DEVELOPMENT OF CHAMBERED PITH IN STEMS OF PHYTOLACCA AMERICANA L. (PHYTOLACCACEAE)

An integrated microscopic (light and electron microscopy) and macroscopic investigation of chambered pith development was made of Phytolacca americana L. Terminal internodes have a solid pith cylinder in contrast to the alternating diaphragms and chambers occurring in subjacent pith. Macroscopically, chambers and diaphragms of any one internode are of equal size. Microscopically, diaphragms vary in height within an internode (from 1–6 cells high). Nevertheless, all diaphragms become thicker circumferentially (5–12 cells high) and connect with long files of intact peripheral pith cells. Diaphragm cells have a large centrally positioned vacuole with a thin, parietal layer of cytoplasm; nuclei, mitochondria, endoplasmic reticulum, and unidentified organelles differentiate in the cytoplasm of diaphragm cells. Although schizogenous activity has most often been implicated as the mechanism by which chambered pith develops in vegetative organs of angiosperms, the results of this study show that cavities in pokeweed result from both schizogenous and lysigenous mechanisms. Schizogeny is suggested by the fact that central pith cells of terminal internodes are longer and thinner walled than peripheral pith cells arranged in vertical files, thus indicating elongation of cells as a possible result of internode elongation. The precise developmental pattern and arrangement of chambers and diaphragms also suggest schizogenous processes. Lysigenous or enzymatic activity is indicated by the fact that cavities are bounded by broken cells, and wall fragments and organelles are often found within enlarging cavities. Chamber formation occurs continuously acropetally and centrifugally in the central pith. A comparison of diaphragms is made with Liriodendron tulipifera and Juglans nigra in an attempt to resolve differences in structure and terminology regarding the differentiation of chambered and diaphragmed pith.     

Comparison of Morphological and Anatomical Characteristics of <i>Taxus chinensis</i> var. <i>mairei </i>Seedlings Root under Waterlogging Stress in Different Substrates

Four different ratios of river sand, ceramic pellets, vermiculite and perlite (1:1), and field soil were selected as the substrates in this experiment, and four gradient levels of root waterlogging, half waterlogging, full waterlogging and normal were set to investigate the effects of different gradients of waterlogging stress on the root morphology of Taxus chinensis var. mairei seedlings under different substrates. In this study, the root anatomical structure of Taxus chinensis var. mairei under different waterlogging stress was observed by the paraffin section method. The roots of T. chinensis var. mairei were diarch, with no pith and resin canals. There was a large number of tannins in the pericycle of the aerial adventitious roots of seedlings adapted to waterlogging. Also, the endodermis has obvious casparian strip thickening, and there were 4-5 layers of large parenchymatous cells in the close to the inner side of the pericycle in the vascular cylinder, which could increase the storage capacity, and transport capacity of the root. Under the treatment of root waterlogging stress, the development of plant roots in the mixed substrate of vermiculite and, perlite was the earliest. Under half waterlogging stress, T. chinensis var. mairei seedlings treated with various substrates all could better adapt to the environment of waterlogging stress. Under the stress of fully waterlogging, the roots of seedlings planted in river sand substrate developed secondary growth.     

EFFECT OF ETHYLENE ON CELLULAR DIFFERENTIATION IN ETIOLATED PEA SEEDLINGS

Ethylene (50 ppm) and 2,4-D (1 mm ) inhibit xylogenesis and completely prevent fiber lignification in the third internode of etiolated pea seedlings (Pisum sativum var. Alaska) during 96–240 hr of treatment. When ethylene is removed after 72 hr, normal differentiation resumes; thus the gas causes no irreversible damage to the cells. A combined treatment with 50 ppm ethylene and 50 μm 2,4-D or treatment with 1 mm 2,4-D alone causes inhibition similar to that observed with 50 ppm ethylene, and 50 μm 2,4-D is partially effective. The inhibition of xylogenesis and fiber lignification caused by 2,4-D is partly or wholly due to auxin-induced ethylene production.     

A localised decrease of GA1 in shoot tips of Salix pentandra seedings precedes cessation of shoot elongation under short photoperiod

By application of a recently developed method allowing analysis of gibberellins (GAs) in mg amounts of tissue, the effect of photoperiod on levels of GAs in shoot tips of individual seedlings of the woody species Salix pentandra was studied. In elongating long day-grown seedlings, maximum levels of GA₁ were found 5–20 mm below the apex, approximately twice the levels in other segments. After exposure of plants to 5 or 15 short days, the levels of GA₁ were about 50% lower within this specific region of the stem, as compared with seedlings grown under long days. Short day-induced cessation of shoot elongation also correlated with overall declines in the levels of GA₅₃, GA₁₉, GA₂₀ and GA₈, Within each photoperiodic treatment the levels of these GAs were generally relatively similar throughout the upper 35 mm of stems. No differences in internode lengths or in lengths of pith or epidermal cells were found in plants grown under long days compared with those exposed to 5 short days. In both cases, cells in mitosis were observed in the subapical stem tissues of shoot tips. After 15 short days, stem elongation was completed, and dividing cells were generally not found in the subapical part of the stem. However, short day exposure did not prevent elongation of internodes and cells differentiated before the treatment was started. Thus, the localised decrease in level of GA₁ in shoot tips under short days precedes the morphological and anatomical changes connected with the short day-induced cessation of elongation growth. This supports the hypothesised role for GA₁ in photoperiodic control of shoot elongation in S. pentandra .     

Localization of a nuclear protein during development in the pea seedling

The subcellular localization of a nuclear antigenic protein has been accomplished through the application of immunogold methodology for immunocytochemistry. Protein extracted from internode tissue of pea seedlings was used for the production of a monoclonal antibody that was applied to tissue sections for electron microscopy. As second antibody, goat anti-mouse IgG conjugated with colloidal gold served as label. The 30 kDa nuclear protein was localized in internode tissue only in the chromatin of differentiating and mature companion cells and parenchyma cells of the phloem. By contrast, virtually all of the meristematic, undifferentiated, cells in the apical dome exhibited nuclear labeling. In young, differentiating internode tissue immediately behind the apex nuclear labeling was found in all cell types except pith parenchyma. The changing localization of this 30 kDa protein with the progression of cell differentiation suggests the possibility that this protein may, in some way, be related to regulatory events of development.     

POLAR TRANSPORT OF GROWTH-REGULATORS IN PITH AND VASCULAR TISSUES OF COLEUS STEMS

Movement of IAA-C¹⁴ and 2,4-D-C¹⁴ through cylinders of known size and histology was compared using liquid scintillation counting. Both auxins showed strongly polar movement, even through pith parenchyma cut from Coleus internode #5, the youngest internode to have ceased elongation. The polar movement was correlated with sizable elongation of the excised cylinders. Velocities of basipetal movement for a given auxin, as determined by the intercept method, showed small or negligible differences between pith and “corner” cylinders. (Corner cylinders comprised mostly vascular tissue, plus some cortical, pith, and epidermal cells.) For IAA, basipetal velocities ranged from 2.1 to 3.3 mm per hr; for 2,4-D, they were 0.6–0.8. For both auxins there was much more net loss into corner than into pith cylinders, a difference associated with the fact that corner cylinders showed 10 times as many cells in transection. More 2,4-D moved basipetally through corner than through pith cylinders and the reverse was true of IAA. By chromatographic evidence, all the radioactivity in the basal receiving blocks was still associated with the auxin molecules.     

A SEASONAL STUDY OF THE VEGETATIVE SHOOT APEX AND THE PATTERN OF PITH DEVELOPMENT IN HIBISCUS SYRIACUS

Tolbert , Robert J. (West Virginia U., Morgantown.) A seasonal study of the vegetative shoot apex and the pattern of pith development in Hibiscus syriacus. Amer. Jour. Bot. 48(3): 249–255. Illus. 1961.—The shoot apex of Hibiscus syriacus L. is described as having a cytohistological zonation superimposed on a tunica-corpus configuration. The apex is flat-topped or may have a saddle-back or concave appearance as seen in median longitudinal section. The metrameristem, consisting of the tunica and corpus initials, is comprised of large, light-staining, vacuolate cells that have thick cell walls and exhibit much dark-staining intercellular substance. Surrounding the metrameristem is the flanking meristem, which is responsible for the outer layers of the shoot, and from which the leaf primordia arise. The pith rib meristem lies below the metrameristem and consists of files of cells that are responsible for the pith. There are no major seasonal changes in the structure of the apex during the yearly cycle. The pith displays a long-shoot type of development with the cells remaining in distinct files during the first flush of growth in the spring. As growth slows and internode elongation is gradually reduced, the pith displays the characteristic short-shoot type of development, consisting of a spongy tissue of rounded cells with many intercellular spaces and no distinct files of cells. A crown is differentiated across the top of the pith at the end of the growth period. This consists of a band of cells with thick, dark-staining cell walls, which separates by the apex from the last year's growth. In contrast to many gymnosperms, this crown is dispersed by renewed cell activity the following spring.     

Nuclear changes accompanying cell differentiation in stems of Pisum sativum L.

Summary Nuclear DNA, nuclear protein and nuclear size have been measured in cells of the cortex, pith and vascular tissue from three successive internodes in the stem of Pisum sativum. New techniques of computer-linked cytophotometry were used to measure these parameters simultaneously in both section and squash preparations. In cortical cells no endoreduplicated nuclei were seen in the internodes measured. In cortical cells from the oldest internode measured, a population of large nuclei with the 2C DNA amount was observed which was not present in the younger internodes. In the oldest pith nuclei measured a few 8C nuclei were present, but maturing pith was most characterized by increasing nuclear size and the population of nuclei accumulating with the 4C DNA amount. Polyploid nuclei were present in all of the vascular tissue measured, including the youngest internode. Maturing vascular tissue was also characterized by increasing nuclear size. Nuclear protein measurements demonstrated a close link between nuclear protein and nuclear size and suggest that increased nuclear size, with constant DNA content, may be due to increased nuclear protein. This raises the question of the nature and function of this nuclear protein, perhaps more characteristic of differentiating cells than dividing cells.To whom offprint requests should be sent     

Effects of gibberellin on the cellular dynamics of dwarf pea internode development

This study analysed the dynamics of cell production and extension, and how these were affected by applied gibberellic acid (GA₃), during internode development in dwarf peas (Pisum sativum L. cv. Meteor). Image analysis was used to obtain cell number and length data for entire cell columns along the epidermis, the two outermost cortical layers, and the pith, from internode 7, over a time period covering the whole of the internode's growth phase. For a few days following the inception of an internode at the shoot apex, little further growth occurred, and there was no significant effect of GA₃ on cell division or cell extension. The subsequent growth of the internode was stimulated more than fourfold by GA₃ as a result of the production of more than twice the number of cells, which were twice as long. At least 96.5% of the cells of the mature internode were actually formed within the internode itself during this period of growth, demonstrating that the internode cells themselves represent the morphogenetic site of response to GA₃. Mitoses and cell extension occurred along the full length of the internode throughout its development. The daily changes in cell numbers were modelled by the Richards function, and manipulations of the fitted functions to reveal time trends of absolute and specific cell production rates were performed for each stem tissue. The increase in cell numbers in the +GA₃ plants was brought about by an increase in the rate of cell production, over a shorter time interval; specific cell production rates declined continuously from initial rapid rates in the +GA₃ epidermis and pith, but declined more slowly in the cortex. The control (−GA₃) epidermis and cortex cells exhibited a constant specific cell production rate (i.e. purely exponential) for several days. Cell extension rates were calculated so as to compensate for the size-reduction effects of concurrent cell division. These calculations confirmed that `real' cell extension rates were higher in the +GA₃ internodes. Models of the cellular controls of internode growth, based on the estimated dynamics of cell division and extension, are discussed. Received: 1 July 1997 / Accepted: 30 July 1997     

Phenotypic plasticity of internode elongation stimulated by deep-seeding and ethylene in wheat seedlings

Deep-seeding and ethylene were found to stimulate extension growth of the first internode of intact wheat (Triticum aestivum L.) seedlings in darkness. Seedlings of Hon Mang Mai emerged from much deeper in the soil than the seedlings of the other varieties used and their first internodes elongated to a much greater extent in response to ethylene. Carbon dioxide slowed elongation of the first internode and inhibited ethylene action. Elongation of the first internode due to deep-seeding and ethylene treatment showed high heritabilities, suggesting a genetic basis underlying those traits.     

Vertical and radial profiles in tracheid characteristics along the trunk of Douglas-fir trees with implications for water transport

The main stems of three young Douglas-fir (Pseudotsuga menziesii var. menziesii (Mirbel) Franco) trees were dissected to obtain samples of secondary xylem from internodes axially along the trunk and radially within each internode. From these samples, measurements were obtained of tracheid diameter, length, the number of inter-tracheid pits per tracheid, and the diameter of the pit membranes. In addition, samples were obtained along the trunks of three old growth trees and also a small sample of roots for measurement of tracheid diameter. A gradient was apparent in all measured anatomical characters vertically along a sequence among the outer growth rings. These gradients arose not because of a gradient vertically along the internodes, but because of the strong gradients present at each internode among growth rings out from the pith. Tracheid characteristics were correlated: wider and longer tracheids had more numerous pits and wider pits, such that total pit area was about 6% of tracheid wall area independent of tracheid size. A stem model combining growth rings in parallel and internodes in series allowed for estimates of whole trunk conductance as a function of tree age. Conductance of the stem (xylem area specific conductivity) declined during the early growth of the trees, but appeared to approach a stable value as the trees aged.     

Pith Autolysis in Herbaceous Dicotyledonous Plants. A Physiological Ecological Study of Pith Autolysis under Native Conditions with Special Attention to the Wild Plant Impatiens capensis Meerb.

Pith autolysis, in the past studied only in herbaceous cultivateddicotyledonous plants, commonly occurs in wild plants. A surveyof pith autolysis in wild plants was conducted in several differentbiomes in both North Carolina, USA and in Belgium. An objectivemethod and a subjective method were developed and used to estimatethe amount of pith autolysis in the internodes of bean (Phaseolusvulgaris L.) and the wild plant jewelweed (Impatiens capensisMeerb.) under experimental conditions. A ranking system alsowas used to quantify pith autolysis in other wild plants. Inthe field, a positive correlation was found between the rateof internode elongation and pith autolysis. Injections of sucrose solution were found to reduce the amountof pith autolysis developed in the upper internodes of jewelweedwhen compared to injections of control solution. Extracts fromthe nodal tissues of jewelweed increased the amount of pithautolysis developed by both jewelweed in the field and beanin the greenhouse. This increase in pith autolysis may be dueto a principle present in the jewelweed nodes that signals orcauses the supertending internodes to become hollow.Copyright1995, 1999 Academic Press Pith, autolysis, Phaseolus vulgaris L., Impatiens capensis Meerb., sucrose, internode, hollow stem     

Sucrose Mobilisation in Sugarcane Stalk Induced by Heterotrophic Axillary Bud Growth

A theoretical high-yield sugarcane biofactory can be idealised as containing culm tissue that functions as a secondary source tissue rather than a sink. To investigate this potential process, heterotrophic axillary bud outgrowth from sugarcane (Saccharum spp. hybrids) setts was used as a model system to demonstrate that sucrose is a mobilisable carbon source. The outgrowth and subsequent biomass accumulation of axillary buds from two-eye setts of mature sugarcane stalks grown in the dark was used to measure carbon mobilisation from sett internode pith tissue. After 42 days growth 99.0 ± 0.72% of sett internode pith sucrose was depleted and 2.66 ± 0.16 g of new tissue accumulated. Comparison with a control treatment in which axillary buds were excised at day zero demonstrated that carbon mobilisation was driven by the accumulation of new biomass. Profiling of soluble carbohydrates (viz. sucrose, glucose and fructose), starch, total soluble protein, total amino nitrogen, free amino acids and total insoluble material showed that the sucrose stored in the sett internode pith was the only available carbon source of sufficient size at day zero for the observed biomass accumulation. Other metabolites mobilised were glucose, fructose and some amino acids, notably isoleucine and leucine that were depleted in shoot treatment setts at day 42.     

On the Mode of Action of Carbamate Herbicides

The hypothesis of Mann et al. that carbamate herbicides act as anti-derepressor substances was tested in two systems: cell enlargement and proliferation in cultured tobacco pith, and auxin-induced Avena first internode extension. Isopropyl-N-phenylcarbamate (IPC) and isopropyl-N-(3-chIorophenyl carbamate (CIPC) both severely inhibited kinetin-induced cell proliferation in tobacco pith at 10^?6M. Auxin-induced tobacco cell enlargement was inhibited to the same degree only at 10^?3M. Avena internode extension was insensitive to these compounds in the range 10^?3 to 10^?7M of IPC and 10^?4 to 10^?8M CIPC. The results support but do not prove Mann's hypothesis. Results of various studies with carbamates are interpreted in terms of this mechanism.     

A comparative histological study between normal and fasciated shoots of <Emphasis Type="Italic">Prunus avium</Emphasis> generated <Emphasis Type="Italic">in vitro</Emphasis>

A combination of Murashige and Skoogs medium and N⁶–benzyladenine (BA) at various concentrations (0, 0.1, 0.25, 0.5, 0.75, 1.0, 1.25 and 1.5 mg l^–1) was supplied to shoot tips from root cuttings of a 50-year-old wild-cherry tree (Prunus avium). The concentration of BA in the growing medium was a determining factor with respect to the number of proliferated shoots per explant in vitro.Normal and fasciated shoots were generated when BA was present at 0.5, 0.75, 1.0 and 1.25 mg l^–1 in the medium and the mean numbers of normal shoots per explant were 3.63, 5.37, 8.93 and 7.30 respectively, and those of the fasciated shoots per explant were 0.03, 0.1, 0.47 and 0.4 respectively. Anatomical analysis by confocal microscopy of sections of paraffin-embedded specimens revealed that the cell structure and organization of the cortex and vascular cylinder in the fasciated shoots was similar to that in normal shoots. However, the cross-sectional area of stem of the fasciations was apparently greater than that of the normal shoots. In particular, the volume of vascular tissues, of pith and of some individual parenchyma cells in the cortex and pith was apparently greater in fasciated shoots than in normal shoots. Increases in cytokinesis and morphogenetic activity, such as the development of callus-like regions and the formation of adventitious shoots, were observed in the cortex and pith throughout the fasciations. The fasciated shoots had numerous buds and initiating new shoots at their apices while normal shoots had a single dominant axial bud.     

Laurina mutation affected Coffea arabica tree size and shape mainly through internode dwarfism

The two varieties—Bourbon (B) and its natural mutant Bourbon pointu (BP)—of Arabica coffee (Coffea arabica L.) differ by an epigenetic, monolocus, and recessive laurina mutation that results in pleiotropic effects, such as tree dwarfism and tree-shape modification. The objective of the study was to search for the origin of the differences in size and shape of the tree both at the macroscopic (length and number of internodes, branching angle) and at the microscopic levels (size and number of cells within the internode pith). At the macroscopic level, the laurina mutation acted only by decreasing the internode size. Neither the angle of branching nor the number of internodes was influenced by the mutation. At the microscopic level, the mutation lowered mainly the number of cells present along the longitudinal axis of the internode, and, at a lesser extent, the cell height. Especially, the internode size decreasing explained both the tree dwarfism and the tree-shape modification. In fact, the laurina mutation strengthened the dwarfism of plagiotropic internodes when compared to that of the orthotropic ones, and such an impact was mainly due to a strong cell number decrease. To summarize, two major pleiotropic effects of the laurina mutation can be explained only by a modification of the trade-off between meresis and auxesis during the internode growth. This opens new perspectives for the characterization of the other effects at the hormonal level, and then, for the identification of the gene at the molecular level.     

Internodes of grain legumes — New location for xylem parenchyma transfer cells

Summary Xylem parenchyma transfer cells were observed in the primary and secondary vascular tissue of stem internodes of 21 in 28 species of grain legumes. Their structural features were similar to those of other transfer cells. The relationships of these cells to transfer cells at nodes were investigated. Non-nodulated seedlings ofPhaseolus vulgaris L. formed internode transfer cells if provided mineral nutrients through their roots, but not if grown in distilled water or fed nutrients entirely through their leaves. Wall ingrowths formed in parenchyma of primary xylem ofPhaseolus just before full extension of an internode. The significance of this new location for transfer cells was discussed.     

SHOOT GROWTH AND HISTOGENESIS OF TREES POSSESSING DIVERSE PATTERNS OF SHOOT DEVELOPMENT

Shoot growth and histogenesis were followed in five unrelated tree taxa possessing inherently diverse patterns of shoot development. Following the resumption of growth in spring, each species differs quantitatively in the number of internodes elongating contemporaneously, in rates and duration of internodal elongation and seasonal periodicity of shoot growth. The basic pattern of internode elongation and histogenesis is qualitatively similar in each of the dicotyledonous species observed irrespective of growth habit or final form of the shoot produced. During the intial phase of internode development, growth is essentially uniform throughout young internodes, corresponding to an active period of cell division during which time pith cells increase in size to about one-third their final length. Subsequently, the pattern of cell division shifts progressively upward concomitant with increased elongation and maturation of pith cells in the basal portion of developing internodes. Thereafter, a wave of cell division accompanied by cell elongation continues to proceed acropetally until growth finally ceases in the distal portion of each internode. As long as internode elongation continues, frequently at distances 15–20 cm below the shoot apex, cell divisions still occur in the distal growing portion. As successive portions of each internode mature acropetally, final length of pith cells becomes relatively uniform throughout the internode. During the process of internode growth and development, cell lengths increase only two- to threefold, whereas cell numbers increase ten- to 30-fold, indicating the dominant role of cell division and increases in cell number to final internode length. Morphological patterns of shoot expression associated with differences in internode lengths along the axis of either preformed or neoformed shoots, as well as sylleptic branches, are due to differences in cell number rather than final cell length. Significant variations in final internode lengths along the axis of episodic shoots, caused by either endogenous or exogenous factors, are also attributed to differences in cell number.     

接骨草的显微结构及其绿原酸组织化学定位研究

运用石蜡切片法和荧光显微镜观察法研究了3个不同接骨草(Sambucus chinensis Lindl.)居群营养器官的显微结构及其绿原酸的分布规律。结果表明:(1)接骨草地上茎厚角组织明显,髓部由大小不等的两类薄壁细胞组成,且有单宁细胞分布;地下根状茎厚角组织细胞小,髓部薄壁细胞大小差异不明显,皮层及髓中有油细胞分布。(2)叶片为异面叶,栅栏组织细胞为短柱状,油细胞不明显。(3)绿原酸分布在根状茎皮层部分细胞、茎厚角组织部分细胞及叶片的海绵组织中,以海绵组织中含量最高。研究认为,髓部薄壁细胞大小的差异可作为接骨草的一个鉴别特征;荧光显微镜观察法可迅速准确显示绿原酸的分布;在所研究的3个接骨草居群中,怀化居群的绿原酸含量最高,若以绿原酸为有效成分来采收接骨草,可以只采收叶。