 for the five kinds of diameters of Sieve pore. Additionally, a simulating test of the pores being partly plugged has been performed. The result shows that the pressure gradients measured are greatly beyond the range that may be kept in plants. This study will be helpful to assess Munch hypothesis.     

Electron-Microscopic Evidence of the Vacuolar Nature of Phloem Exudate

We performed electron-microscopic examination of structural diurnal changes in the lumen of sieve tubes and the vacuolar system of corresponding companion cells and changes induced by the experimental blockage of assimilate export from the leaf by its cold-girdling. For these investigations, Cucurbita pepo L. and Helianthus annuus L. plants were used, that is, plant species from groups of symplastic and apoplastic plants, which differ in the type of companion cells and a mode of phloem terminal loading. The examinations showed the complete identity of changes in the electron texture of the sieve-tube lumens and companion-cell vacuoles in both plant species in the course of a day, when the level of assimilates changed, or after export blockage. Similar changes in the structure of the vacuolar labyrinths were stated in the companion cells under normal conditions and after cold-girdling, as related to the rate of sieve-tube loading with the vacuolar exudate. Vacuolar expansion and starch accumulation developing in response to changes in the assimilate level in the evening and after cold blockage of the assimilate export occurred in different types of cells, as dependent on their position in the symplast domains. However, the rate of the process similarly depended on the balance between assimilate synthesis and export. Synchronous changes in the texture of the sieve-tube lumen and companion-cell vacuoles were observed within each complex, but asynchronous changes occurred in different complexes. We suggested this phenomenon for recognizing the particular complexes, when they are grouped in a bundle. We observed no signs of cytoplasm or protein synthetic machinery in the sieve tubes. We concluded that the sieve-tube lumen and vacuoles of companion cells are common in nature. Similar electron texture of the images of the companion-cell vacuolar labyrinth and tube lumens, their connection through the lateral sieve fields, morphological modifications of the companion-cell vacuolar system as dependent on the activity of sieve tube loading—all of these facts imply the continuity of these transport compartments and fluxes in them and the similarity in the composition of the exudates from companion-cell vacuoles and phloem tubes.     

Protein Kinases in Plants in the Transduction of Abiotic and Biotic Signals

The effects of extrinsic and intrinsic factors upon signaling pathways involving protein kinases are reviewed with particular reference to monomeric GTP-binding proteins and MAP kinases. The possible roles of feedback control and interactions between cascades in plant responses are stressed.     

高等植物开花诱导途径信号整合的分子机制

开花是高等植物从营养生长到生殖生长的重要转折点。花分生组织的形成是开花植物对内外环境信号的响应。近年来在开花诱导方面已获得许多研究成果,我们介绍了高等植物开花诱导的4条主要途径（光周期途径、春化途径、自主途径和赤霉素途径）和复杂的信号整合机制。     

Analysis of Possible Involvement of Local Bioelectric Responses in Chilling Perception by Higher Plants Exemplified by <Emphasis Type="Italic">Cucurbita pepo</Emphasis>

Macroelectrodes and electrophysiological setup were used in experiments with stems of 15-day-old pumpkin (Cucurbita pepo L.) seedlings for computer-assisted data recording. It is shown that local bioelectric responses induced by graded local chilling are similar to the receptor potentials of animals. These responses increase gradually with stimulus strength and trigger the action potential generation when a temperature threshold is attained. The excitation threshold of cells in seedling stems displays the phenomenon of accommodation. Parameters of local bioelectric responses induced by intermittent cooling can undergo changes similar to sensitization-habituation patterns. The results indicate that local electrical responses may be involved in early stages of cooling perception in cells of higher plants devoid of locomotive functions.     

高等植物自交不亲和反应中花粉管信号传导研究进展(综述)

高等植物自交不亲和反应是由基因控制、避免发生自花授粉的一种机制。本文介绍以虞美人为主的高等植物在自交不亲和反应中肌动蛋白骨架的动态变化及Ca2 的时空变化,着重阐述花粉管生长被抑制的最初信号传导。     

A Simple Method for Fixing, Dehydrating and Embedding Pollen Tubes Cultivated in Vitro for Optical and Transmission Electron Microscopy

A simple method to cultivate pollen tubes in a gelatin medium is presented. After the growth of the pollen tubes in the culture medium, they are fixed, dehydrated, and embedded in resin for ultramicrotomy. The method is easy and does not require the purchase of special materials beyond those needed for the usual techniques for studying biological specimens under transmission electron microscopy.     

Electric field-induced breakdown of lipid bilayers and cell membranes: A thin viscoelastic film model

Summary A simple viscoelastic film model is presented, which predicts a breakdown electric potential having a dependence on the electric pulse length which approximates the available experimental data for the electric breakdown of lipid bilayers and cell membranes (summarized in the reviews of U. Zimmermann and J. Vienken, 1982,J. Membrane Biol. 67:165 and U. Zimmermann, 1982,Biochim. Biophys. Acta 694:227). The basic result is a formula for the time of membrane breakdown (up to the formation of pores): =(/C)/( _m ² ₀ ² U ⁴/24Gh ³+T ²/Gh–1), where is a proportionality coefficient approximately equal to ln(h/2₀),h being the membrane thickness and ₀ the amplitude of the initial membrane surface shape fluctuation ( is usually of the order of unity), represents the membrane shear viscosity,G the membranes shear elasticity modules, _m the membrane relative permittivity, ₀=8.85×10^–12 Fm,U the electric potential across the membrane, the membrane surface tension andT the membrane tension. This formula predicts a critical potentialU _c ;U _c =(24Gh ³/ _m ² ₀ ² )^1/4 (for = andT=0). It is proposed that the time course of the electric field-induced membrane breakdown can be divided into three stages: (i) growth of the membrane surface fluctuations, (ii) molecular rearrangements leading to membrane discontinuities, and (iii) expansion of the pores, resulting in the mechanical breakdown of the membrane.     

植物感受盐胁迫及相关钙信号的研究进展

盐胁迫对植物的生长和发育造成严重影响,其危害包括渗透胁迫、离子毒害等,严重损害了农业生产和粮食安全。在盐胁迫下,植物相关感受器接受刺激,使得Ca²⁺通过细胞膜以及细胞内钙库膜上打开的Ca²⁺通道进入细胞质基质,导致细胞质内Ca²⁺浓度升高,产生钙信号。钙离子作为重要的第二信使,在植物细胞内和细胞间传递信号,信号往下游传递,在不同生长和发育阶段引起植物一系列的生理响应来应对盐胁迫影响。钙信号主要通过钙调蛋白（CaM）、钙调素样蛋白（CML）、钙依赖性蛋白激酶（CDPK）、钙调磷酸酶B样蛋白（CBL）和CBL互作蛋白激酶（CIPK）感知并将特异的钙信号信息传递到下游;从而激活植物盐胁迫生理响应。本文主要综述植物如何感知盐胁迫刺激,以及钙信号产生与传导机制,并对该研究领域需解决的问题进行了展望。     

Control of plant development: Signals from without—Signals from within

The powerful technology for transferring functional foreign genes into plants can only express its potential to the extent that our knowledge about signals and signal transmission in plants improves. In higher plants gene expression is regulated by ‘signals from within’ and ‘signals from without’ (e.g., light). Since light-mediated changes provide the basis for much of plant development, photocontrol of gene expression will mainly be considered. The recently discovered ‘plastidic factor’ will serve as the prototype of an intracellular ‘signal from within’. Particular emphasis will be laid on rapid interorgan signal transmission since these novel observations suggest a revision of the presently held concepts about the means of communication within a plant. It will be concluded in the end that the prevailing views about the nature of plants underestimate the degree of sophistication actually exhibited by higher plants.     

Thiamine and Cholinergic Transmission in the Electric Organ of Torpedo

The electric organ of Torpedo marmorata was found to contain as much as 120 +/- 24 nmol of thiamine per g of fresh tissue. The vitamin was distributed as nonesterified thiamine (32%), thiamine monophosphate (22%), thiamine diphosphate (8%), and an important proportion of thiamine triphosphate (38%). A high level of thiamine triphosphate was found in synaptosomes isolated from the electric organ. In contrast, the synaptic vesicles did not show any enrichment in thiamine, whereas they contained a marked peak of acetylcholine (ACh) and ATP. Thus thiamine seems to be very abundant in cholinergic nerve terminals; its localization is apparently extravesicular, either in the axoplasm or in association with plasma membrane. When calcium was reduced and magnesium increased in the external medium, the efficiency of transmission was diminished, owing to inhibition of ACh release; in a parallel manner the degree of thiamine phosphorylation was found to increase--this condition is known to modify the repartition of ACh between vesicular and extravesicular compartments. Electrical stimulation, which causes periodic variations of the level of ACh and ATP, also caused significant changes in thiamine esters. In addition, related changes of the vitamin and the transmitter were observed under other conditions, suggesting a functional link between the metabolism of thiamine and that of ACh in cholinergic nerve terminals.     

Transmission of Zucchini yellow mosaic virus by Colonizing and Non-colonizing Aphids in Greece and New Aphid Species Vectors of the Virus

Nineteen aphid species were tested for their ability to transmit Zucchini yellow mosaic virus (ZYMV) from and to zucchini under laboratory conditions. Sixteen species were found to be new vectors of ZYMV (i.e. Aphis craccae, Aphis fabae, Aphis nerii, Aulacorthum solani, Brachycaudus cardui, Brevicoryne brassicae, Hyalopterus pruni complex, Hyperomyzus lactucae, Macrosiphoniella sanborni, Macrosiphum rosae, Metopolophium dirhodum, Myzus cerasi, Rhopalosiphum maidis, R. padi, Semiaphis dauci and Sipha maydis). Their transmission efficiency by a single aphid was low (0.1–4.2%). Myzus persicae was used as a control and was the most efficient vector (41.1%, one aphid per plant). Hayhurstia atriplicis, Myzus ascalonicus and Sitobion avenae did not transmit the virus. In four out of six new vectors assayed in arena tests for propensity estimation, propensity was higher than efficiency. Data from an experimental zucchini field in northern Greece revealed a high correlation between ZYMV spread and alatae of the vector species. The most abundant aphid vectors during 2 years experimentation were M. persicae, Aphis gossypii and Aphis spiraecola. The possible role of the 16 new and the previously known aphid vectors in the epidemiology of ZYMV was investigated using data of transmission efficiency combined with the captures of their alatae in the Greek net of a Rothamsted type suction trap.     

蛋白质可逆磷酸化在信号传递中的作用

主要介绍了蛋白激酶和蛋白磷酸酶的分类、特征及其催化蛋白质的可逆磷酸化在信号传递中的作用。蛋白激酶和蛋白磷酸酶作为脑内信使的直接或间接的靶酶,通过控制信号传递途径中其它酶类或蛋白质的活性,使细胞对外界信号做出相应的反应。     

高等植物共质体区内与区间的电波传递

本文以白花紫露草(Tradescantia flumi—nensis Vel)为材料,运用胞外、胞内微电极及显微注射等方法,对电波在植物体内的传递方式进行了进一步探讨。结果表明,白花紫露革对于伤害性刺激可以产生强烈的电位波动在周身传递。这种电位波动可以被FAA固定液造成的生理障碍阻断;白花紫露草叶片可分为不同的共质体区,在共质体区内,电波传递很快,远高于液相中物质的扩散速度;而在各共质区问,电波传递出现时间上的延迟,与共质体内电波传递速度相差一个数量级,因此,可以推测,在共质体区内电波是以局部电流的方式传递的,而在共质体区问,电波的传递可能有化学介质的参与。     

Modulation of Root Signals in Relation to Stomatal Sensitivity to Root-sourced Abscisic Acid in Drought-affected Plants

Stomatal sensitivity to root signals induced by soil drying may vary between environments and plant species. This is likely to be a result of the interactions and modulations ámong root signals. As a stress signal, abscisic acid （ABA） plays a central role in root to shoot signaling, pH and hydraulic signals may interact with ABA signals and thus, jointly regulate stomatal responses to changed soil water status, pH itself can be modified by several factors, among which the chemical compositions in the xylem stream and the live cells surrounding the vessels play crucial roles. In addition to the xylem pH, more attention should be paid to the direct modulation of leaf apoplastic pH, because many chemical compositions might strongly modify the leaf apoplastic pH while having no significant effect on the xylem pH. The direct modulation of the ABA signal intensity may be more important for the regulation of stomatal responses to soil drying than the ABA signal per se. The ABA signal is also regulated by the ABA catabolism and the supply of precursors to the roots if a sustained root to shoot communication of soil drying operates at the whole plant level. More importantly, ABA catabolism could play crucial roles in the determination of the fate of the ABA signal and thereby control the stomatal behavior of the root-sourced ABA signal.     

Separation of tonoplast and plasma membrane H+-ATPase from Zucchini hypocotyls by consecutive sucrose and glycerol gradient centrifugation

Summary In order to isolate tonoplast and plasma membrane vesicles involved in ATP-dependent proton transport we devised a preparative procedure with two consecutive centrifugations. Three fractions were obtained on a sucrose step gradient: light microsomes, heavy microsomes, and a mitochondria-rich fraction. The light and heavy microsomal fractions were each recentrifuged on an isopycnic glycerol density gradient. Recentrifugation of light microsomes resulted in two fractions with H⁺-ATPase activity, one equilibrating at a density less than 1.11 g/cm³ and one equilibrating at a density of about 1.17g/cm³. Comparison with marker enzyme activities suggests that the upper fraction was enriched in tonoplast, and the dense fraction with plasma membrane. In addition to marker enzyme content, H⁺ transport in the H⁺-ATPase-containing fractions was further characterized with respect to pH dependence, cation and anion dependence, and uncouplers and inhibitors. H⁺ transport in all fractions was strongly dependent on the presence of halides but no specific stimulation by potassium or any other monovalent cation was found. Of the anions tested, malate and fumarate preferentially stimulated H⁺ transport in the tonoplast-enriched fraction. It is suggested that a Ca²⁺/H⁺ antiporter is present in all fractions. Only H⁺-ATPase in the plasma membrane-enriched fractions was sensitive to nystatin, an uncoupler, and to orthovanadate, an inhibitor. The tonoplast fraction was more sensitive to nitrate than the plasma membrane-enriched fraction, and all fractions showed some sensitivity to high concentrations of oligomycin. Oligomycin sensitivity was not due to the presence of mitochondria.     

微丝骨架的构成及其对花粉管极性生长的调控作用

微丝骨架是细胞骨架的重要组成部分,它由肌动蛋白和肌动蛋白结合蛋白组成,广泛存在于真核细胞中。近年来,大量研究表明植物花粉及花粉管中存在丰富的微丝骨架。目前,在微丝骨架作为信号转导途径的靶标参与对花粉管极性生长的调控、微丝骨架在花粉和花粉管中的分布及其在花粉管生长过程中与其他信号分子之间的相互作用等方面取得了一系列突破性进展。     

Interpretation of the electrical potential on the surface of plant roots

The electrical potential difference (p.d.) between two points on the surface of a plant root is shown to be a measure of the difference between the transmembrane potentials at those two points. More precisely, it is shown that axial differences in electrical potential on the surface of the root, or within several tenths of millimeters of it in the rhizosphere, are primarily a result of axial differences in p.d. across the plasmalemma of cells in the cortex, with an additional small effect from axial differences in p.d. across the plasmalemma of cells in the stele. This conclusion results from a model of the root as a three-conductor electrical transmission line. The model requires the solution of a set of differential equations, but simple algebraic approximations are found to apply over a range of model parameters derived from published data. Given the predictions of the model, it follows that, for many research purposes, microelectrode measurements could be replaced with measurements of p.d. on the surface of the root. Such measurements offer substantial advantages over microelectrode measurements: they are non-invasive; they do not include the potential difference across the tonoplast; they measure a spatial average of many cells, not just a single cell; and the measurement is physically robust. Surface potential measurements do not, however, measure trans-membrane potential at one point, but measure differences in trans-membrane potential.