The action potential of Dionaea muscipula Ellis

The intention of this investigation was to acquire more concise information about the nature of the action potential of Dionaea muscipula Ellis and the different types of cells generating and conducting it. It is shown by microelectrode measurements that, besides the sensory cells, all the major tissues of the trap lobes are excitable, firing action potentials with pronounced after-hyperpolarizations. The action potentials are strictly dependent on Ca²⁺. Their peak depolarizations are shifted 25–27 mV in a positive direction after a tenfold increase in external Ca²⁺ concentration. Perfusions with 1 mM ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid (EGTA) or 1 mM LaCl₃ completely inhibit excitability. Magnesium ions only slightly affect the peak depolarizations but considerably prolong action potentials. Sodium azide and 2,4-dinitrophenol also abolish excitation, probably by reducing the intracellular ATP concentration. Furthermore, it is tested whether the sensory cells can be distinguished from the other cells of the trap by their electrical behaviour. The resting potentials of sensory cells (-161±7 mV) and mesophyll cells (-155±8 mV) are of the same magnitude. Changes in external ion concentrations affect resting and action potentials in both cell types in a similar way. Additional freeze-fracture studies of both cell types reveal similar numbers and distributions of intramembrane particles on the fracture faces of the plasma membrane, which is most likely the mechanosensor. These findings stress the view that the high mechanosensitivity of the sensory hair results from its anatomy and not from a specialized perception mechanism. It is proposed that trap closure is triggered by a rise in the cytoplasmic concentration of Ca²⁺ or a Ca²⁺-activated regulatory complex, which must exceed a threshold concentration. Since the Ca²⁺ influx during a single action potential does not suffice to reach this threshold, at least two stimulations of the trap are necessary to elicit movement.Abbreviations DNP 2,4-dinitrophenol - EF exoplasmic fracture face - EGTA ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid - E_m membrane potential - E_m,r resting potential - PF protoplasmic fracture face Dedicated to Professor Karl-Ernst Wohlfarth-Bottermann on the occasion of his 65th birthdayA preliminary report was presented at the 14th International Botanical Congress, Berlin, July 1987. This work is part of the dissertation (D5) of D.H.     

Evidence for the H+-co-transport of d-alanine by the digestive glands of Dionaea muscipula Ellis

Abstract. The net uptake of ¹⁴C-D-alanine by the adaxial surfaces of the trap lobes of Dionaea muscipula Ellis (Venus's flytrap) is positively correlated with the rate of H⁺ efflux from the trap lobes. Effectors that stimulate H⁺ efflux (secretion elicitor, fusicoccin and auxin) enhance D-alanine uptake whereas effectors that inhibit H⁺ efflux (diethylstilbestrol) retard uptake. It is proposed that amino acid uptake by the digestive glands of the trap lobes proceeds via a mechanism which mediates the co-transport of H⁺ ions and amino acids. A model is presented in which a positively charged ternary complex of carrier plus amino acid plus H⁺ is transported down an inwardly directed H⁺ electrochemical potential difference generated by the secretion-elicitor-provoked discharge of HCl into the trap cavity.     

The nature of the stimuli causing digestive juice secretion in Dionaea muscipula Ellis (venus's flytrap)

Summary An investigation into the stimuli of the secretory system in the carnivorous plant Dionaea muscipula is presented. Secretion of fluid and protein are both stimulated by various nitrogenous small molecules. These secretions are studied as a function of time. A new method is described for the collection of secreted juice. Significant differences are found between the quantities of fluid and protein produced in response to different stimuli. The results are discussed in comparison to the mammalian gastro-intestinal secretory systems.     

Action Potentials Evoked by Light in Traps of Dionaea muscipula Ellis

At low light intensities (less than 50 µmol m^–2s^–1) illumination evokes transient depolarization of membranepotential in mesophyll cells of the leaf-trap of Dionaea muscipulaEllis. Darkening causes hyperpolarization approximately symmetricto the response to illumination. The amplitude as well as therate of potential changes depend on light intensity. After exceedinga definite threshold (usually between 50 and 80 µmol m^–2s^–1)the depolarization plays the role of a generator potential andan all-or-none action potential (AP) is released. Switchinglight off in a depolarization phase of an AP does not changeits shape and the amplitude. When the light intensity is increasedto 80–150 µmol m^–2 s^–1 a single lightstimulus triggers two successive APs. The time interval betweenthe two APs decreases with increasing stimulus strength andreaches the minimum between 300 and 400 µmol m^–2s^–1. At higher light intensities the interval increasesagain, and finally only a single AP is triggered. It was shownthat the effect was evoked by light but not by temperature changeaccompanying illumination. An inhibitor of the photosyn-theticelectron transport chain, DCMU, blocked the generator potentialsmediating between light absorption and APs. Residual responsesto light stimuli in plants treated with DCMU had reverse polarityand strongly reduced the amplitudes. (Received September 16, 1997; Accepted January 16, 1998)     

食虫植物捕蝇草的花序轴组织培养

捕蝇草(ＤｉｏｎａｅａｍｕｓｃｉｐｕｌａＥｌｌｉｓ)是茅膏菜科捕蝇草属食虫植物。该属只有一种,原产美国南、北卡罗莱那州的海岸平原地区,新泽西和弗吉尼亚也有分布,主要生长在半稀树草原的沼泽、湿地周围[1]。捕蝇草植株姿态优雅,叶型奇特,是一种特化程度很高的食虫植...     

Polarity in mechanoreceptor cells of trigger hairs of Dionaea muscipula Ellis

Both the apical and the basal cell poles of the sensory cells in trigger hairs of Dionaea muscipula are structured identically. A complex of concentrically arranged endoplasmic reticulum cisternae occupies each of the poles. One to four vacuoles are enclosed within the central cisterna and contain polyphenols (deposits of “tannin”). Structural polarity, whether symmetric or asymmetric, as well as the occurrence of abundant endoplasmic reticulum and numerous mitochondria are characteristics of the perception cells of most animals and plants.     

Active movements in plants: Mechanism of trap closure by Dionaea muscipula Ellis

The Venus flytrap (Dionaea muscipula Ellis) captures insects with one of the most rapid movements in the plant kingdom. We investigated trap closure by mechanical and electrical stimuli using the novel charge-injection method and high-speed recording. We proposed a new hydroelastic curvature mechanism, which is based on the assumption that the lobes possess curvature elasticity and are composed of outer and inner hydraulic layers with different hydrostatic pressure. The open state of the trap contains high elastic energy accumulated due to the hydrostatic pressure difference between the hydraulic layers of the lobe. Stimuli open pores connecting the two layers, water rushes from one hydraulic layer to another, and the trap relaxes to the equilibrium configuration corresponding to the closed state. In this paper we derived equations describing this system based on elasticity Hamiltonian and found closing kinetics. The novel charge-injection stimulation method gives insight into mechanisms of the different steps of signal transduction and response in the plant kingdom.Key words: hydroelastic model, electrical signaling, electrophysiology, hydroelastic curvature, venus flytrap, ion channels, water channels     

On the mechanism of trap closure of Venus flytrap (Dionaea muscipula Ellis)

The rapid trap closure of Dionaea muscinula Ellis has been explained by either a loss of turgor pressure of the upper epidermis, which should thus become flexible, or by a sudden acid-induced wall loosening of the motor cells. According to our experiments both explanations are doubtful. Objections against the turgor mechanism come from the determination by extracellular measurements from the upper epidermis of action-potential amplitudes before and after trap closure. Neither time course nor amplitude of the action potentials are altered by trap closure. In contrast a rise in the apoplastic concentration of K⁺ or Na⁺, which are the only ions present in the trap in osmotically significant concentrations, from 1 to 10 mM reduces the action-potential amplitudes by 25% and 15%, respectively. Furthermore, after trap closure the upper epidermal cells retain a considerable cell sap osmolality of 0.41 mol·kg^-1 which equals that of the mesophyll cells as determined by incipient plasmolysis. A sudden cell-wall acidification causing movement is improbable since an acidification of the apoplast from pH 6 to pH 4 reduces action-potential amplitudes by 33% whereas the amplitudes measured extracellylarly from the mesophyll and lower epidermis remain unchanged by trap closure. In addition, buffering the apoplast at pH 6 does not prevent movement in traps which have been incised several times from the margin to the midrib to facilitate buffer diffusion into the mesophyll. Even an alkalinization of cell walls of plasmolysed leaf segments to pH 9 does not prevent considerable extensions of the mesophyll and subsequent movement of the specimens during deplasmolysis.These experiments make it very likely that the mesophyll cells are already extensible but are kept compressed in the open trap, thus developing tissue tension. The mechanism which prevents their extension as long as the trap is open can so far only be explained for traps which have been paralysed by a long-term incubation in 1 mM La³⁺. Leaf strips taken from stimulated, closed traps, comprising the lower epidermis and some mesophyll, prove to be highly extensible if they are stretched perpendicular to the midrib on a constant-load extensiometer. By contrast, strips taken from the lower side of paralysed traps are as rigid as those from the upper side of both stimulated and paralysed traps. From observations of semithin cross sections in a polarizing microscope, it is concluded that the extensibilities of these tissue strips are mainly determined by the cell walls of the upper epidermis plus a layer of adjacent mesophyll and by the lower epidermis, respectively, since these are the only cell walls with a preferential microfibril orientation in the direction of the applied stress.Abbreviations E _m membrane potential - E _s surface potential - Mes 2-(N-morpholino)ethanesulfonic acid - Tris 2-amino-2(hydroxymethyl)-1,3-propanediol     

Biomimetic robotic Venus flytrap (Dionaea muscipula Ellis) made with ionic polymer metal composites

The work described in this paper is a novel design of a robotic Venus flytrap (VFT) (Dionaea muscipula Ellis) by means of ionic polymeric metal composite (IPMC) artificial muscles as distributed nanosensors and nanoactuators. Rapid muscular movements in carnivorous plants, such as VFT, which are triggered by antenna-like sensors (trigger hair), present a golden key to study distributed biomolecular motors. Carnivorous plants, such as VFT, possess built-in intelligence (trigger hairs), as a strategy to capture prey, that can be turned on in a controlled manner. In the case of the VFT, the prey that is lured by the sweet nectar in the VFT pair of jaw-like lobes has to flip and move the trigger hairs, which are colorless, bristle-like and pointed. The dynamically moved trigger hairs then electro-elastically send an electric signal to the internal ions in the lobe to migrate outwardly for the jaw-like lobes to close rapidly to capture the prey. The manner in which the VFT lobes bend inward to capture the prey shows a remarkable similarity with typical IPMCs bending in an electric field. Furthermore, the mechano-electrical sensing characteristics of IPMCs also show a remarkable resemblance to mechano-electrical trigger hairs on the lobes of the VFT. The reader is referred to a number of papers in connection with sensing and actuation of IPMCs in particular. Thus, one can integrate IPMC lobes with a common electrode in the middle of one end of the lobes to act like a spine and use IPMC bristles as trigger finger to sense the intrusion of a fly or insect to send a sensing signal to a solid state relay which then triggers the actuation circuit of the IPMC lobes to rapidly bend toward each other and close. The two lobes, which form the trap, are attached to the midrib common electrode which is conveniently termed the spine. The upper surface of each lobe is dished, and spaced along the free margins of the lobes with some 15-20 prong-like teeth. These are tough and pointed, and are inclined at an inward angle so that when the trap is sprung shut they will interlock. We have been experimenting with the VFT closing of its jaw-like lobes that close in about 0.3 s and have gained a lot of knowledge to report on the ionic and electrical mechanisms involved in the operation of such intelligent distributed biomolecular motors.     

Energetics and forces of the Dionaea muscipula trap closing

The Venus flytrap is the most famous carnivorous plant. The electrical stimulus between a midrib and a lobe closes the Venus flytrap upper leaf in 0.3 s without mechanical stimulation of trigger hairs. Here we present results for direct measurements of the closing force of the trap of Dionaea muscipula Ellis after mechanical or electrical stimulation of the trap using the piezoelectric thin film or Fuji Prescale indicating sensor film. The closing force was 0.14 N and the corresponding pressure between rims of two lobes was 38 kPa. We evaluated theoretically using the Hydroelastic Curvature Model and compared with experimental data velocity, acceleration and kinetic energy from the time dependencies of distance between rims of lobes during the trap closing. The Charge Stimulation Method was used for trap electrostimulation between the midrib and lobes. From the dependence of voltage between two Ag/AgCl electrodes in the midrib and one of the lobes, we estimated electrical charge, current, resistance, electrical energy and electrical power dependencies on time during electrostimulation of the trap.     

捕蝇草的组织培养和快速繁殖

1　植物名称　捕蝇草 (Ｄｉｏｎａｅａｍｕｓｃｉｐｕｌａ)。2　材料类别　子叶、叶片。3　培养条件　种子培养基 :(1 )ＭＳ 6 ＢＡ0 .1ｍｇ·Ｌ-1(单位下同 ) ＮＡＡ 0 .1。愈伤组织与不定芽诱导培养基 :(2 )ＭＳ 2 ,4 Ｄ0 .5 6 ＢＡ 0 .5 ;(3)ＭＳ 6 ＢＡ 0 .5 ;     

Digestive Secretion of Dionaea muscipula (Venus's Flytrap)

The digestive fluid of Dionaea muscipula has been studied with respect to its protein content as a function of time after entrapment of protein material and some enzymes of the secretion. Maximum secretion of enzyme occurs within the first 3 days of the digestive cycle and protein reaches its maximum at 4 days. Phosphatase, proteinase, nuclease and amylase have been observed in the secretion. The enzymes have acid pH optima and the proteinase has a molecular weight of about 40,000.     

A cysteine endopeptidase ("dionain") is involved in the digestive fluid of Dionaea muscipula (Venus's fly-trap)

The carnivorous plant Dionaea muscipula (Venus's flytrap) secretes proteinases into the digestive fluid to digest prey proteins. In this study, we obtained evidence that the digestive fluid contains a cysteine endopeptidase, presumably belonging to the papain family, through inhibitor studies and partial amino acid sequencing of the major SDS-PAGE band protein. The name "dionain" is proposed for the enzyme.     

On the mechanism underlying photosynthetic limitation upon trigger hair irritation in the carnivorous plant Venus flytrap (Dionaea muscipula Ellis)

Mechanical stimulation of trigger hairs on the adaxial surface of the trap of Dionaea muscipula leads to the generation of action potentials and to rapid leaf movement. After rapid closure secures the prey, the struggle against the trigger hairs results in generation of further action potentials which inhibit photosynthesis. A detailed analysis of chlorophyll a fluorescence kinetics and gas exchange measurements in response to generation of action potentials in irritated D. muscipula traps was used to determine the 'site effect' of the electrical signal-induced inhibition of photosynthesis. Irritation of trigger hairs and subsequent generation of action potentials resulted in a decrease in the effective photochemical quantum yield of photosystem II (Φ(PSII)) and the rate of net photosynthesis (A(N)). During the first seconds of irritation, increased excitation pressure in photosystem II (PSII) was the major contributor to the decreased Φ(PSII). Within ～1?min, non-photochemical quenching (NPQ) released the excitation pressure at PSII. Measurements of the fast chlorophyll a fluorescence transient (O-J-I-P) revealed a direct impact of action potentials on the charge separation-recombination reactions in PSII, although the effect seems to be small rather than substantial. All the data presented here indicate that the main primary target of the electrical signal-induced inhibition of photosynthesis is the dark reaction, whereas the inhibition of electron transport is only a consequence of reduced carboxylation efficiency. In addition, the study also provides valuable data confirming the hypothesis that chlorophyll a fluorescence is under electrochemical control.     

食虫植物捕蝇草叶片的组织培养

1　植物名称　捕蝇草 (Ｄｉｏｎａｅａｍｕｓｃｉｐｕｌａ)。2　材料类别　叶片。3　培养条件　以 1 /2ＭＳ为基本培养基。 ( 1 )诱导培养基 :1 /2ＭＳ + 6 ＢＡ 2ｍｇ·Ｌ- 1 (单位下同 ) ;( 2 )继代和增殖培养基 :1 /2ＭＳ + 6 ＢＡ 2 +ＮＡＡ 0 .1 ;( 3)生根培养基 :1 /2ＭＳ +ＮＡＡ 0 .1 +肉汁 1 0 %。以上培养基均加 0 .9%琼脂 ,3%蔗糖 ,ｐＨ 5 .8,培养温度 2 5～ 2 7℃ ,光照度 2 0 0 0ｌｘ ,每天光照 1 0～1 2ｈ。4　生长与分化情况4.1　不定芽诱导　取 1年生的捕蝇草植株嫩叶为外殖体 ,用自来水冲洗干净后无菌水泡 1 0ｍｉｎ ,吸干…     

Strategy of nitrogen acquisition and utilization by carnivorous Dionaea muscipula

Plant carnivory represents an exceptional means to acquire N. Snap traps of Dionaea muscipula serve two functions, and provide both N and photosynthate. Using ¹³C/¹⁵N-labelled insect powder, we performed feeding experiments with Dionaea plants that differed in physiological state and N status (spring vs. autumn plants). We measured the effects of ¹⁵N uptake on light-saturated photosynthesis (A _max), dark respiration (R _D) and growth. Depending on N status, insect capture briefly altered the dynamics of R _D/A _max, reflecting high energy demand during insect digestion and nutrient uptake, followed by enhanced photosynthesis and growth. Organic N acquired from insect prey was immediately redistributed, in order to support swift renewal of traps and thereby enhance probability of prey capture. Respiratory costs associated with permanent maintenance of the photosynthetic machinery were thereby minimized. Dionaea’s strategy of N utilization is commensurate with the random capture of large prey, occasionally transferring a high load of organic nutrients to the plant. Our results suggest that physiological adaptations to unpredictable resource availability are essential for Dionaea’s success with regards to a carnivorous life style.     

Isolation and structures of diomuscinone and diomuscipulone from Dionaea muscipula

From the fresh leaves and roots of Dionaea muscipula, two new substances (diomuscinone and diomuscipulone) have been isolated together with the known naphthoquinone plumbagin. The structures of the new compounds have been elucidated on the basis of their spectral data coupled with some chemical evidence.     

The dynamics of H+ efflux from the trap lobes of Dionaea muscipula Ellis (Venus's flytrap)

Abstract. H⁺ efflux from the trap lobes of Dionaea muscipula Ellis (Venus's flytrap) was measured in vitro. FC, IAA, and 2,4-D markedly increase the rate of H⁺ efflux within minutes of their addition to the incubation medium whereas ABA and DES cause the rate to decrease. Consequently, the H⁺ efflux mechanism of Dionaea is considered to be similar to the H⁺ extrusion pumps of other higher plants in this respect. However, the H⁺ extrusion mechanism of Dionaea may be unusual in that long-term exposure of the trap lobes to known secretion elicitors— bactopeptone, NH₄⁺, Na ⁺, urea, thiourea, glycine or xanthine—also causes a large increase in the steady-state rate of H⁺ efflux from the trap lobes. Since the observed H⁺ effluxes primarily correspond to the adaxial surface of the trap lobes and show similar time- and secretion elicitor-dependencies to the responses seen in situ, it appears that the H⁺ effluxes measured in vitro bear a direct relationship to those observed in the intact, actively secreting plant. Three of the secretion elicitors that were tested— K⁺, NH₄⁺, and urea—have rapid effects on the rate of H⁺ extrusion in addition to their long-term effects. K⁺ and NH₄⁺ cause a rapid acceleration of H⁺ efflux whereas urea causes a rapid deceleration or, at high external concentrations, reversal of the net flux. The effect of K⁺ is inferred to result from K⁺ -H⁺ exchange between the tissue and bathing medium. Studies with structural analogues of NH₄⁺ and urea and inhibitors of the assimilation of reduced nitrogen suggest that the effects of NH₄⁺ and urea result from the pH-perturbing consequences of their metabolism subsequent to their absorption. These effects are considered to be auxiliary to the elicitation of secretion. It is proposed that H⁺ efflux from the trap lobes is mediated by a K⁺-H⁺ exchange mechanism, the activity of which is modified by long-term exposure to secretion elicitors and/or short-term exposure to factors which alter the availability of endogenous H⁺ ions.     

Effects of ion channel inhibitors on cold- and electrically-induced action potentials in Dionaea muscipula

Glass microelectrodes were inserted into Dionaea muscipula (Venus flytrap) lobes and the action potentials (APs) were recorded in response to a sudden temperature drop or a direct current (DC) application. The effect of potassium channel inhibitor, tetraethylammonium ion, was the lengthening of the depolarization phase of AP. APs were also affected by the anion channel inhibitor, anthracene-9-carboxylic acid, that made them slower and smaller. Neomycin, which disturbs inositol triphosphate-dependent Ca²⁺ release, caused the visible inhibition of AP, too. Ruthenium red, which blocks cyclic ADP-ribose-dependent Ca²⁺ release, totally inhibited DC-triggered APs and induced the decrease in the amplitudes of cold-evoked APs. Lanthanum ions significantly inhibited both cold- and DC-induced membrane potential changes. It was concluded that during excitation Dionaea muscipula relied upon the calcium influxes from both the extra- and intracellular compartments.