HELIOTROPIC LEAF MOVEMENT RESPONSE TO H+/ATPase activation,H+/ATPase inhibition,AND K+ CHANNEL INHIBITION IN VIVO

The pulvinus, located at the base of soybean leaflets, is both the light perception and motor organ for heliotropic leaf movements. Our objective was to investigate the role of plasma membrane H⁺/ATPase and TEA-sensitive K⁺ channels in mediating pulvinar response to light. The plasma membrane H+/ATPase activator, fusicoccin, plasma membrane H⁺/ATPase inhibitors, vanadate and erythrosin-B, and the K⁺ channel blocker TEA were introduced to the intact pulvinus through the transpiration stream. The pulvinus was illuminated by a vertical light beam of 1,400 μmol m^-2 s^-1 to stimulate leaf movement. Leaf orientation was measured every 5 min for 60 min of illumination. All compounds tested inhibited pulvinar bending, but concentration and uptake time required for inhibition varied: 12.5 μM fusicoccin reduced leaf movement after 3 hr uptake, 2 mM vanadate reduced leaf movement after 6 hr uptake, 100 μM erythrosin-B reduced leaf movement after 3 hr uptake, and 15 mM TEA reduced leaf movement after 6 hr uptake. In all cases final leaf angle was reduced by higher concentrations and/or increased time for uptake of the chemical into the pulvinus. Results support the hypothesis that the proximal mechanism of heliotropic movement is similar to that of nyctinastic movements.     

Migration of Calcium and Its Role in the Regulation of Seismonasty in the Motor Cell of Mimosa pudica L

Volume and conformational changes of the contractile tannin vacuoles of the abaxial motor cells of the primary pulvinus of Mimosa pudica L. parallel the seismonastic leaf movement. Since such changes in cells and organelles of animal systems are often regulated by calcium, we studied Ca²⁺ movement in the motor cells and tissue. By fixation with Lillie's neutral buffered formalin, followed by staining with alizarin red sulfate (ARS), calcium was localized in the tannin vacuoles of the motor cells of the primary pulvinus. After treatment with ethylenediaminetetraacetate, 8-hydroxyquinoline, and several other calcium-complexing or extracting agents, the color reaction due to alizarin red sulfonate was no longer present. By using an analytical method, it was shown that the effluent from stimulated pulvini has significantly more Ca²⁺ than that from unstimulated controls. Ten millimolar LaCl₃ inhibits recovery of the tannin vacuole in vivo in 10 mm CaCl₂ or in distilled water. Quantitative data obtained by microspectrophotometry demonstrated calcium migration during the bending movement of the primary pulvinus. In the adaxial motor cells a small amount of calcium migrates from the tannin vacuole, and calcium on the cell wall moves to the central vacuole. In the abaxial half, a large amount of calcium from the tannin vacuole moves to the central vacuole of the motor cell. It is probable that the calcium binds to the microfibrillar contents of the central vacuole. These observations support the contention that Ca²⁺ migrates between the surface of the tannin vacuole and the inside of the central vacuole. The recovery and maintenance of the tannin vacuole in the spherical form may play a role in maintaining turgor in the motor cells of the abaxial half of the primary pulvinus of Mimosa.     

Circadian rhythm leaf movement of Phaseolus vulgaris and the role of calcium ions

Legume plants, due to their distinctive botanical characteristics, such as leaf movements, physiological characteristics, such as nitrogen fixation, and their abilities to endure environmental stresses, have important roles in sustainable pastures development. Leaf movement of legume plants is turgor regulated and osmotically active fluxes of ions between extensor and flexor of pulvinus cause this movement. To determine the role of calcium ions in circadian leaf movements of Phaseolus vulgaris L., a radiotracer technique experiment using ⁴⁵Ca ions were employed. Measurements were taken during circadian leaf movements, and samples were taken from different parts of the leaflet. The ⁴⁵Ca β-particle activity reduced from leaflet base pulvinus to leaf tip. The pulvinus had the highest activity, while the leaf tip had the lowest. By increase of the ratio of ⁴⁵Ca β-particle activity within flexor to extensor (Fl/Ex) the midrib-petiole angle, as an indicator of leaf movement, increased linearly during circadian leaf movement (r = 0.86). The ⁴⁵Ca β-particle activity of Flex/Ext ratio reduced linearly (r = −0.88) toward midnight. In conclusion, it was found that calcium ions accumulation is opposite to the fluxes of osmatically active ions and water movement. Calcium ions accumulate at less negative water potential side of the pulivnus.Key words: pulvinus, extensor, flexor, leaf movement, rhythm, circadian, calcium, Phaseolus vulgaris, radioactive     

Actin dynamics mediates the changes of calcium level during the pulvinus movement of Mimosa pudica

The bending movement of the pulvinus of Mimosa pudica is caused by a rapid change in volume of the abaxial motor cells, in response to various environmental stimuli. We investigated the relationship between the actin cytoskeleton and changes in the level of calcium during rapid contractile movement of the motor cells that was induced by electrical stimulation. The bending of the pulvinus was retarded by treatments with actin-affecting reagents and calcium channel inhibitors. The actin filaments in the motor cells were fragmented in response to electrical stimulation. Further investigations were performed using protoplasts from the motor cells of M. pudica pulvini. Calcium-channel inhibitors and EGTA had an inhibitory effect on contractile movement of the protoplasts. The level of calcium increased and became concentrated in the tannin vacuole after electrical stimulation. Ruthenium Red inhibited the increase in the level of calcium in the tannin vacuole and the contractile movement of the protoplasts. However, treatment with latrunculin A abolished the inhibitory effect of Ruthenium Red. Phalloidin inhibited the contractile movement and the increase in the level of calcium in the protoplasts. Our study demonstrates that depolymerization of the actin cytoskeleton in pulvinus motor cells in response to electrical signals results in increased levels of calcium.Key words: actin, calcium, pulvinus movement, the tannin vacuole, Mimosa pudica     

Pulvinus activity,leaf movement and leaf water-use efficiency of bush bean (<Emphasis Type="Italic">Phaseplus vulgaris</Emphasis> L.) in a hot environment

Pulvinus activity of Phaseolus species in response to environmental stimuli plays an essential role in heliotropic leaf movement. The aims of this study were to monitor the continuous daily pulvinus movement and pulvinus temperature, and to evaluate the effects of leaf movements, on a hot day, on instantaneous leaf water-use efficiency (WUE_i), leaf gas exchange, and leaf temperature. Potted plants of Phaseolus vulgaris L. var. Provider were grown in Chicot sandy loam soil under well-watered conditions in a greenhouse. When the second trifoliate leaf was completely extended, one plant was selected to measure pulvinus movement using a beta-ray gauging (BRG) meter with a point source of thallium-204 (²⁰⁴Tl). Leaf gas exchange measurements took place on similar leaflets of three plants at an air temperature interval of 33–42°C by a steady-state LI-6200 photosynthesis system. A copper-constantan thermocouple was used to monitor pulvinus temperature. Pulvinus bending followed the daily diurnal rhythm. Significant correlations were found between the leaf-incident angle and the stomatal conductance (R ² = 0.54; P < 0.01), and photosynthesis rate (R ² = 0.84; P < 0.01). With a reduction in leaf-incidence angle and increase in air temperature, WUE_i was reduced. During the measurements, leaf temperature remained below air temperature and was a significant function of air temperature (r = 0.92; P < 0.01). In conclusion, pulvinus bending followed both light intensity and air temperature and influenced leaf gas exchange.     

Age-dependent changes of the ion content and the circadian leaf movement period in thePhaseolus pulvinus

The circadian movement of the lamina of primary leaves ofPhaseolus coccineus L. depends on circadian changes of the K⁺, Cl^- and (depending on the Cl^- availability) malate content in the swelling and shrinking motor cells of the laminar pulvinus. After sowing in soil, the laminar pulvinus develops within about 26 days. When the leaves emerge from the soil (about 6 days after sowing) and the pulvinus starts with the diurnal movement (about 9 days after sowing) the pulvinar dimensions are about half of those of the mature pulvinus. The anatomical structure, however, is basically the same as in the developed pulvinus. In soil-grown plants, the K⁺, Cl^- and malate content as well as the period length of the circadian leaf movement rhythm change in the developing pulvinus. In the embryo of the dry seed, the Cl^- content is low (0.03 mmol g^-1 DW), the K⁺ content, however, 22-fold higher than the Cl^- content. When the leaves emerge from the soil, the pulvinar K⁺ and Cl^- content is the same as in the whole embryo of the dry seed. In the developing pulvinus the K⁺ content increases by a factor of 2 and the Cl^- content by a factor of 41 in the mature pulvinus. The pulvinar malate content increases between the 6th and 10th day from about 40 to 180Μmol g^-1 DW, then decreases until the 17th day and remains thereafter on a low level (around 80 Μmol g^-1 DW). These results indicate that the Cl^- availability increases in the developing pulvinus with age. It explains furthermore why in young leaves malate was found as counterion to K⁺ in the osmotic leaf movement motor, in older ones, however, Cl^-. The circadian leaf movement starts 9 days after sowing. The period length decreases during the development of the pulvinus from 31.3 to 28.6 h in leaves of intact soil-grown plants. In leaves which were cut from the plants and immersed with their petioles in distilled water, the age dependent decrease of the period length is also found. However, the period lengths are shorter by more than 1 h than in the leaves of intact plants. The increasing Cl^- availability in the developing pulvinus does not seem to be the cause for the age dependent shortening of the period length, because the period length in 22 days old Cl^- deprived pulvini is the same as in 22 days old pulvini with a high Cl^- content.     

The Trans-Acting Short Interfering RNA3 Pathway and NO APICAL MERISTEM Antagonistically Regulate Leaf Margin Development and Lateral Organ Separation,as Revealed by Analysis of an argonaute7/lobed leaflet1 Mutant in Medicago truncatula

Leaf shape elaboration and organ separation are critical for plant morphogenesis. We characterized the developmental roles of LOBED LEAFLET1 by analyzing a recessive mutant in the model legume Medicago truncatula. An ortholog of Arabidopsis thaliana ARGONAUTE7 (AGO7), Mt-AGO7/LOBED LEAFLET1, is required for the biogenesis of a trans-acting short interfering RNA (ta-siRNA) to negatively regulate the expression of AUXIN RESPONSE FACTORs in M. truncatula. Loss of function in AGO7 results in pleiotropic phenotypes in different organs. The prominent phenotype of the ago7 mutant is lobed leaf margins and more widely spaced lateral organs, suggesting that the trans-acting siRNA3 (TAS3) pathway negatively regulates the formation of boundaries and the separation of lateral organs in M. truncatula. Genetic interaction analysis with the smooth leaf margin1 (slm1) mutant revealed that leaf margin formation is cooperatively regulated by the auxin/SLM1 (ortholog of Arabidopsis PIN-FORMED1) module, which influences the initiation of leaf margin teeth, and the TAS3 ta-siRNA pathway, which determines the degree of margin indentation. Further investigations showed that the TAS3 ta-siRNA pathway and NO APICAL MERISTEM (ortholog of Arabidopsis CUP-SHAPED COTYLEDON) antagonistically regulate both leaf margin development and lateral organ separation, and the regulation is partially dependent on the auxin/SLM1 module.     

Oscillations of the Membrane Potential of Pulvinar Motor Cells In Situ in Relation to Leaflet Movements of Desmodium motorium

The ultradian rhythmic movement of the lateral leaflets of Desmodiummotorium is accompanied by rhythmic changes of the extra- andintracellular electrical potentials in the pulvinus, which aremeasured in situ in the pulvinus against the bathing solutionof the petiole. Extra- and intracellular potentials oscillatewith 180'b0 phase difference to each other, as shown by simultaneousmeasurements of both types of potentials in the abaxial partof the pulvinus. Light-induced changes of these potentials movein opposite directions. The in situ membrane potential of themotor cells of the pulvinus was calculated from the differencebetween the extra- and intracellular potentials. It was foundto oscillate between –136 and –36 mV, in phase withthe intracellular and inverse to the extracellular potential.The phase relationship between the leaflet movement rhythm andthe in situ membrane potential rhythm was as follows: downwardmovement is preceded and accompanied by a strong depolarization,upward movement by hyperpolarization. Our results suggest that membrane depolarization in pulvinarmotor cells of Desmodium motorium drives and controls potassiumefflux and hyperpolarization potassium influx via potassiumchannels. Key words: Desmodium pulvinus, leaf movement, pulvinar motor cells, electrical potential     

Action Spectrum of Light Pulse-Induced Membrane Depolarization in Pulvinar Motor Cells of Phaseolus

In addition to circadian changes in the membrane potential andleaf movement, light applied to the pulvinus causes changesin both the membrane potential and the pulvinar movement inPhaseolus vulgaris L. Even after a short pulse of light, a transientdepolarization of the membrane occurs and leaf movement is observed.Decreases of turgor pressure of the motor cells are always precededby the depolarization. The direction of the leaf movement canbe explained by the decrease of turgor pressure in the motorcells on the irradiated side of the pulvinus. Using the OkazakiLarge Spectrograph at the National Institute for Basic Biology,we determined the action spectrum of the membrane depolarizationinduced by light pulses (30 s) in motor cells of Phaseolus.The pulvinus was left exposed to air during measurement of themembrane potential with microelectrodes. The action spectrumobtained was in the range of 300 to 730 nm. It had the highestpeak at 460 nm with lower peaks at 380 nm and 420 nm. Almostno sensitivity was observed at wavelengths shorter than 360nm and longer than 520 nm. Red and far-red light had no effecton the depolarization of the motor cell. The features of theaction spectrum are almost the same as those of the Blue-Typeresponse in plants. (Received January 9, 1997; Accepted February 14, 1997)     

Compartmentation of soluble carbohydrates,of starch and of malate in motor organs (pulvini) and other parts of Phaseolus coccineus L. leaves

Quantitative histochemistry was used to investigate the tissue-specific compartmentation of soluble carbohydrates (sucrose, glucose, fructose), starch and malate in the laminar pulvinus, leaf blade and petiole of Phaselous coccineus L. at day and night positions of diurnal leaf movement. Total carbohydrate levels measured in a series of cross sections along individual pulvini of 24-d-old plants showed only small differences between the day and night positions of the respective leaf. In contrast, the level of malate changed during diurnal leaf movement, especially in the central part of a pulvinus. The levels of glucose and fructose in the pulvinus increased towards the transition zones between the pulvinus and lamina, and pulvinus and petiole, and this trend was even more pronounced for starch. By contrast, sucrose levels were highest in the pulvinus proper. The transverse compartmentation of metabolites was studied in distinct, approx. 0.5-mm-thick tissue slices from the central part of a pulvinus. These were dissected further into up to 14 distinct subsamples (bundle, bundle sheath, motor tissues, flanks). Irrespective of the position of the leaf (day or night), the central vascular core and the surrounding bundle sheath had high levels of sucrose (up to 500 mmol-(kg DW)^–1) and low levels of glucose and fructose (below 100 mmol-(kg DW)^–1), while in the cortex the situation was reversed. In the night position the level of sucrose decreased by approx. 30% in the bundle sheath and the central vascular core but not in the other sections. We thus suggest that because of the relatively small diurnal changes in their cortical pools, soluble sugars are not involved in the osmotic processes resulting in leaf movement. In contrast, pulvini from 14-d-old plants showed an interesting diurnal change in starch and malate pools in the outermost layer of the extensor. Here starch increased at night while the malate pool was lowered nearly stoichiometrically. Inverse pool sizes were found in the day position of the respective leaves. Although less significant, the opposite diurnal variation occurred in samples taken from the flexor region. We thus were able to locate areas of different carbohydrate activities in the laminar pulvinus of P. coccineus. The central vascular core, including the bundle sheath, is involved in temporary storage of photoassimilates, and the cortical regions are responsible for osmotically driven leaf movement. The results are discussed with respect to guard-cell physiology.Abbreviations CLP cut-leaf pulvini - ILP intact-leaf pulvini This work was supported by a grant from the Deutsche Forschungsgemeinschaft.     

Comparative Histocytology of the Petiole and the Main Pulvinus in Mimosa pudica L.

In Mimosa pudica, the main pulvinus, which brings about leafmovements, presents unusual structural characteristics in comparisonwith the petiole. Peculiar cellular features which exist inthe cortex, epidermis, parenchyma and endodermal regions includethe shape of the cells, their disposition and the location ofthe organelles. The central cylinder of the petiole is surrounded only by afew parenchyma layers whereas the central cylinder of the pulvinusforms a narrow central core enclosed in numerous cortical parenchymalayers. The phloem of the pulvinus contains collenchymatouscells towards the outside and possesses companion cells withwall ingrowths; these phloem members do not exist in the petiole.Xylem and protoxylem parenchyma cells of the petiole possesswall ingrowths which do not occur in homologous cells of thepulvinus. Moreover the pith of the pulvinus is composed of smallfibriform elements similar to the xylem fibriform elements ofthe organ. The structures observed may facilitate exchanges between cellsin the petiole and in the pulvinus. The predominant functionsof the organs relative to lateral and longitudinal transferof nutrients and conduction of stimuli are discussed. Mimosa pudica L., sensitive plant, pulvinus, ultrastructure, conduction of stimuli, leaf movement     

Vacuolar remodelling mediates touch-induced androgynophore movement in Passiflora (Subg. Decaloba, Sect. Xerogona) flowers

Some species of the genus Passiflora (sect. Xerogona) have flowers that exhibit a mechanically-induced movement of the androgynophore that is probably involved in pollination. Despite the many reports of touch-induced movements of floral parts, the studies concerning anatomical, ultrastructural and molecular aspects of the plant movements are restricted to the vegetative parts. Rapid plant movements are highly dependent on turgor changes of a particular flexible tissue formed by specialized cells capable of losing and gaining water rapidly. Thigmotropic androgynophores of four species of Passiflora from section Xerogona were analyzed at cellular and subcellular levels. Our results show that the movement is due to a vacuolar remodelling in a group of parenchymatous cells at the base of the androgynophore. After the movement, plasmolyzed and multivacuolated cells are present at the stimulated side and turgid and univacuolated cells at the opposite side. The results suggest that the mechanisms leading to the androgynophore movement in Passiflora are, in general, analogous to those reported for the movements of legume pulvinus, motile stamen filaments and guard cells of stomata, pointing to conserved cellular mechanisms of plant movement.     

STRUCTURAL SPECIALIZATION OF THE SITE OF RESPONSE TO VECTORIAL PHOTO-EXCITATION IN THE SOLAR-TRACKING LEAF OF LAVATERA CRETICA

Structural features of the pulvinus of the solar-tracking leaf of Lavatera cretica L. that are involved in its capacity for omnidirectional and fully reversible bending in response to vectorial excitation of the lamina were studied by light- and scanning electron microscopy. Pulvini that had bent in the plane at right angles to the midvein were bisected along that plane and the symmetrical tissues of the expanded and contracted flanks were compared. The pulvinus contains a central vascular core and exhibits a transversely furrowed exterior. These specialized features enable the fully mature tissue of this region of the petiole to bend reversibly. The epidermis, chlorenchyma, peripheral collenchyma, and cortical parenchyma in the pulvinus form concentric, radially symmetric sheaths around the vascular core and exhibit structural features in their cell walls that would allow considerable changes in cell volume and consequently enable the omnidirectional bending of this pulvinus. Thickened wall portions of the pulvinar epidermis and peripheral collenchyma exhibit a highly specialized architecture, consisting of alternating thick and thin strips, that enhances their flexibility, while maintaining mechanical support. Cell walls of the chlorenchyma and the cortical parenchyma are thin and capable of reversible infolding. Those of the cortical parenchyma also exhibit numerous prominent transverse pit fields, indicative of anisotropic orientation of their microfibrillar lattice transverse to the pulvinar axis. This orientation is compatible with elastic deformation of the cortical parenchyma cells along the pulvinar axis. Filament-like cytoplasmic strands were observed along the walls of pulvinar motor cells, predominantly transverse to the pulvinar axis, but their function (if any) in volume changes of these cells is unknown.     

Exogenous application of brassinosteroids regulates tobacco leaf size and expansion via modulation of endogenous hormones content and gene expression

Brassinosteroids (BR) play diverse roles in the regulation of plant growth and development. BR promotes plant growth by triggering cell division and expansion. However, the effect of exogenous BR application on the leaf size and expansion of tobacco is unknown. Tobacco seedlings are treated with different concentrations of exogenous 2,4-epibrassinolide (EBL) [control (CK, 0 mol L⁻¹), T1 (0.5 × 10⁻⁷ mol L⁻¹), and T2 (0.5 × 10⁻⁴ mol L⁻¹)]. The results show that T1 has 17.29% and T2 has 25.99% more leaf area than control. The epidermal cell area is increased by 24.40% and 17.13% while the number of epidermal cells is 7.06% and 21.06% higher in T1 and T2, respectively, relative to control. So the exogenous EBL application improves the leaf area by increasing cell numbers and cell area. The endogenous BR (7.5 times and 68.4 times), auxin (IAA) (4.03% and 25.29%), and gibberellin (GA₃) contents (84.42% and 91.76%) are higher in T1 and T2, respectively, in comparison with control. Additionally, NtBRI1, NtBIN2, and NtBES1 are upregulated showing that the brassinosteroid signaling pathway is activated. Furthermore, the expression of the key biosynthesis-related genes of BR (NtDWF4), IAA (NtYUCCA6), and GA₃ (NtGA3ox-2) are all upregulated under EBL application. Finally, the exogenous EBL application also upregulated the expression of cell growth-related genes (NtCYCD3;1, NtARGOS, NtGRF5, NtGRF8, and NtXTH). The results reveal that the EBL application increases the leaf size and expansion by promoting the cell expansion and division through higher BR, IAA, and GA₃ contents along with the upregulation of cell growth-related genes. The results of the study provide a scientific basis for the effect of EBL on tobacco leaf growth at morphological, anatomical, biochemical, and molecular levels.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-00971-x.     

Osmoregulation of leaf motor cells

"Osmotic Motors"--the best-documented explanation for plant leaf movements--frequently reside in specialized motor leaf organs, pulvini. The movements result from dissimilar volume and turgor changes in two oppositely positioned parts of the pulvinus. This Osmotic Motor is powered by a plasma membrane proton ATPase, which drives KCl fluxes and, consequently, water, across the pulvinus into swelling cells and out of shrinking cells. Light signals and signals from the endogenous biological clock converge on the channels through which these fluxes occur. These channels and their regulatory pathways in the pulvinus are the topic of this review.     

STM/BP-Like KNOXI Is Uncoupled from ARP in the Regulation of Compound Leaf Development in Medicago truncatula

Class I KNOTTED-like homeobox (KNOXI) genes are critical for the maintenance of the shoot apical meristem. The expression domain of KNOXI is regulated by ASYMMETRIC LEAVES1/ROUGHSHEATH2/PHANTASTICA (ARP) genes, which are associated with leaf morphology. In the inverted repeat-lacking clade (IRLC) of Fabaceae, the orthologs of LEAFY (LFY) function in place of KNOXI to regulate compound leaf development. Here, we characterized loss-of-function mutants of ARP (PHAN) and SHOOTMERISTEMLESS (STM)- and BREVIPEDICELLUS (BP)-like KNOXI in the model IRLC legume species Medicago truncatula. The function of ARP genes is species specific. The repression of STM/BP-like KNOXI genes in leaves is not mediated by PHAN, and no suppression of PHAN by STM/BP-like KNOXI genes was observed either, indicating that STM/BP-like KNOXI genes are uncoupled from PHAN in M. truncatula. Furthermore, comparative analyses of phenotypic output in response to ectopic expression of KNOXI and the M. truncatula LFY ortholog, SINGLE LEAFLET1 (SGL1), reveal that KNOXI and SGL1 regulate parallel pathways in leaf development. We propose that SGL1 probably functions in a stage-specific manner in the regulation of the indeterminate state of developing leaves in M. truncatula.     

INTERNAL PHLOEM IN THE PULVINUS OF SOYBEAN PLANTS

Glycine max, like many species of Fabaceae, has pulvini at the base of the petiole. In this structure, the vascular cylinder is constricted and consists of a ring of phloem surrounding a ring of xylem. A combination of light and transmission electron microscopy and histochemistry showed that, in addition, there are groups of internal phloem strands in the pulvinar pith. This was confirmed by direct observation of sieve plates and crystalline P-protein inclusions typical of leguminous sievetube members. Serial sections through the stem–pulvinus–petiole revealed that a spatial reorientation of the vascular tissue in the pulvinus resulted in the formation of internal phloem strands, which are continuous with the external phloem bundles above and below the pulvinus. Using 6(5)carboxyfluorescein (6CF) as a fluorescent tracer of phloem transport, we have shown that the internal phloem was active. In most of the experiments, when 6CF was applied to a source leaf, the internal phloem was not stained when the stem was girdled between the source leaf and the roots. Thus, we suggest that the internal phloem of the pulvinus of soybean is specialized for transport toward the root.