SIEVE TUBES IN FOLIAR VEIN ENDINGS: REVIEW AND QUANTITATIVE SURVEY OF RUDBECKIA LACINIATA (ASTERACEAE)

Sieve tube distribution in foliar vein endings of angiosperms is mentioned in 33 publications. Their collective judgment is that sieve tubes may be lacking, may end partway along, or may extend to the vein tip. Most reports conclude that all vein endings are the same within a species, but a few studies mention variation. Only three studies include quantitative data, all based on reconstruction from microtome sections. The present quantitative study, the most extensive one to date, surveyed sieve tubes in vein endings from cleared and stained leaf pieces of Rudbeckia laciniata (Asteraceae, tribe Heliantheae), a North American perennial herb. Of 203 areoles sampled, 32 (15.8%) lacked vein endings whereas 171 (84.2%) had one to six endings per areole. There were 385 individual vein endings in the 171 areoles: 115 lacked sieve tubes, 38 had them to the vein tip, but in most (232) they ended at some intermediate point. Serial cross sections of the latter showed phloem parenchyma continuing beyond the sieve tubes for some distance, beyond which occurred a distal zone consisting solely of tracheary elements. R. laciniata vein endings exhibit the same range of variation reported for dicots in general. More studies combining clearings with selected sections are needed to establish the range of vein ending patterns among angiosperms.     

STUDIES ON THE LEAF OF POPULUS DELTOIDES (SALICACEAE): QUANTITATIVE ASPECTS,AND SOLUTE CONCENTRATIONS OF THE SIEVE-TUBE MEMBERS

The vascular system of the leaf of Populus deltoides Bartr. ex Marsh, was examined quantitatively, and plasmolytic studies were carried out to determine the solute concentrations of sieve-tube members at various locations in the leaf. Both the total number and total crosssectional area of each cell type decreases with decreasing vein size. Although the proportion of phloem occupied by sieve tubes varies considerably from location to location, a linear relationship exists between cross-sectional area of the vascular bundles and both total and mean cross-sectional area of sieve tubes. Collectively, the cross-sectional area of all tertiary and minor veins feeding into a secondary exceeds the total cross-sectional area of sieve tubes at the base of that secondary. Moreover, the total volume of sieve tubes in the “catchment area” of a secondary vein is much greater than the total sieve tube volume of the secondary itself. Both tracheary elements and sieve-tube members undergo a reduction in both total and mean crosssectional area in the constricted zone at the base of the leaf. The plasmolytic studies revealed the presence of positive concentration gradients in sieve tubes of the lamina from the minor veins and tips of the secondaries to the bases of the secondaries and their associated subjacent midvein bundles and from the upper to lower portions of the median bundle of the midvein.     

STUDIES ON THE LEAF OF AMARANTHUS RETROFLEXUS (AMARANTHACEAE): QUANTITATIVE ASPECTS,AND SOLUTE CONCENTRATION IN THE PHLOEM

The vascular system of the leaf of Amaranthus retroflexus L. was examined quantitatively, and plasmolytic studies were carried out on it to determine the solute concentration in cells of the phloem at various locations in the leaf. The proportion of phloem occupied by sieve tubes varies considerably with vein size and leaf size. Collectively, the cross-sectional area of sieve tubes of all tributaries at their points of entry into either a secondary or midvein far exceeds the total cross-sectional area of sieve tubes at the bases of those major veins. In addition, the total volume of sieve tubes in the “catchment area” of a secondary vein is much greater than total sieve-tube volume of the secondary vein itself. The plasmolytic studies revealed the presence of positive concentration gradients in the sieve tubes of the lamina from the minor veins and tips of the secondaries to the bases of the secondaries and from the tip to the base of the midvein. The C₅₀ (the estimated mannitol concentration plasmolyzing, on the average, 50% of the sieve-tube members) was 1.5 m for minor veins and tips of secondary veins and 1.1 m for the bases of secondaries; 1.3 m for the tip of the midvein and 0.6-0.7 m for the midvein in the basal third of the lamina.     

Leaf structure in relation to solute transport and phloem loading in Zea mays L.

Small and intermediate (longitudinal) vascular bundles of the Zea mays leaf are surrounded by chlorenchymatous bundle sheaths and consist of one or two vessels, variable numbers of vascular parenchyma cells, and two or more sieve tubes some of which are associated with companion cells. Sieve tubes not associated with companion cells have relatively thick walls and commonly are in direct contact with the vessels. The thick-walled sieve tubes have abundant cytoplasmic connections with contiguous vascular parenchyma cells; in contrast, connections between vascular parenchyma cells and thin-walled sieve tubes are rare. Connections are abundant, however, between the thin-walled sieve tubes and their companion cells; the latter have few connections with the vascular parenchyma cells. Plasmolytic studies on leaves of plants taken directly from lighted growth chambers gave osmotic potential values of about-18 bars for the companion cells and thin-walled sieve tubes (the companion cell-sieve tube complexes) and about-11 bars for the vascular parenchyma cells. Judging from the distribution of connections between various cell types of the vascular bundles and from the osmotic potential values of those cell types, it appears that sugar is actively accumulated from the apoplast by the companion cell-sieve tube complex, probably across the plasmalemma of the companion cell. The thick-walled sieve tubes, with their close spatial association with the vessels and possession of plasmalemma tubules, may play a role in retrieval of solutes entering the leaf apoplast in the transpiration stream. The transverse veins have chlorenchymatous bundle sheaths and commonly contain a single vessel and sieve tube. Parenchymatic elements may or may not be present. Like the thick-walled sieve tubes of the longitudinal bundles, the sieve tubes of the transverse veins have plasmalemma tubules, indicating that they too may play a role in retrieval of solutes entering the leaf apoplast in the transpiration stream.     

Leaf development and phloem transport in Cucurbita pepo: Maturation of the minor veins

Summary Young leaves of Cucurbita pepo L. were examined by whole-leaf autoradiography and serial paradermal sections were examined by light microscopy to determine whether commencement of sugar export depends upon the minor vein phloem achieving structural maturity. Maturation of these veins develops progressively from the largest toward the smallest elements with the minor veins in the distal region of the leaf maturing before those in the proximal region. Commencement of sugar export is coincident with maturation of the abaxial phloem of the minor veins delimiting the areoles. The abaxial phloem elements of the larger minor veins, which are probably capable of vein loading too but border only relatively few areoles, mature before export starts. The adaxial phloem surrounding the areoles and the xylem elements, mature in advance of the abaxial phloem and well before the beginning of sugar export. It is therefore considered unlikely that structural development alone directly governs the initiation of export. The results suggest that some other rate controlling step is involved.     

Leaf Architecture of Some Monocotyledons with Reticulate Venation

Twelve species belonging to seven monocotyledonous families:Hydrocharitaceae, Taccaceae, Dioscoreaceae, Smilacaceae, Araceae,Alismataceae and Aponogetonaceae show reticulate venation typicalof dicotyledons. A study of the leaves of these species showsthat venation patterns are usually curvipalmate-convergate,occasionally rectipalmate or collimate, and rarely pinnate lyratetype. Number, size and shape of areoles, number of primaries,number of secondaries along one side of the primaries, anglebetween 1 and 2° veins and number of vein endings per areoleare given for each species. Intesecondary veins, isolated tracheids,loops, extension cells, raphide and raphide idioblasts and terminaltracheids were observed. Marginal ultimate venation is mostlyarcuate. Major and minor veins are jacketed by parenchymatousbundle sheath cells. The lamina of Tacca leontopetaloides, Colocasiaesculenta and Scindapsus aureus show a single midrib-like centralregion similar to that of dicotyledonous leaves, and it is multistrandedin Aponogeton natans, Limnophyton obtusifolium and Ottelia alismoides.The degree of vein order is most commonly up to fourth or fifthand rarely up to sixth in Dioscorea hispida. Monocotyledons, leaf architecture, vein endings, venation, areoles     

Sink Effect of Cytokinin in Detached Leaves Is Not Caused by Structural Rearrangements of Phloem

The sink effect of cytokinin is manifested as a decrease in source capacity and the induction of sink activity in the phytohormone-treated region of a mature excised leaf. In order to find out whether this effect was due to the direct action of cytokinin on the phloem structure, two types of phloem terminals were examined. In pumpkin (Cucurbita pepo L.) leaves, the phloem terminals are open; i.e., they are linked to mesophyll by numerous symplastic connections, which are located in narrow areas called plasmodesmal pit fields. In broad bean (Vicia faba L.) leaves, the phloem terminals belong to the closed type and have no symplastic links with mesophyll. The electron microscopic study of terminal phloem did not reveal any structural changes in the companion cells, which could account for the suppression of assimilate export. The treatment of leaves with cytokinin neither disturbed the structure of plasmodesmal pit fields in pumpkin leaves nor eliminated the wall protuberances (the ingrowths promoting phloem loading) in bean leaves. No evidence was obtained that the cytokinin-induced import of assimilates in mature leaves is caused by the recovery of meristematic activity, i.e., by either formation of new phloem terminals having immature sieve elements capable of unloading or by the development of new sieve elements within the existing veins. Cytokinin did not induce de novo formation of phloem elements. Structural characteristics of the leaf phloem, such as the number of branching orders in the venation pattern, the number of vein endings per areole, the number of areoles per leaf, the area of one areole, and the number of sieve elements per bundle remained unaltered. It is concluded that the sink effect of cytokinin in excised leaves cannot be determined by alteration of the phloem structure.     

Evidence for two pathways of phloem loading

The minor veins of small leaf discs, punched out of mature leaves and incubated in ¹⁴C-sucrose solution, appear labeled in macro- and microautoradiographs. Discs with a labeled vein pattern and with labeled sieve tubes in microautoradiographs were found in Beta vulgaris, Vicia faba, Gomphrena globosa and Antirrhinum majus . However, in several other plant species, minor veins appeared unlabeled in macroautoradiographs when the discs were incubated in ¹⁴C-sucrose. Mesophyll cells ( Acer pseudoplatanus, Juglans regia, Fagia, sylvatica, Syringa vulgaris, Laburnum anagyroides ), bundle-sheath cells of major veins ( Salix viminalis, Robinia pseudoacacia, Commelina communis ) or epidermal layers ( Ginkgo biloba, Chlorophytum comosum ) appeared labeled. Lack of radioactivity in sieve tubes of this latter group was confirmed by microauto-radiography. Using ¹⁴C-glucose instead of ¹⁴C-sucrose, leaf discs of Beta vulgaris showed no labeled vein pattern and in microautoradiographs the sieve tubes appeared unlabeled. In view of the by-pass phloem loading, this study provides evidence for two pathways of phloem loading.     

Foliar architecture and anatomy of <Emphasis Type="Italic">Bernardia</Emphasis> and other genera of Acalyphoideae (Euphorbiaceae)

A comparative study of leaf architecture and anatomy of 42 species of Bernardia and other genera related of Acalyphoideae was undertaken to identify characters that support infrageneric and specific delimitation. Thirty variable foliar architectural and anatomical characters were studied. Some characters are consistent (e.g., venation pattern, secondary and tertiary vein arrangement, presence or absence of agrophic veins, type of areoles, bundle sheath cell type, presence or absence of fibers in the mesophyll, trichome type, stomata location, and type of crystals), and characterize genera. In addition, foliar character distribution within Bernardia supports the most recent infrageneric classification.     

Bundle sheath lignification mediates the linkage of leaf hydraulics and venation

The lignification of the leaf vein bundle sheath (BS) has been observed in many species and would reduce conductance from xylem to mesophyll. We hypothesized that lignification of the BS in lower‐order veins would provide benefits for water delivery through the vein hierarchy but that the lignification of higher‐order veins would limit transport capacity from xylem to mesophyll and leaf hydraulic conductance (K_leaf). We further hypothesized that BS lignification would mediate the relationship of K_leaf to vein length per area. We analysed the dependence of K_leaf, and its light response, on the lignification of the BS across vein orders for 11 angiosperm tree species. Eight of 11 species had lignin deposits in the BS of the midrib, and two species additionally only in their secondary veins, and for six species up to their minor veins. Species with lignification of minor veins had a lower hydraulic conductance of xylem and outside‐xylem pathways and lower K_leaf. K_leaf could be strongly predicted by vein length per area and highest lignified vein order (R² = .69). The light‐response of K_leaf was statistically independent of BS lignification. The lignification of the BS is an important determinant of species variation in leaf and thus whole plant water transport.     

Phloem Unloading in Developing Leaves of Sugar Beet : II. Termination of Phloem Unloading

Phloem unloading in developing leaves of Beta vulgaris L. (`Klein E' multigerm) occurred from successively higher order branches of veins as leaves matured. Phloem unloading was studied in autoradiographs of leaf samples taken at various times during the arrival of a pulse of ¹⁴C-labeled photoassimilate. Extension of mass flow of sieve element contents into leaf vein branches was determined from the high level of radiolabel in veins soon after first arrival of the pulse. Rapid entry, indicative of mass flow through open sieve pores, occurred down to the fourth division of veins in young, importing leaves and to the fifth or terminal branch in importing regions near the zone of transition from sink to source. The rate of unloading decreased with leaf age, as evidenced by the increased time required for the vein-mesophyll demarcation to become obscured. The rate of import per unit leaf area, measured by steady state labeling with ¹⁴CO₂ also decreased as a leaf matured. The decline in import appeared to result from progressive changes that increased resistance to unloading of sieve elements and eventually terminated phloem unloading.     

VASCULAR SYSTEM OF LODICULES OF DACTYLIS GLOMERATA L.

The vascular system for the two lodicules in a floret of Dactylis glomerata L. was studied in serial sections. The floret stele contained a few modified tracheary elements and xylem transfer cells enveloped by a phloem of squat sieve-tube members and intermediary cells. A single sieve tube and associated phloem parenchyma exited the right and left sides of the stele and upon nearing the base of each lodicule branched and formed the minor veins of the lodicule. The minor veins underwent limited branching and anastomosing to form a small three-dimensional system which described an arc during its ascent in the adaxial portion of each lodicule. The sieve tubes in the minor veins extended halfway up the lodicule and contained short sieve-tube members with transverse, slightly oblique, or lateral simple sieve plates. The associated phloem parenchyma cells were intermediary cells, companion cells, and less intimate parenchyma cells. Intermediary cells terminated the minor veins and touched the distal ends of the terminal sieve-tube members, which lacked distal sieve plates. Although the transverse area of the sieve-tube members remained constant up the lodicule, the transverse area of the associated phloem parenchyma fluctuated.     

Symplastic Transfer of Fluorescent Dyes from Mesophyll to Sieve Tube in Stripped Leaf Tissue and Partly Isolated Minor Veins of Commelina benghalensis

We have stripped small (3 × 3 mm) fields of the upper and the opposite lower epidermis of Commelina benghalensis leaves. Pectinase treatment of the resulting chlorenchyma windows produced free-lying viable minor veins with small lumps of mesophyll cells attached. These veins were still connected with the intact remainder of the leaf. Fluorescent dyes were injected into mesophyll cells or mestome sheath cells. Continuous following of the dye from the moment of injection and use of the simple vein system allowed an unhindered and precise assessment of the cell-to-cell route of dye transfer. Disodium fluorescein and Lucifer Yellow CH injected into mesophyll or mestome sheath cells readily moved to the sieve tube. This symplastic dye transfer from mesophyll to sieve tube was also observed after injection into unmacerated stripped leaf tissue. The displacement of fluorescent dyes substantiates a symplastic continuity between mesophyll and sieve tube and therefore supports the possibility of symplastic phloem loading.     

Studies on the leaf anatomy of Euphorbia: II. Venation patterns*

Leaf venation patterns of 150 species of Euphorbia are presented and their value as a diagnostic feature in the genus assessed. Based on the gross venation patterns, the species have been grouped into uni-, bi-, tri-veined and special categories. The majority of the species studied belonged to the tri-veined category, in which ornamentation of the veins and the course of traces in the lamina proved useful additional characters. Features such as the size of the areoles, the number of vein endings, and their further ramifications and composition in each areole varied in the same leaf or in different leaves of a given species. Furthermore, no direct correlation could be established between the areole size and the numbers of vein endings and tips per areole. Forty species possess a parenchymatous vein sheath. Globular chloroplasts showed up conspicuously in the sheath cells of a few species. Dilated tracheidal elements, localized on the venules or at the tips of vein endings, are characteristic of the xerophytic species. In some instances, correlation was apparent between plants grouped according to their venation patterns and their habit.     

贵州小檗属植物(小檗科)叶脉序研究

采用制作叶脉标本和透明叶标本的方法,对贵州产28种2变种小檗属植物叶脉特征进行比较研究。结果表明:贵州小檗属植物的脉序类型有5种:半达缘羽状脉、花环状半达缘羽状脉、简单弓形羽状脉、花环状弓形羽状脉和混合型。叶脉分支一般有五级:1一级脉构架均为羽状脉,粗度有很粗、粗、中等粗细和纤细四种类型,分支方式包括单轴分支和合轴分支;2粗二级脉构架中有分支达缘或分支均不达缘,与中脉夹角变化各异,内二级脉存在或缺失,细二级脉半达缘、真曲行或简单弓形,间二级脉类型复杂多变但频度种间有差异;3三级脉贯串型、结网型或分支型;4四、五级脉网状或自由分支且常混合在一起。脉间区从发育差到良好,小脉从不分支到不均等分支等各种类型均有,叶缘末级脉缺失、不完整、钉状和环状。大部分种类叶缘具齿,每1cm齿数目和齿内腺点的特性等特征在不同种类间有区别,具有鉴定价值,但齿其它特征复杂多变或种间区别较小,同时齿内脉性状也不稳定。此外,齿的有无会对脉序类型产生影响。小檗属植物叶脉类型存在种间差异,具有重要的分类学价值,叶脉类型的变化和复杂程度显示了该属植物的进化特点;叶齿的有无和齿特征具有分类学和系统学意义。基于叶脉特征的研究结果并结合重要的外部形态学特征编制了贵州小檗属植物的分种检索表。研究结果可为小檗属植物分类寻找新的依据并探讨其系统学意义。     

Changes in the amino acid composition and protein modification in phloem sap of rice

Pure phloem sap was collected from insects feeding on rice (Oryza sativa L.) leaves by a laser technique similar to the aphid stylet technique. Rapid circulation of nitrogen in the sieve tubes was demonstrated directly using ¹⁵N as a tracer. Application to the roots of the metabolic inhibitors of amino acids, aminooxyacetate and methioninesulfoximine, changed the amino acid composition in the sieve tubes. Feeding methionine to leaf tips resulted in its bulk transfer into the sieve tubes. In vitro experiments confirmed the existence of protein kinases in the pure rice phloem sap. The phosphorylation status of the sieve tube sap proteins was affected by the light regime. The possibility that changes in chemical composition or protein modification such as phosphorylation in the sieve tubes might affect plant growth are discussed.Analysis of pure phloem sap collected from rice plants by insect laser technique has shown dynamic changes in the chemical composition and the quality of proteins in the sap.     

A QUANTITATIVE STUDY OF SIEVE-TUBE DIFFERENTIATION IN VEGETATIVE SHOOT APICES OF COLEUS

Quantitative methods, with round-the-clock collecting of large samples in successive years, have uncovered several new phenomena of sieve-tube differentiation in young leaves of Coleus vegetative shoots. In small leaves (1–350 μ), there are no sieve tubes in the leaf itself, but they differentiate acropetally in the two traces to each leaf. Regression lines fitted to the data for leaf length vs. most distal position of sieve tubes in the traces support the view that differentiation is steady and acropetal, but they also reveal that differentiation in the traces falls steadily farther behind elongation of the leaf. Leaves more than 500 μ long have sieve tubes close to their tips. An intensive search of leaves of intermediate lengths revealed an isolated locus of sieve-tube differentiation. These relationships were reproducible year after year. Every plant with discontinuous strands of sieve tubes in the second leaf pair had discontinuous xylem in the third. This isolated locus was not seen before, probably because of small samples and daytime collections; most of our cases were from night collections.     

Microautoradiographic studies of phloem loading and transport in the leaf of Zea mays L.

Microautoradiographs showed that [¹⁴C]sucrose taken up in the xylem of small and intermediate (longitudinal) vascular bundles of Zea mays leaf strips was quickly accumulated by vascular parenchyma cells abutting the vessels. The first sieve tubes to exhibit ¹⁴C-labeling during the [¹⁴C]sucrose experiments were thick-walled sieve tubes contiguous to the more heavily labeled vascular parenchyma cells. (These two cell types typically have numerous plasmodesmatal connections.) With increasing [¹⁴C]sucrose feeding periods, greater proportions of thick- and thin-walled sieve tubes became labeled, but few of the labeled thin-walled sieve tubes were associated with labeled companion cells. (Only the thin-walled sieve tubes are associated with companion cells.) When portions of leaf strips were exposed to ¹⁴CO₂ for 5 min, the vascular parenchyma cells-regardless of their location in relation to the vessels or sieve tubes-were the most consistently labeled cells of small and intermediate bundles, and label (¹⁴C-photosynthate) appeared in a greater proportion of thin-walled sieve tubes than thick-walled sieve tubes. After a 5-min chase with ¹²CO₂, the thin-walled sieve tubes were more heavily labeled than any other cell type of the leaf. After a 10-min chase with ¹²CO₂, the thin-walled sieve tubes were even more heavily labeled. The companion cells generally were less heavily labeled than their associated thin-walled sieve tubes. Although all of the thick-walled sieve tubes were labeled in portions of leaf strips fed ¹⁴CO₂ for 5 min and given a 10-min ¹²CO₂ chase, only five of 72 vascular bundles below the ¹⁴CO₂-exposed portions contained labeled thick-walled sieve tubes. Moreover, the few labeled thick-walledsieve tubes of the transport region always abutted ¹⁴C-labeled vascular parenchyma cells. The results of this study indicate that (1) the vascular parenchyma cells are able to retrieve at least sucrose from the vessels and transfer it to the thick-walled sieve tubes, (2) the thick-walled sieve tubes are not involved in long-distance transport, and (3) the thin-walled sieve tubes are capable themselves of accumulating sucrose and photosynthates from the apoplast, without the companion cells serving as intermediary cells.     

Sieve element specific labeling with an anti-idiotypic monoclonal antibody mimic of the phytotoxin dothistromin

A monoclonal antibody, 12C9, an anti-idiotypic mimic of dothistromin, a toxin produced by Dothistroma pini, was found to label the cell wall of sieve elements in a number of different plant tissues and species. The antibody labeled apple leaf tissue, tobacco leaf mid vein, leaf and meristem, and Coprosma robusta leaf mid vein. Labeling was restricted to cell walls of sieve elements and did not label the companion cells or the lumen of the cells. The antibody labeled over a wide range of dilutions. This antibody could be used to differentiate sieve elements from other types of phloem. It could also be used to co-localize sieve elements and microorganisms such as phytoplasmas stained with DAPI.     

Transmission of Electric Signals in Sieve Tubes of Zucchini Plants

Membrane potentials of ?;160 to ?210 mV were recorded with microelectrodes inserted into meta-phloem sieve tubes of intact zucchini plants (Cucurbita pepo L. var. medullosa Alef.). The effects of darkness, white light and colored light on membrane potential were studied. Reference electrodes were in contact with the apoplast via fluid-filled cavities or “drinks”. Electrolyte solutions (100 mM) in the cavities could be quickly replaced by flushing with 100 mM solutions of sucrose, KCl, sorbitol, or EDTA without altering osmolarity. KCl and EDTA caused depolarization of the sieve tube membrane potential, while sucrose caused depolarization or hyperpolarization of the sieve tube membrane potential in mature or growing plant parts respectively. Recovery of the original voltage was recorded when rapid (sucrose) or slow (sorbitol) transients occurred. When two measuring circuits were installed, one in a growing fruit and the other in the petiole of the subtending mature leaf, the alteration of the sieve tube membrane potential at one site was accompanied by an alteration of the potential at the other site after a few seconds. The responses were opposite in the exporting leaf and importing fruit when sucrose was applied. The signal, transmitted via the sieve tubes, reached maximum velocities of 10 cm per second.