Adenosine triphosphatase in the phloem of Cucurbita

Summary The distribution of adenosine triphosphatase (ATPase) activity in the phloem of petioles and minor veins of Cucurbita maxima has been studied using a lead phosphate precipitation procedure. ATPase activity was localized in sieve elements, companion cells and parenchyma cells. Activity was found at the cell surfaces, associated with the dispersed P-protein of mature sieve elements, in mitochondria, sieve-element reticulum, and at specific regions of the cell walls. It is suggested that the ATPase at the phloem cell surfaces may function in intercellular transport of assimilates or ions, and that the ATPase activity associated with the P-protein may function in the translocation process or in callose deposition.     

Histochemical localization of nucleoside triphosphatase activity in assimilate conducting plant tissue

Summary For the histochemical localization of nucleoside triphosphatases at the electron microscopic level, prefixed tissues were incubated with lead nitrate in addition to substrate (GOMORI reaction). While ATP and UTP as substrates gave electron-dense reaction products at the plasmalemma of sieve tubes, companion cells and phloem parenchyma cells, and at plasmodesmata in primary pitfields, AMP gave reaction products only at the tonoplast of parenchyma cells. Since electron-dense deposits also occur in cell walls and vacuoles, energy dispersive X-ray microanalysis was used to distinguish between lead deposits and lead-phosphate deposits. The latter were restricted to the symplast. Among the three plant species used, the leaf bundle phloem ofHordeum distichon showed ATPase activity largely restricted to the phloem cells, except for the thickwalled sieve tubes. Some activity also bordered the chloroplasts of the bundle sheath cells. In the C₄ plantGomphrena globosa, ATPase and UTPase activities appeared to be the greater in phloem parenchyma cells than in sieve tubes. In the phloem of youngMonstera deliciosa roots, ATPase occurred not only at the plasmalemma of sieve tubes, but also around sieve-tube plastids. When compared with AMP as substrate, it appears that nucleoside triphosphates are the natural substrates of the enzyme(s) in the plasmalemma of sieve tubes and phloem parenchyma cells.     

Zur Lokalisation von Dehydrogenasen des Energiestoffwechsels im Phloem von Cucurbita pepo L.

Summary In the stem phloem of Cucurbita pepo the enzymes GAPDH, ADH, MDH, NADP-IDH, NAD-IDH, G₆PDH and SDH were localized histochemically with the aid of tetrazolium salt (NBT). When the stems were deep-frozen the most intense formation of formazan was found in companion cells, less in phloem parenchyma cells, and very little in sieve tubes.The distribution of enzymes in phloem markedly changes when stems were cut 2 minutes before freezing: 2,5 cm behind the sectional area little formazan was found. Companion cells and parenchyma cells had lost nearly all activity. 15 cm behind the sectional area there was again a higher concentration of formazan in the companion cells and parenchyma cells. In this region an even higher activity was detected in sieve tubes. 25–30 cm behind the sectional area the distribution of formazan was nearly the same as in the intact stems.Apparently activities of the enzymes tested primaily occur in the companion cells and parenchyma cells only. After wounding they are translocated into sieve tubes or exudate.     

Minor vein structure and sugar transport in Arabidopsis thaliana

Leaf and minor vein structure were studied in Arabidopsis thaliana (L.) Heynh. to gain insight into the mechanism(s) of phloem loading. Vein density (length of veins per unit leaf area) is extremely low. Almost all veins are intimately associated with the mesophyll and are probably involved in loading. In transverse sections of veins there are, on average, two companion cells for each sieve element. Phloem parenchyma cells appear to be specialized for delivery of photoassimilate from the bundle sheath to sieve element-companion cell complexes: they make numerous contacts with the bundle sheath and with companion cells and they have transfer cell wall ingrowths where they are in contact with sieve elements. Plasmodesmatal frequencies are high at interfaces involving phloem parenchyma cells. The plasmodesmata between phloem parenchyma cells and companion cells are structurally distinct in that there are several branches on the phloem parenchyma cell side of the wall and only one branch on the companion cell side. Most of the translocated sugar in A. thaliana is sucrose, but raffinose is also transported. Based on structural evidence, the most likely route of sucrose transport is from bundle sheath to phloem parenchyma cells through plasmodesmata, followed by efflux into the apoplasm across wall ingrowths and carrier-mediated uptake into the sieve element-companion cell complex. Received: 5 October 1999 / Accepted: 20 November 1999     

Wood Anatomy and the Development of Interxylary Phloem of Ipomoea hederifolia Linn. (Convolvulaceae)

In Ipomoea hederifolia Linn., stems increase in thickness by forming successive rings of cambia. With the increase in stem diameter, the first ring of cambium also gives rise to thin-walled parenchymatous islands along with thick-walled xylem derivatives to its inner side. The size of these islands increases (both radially and tangentially) gradually with the increase in stem diameter. In pencil-thick stems, that is, before the differentiation of a second ring of cambium, some of the parenchyma cells within these islands differentiate into interxylary phloem. Although all successive cambia forms secondary phloem continuously, simultaneous development of interxylary phloem was observed in the innermost successive ring of xylem. In the mature stems, thick-walled parenchyma cells formed at the beginning of secondary growth underwent dedifferentiation and led to the formation of phloem derivatives. Structurally, sieve tube elements showed both simple sieve plates on transverse to slightly oblique end walls and compound sieve plates on the oblique end walls with poorly developed lateral sieve areas. Isolated or groups of two to three sieve elements were noticed in the rays of secondary phloem. They possessed simple sieve plates with distinct companion cells at their corners. The length of these elements was more or less similar to that of ray parenchyma cells but their diameter was slightly less. Similarly, in the secondary xylem, perforated ray cells were noticed in the innermost xylem ring. They were larger than the adjacent ray cells and possessed oval to circular simple perforation plates. The structures of interxylary phloem, perforated ray cells, and ray sieve elements are described in detail.     

西瓜小叶脉超微结构研究

用透射电子显微技术研究了西瓜叶片小叶脉,结果表明,小叶脉是由大型维管束鞘细胞包围的维管束,维管束呈现大的头部和线形的柄部,柄部是单列细胞的木质部,由维管薄壁细胞和导管分子组成;头部是韧皮部,由维管薄壁细胞、伴胞和筛管分子组成。同一小叶脉内常见有超微结构特征显著不同的两种伴胞:一种伴胞体积小,与维管束鞘细胞接触面较小或不接触,细胞内有大液泡,细胞壁上没有胞间连丝或只有少数不分枝的胞间连丝,这种伴胞为2a型;另一种伴胞体积大,通常位于韧皮部两翼,不含大液泡而含大量小泡,与维管束鞘细胞接触面较大,接触面上有大量具分枝的胞间连丝,分枝部分比未分枝部分直径小,这种伴胞为中间细胞类型。显然,西瓜是小叶脉内兼具两种类型伴胞的植物。     

西瓜小叶脉超微结构研究

用透射电子显微技术研究了西瓜叶片小叶脉,结果表明,小叶脉是由大型维管束鞘细胞包围的维管束,维管束呈现大的头部和线形的柄部,柄部是单列细胞的木质部,由维管薄壁细胞和导管分子组成;头部是韧皮部,由维管薄壁细胞、伴胞和筛管分子组成。同一小叶脉内常见有超微结构特征显著不同的两种伴胞：一种伴胞体积小,与维管束鞘细胞接触面较小或不接触,细胞内有大液泡,细胞壁上没有胞间连丝或只有少数不分枝的胞间连丝,这种伴胞为2a型;另一种伴胞体积大,通常位于韧皮部两翼,不含大液泡而含大量小泡,与维管束鞘细胞接触面较大,接触面上有大量具分枝的胞间连丝,分枝部分比未分枝部分直径小,这种伴胞为中间细胞类型。显然,西瓜是小叶脉内兼具两种类型伴胞的植物。     

Symplasmic Constriction and Ultrastructural Features of the Sieve Element/Companion Cell Complex in the Transport Phloem of Apoplasmically and Symplasmically Phloem-Loading Species

The ultrastructural features of the sieve element/companion cell complexes were screened in the stem phloem of two symplasmically loading (squash, [Cucurbita maxima L.] and Lythrum salicaria L.) and two apoplasmically loading (broad bean [Vicia faba L.] and Zinnia elegans L.) species. The distinct ultrastructural differences between the companion cells in the collection phloem of symplasmically and apoplasmically phloem-loading species continue to exist in the transport phloem. Plasmodesmograms of the stem phloem showed a universal symplasmic constriction at the interface between the sieve element/companion cell complex and the phloem parenchyma cells. This contrasts with the huge variation in symplasmic continuity between companion cells and adjoining cells in the collection phloem of symplasmically and apoplasmically loading species. Further, the ultrastructure of the companion cells in the transport phloem faintly reflected the features of the companion cells in the loading zone of the transport phloem. The companion cells of squash contained numerous small vacuoles (or vesicles), and those of L. salicaria contained a limited number of vacuoles. The companion cells of broad bean and Z. elegans possessed small wall protrusions. Implications of the present findings for carbohydrate processing in intact plants are discussed.     

ONTOGENY AND STRUCTURE OF THE SECONDARY PHLOEM IN PYRUS MALUS

Evert , Ray F. (U. Wisconsin, Madison.) Ontogeny and structure of the secondary phloem in Pyrus malus. Amer. Jour. Bot. 50(1): 8–37. Illus. 1963.—The secondary phloem of apple consists of sieve-tube elements, companion cells, phloem parenchyma cells, fiber-sclereids, and ray parenchyma cells. The sieve-tube elements are generally long, slender cells with very oblique end walls and much-compounded sieve plates. All sieve-tube elements initially possess nacreous thickenings. Similar wall thickenings were observed in the differentiating fiber-sclereids and xylem elements. Of the 245 sieve-tube elements critically examined, 242 were associated with companion cells. All of the companion cells were shorter than their associated sieve-tube elements. Young companion cells possess slime bodies which later become dispersed. Callose is often found on the sieve-tube element side of the common wall between sieve-tube element and companion cell. In several collections, callose was found on both sides of that wall. The parenchyma cells are of 3 types: crystal-containing cells; tannin-and/or starch-containing cells; and those with little or no tannins or starch. Any type parenchyma cell may be on to genetically related to a sieve-tube element, that is, may be derived from the same phloem initial as the sieve-tube element. Morphologically, the phloem parenchyma cells intergrade with the companion cells, the tannin- and starch-free parenchyma cells often being difficult to distinguish from companion cells. Most of the tannin- and starch-free parenchyma cells collapse when the contiguous sieve-tube elements become nonfunctional. The fiber-sclereids arise from parenchyma cells which overwinter on the margin of the cambial zone and differentiate in nonfunctional phloem.     

Effects of low light on phloem ultrastructure and subcellular localization of sucrose synthase in <Emphasis Type="Italic">Prunus persica</Emphasis> L. var. <Emphasis Type="Italic">nectarina</Emphasis> Ait. fruit

Using electron microscopy, the ultrastructure of phloem unloading zone was examined in the Prunus persica L. var. nectarina Ait. fruit. Our study showed that, in the SE/CC (sieve element/companion cell) complexes, CC developing under low light had a thin cytoplasm layer with few mitochondria and numerous small vacuoles, and not clearly seen nuclei. The cytoplasm vacuolation indicated that the cytoskeleton was destroyed at low light. The effects of low light on CC development suggest that unloading evidently linked to the low accumulation of soluble sugars by fruit. At the young fruit stage, flesh parenchyma around the phloem tissue had no starch grains in the plastids in fruit developing under low light. This is a further indication that less photoassimilates was translocated from source leaves to fruit sinks under low light during the young fruit developmental stage. The activity of sucrose synthase (SuSy), the key enzyme of sucrose metabolism in fruit, increased dramatically during fruit maturation. The highest SuSy activity during the rapid fruit growth phase suggests that sink strength could be correlated with the SuSy activity. The high SuSy activity under normal light possibly indicates that fruit had a capacity to utilize sucrose irrespective of their site of phloem unloading. Immunogold electron microscopy showed that SuSy was localized mainly in the vacuole of flesh parenchyma cells. The vacuole-localized SuSy can hydrolyze sucrose imported from the phloem, which may explain the apparent correlation between SuSy activity and phloem unloading. The double sieve element (SE/SE) complexes occurred in a greater number and had thicker cell walls under normal light intensity than under low light intensity. These data demonstrate clearly that low light decreased SuSy activity in the control of phloem unloading. Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 4, pp. 509–517. This text was submitted by the authors in English.     

A study of the source of virus in the Pholem exudate appearing on leaves of Amscinckia infected with the beet curly top virus

Abstract Leaves of Amsinckia douglasiana discharging phloem exudate after infection with the beet curly top virus (BCTV) were studied with the electron microscope. Infected tissue differed from the noninfected in having much hyperplastic phloem characterized by abnormally high proportion of sieve elements, scarcity of companion cells, degenerating parenchyma cells, and some unusually large intercellular spaces. Many spaces contained amorphous debris. Particles resembling BCTV were discernible within the debris. Such particles were encountered also in the debris trapped between stomatal guard cells. Since the phloem exudate excreted from leaves of BCTV-infected plants contains virus particles, and since the virus is found extremely rarely in sieve elements, we suggest (1) that most of BCTV particles apparently released into intercellular spaces are derived from degenerating parenchyma cells in which the virus had multiplied; (2) that the exudate is derived from sieve elements of the hyper-plastic phloem in which the normal functional control by companion cells is lacking; (3) that the exudate leaks from the nontransporting sieve elements through cell walls into intercellular spaces and carries the virus to the outside. Initially, stomata may serve as exits for the infectious exudate, but subsequently ruptures in the epidermis are involved.     

The Phloem of Nelumbo nucifera Gaertn

In common with other aquatic angiosperms, Nelumbo nucifera Gaertn.has a relatively strongly developed phloem tissue. The vascularsystem consists of discrete collateral bundles in which no cambiumdevelops and the phloem and xylem are separated by a narrowlayer of parenchyma cells. The phloem consists of sieve elements,companion cells, and phloem parenchyma cells. The sieve elementshave transverse end walls with simple sieve plates. The cellsattain considerable width in the late phloem (metaphloem). Thecompanion cells are in vertical strands. In the early phloem(protophloem) of large bundles the sieve tubes and companioncells are eventually obliterated. The parenchyma cells alsoform vertical strands which may contain tannin cells. Some parenchymacells and companion cells are binucleate. The sieve elementsshow ultrastructural features common for these cells in dicotyledons.At maturity, they lack nuclei, ribosomes, and tonoplasts, butretain a plasmalemma, mitochondria, and plastids. The latterare poorly differentiated and form starch. The endoplasmic reticulumis in part stacked, in part it forms a network next to the plasmalemma.The P-protein occurs in two forms. One consists of tubules notassembled in any specific type of array. The other, possiblycomposed of much extended tubules, is assembled in crystallineaggregates which are retained as such in mature cells. The sieveplate pores are lined with callose and plasmalemma. The lateralwalls are relatively thin and the nacreous layer varies in degreeof distinctness.     

Fine structure,pattern of division,and course of wound phloem in Coleus blumei

The wound phloem bridges which have developed six days after interrupting an internodal vascular bundle contain wound sieve-elements, companion cells, and phloem parenchyma cells. An analysis of the meristematic activity responding to the wounding clearly demonstrates that three consecutive divisions are prerequisite to the formation of phloem mother-cells. Companion cells are obligatory sister cells of wound sieve-elements, connected to the latter by specific plasmatic strands and provided with a dense protoplast. Six days after wounding most of the wound sieve-elements are still at a nucleate state of development, but already have characteristic P-protein bodies and plastids containing sieve-element starch. Their cytoplasmic differentiation corresponds to the changes recorded during maturation of ordinary sieve elements. Sieve-plate pores penetrate through preexisting parenchyma cell walls, only, and develop from primary pitfield-plasmodesmata. Wound sieve-elements do not connect to preexisting bundle sieve-elements, they open a new tier of young sieve elements produced by cambial activity.     

杨柳科(Salicaceae)7种植物次生韧皮部的比较研究

本文研究和比较了杨柳科2属7种植物次生韧皮部解剖结构。结果表明:(1)杨属和柳属植物在次生初皮部解剖上有某些共同特征:次生韧皮部具有明显分层现象;韧皮纤维和含晶细胞与筛管分子、伴胞和韧皮薄壁组织细胞是切向带相间排列;筛管分子均为复筛板,端壁倾斜平均含有7-8个筛域。(2)两属植物在射线和晶体类型上有明显区别:柳属植物次生韧皮部无石细胞;杨属植物不具功能韧皮部中含有石细胞。(3)两属植物均有一些较为原始的韧皮部解剖特征。     

Cambial Activity and Distribution of Rubber in Guayule (Parthenium argentatum)

Vascular cambium in Guayule, a rubber producing Mexican shrubof Asteraceae family is non-storied. Cambial activity variesperiodically, and the vascular cambium and its immediate derivativesdo not contain rubber. However, as the xylem and phloem parenchymacells derived from the vascular cambium age, rubber depositionstarts from the cell periphery along the walls and later towardstheir cell lumen. Though the sieve tubes and companion cellsof phloem contain no rubber, all parenchyma cells of xylem andphloem, show the presence of rubber, though its amount varies.However, certain lignified xylem ray cells and lignified pithcells are devoid of rubber accumulation. Microfluorescence studiesshow that the epithelial, phloem ray parenchyma, cortical andpith cells, in descending order, have the highest to lowestrubber content. The size and number of rubber particles observedin the parenchyma cells are greatest during the period of cambialdormancy than in an active cambial period Cambium, guayule, rubber     

HYPERPLASTIC PHLOEM IN SUGARBEET LEAVES INFECTED WITH THE BEET CURLY TOP VIRUS

Electron microscopy of sugarbeet leaves infected with the beet curly top virus confirmed earlier findings by light microscopy that the hyperplastic phloem consists mainly of sieve elements that are more or less abnormal in structure. Some parenchyma cells and occasional companion cells may be present. The hyperplastic phloem develops in the place of normal phloem and sometimes in the adjacent ground tissue and the xylem. The sieve elements vary in shape and may be haphazardly arranged. The protoplasts of the sieve elements have the usual characteristics of this type of cell. The sieve element plastids develop from chloroplasts if the hyperplasia occurs in chloroplast-containing parenchyma cells. The cell walls have sieve areas that often are less well differentiated than those of normal sieve elements. The hyperplastic growth in the phloem of curly top diseased plants is discussed with reference to plant tumors induced by certain other plant viruses.     

Anatomical Studies on Secondary Phloem of Euonymus bungeanus

The anatomical structures, especially the type, distribution and arrangement of the constituent elements in the secondary phloem of Euonymus bungeanus Maxim. have been studied. The results showed that the secondary phloem was thicker, consisted of sieve-tube elements, companion ceils ,phloem parenchyma cells ,secretory ceils and rays. Sieve-tube elements, phloem parenchyma cells and secretory cells were alternately arranged in tangential bands, forming a conspicuous zone-like constitution. There was no obvious boundary between the functional phloem and the non-functional phloem. Sieve-tube elements were long, slender cells with very oblique end walls and compound sieve plates. Sieve areas on lateral wall were highly differentiated. Companion cells were triangular in transection and slender in radial section. Mostly,two or three companion cells stayed along with one sieve-tube element. In the functional phloem, phloem parenchyma cells were also slender, containing a few starch grains;but in the nonfunctional phloem they enlarged and contained abundant starch grains. Secretory cells were longer than sieve-tube elements, consisting of rubber-like material. Rays were uniseriate. Finally, the authors also discussed the phylogenetic position of E. bungeanus, which may provide some references for further study of the classification of different genera of Celastraceae.     

桑叶细脉中伴胞的超微结构研究

为了解桑叶细脉中伴胞的超微结构,采用透射电子显微技术对桑叶细脉中伴胞进行观察,着重伴胞与相邻细胞界面上胞间连丝发生频率.结果表明,(1)伴胞含丰富细胞器,细胞壁光滑,无壁内突;(2)伴胞细胞壁上具有大量胞间连丝,胞间连丝通常聚集,并常发生分枝;(3)伴胞与不同类型细胞界面上的胞间连丝发生频率有差异,伴胞-维管束鞘细胞界面上发生频率为25.12±1.83个/μm²,伴胞-伴胞界面上20.18±1.7个2/μm²,伴胞-维管薄壁细胞界面上5.42±0.6个/μm².基于上述观察,认为桑叶细脉中的伴胞属于1-2a型,韧皮部装载途径属于共质体类型.     

Glass microelectrode measurements of sieve tube membrane potentials in internode discs and petiole strips of tomato (Solanum lycopersicum L.)

Summary In tissue slices of tomato (Solanum lycopersicum L.) sieve tube membrane potentials (E_m) were measured by use of glass microelectrodes. In internode discs, the potential differences (pd) of phloem cells near the cut surface fell into two distinct categories with average values of –66 and –109 mV. More distant from the cut surface the values decreased to averages of –71 and –140 mV. These pds were associated with phloem parenchyma cells and sieve tube/companion cell complexes, respectively. In petiole strips, pds were recorded from cells which were identified by iontophoretic injection of fluorescent dye. Averages in two different bathing media, were –140/–146mV, –149/–152mV, and –70/–68mV for sieve tubes, companion cells, and phloem parenchyma cells, respectively. The membrane potentials recorded from sieve tubes were transiently reduced upon sucrose addition. Reduction by CCCP and KCN was more permanent. Sieve tube E_ms recovered more slowly from potassium than from sucrose-induced depolarizations. Light/ dark (L/D) responses were minute (±3 mV). The limitations of the present experimentation are evaluated with special reference to the question as whether the recorded E_ms represent sieve tube membrane potentials occurring in the intact plant.Abbreviations CCCP carbonyl cyanide m-chlorophenylhydrazone - D dark(ness) - E_m membrane potential - L light - LYCH Lucifer yellow CH - pd potential difference - SE standard error     

Symplastic isolation of the sieve element-companion cell complex in the phloem of Ricinus communis and Salix alba stems

The anatomical and physiological isolation of the sieve element-companion cell complex (se-cc complex) was investigated in stems of Ricinus communis L. and Salix alba L. In Ricinus, the plasmodesmatal frequencies were in the proportions 8∶1∶2∶30, in the order given, at the interfaces between sieve tube-companion cell, sieve tube-phloem parenchyma cell, companion cellphloem parenchyma cell, and phloem parenchyma cellphloem parenchyma cell. The membrane potentials of the se-cc complex and the surrounding phloem-parenchyma cells sharply contrasted: the membrane potential of the se-cc complex was about twice as negative as that of the phloem parenchyma. Lucifer Yellow CH injected into the sieve element or into the companion cell remained within the se-cc complex. Dye introduced into phloem parenchyma only moved (mostly poorly) to other phloem-parenchyma cells. The distribution of the plasmodesmatal frequencies, the differential dye-coupling and the sharp discontinuities in membrane potentials indicate that the se-cc complexes constitute symplast domains in the stem phloem. Symplastic autonomy is discussed as a basic necessity for the functioning of the se-cc complex in the stem.