Physiological studies on pea tendrils. XII. Effects of temperature on contact coiling and uncoiling

When excised tendrils of pea ( Pisum sativum L. cv. Alaska 2B) are mechanically perturbed and allowed to coil at different constant temperatures, the greatest amount of coiling occurs between 27°C and 33°C. Coiling of tendrils continues for about 2 h after mechanical perturbation at which time uncoiling usually begins. The temperature at which the rate of uncoiling is greatest appears to be influenced, at least in part, by the temperature at which the tendrils coiled. For example, when tendrils coil at 20°C their rate of uncoiling at 20°C is less than if they had coiled at 23°C. Estimated activation energies for the uncoiling process are greater than for coiling, with 35 J/mol × s and 97 J/mol × s for uncoiling in the temperature ranges 18°C to 23°C and 10°C to 18°C, respectively. The estimated activation energy for coiling is 5.4 J/mol × s. It is suggested that the process of tendril uncoiling, as well as tendril coiling, might be an active, energy requiring process.
When mechanically perturbed tendrils are placed in the cold (5°C) they do not coil. But this interruption of the coiling process with a cold (5°C) treatment, either immediately after mechanical perturbation or after coiling has begun, does not prevent coiling from continuing after tendrils are again given a more suitable temperature. It is concluded that the cessation of coiling during the cold period may be due to a slowdown in metabolism. It is suggested that there may be a factor which is responsible for the motor response and which is retained during the cold treatment.     

Sink to Source Transition in Tendrils of a Semileafless Mutant, Pisum sativum cv Curly

Sink to source transition parallels loss of thigmotropic capacity in tendrils of a semileafless mutant, Pisum sativum cv Curly. Macroscopic tendril development is subdivided based on thigmotropic capacity. Stage I is the elongation stage and, although the rate of photosynthesis is similar to that of stage II and III tendrils, dark respiration rates are higher in stage I. During stage II, tendrils are thigmotropic and act as a sink. Even though stage II tendrils have CO₂ exchange characteristics similar to those of stage III tendrils, which are coiled, our fluorescein, ¹⁴C-partitioning, and ¹¹C-translocation experiments suggest that stage I and II tendrils do not export carbon. Only stage III tendrils act as sources of newly fixed carbon. Export from them is blocked by cold, heat girdling of the petiole, or anoxia treatment of the tendrils. A late stage II tendril complex, in which coiling is occurring, may be exporting photoassimilates; however, this phenomenon can be attributed to the fact that the pea leaf is a compound structure and there may be one or more stage III tendrils, no longer thigmotropic, within the tendril complex. Photosynthetic maturity in pea tendrils occurs at stage III and is characterized by the ability of these tendrils to export photoassimilates.     

Tendril Coiling in Grapevine: Jasmonates and a New Role for GABA?

Grapevine (Vitis vinifera L., Vitaceae) belongs to the genus Vitis, and is characterized as a vine due to the presence of tendrils, which are located opposite to leaves. Tendrils are thigmo-responsive organs, able to carry out delicate mechanosensory responses upon touch and related stimuli. These organs are an adaptation of the plant to climb with the help of support to higher places and finally remain at a position with favorable light quality. In previous studies on Bryonia dioica (Cucurbitaceae), phytohormones of the jasmonate class were identified as the endogenous hormone signals to initiate coiling of the tendrils. Strikingly, this is still the only example for jasmonate-induced tendril coiling. In grapevine, three compounds (12-oxo-phytodienoic acid, jasmonic acid (JA), and JA isoleucine conjugate) of the jasmonate class were found at higher concentrations in non-coiled tendrils when compared with coiled ones. Upon treatment with phytohormones, we could confirm the activity of jasmonates on tendril coiling in grapevine. However, not jasmonates but a non-proteinogenic amino acid, γ-aminobutyric acid (GABA), was detected to accumulate in grapevine tendrils at significantly higher levels than in all other tissues, independent of their coiling status. For GABA we detected a significant, transient positive effect on tendril coiling. Use of a GABA synthesis blocker, 3-mercaptopropionic acid, caused reduced GABA- but not JA-induced coiling scores. No additive effect of JA plus GABA was detectable on the tendrils’ coiling score. Thus, for grapevine, our data demonstrate a strong activity of jasmonates in tendril coiling induction even without mechanical stimuli and, furthermore, the data also suggest that GABA can independently promote tendril coiling.

     

Reciprocal influence of ethylene and gibberellins on response-gene expression in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

A cortical band of fiber cells originate de novo in tendrils of redvine [Brunnichia ovata (Walt.) Shiners] when these convert from straight, supple young filaments to stiffened coiled structures in response to touch stimulation. We have analyzed the cell walls of these fibers by in situ localization techniques to determine their composition and possible role(s) in the coiling process. The fiber cell wall consists of a primary cell wall and two lignified secondary wall layers (S₁ and S₂) and a less lignified gelatinous (G) layer proximal to the plasmalemma. Compositionally, the fibers are sharply distinct from surrounding parenchyma as determined by antibody and affinity probes. The fiber cell walls are highly enriched in cellulose, callose and xylan but contain no homogalacturonan, either esterified or de-esterified. Rhamnogalacturonan-I (RG-I) epitopes are not detected in the S layers, although they are in both the gelatinous layer and primary wall, indicating a further restriction of RG-I in the fiber cells. Lignin is concentrated in the secondary wall layers of the fiber and the compound middle lamellae/primary cell wall but is absent from the gelatinous layer. Our observations indicate that these fibers play a central role in tendril function, not only in stabilizing its final shape after coiling but also generating the tensile strength responsible for the coiling. This theory is further substantiated by the absence of gelatinous layers in the fibers of the rare tendrils that fail to coil. These data indicate that gelatinous-type fibers are responsible for the coiling of redvine tendrils and a number of other tendrils and vines.     

Physiological studies on pea tendrils

When tendrils which have been dark adapted for 3 days are mechanically stimulated, they will only coil appreciably if they are irradiated with light. A spectral response curve suggests that this is a blue light effect. Red or far red light do not modulate this response, but a high intensity flash of blue white light given before the actinic blue light blocks coiling for 30 min. The dark-adapted tendril, which has not been previously rubbed, has the ability to store the tactile sensory information for about 60 min, but loses it if it is not irradiated within 120 min after mechanical perturbation.     

Physiological Studies on Pea Tendrils: VI. The Characteristics of Sensory Perception and Transduction

Tendrils may be said to possess a sense of touch, and the direction and amplitude of the coiling response can be used to define the characteristics of this sense. These characteristics are, first, that the tendril will coil only in response to ventral mechanical stimulation, that this coiling can be inhibited by subsequent dorsally presented stimulation, and that dorsal stimulation alone causes no coiling. This phenomenon seems to be due to some asymmetry in the response system. Second, the nature of the response is always determined by the location (i.e., dorsal or ventral) of the last stimulation the tendril experienced. Third, the ability of dorsally presented stimulation to reverse ventrally stimulated coiling is gradually lost. Complete escape from reversibility is attained if the interval between ventral and dorsal stimulation reaches 9 minutes. Fourth, the magnitude of response is determined by both the number and the frequency of the stimuli. Both ventrally stimulated coiling and dorsally stimulated inhibition of coiling can be temporarily stopped by a 9-minute cold break at 10 C, given immediately after stimulation. As soon as the tendrils are restored to room temperature, they proceed to respond to the stimulus.     

Interaktionen von Auxin und Äthylen bei der thigmotropen Bewegung der Ranken von Cucumis sativus

Summary Coiling of intact or excised cucumber (Cucumis sativus) tendrils can be induced by IAA or ethylene. The velocity of coiling in different regions of the tendrils correlates with the capacity for auxin-stimulated ethylene synthesis. Ethylene (Ethephon) induces an increase in membrane permeability of tendrils, and as a result the efflux of substances previously taken up (glucose) is stimulated. It is assumed that this may contribute to the contraction of the ventral side of the tendril. The excretion of glucose after ethylene treatment can be reduced by Ca²⁺, and calcium also inhibits coiling of tendrils following incubation in ethephon solution. Auxin stimulated ethylene synthesis in the ventral half of the tendril is several times higher than in the dorsal half and it is hypothesized that this may be a cause for the different reactions of the two sides of a tendril following a mechanical stimulus.

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Abkürzungen: IAA=Indol-3-essigsäure; ABA=Abscisinsäure     

Touch- and Methyl Jasmonate-induced Lignification in Tendrils of Bryonia dioica Jacq.

The touch-induced free coiling response of Bryonia dioica tendrils is accompanied by the differentiation of supporting tissue at the ventral side of the organ, becoming the inner (concave) side of the coiled tendril. As part of this process, the Bianconi plate, a continuous sclerenchyma sheath stretching along the ventral face of the five bicollateral vascular bundles, becomes strongly lignified. During this reaction, the extractable activity of phenylalanine ammonia-lyase in the tendrils increases four- to five-fold, while the amount of PAL, as demonstrated by immunoblotting, remains unchanged. This touch-induced process can also be elicited by airborne application of methyl jasmonate. The PAL inhibitor, 2-aminoindan-2-phosphonic acid (AIP) completely inhibits both the touch- and methyl jasmonate-induced deposition of lignin-like material in the Bianconi plate, but has no effect on coiling. From these results, it can be concluded that the cessation of growth at the ventral side of a free-coiling tendril is not due to induced lignification but rather, lignification seems to serve the function of increasing the mechanical strength of the coiled tendril.     

Physiological Studies on Pea Tendrils. II. The Role of Light and ATP in Contact Coiling

Excised pea (Pisum sativum L.) tendrils incubated in the light coil more than those incubated in the dark. This light effect, which displays spectral responses characteristic of chlorophyll-mediated mechanisms, is increased by at least 8 hours of prior dark incubation of plants from which the tendrils were derived. Considerable evidence indicates a major role of ATP in coiling. For example, inhibitors of ATP production decrease contact coiling. Exogenous ATP increases curvature in the dark, whereas exogenous adenosine, AMP and ADP are practically without effect. The ATP effect can be reversed by the addition of sucrose to the bathing solution. Tendrils of plants placed in the dark overnight have lower ATP levels than those held in the light. One half hour after stimulation, the endogenous ATP level of tendrils on plants kept in the light decreased fourfold. In the same period, the endogenous inorganic phosphate level increased markedly, indicating high adenosine triphosphatase activity.     

Evolutionary,genetic, environmental and hormonal-induced plasticity in the fate of organs arising from axillary meristems in Passiflora spp.

Tendrils can be found in different plant species. In legumes such as pea, tendrils are modified leaves produced by the vegetative meristem but in the grape vine, a same meristem is used to either form a tendril or an inflorescence. Passiflora species originated in ecosystems in which there is dense vegetation and competition for light. Thus climbing on other plants in order to reach regions with higher light using tendrils is an adaptive advantage. In Passiflora species, after a juvenile phase, every leaf has a subtending vegetative meristem, and a separate meristem that forms both flowers and a tendril. Thus, flowers are formed once a tendril is formed yet whether or not this flower will reach bloom depends on the environment. For example, in Passiflora edulis flowers do not develop under shaded conditions, so that tendrils are needed to bring the plant to positions were flowers can develop. This separate meristem generally forms a single tendril in different Passiflora species yet the number and position of flowers formed from the same meristem diverges among species. Here we display the variation among species as well as variation within a single species, P. edulis. We also show that the number of flowers within a specific genotype can be modulated by applying Cytokinins. Finally, this separate meristem is capable of transforming into a leaf-producing meristem under specific environmental conditions. Thus, behind what appears to be a species-specific rigid program regarding the fate of this meristem, our study helps to reveal a plasticity normally restrained by genetic, hormonal and environmental constraints.     

Physiological Studies on Pea Tendrils VIII. The Relationship of Circumnutation to Contact Coiling. — With a Description of a Laboratory Intervalometer Using Integrated Digital Circuits

The average rate of rotation of circumnutating tendrils of Pisum sativum L. ev. Alaska, was 1.57 ± 0.29 mm/min. 53% of the tendrils rotated clockwise and 47% counterclockwise. Circumnutation is apparently dependent on the maintenance of sufficient turgor as it stopped when either the roots or all the shoot appendages except the terminal tendril were excised, but resumed when the aerial wounds were covered with petroleum jelly. Both circumnutation and contact coiling were similarly retarded when the plant was cut in the middle of the top inter-node, or by the use of either juvenile or senescent organs. As the tendril circumnutated rapidly during the sweeping portion of its circuit, it was capable of coiling at only about 57% of the rate of which it could coil if stimulated during the relatively slow moving turn, Conversely, when the tendril was mechanically stimulated to coil, its rate of circumnutation decreased markedly and remained retarded as long as the tendril continued to coil. On the basis of these observations, it is concluded that contact coiling does not seem to be simply a modified form of circumnutation, but the two modes of movement might be related through a negative feedback system.     

Actin and Myosin in pea tendrils

We demonstrate here the presence of actin and myosin in pea (Pisum sativum L.) tendrils. The molecular weight of tendril actin is 43,000, the same as rabbit skeletal muscle actin. The native molecular weight of tendril myosin is about 440,000. Tendril myosin is composed of two heavy chains of molecular weight approximately 165,000 and four (two pairs) light chains of 17,000 and 15,000. At high ionic strength, the ATPase activity of pea tendril myosin is activated by K⁺-EDTA and Ca²⁺ and is inhibited by Mg²⁺. At low ionic strength, the Mg²⁺-ATPase activity of pea tendril myosin is activated by rabbit skeletal muscle F-actin. Superprecipitation occurred after incubation at room temperature when ATP was added to the crude actomyosin extract. It is suggested that the interaction of actin and myosin may play a role in the coiling movement of pea tendril.     

Gelatinous fibers are widespread in coiling tendrils and twining vines

Although the coiling of tendrils and the twining of vines has been investigated since Darwin's time, a full understanding of the mechanism(s) of this coiling and twining ability has not yet been obtained. In a previous study (Planta 225: 485-498), gelatinous (G) fibers in tendrils of redvine occurred concomitantly with the ability to coil, strongly indicating their role in the coiling process. In this study, tendrils and twining vines of a number of species were examined using microscopic and immunocytochemical techniques to determine if a similar presence and distribution of these fibers exists in other plant species. Tendrils that coiled in many different directions had a cylinder of cortical G fibers, similar to redvine. However, tendrils that coiled only in a single direction had gelatinous fibers only along the inner surface of the coil. In tendrils with adhesive tips, the gelatinous fibers occurred in the central/core region of the tendril. Coiling occurred later in development in these tendrils, after the adhesive pad had attached. In twining stems, G fibers were not observed during the rapid circumnutation stage, but were found at later stages when the vine's position was fixed, generally one or two nodes below the node still circumnutating. The number and extent of fiber development correlated roughly with the amount of torsion required for the vine to ascend a support. In contrast, species that use adventitious roots for climbing or were trailing/scrambling-type vines did not have G fibers. These data strongly support the concept that coiling and twining in vines is caused by the presence of G fibers.     

Physiological studies on pea tendrils. V. Membrane changes and water movement associated with contact coiling

The coiling of excised pea tendrils in response to mechanical stimulation is accompanied by an increased efflux from their cut bases of electrolytes and label from previously absorbed ¹⁴C-acetate and ¹⁴C-sucrose. The major excreted cation is H⁺; H⁺ loss is potentiated by pretreatment with benzoic acid, which also leaves the tendrils during coiling.

Label from previously absorbed tritiated water is excreted during coiling, mainly from the ventral side of the tendril, which contracts in the initial phase of coiling. Such label does not pass from the ventral to the dorsal side. Similarities between this and other rapidly moving systems in plants are surveyed and a hypothesis to explain turgor movements is advanced.

     

Physiological studies on pea tendrils. XIII. Respiration is necessary for contact coiling

When 9 day old light grown pea ( Pisum sativum L. cv. Alaska 2B) plants are irrigated for 4–6 days with 50 μ M 4-chloro-5-(dimethylamino)-2-(α, α, α-trifluoro- m -totyl)-3(2H)-pyridazinone, designated San-6706, the new leaves and tendrils grow morphologically normal, but with neither chlorophyll nor the ability to carry out photosynthesis. Excised tendrils from these plants coil in response to rubbing as well as those from water irrigated controls. Tendrils from San-6706 irrigated plants, which have been dark adapted for 2 or 3 days, proceed to coil when illuminated, just as do those from water irrigated plants. Rubbing of dark adapted tendrils results in an increased respiration rate over the first hour or two, when most of the coiling response occurs. Inhibitor studies indicate that blockage of oxidative phosphorylation, but not of the alternative, cyanide-insensitive, path of respiration, results in a failure of tendrils to coil in response to mechanical perturbation. It is concluded that the normal path of respiration, perhaps via ATP production, may be necessary for thigmosensory perception leading to contact coiling in pea tendrils.     

Self-discrimination in the tendrils of the vine Cayratia japonica is mediated by physiological connection

Although self-discrimination has been well documented, especially in animals, self-discrimination in plants has been identified in only a few cases, such as self-incompatibility in flowers and root discrimination. Here, we report a new form of self-discrimination in plants: discrimination by vine tendrils. We found that tendrils of the perennial vine Cayratia japonica were more likely to coil around neighbouring non-self plants than neighbouring self plants in both experimental and natural settings. The higher level of coiling around a physiologically severed self plant compared with that around a physiologically connected self plant suggested that self-discrimination was mediated by physiological coordination between the tendril and the touched plant as reported for self-discrimination in roots. The results highlight the importance of self-discrimination for plant competition not only underground, but also above-ground.     

Development of the axillary bud complex in Echinocystis lobata (Cucurbitaceae): interpreting the cucurbitaceous tendril

In the Cucurbitaceae, the tendrils, coiling organs used for climbing and mechanical support, are part of an axillary bud complex (ABC). Although the morphological nature of tendrils and the branching pattern of the ABC in the Cucurbitaceae have been much studied, their homology remains unresolved, with hypothesized candidates being the leaf, flower, stem, or stem-leaf combination. We used Echinocystis lobata as a model to study the early ontogeny of the ABC with epi-illumination microscopy and serial resin sections. The ABC produces four structures (proximal to distal, relative to the subtending leaf) as the result of two successive subdivisions: an inflorescence of staminate flowers, a solitary pistillate flower, a lateral bud, and a tendril. The first separates the tendril primordium from the continuation of the ABC, and the second separates the staminate inflorescence and the ABC. The pistillate flower apparently forms between the staminate inflorescence and the lateral bud. Because there is no subtending leaf during these subdivisions and the first lateral appendages in the resulting primordia arise in the same plane, we conclude that the tendril and other organs formed by the ABC are lateral branches of equal morphological value. This study is the basis for continuing comparative and functional morphological studies.     

Touch-Sensitive NADH Oxidase Activity of Pea and Cucumber Tendrils and Soybean Hypocotyl Sections

The cell surface reduced nicotinamide adenine dinucleotide (NADH) oxidase activity of soybean stems and of pea and cucumber tendrils responded to touch with a several-fold increase in activity. The increase in NADH oxidase persisted for 20 min or longer, and further touch stimulation during this period did not alter activity. With soybean sections, the specific activities in response to touch approximated those achieved maximally by auxin. Where the NADH oxidase was fully stimulated by 2,4-d, the NADH oxidase failed to respond further to touch. The findings indicate that the NADH oxidase of the plant cell surface is involved in the growth response to touch and in tendril coiling.     

LEAF DEVELOPMENT IN DOXANTHA UNGUIS-CATI (BIGNONIACEAE)

Leaf structure in Doxantha unguis-cati is polymorphic. The usual mature compound leaf is composed of two lanceolate leaflets and a terminal tripartite spine-tendril. Leaf primordia are initiated simultaneously in pairs on opposite flanks of the shoot apical meristem by periclinal cell divisions in the third subsurface layer of the peripheral flank meristem. Two leaflet primordia are the first lateral appendages of the compound leaf. Initiation of these leaflet primordia occurs on the adaxial side of a compound leaf primordium 63–70 μm long. Lamina formation is initiated at the base of a leaflet primordium 70–90 μm long and continues acropetally. Mesophyll differentiation occurs in later stages of development of leaflets. The second pair of lateral appendages of the leaf primordium differentiate as prongs of the tendril. Initiation of the second pair of lateral appendages occurs on the adaxial side of a primordium approximately 168 μm long. Acropetal procambialization and vacuolation of cells extend to the apex of tendrils about 112 μm long, restricting the tendril meristem to the adaxial side of the primordium and resulting in curvature of the tendril. The tendril meristem is gradually limited to a more basipetal position as elongation of apical cells continues. Initiatory divisions and early ontogenetic stages of leaflets and tendrils are similar. Their ontogeny differs when the lateral primordia are approximately 70 μm long. Marginal and submarginal initials differentiate within leaflets but not in tendrils. Apical growth of tendrils ceases very early in ontogeny as compared with leaflets.