Parenchyma cells of secondary phloem in Pinus strobus

Summary Parenchyma cells of the secondary phloem in Pinus strobus have all the cellular organelles common in other plant cells. They have mitochondria, endoplasmic reticulum, ribosomes, dictyosomes, and plastids. Parenchyma cells are very conspicuous because of their organic inclusions, starch and lipids. Plasmodesmata in transverse and tangential walls of axial parenchyma cells and in end walls of ray parenchyma cells are regularly distributed and of uniform size, about 500 Å in diameter. In radial walls of axial parenchyma cells and horizontal walls of ray parenchyma cells plasmodesmata are located in primary pit-fields; there they are of variable size and often divided into several branches. The branches are confluent into a median nodule. Perforation of the transverse wall between two axial parenchyma cells and the resultant union of the cellular material of the two connected cells is reported.This research has been supported by NSF Grant GB 3193.     

Vacuole proteins in parenchyma cells of secondary phloem and xylem of Dalbergia odorifera

Summary Light- and electron-microscopic observations were made on the stem parenchyma cells of Dalbergia odorifera T. Chen (Papilionaceae), a tropical deciduous tree. In the secondary phloem of branchlet and trunk, all of the parenchyma cells except companion cells contain vacuole proteins. Only the outer secondary xylem of branchlets, but not trunk secondary xylem, has proteins in the ray parenchyma and the vasicentric parenchyma. The xylem vacuole proteins begin to accumulate at the end of the growing period and they disappear after the first flush of growth in spring. The vacuole proteins in phloem cells, particularly in the cells near the cambium, also show seasonal fluctuations. Under the electron microscope, the vacuole proteins appear as fibrous materials in aggregation or in more or less even dispersion, and they occur in the large central vacuoles during both the growth and dormant periods. According to the published studies, the stem storage proteins in the temperate trees appear as small protein-storage vacuoles or protein bodies, and the proteins in the tropical trees occur in large central vacuoles. This distinction is assumed to be related to the differences in the nature of dormancy between temperate and tropical trees.     

Patterned cell determination in a plant tissue: the secondary phloem of trees

The secondary vascular tissues (xylem and phloem) of woody plants originate from a vascular cambium and develop as radially oriented files of cells. The secondary phloem is composed of three or four cell types, which are organised into characteristic recurrent cellular sequences within the radial cell files of this tissue. There is a gradient of auxin (indole acetic acid) across both the cambium and the immediately postmitotic cells within the xylem and phloem domains, and it is believed that this morphogen, probably in concert with other morphogenic factors, is closely associated with the determination and differentiation of the different cells types in each tissue. A hypothesis is developed that, in conjunction with the positional values conferred by the graded radial distribution of morphogen, cell divisions at particular positions within the cambium are sufficient to determine not only each of the phloem cell types but also their recurrent pattern of differentiation within each radial cell file.     

Characterization of a lignified secondary phloem fibre-deficient mutant of jute (Corchorus capsularis)

BACKGROUND AND AIMS: High lignin content of lignocellulose jute fibre does not favour its utilization in making finer fabrics and other value-added products. To aid the development of low-lignin jute fibre, this study aimed to identify a phloem fibre mutant with reduced lignin. METHODS: An x-ray-induced mutant line (CMU) of jute (Corchorus capsularis) was morphologically evaluated and the accession (CMU 013) with the most undulated phenotype was compared with its normal parent (JRC 212) for its growth, secondary fibre development and lignification of the fibre cell wall. KEY RESULTS: The normal and mutant plants showed similar leaf photosynthetic rates. The mutant grew more slowly, had shorter internodes and yielded much less fibre after retting. The fibre of the mutant contained 50 % less lignin but comparatively more cellulose than that of the normal type. Differentiation of primary and secondary vascular tissues throughout the CMU 013 stem was regular but it did not have secondary phloem fibre bundles as in JRC 212. Instead, a few thin-walled, less lignified fibre cells formed uni- or biseriate radial rows within the phloem wedges of the middle stem. The lower and earliest developed part of the mutant stem had no lignified fibre cells. This developmental deficiency in lignification of fibre cells was correlated to a similar deficiency in phenylalanine ammonia lyase activity, but not peroxidase activity, in the bark tissue along the stem axis. In spite of severe reduction in lignin synthesis in the phloem cells this mutant functioned normally and bred true. CONCLUSIONS: In view of the observations made, the mutant is designated as deficient lignified phloem fibre (dlpf). This mutant may be utilized to engineer low-lignin jute fibre strains and may also serve as a model to study the positional information that coordinates secondary wall thickening of fibre cells.     

Chromoplasts ultrastructure and estimated carotene content in root secondary phloem of different carrot varieties

There have been few studies on quantifying carotenoid accumulation in carrots, and none have taken the comparative approach. The abundance and distribution of carotenes in carrot roots of three varieties, white, orange, and high carotene mass (HCM) were compared using light and transmission electron microscopy (TEM). Light microscopy has indicated that, in all three varieties, carotenes were most abundant in the secondary phloem and this area was selected for further TEM analysis. While carotenes were extracted during the fixation process for TEM, the high-pressure freezing technique we employed preserved the spaces (CS) left behind by the extracted carotene crystals. Chromoplasts from the HCM variety contained significantly (P < 0.05) more CS than chromoplasts from the orange variety. Chromoplasts from the white variety had few or no CS. There was no significant difference between the HCM and orange varieties in the number of chromoplasts per unit area, but the white variety had significantly (P < 0.05) fewer chromoplasts than the other two varieties. A large number of starch-filled amyloplasts was observed in secondary phloem of the white variety but these were not found in the other two varieties. The results from this comparative approach clearly define the subcellular localization of carotenoids in carrot roots and suggest that while the HCM genotype was selectively bred for increased carotene content, this selection did not lead to increased numbers of carotene-containing chromoplasts but rather greater accumulation of carotene per chromoplast. Furthermore, the results confirm that roots of the white carrot variety retain residual amounts of carotene.     

The evolution of minor vein phloem and phloem loading

Phylogenetic analysis provides a rational basis for comparative studies of phloem structure and phloem loading. Although several types of minor vein companion cell have been identified, and progress has been made in correlating structural features of these cells with loading mechanisms, little is known about the phylogenetic relationships of the different types. To add to the available data on companion cells, we analyzed the ultrastructure of minor veins in Euonymus fortunei and Celastrus orbiculatis (Celastraceae) leaves and determined that in these species they are specialized as intermediary cells. This cell type has been implicated in symplastic phloem loading. The data were added to published data sets on minor vein phloem characteristics, which were then mapped to a well-supported molecular tree. The analysis indicates that extensive plasmodesmatal continuity between minor vein phloem and surrounding cells is ancestral in the angiosperms. Reduction in plasmodesmatal frequency at this interface is a general evolutionary trend, punctuated by instances of the reverse. This is especially true in the case of intermediary cells that have many plasmodesmata, but other distinguishing characteristics as well, and have arisen independently at least four, and probably six, times in derived lineages. The character of highly reduced plasmodesmatal frequency in minor vein phloem, common in crop plants, has several points of origin in the tree. Thus, caution should be exercised in generalizing results on apoplastic phloem loading obtained from model species. Transfer cells have many independent points of origin, not always from lineages with reduced plasmodesmatal frequency.     

Patterned cell development in the secondary phloem of dicotyledonous trees: a review and a hypothesis

The secondary phloem of dicotyledonous trees and shrubs is constructed of sieve tube cells (S) and their companion cells, as well as parenchyma (P) and fibre (F) cells. Different species have characteristic sequences of these S, P and F cells within the radial files of their phloem. The sequences are recurrent, and are evidence of rhythmic cell determination and differentiation. A model was devised to account for the sequences found in various dicot tree species. It is based on the pattern of radial displacement of cells through a gradient of morphogen which supports secondary phloem development. According to this model, each tree species shows a particular pattern of post-mitotic cellular displacement along each radial file as a result of a corresponding sequence of periclinal division in the cambial initial and its descendents. The divisions and displacements ensure that at each timestep (equivalent to an interdivisional interval) each cell resides in a specific location within the morphogenic gradient. Cells then emerge from the post-mitotic zone of cell determination, having acquired different final positional values. These values lie above a series of thresholds that permit the respective determination and subsequent differentiation of one or other of the three cell types S, P and F. The recurrent nature of the sequences of the three cell types within each radial cell file, as well as their tangential banding, are a consequence of a shared rhythmic spatio-temporal pattern of periclinal cambial divisions. With a single set of morphogen parameters required for cell determination, and using three positions for cambial cell divisions, all the cellular sequences of secondary phloem illustrated in the literature can be accounted for.This is an invited article.     

桉树树干维管形成层和次生韧皮部热致细胞坏死的定量试验

桉树树干维管形成层和次生韧皮部热致细胞坏死的定量试验 桉树(Eucalyptus)树干暴露在森林火灾辐射热中会杀死形成层细胞及其内嵌的再生分生组织,阻止树木萌枝和恢复。目前尚无组织水平的方法来量化热处理对桉树形成层细胞活力的影响。本研究的目 标包括：(1)采用并验证四氮唑还原法检测桉树细胞活力;(2)应用该方法确定斜叶桉(Eucalyptus oblique)形成层细胞活力的阈值水平,进而确定临界温度。采用四氮唑还原法量化该桉树韧皮部-形成层细胞活力。 从斜叶桉成树上切下带有形成层和韧皮部的圆形树皮切片,将质量为1-30 mg不等的样品在20–85°C 的温 度处理中放置1分钟,并在室温条件下在0.8%的2,3,5-氯化三苯基四氮唑(TTC)中保存20–22小时以检测 细胞活力。用乙醇冷萃取得到1,3,5-三苯基四唑甲臜(TPF),在485 nm处测定吸光度。结果表明,TTC还原 法准确地量化了组织切片(包括维管形成层)中细胞活力随温度升高而下降的情况,并确定60°C 为桉树物种形成层-韧皮部细胞的临界温度。细胞活力按[TPF 处理温度]/[TPF 20°C]计算,在20-85°C之间下降90%。细胞活力结果证实,在50-70°C的温度区间,经过1分钟体外组织加热,桉树的组织坏死显著 增加。TTC 方法显示细胞活力随温度升高而下降,这与温度处理和中性红染色处理后独立获得的活细胞计 数一致。     

Homofusion of Golgi secretory vesicles in flax phloem fibers during formation of the gelatinous secondary cell wall

The gelatinous type of secondary cell wall is present in tension wood and in phloem fibers of many plants. It is characterized by the absence of xylan and lignin, a high cellulose content and axially orientated microfibrils in the huge S2 layer. In flax phloem fiber, the major non-cellulosic component of such cell walls is tissue-specific galactan, which is tightly bound to cellulose. Ultrastructural analysis of flax fiber revealed that initiation of gelatinous secondary cell wall formation was accompanied by the accumulation of specific Golgi vesicles, which had a characteristic bicolor (dark-light) appearance and were easily distinguishable from vesicles made in different tissues and during the other stages of fiber development. Many of the bicolor vesicles appeared to fuse with each other, forming large vacuoles. The largest observed was 4 mum in diameter. Bicolor vesicles and vacuoles fused with the plasma membrane and spread their content in a characteristic "syringe-like" manner, covering a significant area of periplasm and forming "dark" stripes on the inner wall surface. Both Golgi derivatives and cell wall layers were labeled by LM5 antibody, indicating the presence of tissue- and stage-specific (1-->4)-beta-galactan. We suggest that this specific type of galactan secretion, which allows coverage of a large area of periplasm, is designed to increase the chance of the galactan meeting the cellulose microfibrils while they are still in the process of construction. The membrane fusion machinery of flax fiber must possess special components, which may be crucial for the formation of the gelatinous type cell wall.     

Systemic induction of phloem secondary metabolism and its relationship to resistance to a canker pathogen in Austrian pine

The mechanisms and conditions affecting expression of systemic induced resistance (SIR) in pine are not clearly understood. Two hypotheses were tested here: that SIR against a pathogen induced by either a pathogen or an insect involves coordinated shifts in phloem secondary metabolism; and that fertility affects the production of these compounds. To test these hypotheses, a tripartite system was used comprising Austrian pine (Pinus nigra) grown under three different fertility regimes, the fungal pathogen Diplodia pinea, and the defoliator Neodiprion sertifer. Fungal induction led to systemic accumulation of lignin, phenolic glycosides and stilbenes, whereas insect defoliation led to an increase in germacrene D concentration in branch phloem. Fertility affected the concentrations of only the phenolic glycosides. Multivariate analyses showed coregulation of compounds within at least three consistent groupings: phenolic glycosides, stilbenes and monoterpenes. As groups and as individual compounds, accumulation of phenolic glycosides and stilbenes was negatively correlated with disease susceptibility. The experimental manipulation of the phenolics and terpenoids metabolic networks achieved in this study by biotic induction and changes in nutrient availability suggests that lignin, phenolic glycosides and stilbenes are important biochemical factors in the expression of SIR against the pathogen in this system.     

Wound phloem in transition to bundle phloem in primary roots ofPisum sativum L.

Summary 48 hours after interrupting the root stele ofPisum, wound phloem initiated (proximally or distally to the wound) to reconnect the vascular stumps was found to contain some nucleate wound-sieve elements. At the elongating end of an incomplete wound-sieve tube these elements exhibit a sequence of ultrastructural changes as known from protophloem-sieve tubes. Elongation occurs by the addition of newly divided (wound-) sieve-element/companion-cell complexes. In order to dedifferentiate and assume a new specialization formerly quiescent stelar or cortical cells require at least one (mostly more) preliminary division. Companion cells are consequently obligatory sister cells to wound-sieve elements.By reconstruction using serial sections it could be shown that wound-sieve tubes elongate bidirectionally, starting in an early activated procambial cell of the stele. The elongation is directed by the existence of plasmodesmata, preferably when lying in primary pit fields, and by the plane of preceding divisions. Thus, the developing wound-sieve tube can deviate from the damaged bundle and radiate into the cortex as soon as the plane of the preceding divisions is favourable. In the opposite direction, elongating wound-sieve tubes run parallel to pre-existing phloem traces, thus broading their base at the bundle for the deviating part of the wound-sieve tube. Frequently an individual wound-sieve tube is supplemented at the bundle by a further wound-sieve tube which is partly running parallel to it. Both sieve tubes are interlinked with sieve plates by three-poled sieve elements.Ultrastructurally, the developmental changes of nucleate wound-sieve elements follow the known pattern. In spite of its contrasting origin and odd shape a mature wound-sieve element eventually has the same contents as regular sieve elements: sieve-element plastids, mitochondria, stacked ER and small amounts of P-protein within an electronlucent cytoplasm.     

Effects of the hydraulic coupling between xylem and phloem on diurnal phloem diameter variation

Measurements of diurnal diameter variations of the xylem and phloem are a promising tool for studying plant hydraulics and xylem-phloem interactions in field conditions. However, both the theoretical framework and the experimental verification needed to interpret phloem diameter data are incomplete. In this study, we analytically evaluate the effects of changing the radial conductance between the xylem and the phloem on phloem diameter variations and test the theory using simple manipulation experiments. Our results show that phloem diameter variations are mainly caused by changes in the radial flow rate of water between the xylem and the phloem. Reducing the hydraulic conductance between these tissues decreases the amplitude of phloem diameter variation and increases the time lag between xylem and phloem diameter variation in a predictable manner. Variation in the amplitude and timing of diameter variations that cannot be explained by changes in the hydraulic conductance, could be related to changes in the osmotic concentration in the phloem.     

Diversity of the superfamily of phloem lectins (phloem protein 2) in angiosperms

Phloem protein 2 (PP2) is one of the most abundant and enigmatic proteins in the phloem sap. Although thought to be associated with structural P-protein, PP2 is translocated in the assimilate stream where its lectin activity or RNA-binding properties can exert effects over long distances. Analyzing the diversity of these proteins in vascular plants led to the identification of PP2-like genes in species from 17 angiosperm and gymnosperm genera. This wide distribution of PP2 genes in the plant kingdom indicates that they are ancient and common in vascular plants. Their presence in cereals and gymnosperms, both of which lack structural P-protein, also supports a wider role for these proteins. Within this superfamily, PP2 proteins have considerable size polymorphism. This is attributable to variability in the length of the amino terminus that extends from a highly conserved domain. The conserved PP2 domain was identified in the proteins encoded by six genes from several cucurbits, celery (Apium graveolens), and Arabidopsis that are specifically expressed in the sieve element-companion cell complex. The acquisition of additional modular domains in the amino-terminal extensions of other PP2-like proteins could reflect divergence from its phloem function.