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.

     

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     

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.     

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.

     

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.     

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.     

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: 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.     

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.     

MORPHO-HISTOGENIC STUDIES ON TENDRILS OF VITACEAE

The origin and development of the tendrils were studied in 16 species of the Vitaceae: Ampelopsis (7 sp.), Parthenocissus (4 sp.), Vitis (3 sp.), and Tetrastigma (1 sp.). Two types of arrangement of leaf and tendril occur: (a) two successive nodes have leaf-opposed tendrils alternating with each other, followed by a third node, with a leaf unopposed by the tendril; (b) all the nodes have leaf-opposed tendrils. The tendril, like a leaf, is a lateral product of the apical meristem of the shoot. A leaf opposite a tendril is initiated earlier than the tendril. Anticlinal and periclinal divisions in the second and/or third layer of the peripheral meristem of the shoot apex initiate the tendril. The procambium of the tendril first appears towards its abaxial side. Vascularization of the tendril is independent of the axillary bud of the next node below. The positional relationship of the nodal plate vis-à-vis the leaf-opposed tendril shows that the tendril and the leaf belong to the same node. Histological evidence does not show the uplifting of the tendril to the next node above during internodal differentiation. Ontogenetic and morphologic correlation and homology between the inflorescence and the tendril do not substantiate that the tendril in the Vitaceae is an organ sui generis. All available evidence indicates that the tendril is an extra-axillary lateral branch.     

Ultraviolet-B radiation causes tendril coiling in Pisum sativum

Low dose UV-B radiation (UV-B(BE,300) = 0.1 W m(-2)), but neither UV-A radiation, ozone and NaCl stress, nor wounding, caused tendril coiling in Pisum sativum. This coiling occurred with both attached and detached tendrils and can be used as a specific UV-B stress marker in pea.     

Physiological Studies on Pea Tendrils : XIV. Effects of Mechanical Perturbation, Light, and 2-Deoxy-d-Glucose on Callose Deposition and Tendril Coiling

When excised tendrils of pea (Pisum sativum L. cv Alaska) are mechanically perturbed there is an immediate and transient increase in callose deposition in the sieve cells. Mechanical perturbation (MP) results in a coiling response in light-grown tendrils and in dark-adapted tendrils, provided, in the latter case, that they receive adequate illumination within a limited period of time after MP. In nonperturbed tendrils the number of callose deposits decreases to some minimum with increasing time in the dark, and their ability to coil in the dark in response to MP diminishes with time in the dark. The transient increase of callose deposition due to MP, however, occurs whether or not tendrils are dark adapted, and whether they receive light or are retained in the dark after MP. This indicates that if callose is directly involved in tendril coiling, then it exerts its effect on the sensory perception of the mechanical stimulus. In the present investigation, there is never tendril coiling without the transient increase in callose, and the time after MP at which the peak of callose deposition occurs precedes the time of the peak amount of 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.     

Methyljasmonate and α-linolenic acid are potent inducers of tendril coiling

A coiling-inducing factor was isolated from tendrils of Bryonia dioica Jacq. and identified by infrared, ¹H-, ¹³C-nuclear magnetic resonance and mass spectrometry as -linolenic acid. When applied to detached tendrils, exogenous -linolenic acid, but not linoleic acid or oleic acid, induced tendril coiling. Further investigations showed that metabolites of -linolenic acid, jasmonic acid and, even more so, methyljasmonate, are highly effective inducers of tendril coiling in B. dioica. Methyljasmonate was most active when administered by air and, in atmospheric concentrations as low as 40–80 nM, induced a full free-coiling response with kinetics similar to mechanical stimulation. Even atmospheric levels as low as 4–5 nM methyljasmonate were still found to be significantly active. Methyljasmonate could be one of the endogenous chemical signals produced in mechanically stimulated parts of a tendril and, being highly volatile, act as a diffusible gaseous mediator spreading through the intracellular spaces to trigger free coiling of tendrils.Abbreviations EI-MS electron impact-mass spectrometry - HPLC high-performance liquid chromatography - IAA indole-3-acetic acid - NMR nuclear magnetic resonance - TFA trifluoroacetic acid We are indebted to the Deutsche Forschungsgemeinschaft, Bonn and the Fonds der Chemischen Industrie, Frankfurt (literature provision) for their support and to Dr. C. Brückner, Halle, for jasmonic-acid determinations.     

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.     

Thigmonastic Coiling of Tendrils of Passiflora quadrangularis is Not Caused by Lateral Redistribution of Auxin

The effect of thigmotropic stimuli on the distribution of exogenously applied indoleacetic acid in intact tendrils of Passiflora quadrangularis L. has been investigated. Tips of tendrils were dipped in solutions of IAA-2-¹⁴C for 12 h and subsequently stimulated by exposure to carbon dioxide or mechanically by rubbing. The content of ¹⁴C in dorsal and ventral halves was analysed before, during and after coiling. Our experiments failed to detect any difference from the initial dorsal: ventral ratio of ¹⁴C (44:56) as a possible consequence of stimulus or of coiling. This suggests that thigmotropic curvature is not dependent on lateral movement of auxin, supporting a theory of a built-in asymmetrical reaction of the tissues to equal amounts of auxin and CO₂, respectively.     

Physiological Studies on Pea Tendrils. III. ATPase Activity and Contractility Associated with Coiling

Extracts of the tendrils of Pisum sativum, Var. Alaska, exhibit adenosine triphosphatase activity which is inversely proportional to the amount the tendrils have coiled. The specific viscosity of the extract decreases when ATP is added. This evidence indicates a possible role of a contractile adenosine triphosphatase in coiling.     

Kinetics of thigmocurvature in two tendril-bearing climbers

Abstract. Growth of marked zones of watermelon and passion fruit tendrils was measured following brief thigmostimulation. In each case, curvature involved growth promotion on the non-contact side of the tendril. Substantial shrinkage also occurred on the contact side of the passion fruit tendril but very little shrinkage took place in the watermelon tendril. Curvature was detected 10s and 1 min after the completion of stimulation in watermelon and passion fruit, respectively. The rapidity of this curvature argues against the mediation of the response by plant growth substances.     

Physiological Studies on Pea Tendrils: VII. Evaluation of a Technique for the Asymmetrical Application of Ethylene

A technique has been devised for the asymmetrical application of ethylene to specific surfaces of plant tissue. 2-Chloroethylphosphonic acid (CEPA) was dissolved in a phosphate buffer (pH 6.5) containing Tween-20 or dimethylsulfoxide as adjuvant. Ethylene evolution from tendrils of Pisum sativum cv. Alaska was greater during coiling than when they were at rest; and via topical application to the ventral surface, CEPA was able to stimulate contact coiling. Within 1 day of application of CEPA, the tendrils showed symptoms of senescence. It is concluded that ethylene participates in the control of contact coiling stimulated by touch, and it is suggested that this control may be exerted via permeability changes in the membranes of the motor cells.     

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.