The Mechanical Significance of Clasping Leaf Sheaths in Grasses: Evidence from Two Cultivars of Avena sativa

The elastic (Young's) modulus and flexural rigidity of internodeswith and without their clasping leaf sheaths were determinedfor culms from two cultivars (‘Astro’) and (‘Garry’)of Avena sativa L. Data indicate that early in the developmentof culms, leaf sheaths can have a higher elastic modulus thanthe internodes they envelope, and by virtue of their location,leaf sheaths contribute significantly to the flexural rigidity(hence, resistance to bending) of internodal segments. As culmsmature, the elastic modulus of leaf sheath and internodal tissuesreach parity. However, because of the acropetal pattern by whichnew internodes are produced by shoot apices, sheaths continueto provide mechanical support to distal internodes, particularlythe peduncle. Data for the two cultivars indicate that the elasticmodulus and flexural rigidity of culms can vary significantlywithin the species. Comparisons between the flexural rigidityof the two cultures and the resistance of stems to lodging indicatethat flexural rigidity is not significant to lodging. The engineeringprinciples relevant to the mechanical advantages conferred byclasping leaf sheaths are discussed within the context of grassshoot morphology. Biomechanics, leaf sheath, Avena, elastic modulus     

The biophysical basis of elongation growth in internodes of deepwater rice

Partial submergence induces rapid internodal elongation in deepwater rice (Oryza sativa L., cv Habiganj Aman II). We measured in vivo extensibility, tissue tension, hydraulic conductance and osmotic potential in the region of cell elongation in the uppermost internode. The in vivo extensibility of the internode, measured by stretching of living tissue with a custom-made constant stress extensiometer, rose rapidly following submergence of the plant. Both the elastic (E_el) and plastic (E_pl) extensibility increased when growth of the internode was induced. The submerged internode displayed tissue tension (elastic outward bending of longitudinally split internode sections); in air-grown control internodes, no such bending occurred. The hydraulic conductance, estimated from the kinetics of tissue shrinkage in 0.5 molar mannitol and subsequent swelling in distilled water, was not changed by submergence. The osmotic potential, measured with a dew-point hygrometer using frozen-thawed tissue, was only 18% less negative in the submerged internode than in the air-grown control. This indicates that osmoregulation takes place in rapidly elongating rice internodes. We suggest that the rapid expansion of the newly formed internodal cells of submerged plants is controlled by the yielding properties (E_pl) of the cell walls. Experiments with excised stem sections indicate that gibberellin is involved in increasing the E_pl of the elongating cell walls.     

The Internode of Sambucus racemosa L. Originates from a Single Cell Layer

The internodes of Sambucus racemosa L. shoot apices originatefrom a single layer of cells in which mononucleate mother tannincells are present. The mother tannin cells increase in lengthand differentiate together with the internodal growth. Sambucus racemosa L., shoot apices, internode, mother tannin cells, tannin coenocytes     

Modes of Mechanical Failure of Hollow, Septate Stems

Three general modes of mechanical failure were observed whenhollow, septate stem segments ofArundinaria técta(Poaceae)were axially compressed and caused to fail: (1) rupture of tissuesat the opposing ends of the nodal transverse diaphragm parallelto the plane of stem flexure; (2) localized catastrophic transverseinvagination of internodes (Brazier buckling); and (3) longitudinalrupture of internodal walls along the convex surface of stemflexure attended by nodal tissue shearing. The frequencies ofoccurrence of these three modes were not equivalent among the100 stem segments examined; stem failure was dominated by nodaltissue shearing (i.e. 67% of the nodes failed by shearing; 52%of the internodes longitudinally invaginated; 27% of the internodesfailed in Brazier buckling; 21% of the internodes failed byrupturing). Computer simulations of axially compressed stemsegments consisting of one node and one internode composed ofnon-linearly elastic, anisotropic materials (tissues) successfullypredicted observed strain patterns and revealed that the pathwayto stem failure contains a bifurcation point below which deformationpatterns coalesce on a single configuration, and above whichstem failure by Brazier buckling or nodal tissue shearing areessentially mutually exclusive responses to excessive stem flexure.The patterns of actual and simulated stem flexure were consistentwith the hypothesis that nodes store strain energy as stemsflex, and release this energy to restore stem shape when bendingforces abate (i.e. nodes operate as spring-like joints). However,strain energy ‘sinks’ identified by simulationswere also located in bending internodal walls which can storeelastic strain energy to do work. Nodal diaphragms and internodalwalls likely comprise a complex and global rather than a simpleand local elastic system for the recovery of stem shape.Copyright1998 Annals of Botany Company Stem flexure; mechanical failure; stem nodes; internodes; plants; biomechanics; finite element analyses; Brazier buckling.     

Antagonistic and Synergistic Interactions between Ancymidol and Gibberellins in Shoot Growth of Cucumber (Cucumis sativus L. )

Effects of the plant growth retardant, ancymidol, on the growthand morphology of the shoot system of cucumber (Cucumis sativusL. ) were investigated. Ancymidol inhibited stem elongation,reducing both number and length of internodes. Reduction inleaf area, attributable to a reduction in both cell size andnumber, was accompanied by an increase in chlorophyll per unitarea. The retardant decreased apical dominance and delayed anthesis.Gibberellic acid fully reversed ancymidol-induced inhibitionof stem elongation, internode length and production, and leafexpansion. GA_4/7 and ancymidol gave a synergistic promotionof stem elongation by increasing elongation of younger internodesand increasing internode production. Synergistic promotion ofpetiole elongation by this combination was also observed. Ancymidol,applied 7 d previously either to the shoot or root, severelyreduced the level of gibberellin-like activity in bleeding sapcollected from decapitated plants.     

Electron-probe Microanalysis Studies of Silica Distribution in Barley (Hordeum sativum L.)

Silicon occurence has been investigated by means of epidermalpeels, cryostat, and ultrathin sections of the internode, nodes,leaves, inflorescence bracts, and caryopsis of Hordeum sativumL. (cultivar Deba Abed) using the electron probe microanalyser.Analyses were made on growth stages during ear emergence andat maturity. The results indicate that silicon is present inthe internode with the highest concentration associated withthe opaline deposits. Detectable quantities are also found inthe outer tangential walls of the long cells, in the walls ofstomata, the sclerenchyma, and all vascular bundle regions.In mature upper internodes, silicifiation is confined to theupper third region, but this limit extends closer to the basalmeristem with increasing age of internode. The nodes have agreater concentration in the radial than in outer tangentialwalls. Heavy deposits are found in the leaves but with considerablevariation between blade and sheath, abaxial and adaxial surfaces,and the leaf position. The flag leaf contained the highest accumulations. In the inflorescence bracts (lemma and palea), silicon is detectableonly in the abaxial epidermis and hypodermis. Awns are alsoheavily silicified with the highest concentrations in the sclerenchymaand trichomes.     

Effects of CCC and GA on internodal development of barley

Summary The effect of applying (2-chloroethyl)trimethylammonium chloride (CCC) or gibberellic acid (GA) as foliar sprays on internodal development of barley was studied. CCC applied to the whole plant at main tiller leaf stages 1, 2 or 3 decreased shoot elongation, and prevented elongation of internode 6 (internode 5 subtended the head). CCC at all stages delayed senescence of the lower leaves. CCC sprayed at all 6 leaf stages and GA sprayed at main tiller leaf stages 1, 2, 3 or 4 reduced plant height at maturity. GA treatment at leaf stages 2, 3 or 4 initially stimulated internodal elongation; elongation of later developed internodes was inhibited resulting in shorter plants at maturity. Only the treatment with GA at leaf stages 5 and 6 resulted in increased plant height.     

STRUCTURE OF GRAVITY-SENSITIVE SHEATH AND INTERNODAL PULVINI IN GRASS SHOOTS

The location and some morphological, anatomical, and functional aspects of the gravity-sensitive pulvini of a selected number of grass shoots are examined. There are two distinct gravity-sensitive regions near the nodal regions of Gramineae. One, the leaf sheath pulvinus, is located at the base of the sheathing leaf bases, and is characteristic of the subfamily Festucoideae. The other, the internodal pulvinus, is located at the base of the internode, a little above the nodal joint. Most members of the Panicoideae possess internodal pulvini, in addition to more or less developed leaf sheath pulvini. Three members of the Oryzoideae examined possess leaf sheath pulvini only, while Phragmites australis (Arundinoideae) possesses both leaf sheath and internodal pulvini. Leaf sheath pulvini of some grasses develop hairs, cork-silica cell pairs or stomatal apparatuses over the epidermis while many others are devoid of any such idioblasts. Both the leaf sheath and internodal pulvini of all grasses examined preferentially exclude, or accumulate very little silica, whereas the regions of the shoot immediately above and below the pulvini in these same grasses accumulate large quantities of silica. Pulvini remain unsilicified or poorly silicified throughout their life and even after several days following geotropic bending. Pulvini are also distinguished from the regions above and below them by the lack of lignin in the bundle cap cells. Lignin is found only in the xylem vascular tissue, and this consists of annular and helical vessel elements only. The bundle cap cells are rich in pectin and are described as collenchymatous. All pulvini possess specialized zones of cells containing starch statoliths. In response to horizontal displacement of the shoots, the lower side of the pulvini grows by cell elongation only. The collenchymatous cells stretch in a manner that results in alternately thin and thick regions of cell wall.     

Determination of Microgram Quantities of Stevioside from Leaves of Stevia rebaudiana Bert. by Two-Dimensional Thin Layer Chromatography

A method is described for the determination of stevioside fromleaves of Stevia rebaudiana Bert. Separation upon non-activatedsilica gel impregnated with boric acid in a two-dimensionalchromatographic system followed by clution and colorimetricassay is reported. Recuperation from chromoplates is over 95%.The observed ratio of the optical density of glucose: steviosidematches that expected theoretically so that glucose may be employedas a standard where pure stevioside is not available. Effects of the plant growth retardant, ancymidol, on the growthand morphology of the shoot system of cucumber (Cucumis sativusL. ) were investigated. Ancymidol inhibited stem elongation,reducing both number and length of internodes. Reduction inleaf area, attributable to a reduction in both cell size andnumber, was accompanied by an increase in chlorophyll per unitarea. The retardant decreased apical dominance and delayed anthesis.Gibberellic acid fully reversed ancymidol-induced inhibitionof stem elongation, internode length and production, and leafexpansion. GA_4/7 and ancymidol gave a synergistic promotionof stem elongation by increasing elongation of younger internodesand increasing internode production. Synergistic promotion ofpetiole elongation by this combination was also observed. Ancymidol,applied 7 d previously either to the shoot or root, severelyreduced the level of gibberellin-like activity in bleeding sapcollected from decapitated plants.     

Origin of Nodes and Internodes in Plant Shoots. II. Models of Node and Internode Origin from One Layer of Cells

In the promeristem of some plant apices there are two layers'ofcells giving rise to the plant shoot. The first layer on thesurface forms the protoderm, and developing from it the epidermis;it is easily distinguished when there are phenolic compoundsexclusively in that layer, e.g. Crataegus, Vitis. The secondlayer may divide periclinally (a process here called differentiatingdivision) producing, successively, node and internode mothercell layers. Mitoses can start from the central part and continueradially or, more commonly, from leaf primordia and continuecentripetally. The number of periclinal divisions in that layerdetermines how many nodes and internodes the shoot will have.Differences in the number of cell divisions in these two motherlayers lead to the formation of long internodes and short nodes Nodes, internodes, idioblasts, phenolics, development, Crataegus, Vitis     

Silicification of developing internodes in the perennial scouring rush (Equisetum hyemale var. affine)

An electron microprobe (EMP) analysis of silica (SiO₂) deposition in the epidermis of developing internodes of the perennial scouring rush (Equisetum hyemale var. affine) indicates that SiO₂ is first detected in the stomatal apparatus beginning with internode 3, then the epidermal papillae (internode 8), and finally in radial cell walls of the long epidermal cells (internode 10). This process is initiated in the intercalary growth regions at the bases of the elongating internodes. The deposition of SiO₂ in long epidermal cell walls occurs after internodal extension has ceased and should therefore be considered as one of the final stages in internodal differentiation that involves strengthening the cellulosic framework of the cell wall. EMP measurements indicate that SiO₂ in stomata is equivalent to 30% of a pure SiO₂ standard and that SiO₂ in the radial walls of long epidermal cells averages twice that measured on the tangential walls of these same cells. This study supports the view that silicification plays a major role in strengthening the developing perennial scouring rush internodal system and that regulation of this process in this and other species of Equisetum, whose SiO₂ deposition patterns are markedly different, deserves further study.     

Expansins and Internodal Growth of Deepwater Rice

The distribution and activity of the cell wall-loosening protein expansin is correlated with internodal growth in deepwater rice (Oryza sativa L.). Acid-induced extension of native cell walls and reconstituted extension of boiled cell walls were confined to the growing region of the internode, i.e. to the intercalary meristem (IM) and the elongation zone. Immunolocalization by tissue printing and immunoblot analysis, using antibody against cucumber expansin 29 as a probe, confirmed that rice expansin occurred primarily in the IM and elongation zone. Rice expansin was localized mainly around the vascular bundles at the base of the IM and along the inner epidermal cell layer surrounding the internodal cavity. Submergence greatly promoted the growth of rice internodes, and cell walls of submerged internodes extended much more in response to acidification than did the cell walls of air-grown internodes. Susceptibility of cell walls to added expansin was also increased in submerged internodes, and analysis by immunoblotting showed that cell walls of submerged internodes contained more expansin than did cell walls of air-grown internodes. Based on these data, we propose that expansin is involved in mediating rapid internodal elongation in submerged deepwater rice internodes.     

Changes in total nitrogen, soluble protein, and peroxidases in the expanding leaf zone of eastern cottonwood

Nitrogen content and soluble protein and anodal peroxidase banding in acrylamide gel changed with leaf and internode development in the expanding leaf zone of eastern cottonwood (Populus deltoides Bartr.). Nitrogen per unit leaf area was high near the apex and decreased to a constant value at the sixth node below it. Soluble protein banding was qualitatively similar for leaves and internodes in this zone, but anodal peroxidases differed between leaves and internodes. The major leaf peroxidase band was absent from the second leaf below the apex but present in the fourth and sixth leaves; its appearance and intensification seemed to parallel the development of photosynthetic activity. The major internode peroxidase band was present in the apex, second, fourth, and sixth internodes, and intensified during internodal development. It is suggested that these two “isoenzymes” may have different functions in vivo.     

Dynamics of the Elongation of Internodes in Maize (Zea mays L.): Analysis of Phases of Elongation and their Relationships to Phytomer Development

The kinetics of elongation of individual internodes of maizestems were studied under field conditions. Thermal time coursesof internode length were recorded using non-destructive methods,based on direct measurement of X-ray photographs or on indirectestimates using heights of leaf collars. These data were complementedby serial dissections of maize stems, and by precise observationof the process of sheath emergence, to specify its role in thekinetics of internode elongation. The kinetics of elongationwere found to be composed of four phases. The rate of elongationrises exponentially during phase I, and then increases sharplyduring a short period (phase II), which is followed by a majorperiod of constant growth rate (phase III) and a shorter periodin which the rate declines (phase IV). During phase I, elongationappears to be integrated at the level of the whole apical cone.From phase II onwards, elongation becomes determined at thelevel of the phytomer. The emergence of the sheath attachedto the internode appears to be a possible trigger for the transitionbetween phase I and phase II, and it may also be involved invariation in final length among phytomers. Copyright 2000 Annalsof Botany Company Zea mays L., internode, elongation, modelling, dynamics, X-rays, collar, phytomer, stem, thermal time, phasic development     

Particles Associated with the Outer Epidermal Wall in Internodes of Deepwater Rice

Partial submergence induces rapid internodal growth in deepwaterrice (Oryza saliva L., cv. Habiganj Aman II). The infrastructureof the cell wall/plasmalemma interface of air-grown and submergedinternodes was investigated in the region where cell elongationtakes place. In submerged internodes, electron-dense particlesof about 100 nm diameter were found. These particles were detectableonly at the thick outer wall of the outer epidermis but notat the inner walls. In air-grown control plants, no such granuleswere visible. We suggest that these particles are related tothe process of cell wall growth. The wall weight per unit lengthwas 75% lower in the submerged internode as compared to thatof the air-grown control. This indicates that secondary wallformation is suppressed during submergence of the plant Oryza saliva, deepwater rice, intemodal growth, electron-dense particles     

Levels of {beta}-Glucan and Lignin in Elongating Internodes of Deepwater Rice

Partial submergence or treatment with either ethylene or gibberellicacid (GA₃ induces rapid growth in deepwater rice (Oryza sativaL.). We correlated the synthesis of two cell wall componentswith two phases of internodal elongation, namely (13,14)-ß-glucanformation with cell elongation and lignification with differentiationof the secondary cell wall and cessation of growth. The contentof ß-glucan was highest in the zone of cell elongationin internodes of air-grown plants and plants that were inducedto grow rapidly by submergence. In the intercalary meristemand in the differentiation zone of the internode, ß-glucanlevels were ca. 70% lower than in the zone of cell elongation.The outer cell layers, enriched in epidermis, contained moreß-glucan in submerged, rapidly growing internodesthan in air-grown, control internodes. The ß-glucancontent of the inner, parenchymal tissue was unaffected or slightlylowered by submergence. The epidermis appears to be the growth-limitingstructure of rapidly growing rice internodes. We hypothesizethat elevated levels of ß-glucan contribute to elongationgrowth by increasing the extensibility of the cell wall. Lignificationwas monitored by measuring the content of lignin and the activitiesof two enzymes of the lignin biosynthetic pathway, coniferylalcohol dehydrogenase (CAD) and phenylalanine ammonia-lyase(PAL), in growing and non-growing regions of the internode.Using submerged whole plants and GA₃-treated excised stem segments,we showed that lignin content and CAD activity were up to sixfoldlower in newly formed internodal tissue of rapidly growing ricethan in slowly growing tissue. No differences were observedin parts of the internode that had been formed prior to inductionof growth. PAL activity was reduced throughout the internodeof submerged plants. We conclude that lignification is one ofthe processes that is suppressed to permit rapid growth. ¹ This work was supported by the National Science Foundationthrough grants No. DCB-8718873 and DCB-9103747 and by the Departmentof Energy through grant No. DE-FGO2-90ER20021. M.S. was therecipient of a fellowship from the Max Kade Foundation.     

Complementary patterns of stiffness in stem and leaf sheaths of Triticale

The variation in the stiffness of stem and leaf sheaths along the shoot axis of Triticale (Triticosecale W., cv. Jago) was examined, using an ultrasonic method, at two stages of development, (i) at the stage of high stem mechanical instability when upper internodes are forming (heading), and (ii) at milk maturity when development of strengthening tissues is completed (three weeks after anthesis). The squared velocity of low-frequency longitudinal pulse waves was used as a measure of the specific modulus of elasticity, averaged over the whole cross section of the structures and related to the unit density of the material. Structural material of varying effective stiffness was found to be utilized along leaf sheaths with a pattern complementary to that in growing stems. The stiffness increased basipetally along leaf sheaths in the direction of increasing flexibility of internodes. Maximum values of the specific modulus of elasticity in particular leaf sheaths were enhanced acropetally, ensuring the stronger mechanical protection of those meristematic zones which were actively elongating and were located at the upper internodes. The stiffest material present in leaf sheaths covered only those stem sections which could be the most critical for plant safety during stem elongation. This characteristic and very regular pattern of stiffness alteration along the shoot axis was structurally determined as it remained similar after air drying the specimens. It is concluded that adaptation of cereals to withstand environmental loads is realized not only on morphological and anatomical levels but is also reflected in a specific heterogeneity in the material properties of the cell walls which support the plant.This research was supported by the Ministry of Agriculture, Poland (Project CPBR 10.1 C 29).     

Changes in Endogenous Gibberellins and the Metabolism of [H]GA(4) after Geostimulation in Shoots of the Oat Plant (Avena sativa)

The recovery from “lodging,” or bending over, by shoots of 42-day-old Avena sativa plants is controlled primarily by a negatively geotropic differential growth of the lower halves of the p-1 node-pulvinus and the base of the p-1 internode, relative to the upper halves. Although geostimulation causes a significant reduction in p-1 internode length, dry matter accumulation in the p-1 node-pulvinus is increased, apparently at the expense of the sheath. Recovery to an angle of 30° is associated with changes in endogenous gibberellin-like substances (GAs), and in differential metabolism of applied [³H]GA₄ (1.4 Curie per millimole). Although geostimulation depressed total GAs (relative to upright plant parts) to 0.40 and 0.13 for node-pulvini and sheaths, respectively, it increased them 2-fold for internodes. Within the plant part geostimulation increased GAs (relative to upper halves) 29- and 7-fold in lower halves of node-pulvini and internodes, respectively, but reduced GAs to 0.3 in lower halves of sheaths. At age 42 days a GA_4/7-like (nonpolar) substance predominates, with lesser amounts of a GA₃-like (polar) substance. Native GAs of Avena include GA₃, GA₄, and GA₇. Geostimulation enhanced the ratio of nonpolar to polar GAs for both halves of internodes, but tended to depress it for sheaths and nodepulvini.     

Morphological Responses and Molecular Modifications in Tomato Plants after Mechanical Stimulation

Study of the growth responses of Lycopersicon escu-lentum (Mill.cv. VFN8) to mechanical stimulation applied to a single younginternode showed a rapid and sharp decrease in stem elongationand an inhibition of elongation of several internodes, indicativeof information transmission in the plant. A new tomato cDNApartial clone encoding calmodulin was isolated and used to studythe time course of the gene induction in response to the rubbingtreatment. Northern blot analysis showed a maximum accumulationof calmodulin mRNA 2 h after mechanical stimulation, not onlyin the rubbed internode, but also in upper and lower internodesand in young leaves. Treatment of the plant with calcium andEGTA showed the involvement of calcium and, in particular, intracellularcalcium in calmodulin gene expression and cellular response. (Received February 10, 1997; Accepted July 11, 1997)     

The Effect of Light Quality on Shoot Extension Growth in Three Species of Grasses

The effect of light quality on the extension growth of vegetativeshoots and on the final size of their leaves was investigatedin plants of Lolium multiflorum, Sporobolus indicus and Paspalumdilatatum. Three experimental approaches were used, (a) redor far-red end-of-day irradiations of sunlight-grown plants,(b) different red/far-red ratios of white light in a growthroom and (c) sunlight enrichment with radiation of differentred/far-red ratios or with different amounts of far-red lightduring the photoperiod. Plants treated with end-of-day far-redor low red/far-red ratios throughout the photoperiod developedlonger leaves and, as a result, longer shoots. This effect wasmore marked in leaf sheaths than in blades. Tiller extensionand leaf sheath length increased with the amount of far-redadded to sunlight in a simple hyperbolic relationship. Theseresults show that vegetative grass shoots respond to light qualityin a way similar to internodes of dicotyledonous plants. Lolium multiflorum Lam., Sporobolus indicus (L.), Paspalum dilatatum (Poir.), leaf growth, tiller growth, photomorphogenesis