毛白杨维管形成层带细胞超微结构的季节性变化

木材(次生木质部)是树木形成层细胞分化的产物,形成层的活动方式不仅影响木材的产量,而且影响木材的结构和性质.利用透射电子显微镜观察了生长在北京地区的毛白杨(Populus tomentosa Carr.)枝条形成层带细胞一个完整活动周期的超微结构变化.在木质部母细胞完全恢复活动之前,形成层纺锤状原始细胞的分裂和韧皮部细胞的分化已经开始.枝条上芽的展开和幼叶的生长可能决定了形成层带细胞的这种活动方式.透射电镜观察更清楚地揭示了树木形成层细胞在活动初期的分化特点.活动期形成层细胞中的大液泡在进入休眠期后逐渐分成许多小液泡分散在细胞质中.随着液泡融合逐渐消失的深色蛋白类物质又重新充满了大部分液泡.油滴和淀粉颗粒的年变化情况同液泡中的蛋白类物质基本相似.无论在活动期还是休眠期,形成层纺锤形细胞的质膜上都发现有许多可能与物质运输有关的小泡状内折.由核膜、内质网和高尔基体及其分泌小泡组成的细胞内膜系统,在形成层活动周期的不同阶段,其形态和分布明显不同,尤其在形成层细胞的恢复活动及其衍生木质部细胞次生壁的沉积过程中发挥着重要作用.整个活动周期中,形成层纺锤形细胞的径向壁都比弦向壁厚,处在休眠期的形成层带细胞,其径向壁与弦向壁的差别则更明显.形成层恢复活动时,径向壁上特别是与弦向壁相连的角隅处出现部分自溶现象.细胞壁特别是径向壁的变薄是形成层细胞恢复活动的重要特征.     

Nucleolar activity in the fusiform cambial cells of Abies balsamea (Pinaceae): effect of season and age

Nucleolar involvement in the regulation of the activity-rest-quiescence cycle of the vascular cambium was assessed by determining the seasonal variation in number, diameter, and volume of nucleoli in fusiform cells of Abies balsamea (L.) Mill. The cells were isolated from 1- and 19-yr-old cambia and stained with either silver nitrate or Feulgen + naphthol yellow-S. The ability of fusiform cells to incorporate [5-³H]-uridine into nuclei and nucleoli was also determined. In the 1-yr-old cambium, the activity of the nucleoli, as evidenced by their diameter, total volume per cell, and intensity of staining with silver nitrate, exhibited two maxima during the year—a large one during cambial reactivation in April-May and a small one during the rest-quiescence transition in October. Incorporation of radiolabeled uridine at 20 C was low at the end of the active period and increased during the rest–quiescence transition, suggesting that the quiescent, but not the resting, cambium can rapidly resume nucleolar activity when the temperature is permissive. The number of nucleoli per cell varied between two and eight, and was higher during the dormant than the active period. The increase in number took place during the autumnal activity–rest–quiescence transition, when cambial cells were arrested in the G₁ phase of the cell cycle. Similar seasonal changes in nucleolar morphology were observed in the 19-yr-old cambium. Nucleolar diameter and total nucleolar volume were larger in the 19-yr-old cambium than in the 1-yr-old cambium, whereas nucleolar number was lower. Th results suggest that repression of rRNA genes underlies the development of rest when the cambium will not produce new cells.     

Cambial activity,annual rhythm of xylem production in relation to phenology and climatic factors and lignification pattern during xylogenesis in drum-stick tree (Moringa oleifera)

The interrelationship among seasonality of cambium, wood formation, cell size variation, lignification, tree phenology and climatic factors has been examined in Moringa oleifera, a tropical evergreen tree. The vascular cambium in Moringa is a storied with a distinct seasonal variation in its structure due to dimensional changes in rays. Though cambium remains active throughout the year it is sensitive to water availability. Peak cambial cell division and rate of xylem differentiation are influenced by average rainfall during the monsoon period. Cambial cell division reaches higher up in the tree trunk when it is supporting a high number of branches and leaves. Statistical analysis of cell size variation and climate factors revealed that xylem cell development is greatly influenced by rainfall and rarely by temperature. Lengths of fusiform initials and vessel elements are positively correlated. The pattern of lignification during xylogenesis shows that the vessels are the first element to develop lignified walls and ray cells are the last elements to become lignified. Fiber cell walls show more syringyl lignin, while the cell walls of other xylem elements are characterized by relatively more guaiacyl lignin units.     

On the Cytochemistry of Cell Wall Formation in Poplar Trees

Abstract: The ultrastructure of cell walls and the mechanisms of cell wall formation are still not fully understood. The objective of our study was therefore to obtain additional fine structural details on the deposition of cell wall components during the differentiation of xylem cells in hybrid aspen ( Populus tremula L. × P. tremuloides Michx.) we used as a model tree. At the electron microscope level, PATAg staining revealed a successive deposition of polysaccharides with increasing distance from the cambium. Staining with potassium permanganate and UV microspectrophotometry showed that the cell walls were lignified, with some delay to the deposition of polysaccharides. Immunogold labelling of three lignin types in developing cell walls varied with progressive deposition of cell wall layers. Condensed lignin subunits were localized in corners of cells adjacent to the cambium prior to S1 formation, whereas non-condensed lignin subunits became labelled only in later stages - in secondary walls near cell corners and simultaneously with the completion of S1 formation. As S2 polysaccharide deposition progressed, the labelling extended towards the lumen. Labelling of peroxidases revealed their presence in cell corner regions of young xylem cells, still lacking a secondary wall, implying that peroxidases are incorporated into the developing cell wall at early developmental stages. A weak labelling of middle lamella regions and secondary walls could also be seen at later stages. The results are discussed in relation to current knowledge on the succession of polysaccharide and lignin deposition in woody cell walls.     

Decomposition of Microbial Cell Components in a Semi-Arid Grassland Soil

Cell component fractions (¹⁴C-labeled) were prepared from bacterial and fungal cultures isolated from the Pawnee National Grassland in northeastern Colorado and tested for seasonal changes in degradability. The decomposition of cell component fractions was monitored from May to December of 1977 and during March of 1978, using soil samples taken at 2- to 3-week intervals. The release of ¹⁴CO₂ from bacterial and fungal cell walls was inversely related (P < 0.01) to the soil moisture content. Except for cytoplasm isolated from an Aspergillus sp., all other cytoplasmic and polysaccharide fractions did not demonstrate a significant relationship between soil moisture and decomposability. In general, bacterial cell walls and polysaccharides were more susceptible to decomposition than fungal cell walls, although the seasonal changes in decomposability for both fractions were similar. These patterns of cell component utilization indicate that the decomposition of cell wall material may be more closely linked, on an inverse basis, to the availability of soil moisture and release of soluble, low-molecular-weight organics resulting from primary production events.     

Changes in cell wall constituents during the cell cycle in a synchronous culture of Catharanthus roseus

Changes in cell wall constituents during the cell cycle were investigated using a synchronous culture of Catharanthus roseus (L.) G. Don which was obtained by the double phosphate starvation method (S. Amino et al. 1983. Physiol. Plant. 59: 393–396). Cell walls isolated from the cells in each phase of the cell cycle were fractionated into EDTA-soluble (pectin), 5 and 24% KOH-soluble (hemicellulose) and 24% KOH-insoluble (cellulose) fractions. Their sugar compositions were investigated by gas chromatography and methylation analysis. The following changes were observed: (1) a significant increase in total cell walls in the G1 phase after cell division, (2) a temporary increase in the relative amount of the EDTA-soluble fraction during cytokinesis, (3) an increase in the relative amount of galactose, probably 4-linked galactose, in the EDTA-soluble fraction prior to cytokinesis, (4) a temporary increase in the relative amount of 3-linked glucose during cytokinesis, (5) little change in the composition of polysaccharides throughout the cell cycle in the 24% KOH-soluble fraction, which consisted mainly of xyloglucan. The changes observed are discussed in relation to the progression and physiological significance of each phase of the cell cycle.     

Cold stability of microtubules in wood-forming tissues of conifers during seasons of active and dormant cambium

The cold stability of microtubules during seasons of active and dormant cambium was analyzed in the conifers Abies firma, Abies sachalinensis and Larix leptolepis by immunofluorescence microscopy. Samples were fixed at room temperature and at a low temperature of 2–3°C to examine the effects of low temperature on the stability of microtubules. Microtubules were visible in cambium, xylem cells and phloem cells after fixation at room temperature during seasons of active and dormant cambium. By contrast, fixation at low temperature depolymerized microtubules in cambial cells, differentiating tracheids, differentiating xylem ray parenchyma and phloem ray parenchyma cells during the active season. However, similar fixation did not depolymerize microtubules during cambial dormancy in winter. Our results indicate that the stability of microtubules in cambial cells and cambial derivatives at low temperature differs between seasons of active and dormant cambium. Moreover, the change in the stability of microtubules that we observed at low temperature might be closely related to seasonal changes in the cold tolerance of conifers. In addition, low-temperature fixation depolymerized microtubules in cambial cells and differentiating cells that had thin primary cell walls, while such low-temperature fixation did not depolymerize microtubules in differentiating secondary xylem ray parenchyma cells and tracheids that had thick secondary cell walls. The stability of microtubules at low temperature appears to depend on the structure of the cell wall, namely, primary or secondary. Therefore, we propose that the secondary cell wall might be responsible for the cold stability of microtubules in differentiating secondary xylem cells of conifers.     

Structural Changes in the Vascular Cambium of Pinus strobus L. during an Annual Cycle

The changes in the ultrastructure of cambial cells of easternwhite pine (Pinus strobus L.) during an annual cycle are observedand recorded as are relationships of cambial cells during dormancyand at resumption of cambial activity. Cambial activity wasresumed late in March or early in April, when a few cells dividedpericlinally. Cambial activity reached a maximum during thelatter part of May with 15 to 20 undifferentiated cells present.In July it declined markedly, and the number of undifferentiatedcells equalled that of the dormant period. The xylem and phloemtissue cells produced late in the annual cycle overwinteredat varying developmental stages. In October cambial cells structurallyresembled dormant cells. The number of dormant cells in easternwhite pine cambium varied from 6 to 10. Active cells were characterizedby a large central vacuole, by an abundance of all cell organelles,and by thin cell walls. Dormant cells were characterized bynumerous small vacuoles, by structurally and quantitativelymodified cell organelles, and by relatively thick cell walls.     

A seasonal cycle of cell wall structure is accompanied by a cyclical rearrangement of cortical microtubules in fusiform cambial cells within taproots of Aesculus hippocastanum (Hippocastanaceae)

Aspects of the structure and ultrastructure of the fusiform cambial cells of the taproot of Aesculus hippocastanum L. (horse chestnut) are described in relation to the seasonal cycle of cambial activity and dormancy. Particular attention is directed at cell walls and the microtubule and microfilament components of the cytoskeleton, using a range of cytochemical and immunolocalization techniques at the optical and electron-microscopical levels. During the dormant phase, cambial cell walls are thick and multi-layered, the cells possess a helical array of cortical microtubules, and microfilament bundles are oriented axially. In the early stages of reactivation, vesicle-like profiles are associated with the cell walls, whereas arrangement of the cytoskeletal elements remains unchanged. In the succeeding active phase, the cell walls are thin, and cortical microtubules form a random array, although microfilament bundles maintain a near-axial orientation. The observations are discussed in relation to the seasonal cycle of wall structure and cortical microtubule rearrangement within the vascular cambium of hardwood trees. It is suggested that the cell-wall thickening at the onset of cambial dormancy, which is associated with the presence of a helical cortical microtubule array, should be considered to be secondary wall thickening, and that selective lysis of this secondary wall layer during cambial reactivation restores the thinner, primary wall found around active cambial cells.     

Changes in Cell Wall Composition during Ripening of Grape Berries

Cell walls were isolated from the mesocarp of grape (Vitis vinifera L.) berries at developmental stages from before veraison through to the final ripe berry. Fluorescence and light microscopy of intact berries revealed no measurable change in cell wall thickness as the mesocarp cells expanded in the ripening fruit. Isolated walls were analyzed for their protein contents and amino acid compositions, and for changes in the composition and solubility of constituent polysaccharides during development. Increases in protein content after veraison were accompanied by an approximate 3-fold increase in hydroxyproline content. The type I arabinogalactan content of the pectic polysaccharides decreased from approximately 20 mol % of total wall polysaccharides to about 4 mol % of wall polysaccharides during berry development. Galacturonan content increased from 26 to 41 mol % of wall polysaccharides, and the galacturonan appeared to become more soluble as ripening progressed. After an initial decrease in the degree of esterification of pectic polysaccharides, no further changes were observed nor were there large variations in cellulose (30–35 mol % of wall polysaccharides) or xyloglucan (approximately 10 mol % of wall polysaccharides) contents. Overall, the results indicate that no major changes in cell wall polysaccharide composition occurred during softening of ripening grape berries, but that significant modification of specific polysaccharide components were observed, together with large changes in protein composition.     

Enhancement of secondary xylem cell proliferation by Arabidopsis cyclin D overexpression in tobacco plants

Secondary xylem is composed of daughter cells produced by the vascular cambium in the stem. Cell proliferation of the secondary xylem is the result of long-range cell division in the vascular cambium. Most xylem cells have a thickened secondary cell wall, representing a large amount of biomass storage. Therefore, regulation of cell division in the vascular cambium and differentiation into secondary xylem is important for biomass production. Cell division is regulated by cell cycle regulators. In this study, we confirm that cell cycle regulators influence cell division in the vascular cambium in tobacco. We produced transgenic tobacco that expresses Arabidopsis thaliana cyclin D2;1 (AtcycD2;1) and AtE2Fa-DPa under the control of the CaMV35S promoter. Each gene is a positive regulator of the cell cycle, and is known to influence the transition from G1 phase to S phase. AtcycD2;1-overexpressing tobacco had more secondary xylem cells when compared with control plants. In order to evaluate cell division activity in the vascular cambium, we prepared a Populus trichocarpa cycB1;1 (PtcycB1;1) promoter containing a destruction box motif for ubiquitination and a β-glucuronidase-encoding gene (PtcycB1;1pro:GUS). In transgenic tobacco containing PtcycB1;1pro:GUS, GUS staining was specifically observed in meristem tissues, such as the root apical meristem and vascular cambium. In addition, mitosis-monitoring plants containing AtcycD2;1 had stronger GUS staining in the cambium when compared with control plants. Our results indicated that overexpression of AtcycD enhances cell division in the vascular cambium and increases secondary xylem differentiation in tobacco. Key message We succeeded in inducing cell proliferation of cambium and enlargement of secondary xylem region by AtcycD overexpression. We also evaluated mitotic activity in cambium using cyclin-GUS fusion protein from poplar.     

白皮松形成层的活动周期及其多糖贮量和淀粉酶同工酶的变化

4月初,白皮松(Pinus bungeana Zucc.)形成层带细胞开始增大,未成熟的木质部和韧皮部细胞增多,下旬出现成熟的木质部细胞。5月以后,木质部和韧皮部的形成速度加快,6月初进入晚材形成期。8月初停止产生木质部,9月中旬停止产生韧皮部。多糖颗粒的消长与形成层活动有较强的相关性,恢复活动前后颗粒含量持续增长,6月进入晚材形成期才持续减少,至翌年1月初完全消失,3月又重新积累,并迅速达到高峰。淀粉酶同工酶在活动期只有一条酶带,形成层停止产生木质部后出现了3条特异酶带,12月初又出现了2条特异酶带,这5条酶带都一直存在到形成层恢复活动。     

Degradation of pectic polysaccharides extracted from suspension cultures of carrot by purified exo-polygalacturonase

Highly purified exo-polygalacturonase was obtained from suspension cultures of carrot ( Daucus carota L. cv. Kintoki) by dialysis at pH 5.2, chromatography on DEAE-Sephadex A-50 and on Sephadex G-150, and preparative polyacrylamide disc gel electrophoresis. The enzyme did not attack the isolated carrot cell walls directly, but it had some effect on pectic polysaccharides extracted from the walls. The extracted polysaccharides were fractionated by DEAE-Sephadex A-50 column chromatography yielding four carbohydrate fractions. The major fraction (P-3) was then reacted with the exo-polygalacturonase. The enzyme treatment resulted in hydrolysis of approximately 18% of the glycosyl linkages of fraction P-3 with the release of galacturonic acids. The molecular size estimated by Bio-Gel A-5m gel filtration was not markedly affected by the enzyme action, but the percentage of galacturonosyl residues was clearly reduced. The specific activity of exo-polygalacturonase changed during the growth cycle, in relation to the cell growth.     

Structure of Plant Cell Walls : XIX. Isolation and Characterization of Wall Polysaccharides from Suspension-Cultured Douglas Fir Cells

The partial purification and characterization of cell wall polysaccharides isolated from suspension-cultured Douglas fir (Pseudotsuga menziesii) cells are described. Extraction of isolated cell walls with 1.0 m LiCl solubilized pectic polysaccharides with glycosyl-linkage compositions similar to those of rhamnogalacturonans I and II, pectic polysaccharides isolated from walls of suspension-cultured sycamore cells. Treatment of LiCl-extracted Douglas fir walls with an endo-α-1,4-polygalacturonase released only small, additional amounts of pectic polysaccharide, which had a glycosyl-linkage composition similar to that of rhamnogalacturonan I. Xyloglucan oligosaccharides were released from the endo-α-1,4-polygalacturonase-treated walls by treatment with an endo-β-1,4-glucanase. These oligosaccharides included hepta- and nonasaccharides similar or identical to those released from sycamore cell walls by the same enzyme, and structurally related octa- and decasaccharides similar to those isolated from various angiosperms. Finally, additional xyloglucan and small amounts of xylan were extracted from the endo-β-1,4-glucanase-treated walls by 0.5 n NaOH. The xylan resembled that extracted by NaOH from dicot cell walls in that it contained 2,4- but not 3,4-linked xylosyl residues. In this study, a total of 15% of the cell wall was isolated as pectic material, 10% as xyloglucan, and less than 1% as xylan. The noncellulosic polysaccharides accounted for 26% of the cell walls, cellulose for 23%, protein for 34%, and ash for 5%, for a total of 88% of the cell wall. The cell walls of Douglas fir were more similar to dicot (sycamore) cell walls than to those of graminaceous monocots, because they had a predominance of xyloglucan over xylan as the principle hemicellulose and because they possessed relatively large amounts of rhamnogalacturonan-like pectic polysaccharides.     

Polysaccharide composition of unlignified cell walls of pineapple [Ananas comosus (L.) Merr.] fruit.

The polysaccharides of cell walls isolated from the fleshy, edible part of the fruit of the monocotyledon pineapple [Ananas comosus (L.) Merr.] (family Bromeliaceae) were analyzed chemically. These cell walls were derived mostly from parenchyma cells and were shown histochemically to be unlignified, but they contained ester-linked ferulic acid. The analyses indicated that the noncellulosic polysaccharide composition of the cell walls was intermediate between that of unlignified cell walls of species of the monocotyledon family Poaceae (grasses and cereals) and that of unlignified cell walls of dicotyledons. Glucuronoarabinoxylans were the major non-cellulosic polysaccharides in the pineapple cell walls. Xyloglucans were also present, together with small amounts of pectic polysaccharides and glucomannans (or galactoglucomannans). The large amounts of glucuronoarabinoxylans and small amounts of pectic polysaccharides resemble the noncellulosic polysaccharide composition of the unlignified cell walls of the Poaceae. However, the absence of (1-->3,1-->4)-beta-glucans, the presence of relatively large amounts of xyloglucans, and the possible structure of the xyloglucans resemble the noncellulosic polysaccharide composition of the unlignified cell walls of dicotyledons.     

Relationships between cambial activity,cell differentiation and the localization of starch in storage tissues around the cambium in locally heated stems of<Emphasis Type="Italic"> Abies sachalinensis</Emphasis> (Schmidt) Masters

A study was made, in a cool-temperate zone, of the extent of cell division in the cambium, the extent of differentiation of cambial derivatives, and the localization of storage starch around the cambium in locally heated (22–26°C) stems of the evergreen conifer Abies sachalinensis (Schmidt) Masters during cambial dormancy and immediately after natural reactivation of the cambium. In locally heated regions of stems during cambial dormancy, heating induced localized reactivation of the cambium. However, the cells in the heated and reactivated cambium stopped dividing soon after only a few cells had been generated. In addition, no differentiation of the xylem and the disappearance of starch from storage tissues around the cambium were observed. In regions of stem that had been locally heated after natural reactivation of the cambium, cell division continued in the cambium and earlywood tracheids with a large radial diameter and secondary walls were formed, with abundant starch in the storage tissues around the cambium. Our results suggest that the extent of both cell division in the cambium and cell differentiation depends on the amount of starch in storage tissues around the cambium in the locally heated stems of an evergreen conifer growing in a cool-temperate zone.