Cell division patterns of the protoderm and root cap in the “closed” root apical meristem ofArabidopsis thaliana

Summary The peripheral root cap and protoderm inArabidopsis thaliana are organized into modular packets of cells derived from formative T-divisions of the root cap/protoderm (RCP) initials and subsequent proliferative divisions of their daughter cells. Each module consists of protoderm and peripheral root cap packets derived from the same periclinal T-division event of an RCP initial. Anatomical analyses are used to interpret the history of extensively coordinated cell divisions producing this modular construction. Within a given layer of root cap, the columella and RCP initials divided in a centrifugal sequence from the innermost columella initials toward the RCP initials. All RCP initials in the lineages around the circumference of the root divided nearly simultaneously in waves to form one module prior to the next wave of initial divisions forming a younger module. The peripheral root cap and protoderm packets within each module completed four rounds of proliferative divisions in the axial plane to produce, on average, 16 cells per packet in the basalmost modules in axial view. Peripheral root cap and protoderm cells predominantly in the T-type (trichoblast) lineages also underwent radial divisions as they were displaced basipetally. The regularity in the cellular pattern within the modules suggests a timing mechanism controlling highly coordinated cell division in the initials and their daughter cells.Abbreviations RAM root apical meristem - RCP root cap protoderm - prc peripheral root cap     

Aluminium toxicity expression nutrient uptake,growth and root morphology ofTrifolium repens L. cv. ‘Grasslands Huia’

Summary Hydroponic experiments were undertaken to examine the effect of increasing aluminium levels on the mineral nutrition and root morphology ofT. repens growing in nutrient solution. Toxicity symptoms appear between 27.8 and 47.5 M Al³⁺ activity (148 to 297 M total aluminium). The threshold level corresponding to a 10% reduction in leaf fresh weight is estimated to be approximately 20 M Al³⁺ activity.The concentration of aluminium in the leaves of white clover increases exponentially with aluminium activity in the nutrient solution. The uptake of divalent cations was inhibited but aluminium enhanced potassium and nitrogen concentrations in both leaves and roots.At high pH (pH 6.0) the speciation of aluminium is controlled by the formation of solid aluminium phosphate and aluminium hydroxide except at the lowest aluminium level (37 M) where 99.9 per cent is present as the DTPA complex. As the concentration of total aluminium increases, the percentage of Al-DTPA and soluble aluminium hydroxide decreases whilst solid Al(OH)₃ increases rapidly to reach a maximum of 91.6 percent (of the total aluminium) in the 1180 M aluminium treatment. At pH 4.5 the dominant forms of aluminium are free aluminium ion Al-DTPA, AlSO ₄ ⁺ and AlOH²⁺.The roots of aluminium stressed plants showed symptoms typical of aluminium toxicity.     

In vivo analysis of local wall stiffness at the shoot apical meristem in Arabidopsis using atomic force microscopy

Whereas the morphogenesis of developing organisms is relatively well understood at the molecular level, the contribution of the mechanical properties of the cells to shape changes remains largely unknown, mainly because of the lack of quantified biophysical parameters at cellular or subcellular resolution. Here we designed an atomic force microscopy approach to investigate the elastic modulus of the outer cell wall in living shoot apical meristems (SAMs). SAMs are highly organized structures that contain the plant stem cells, and generate all of the aerial organs of the plant. Building on modeling and experimental data, we designed a protocol that is able to measure very local properties, i.e. within 40-100 nm deep into the wall of living meristematic cells. We identified three levels of complexity at the meristem surface, with significant heterogeneity in stiffness at regional, cellular and even subcellular levels. Strikingly, we found that the outer cell wall was much stiffer at the tip of the meristem (5 ± 2 MPa on average), covering the stem cell pool, than on the flanks of the meristem (1.5 ± 0.7 MPa on average). Altogether, these results demonstrate the existence of a multiscale spatialization of the mechanical properties of the meristem surface, in addition to the previously established molecular and cytological zonation of the SAM, correlating with regional growth rate distribution.     

Living plant cells released from the root cap: A regulator of microbial populations in the rhizosphere?

Barley (Hordeum vulgare L. cv. ‘Onda’) plants were grown in nutrient solutions supplied either 0 (no Ni added), 0.6, or 1.0 μM NiSO₄. Plants supplied 0 μM Ni developed Ni deficiency symptoms; Ni deficiency resulted in the disruption of nitrogen metabolism, and affected the concentration of malate and various inorganic anions in roots, shoots, and grain of barley. The concentrations of 10 of the 11 soluble amino acids determined were 50–200% higher in 30-day-old shoots of plants supplied inadequate Ni levels than in shoots of Ni-supplied plants. The total concentration of all amino acids determined was higher in roots and grain of Ni-deficient plants. Concentrations of NO₃ ^- and Cl^- were also higher in Ni-deficient barley shoots than in Ni-sufficient barley shoots. In contrast, the concentration of alanine in shoots of Ni-deficient barley was reduced to one-third of the concentration in Ni-sufficient plants. The shoot concentrations of malate and SO₄ ^2- were also depressed under Ni-deficient conditions. Total nitrogen concentration in grain, but not in shoots, of Ni-deficient plants was significantly increased over that found in Ni-adequate plants. Nickel deficiency results in marked disruptions of N metabolism, malate and amino acid concentrations in barley. These results are discussed in view of the possible roles of Ni in plants. Supported, in part, by an ITT International Fellowship awarded to PHB and administered by the Institute for International Education, United Nations Plaza, New York, NY. This research was part of the program of the Center for Root-Soil Research. Supported, in part, by an ITT International Fellowship awarded to PHB and administered by the Institute for International Education, United Nations Plaza, New York, NY. This research was part of the program of the Center for Root-Soil Research.     

The effect of herbicide "Chwastox p?ynny 30" on the cell cycle parameters in root meristem of Vicia faba L. subsp. minor

The duration of the cell cycle and its phases after the treatment with herbicide "Chwastox p?ynny 30" was calculated using 3H-thymidine labelling method. Inhibition of DNA synthesis and marked prolongation of G2 + 1/2 M phase were observed. Tested herbicide caused a significant lowering in the mitotic activity and accumulation of metaphase cells.     

Linamarase and other β-glucosidases are present in the cell walls of Trifolium repens L. leaves

Linamarase (EC 3.2.1.21) is a specialized -glucosidase that hydrolyses the cyanogenic glucoside linamarin. Two clones of Trifolium repens L. derived from natural populations, of which one clone exhibited linamarase activity, were used in a comparative study to try to establish the localization of linamarase and other -glucosidases. Two methods were used: the first one was vacuum infiltration of intact leaf cells, followed by centrifugation. A significant amount of linamarase and -glucosidase activity could be extracted from intact tissue by a 0.25 M NaCl solution, indicating that these activities are localized in the apoplast. The second method, immuno-cytofluorescense of microtome sections, confirmed this. It was found that linamarase and other -glucosidases are present in the cell walls, especially those of the epidermal cells, and in the cuticle. However their presence in the cell walls of other tissues i.e. walls of the vessels, could not be excluded. No difference in distribution could be detected between linamarase and other -glucosidases.     

Lepidoptera rhopalocera in the area of the planned construction of the “Altai” gas pipeline

Long-term route surveys of butterflies revealed the species composition and distribution within the area of the planned construction of the “Altai” gas pipeline. The distribution of multispecific communities among the altitudinal belts was characterized. The definite finding localities of 99 species and quantitative assessment of their abundance were reported for the first time. The obtained estimates of butterfly abundance in the territory studied can serve as the basis for the “Biodiversity Damage Cost Estimation.” The cadastral data on the butterflies, obtained for the first time, may be used for further eco- and zoogeographic analysis.     

Formation of biphenyl and dibenzofuran phytoalexins in the transition zones of fire blight-infected stems of Malus domestica cv. ‘Holsteiner Cox’ and Pyrus communis cv. ‘Conference’

In the rosaceous subtribe Pyrinae (formerly subfamily Maloideae), pathogen attack leads to formation of biphenyls and dibenzofurans. Accumulation of these phytoalexins was studied in greenhouse-grown grafted shoots of Malus domestica cv. ‘Holsteiner Cox’ and Pyrus communis cv. ‘Conference’ after inoculation with the fire blight bacterium, Erwinia amylovora. No phytoalexins were found in leaves. However, both classes of defence compounds were detected in the transition zone of stems. The flanking stem segments above and below this zone, which were necrotic and healthy, respectively, were devoid of detectable phytoalexins. The transition zone of apple stems contained the biphenyls 3-hydroxy-5-methoxyaucuparin, aucuparin, noraucuparin and 2′-hydroxyaucuparin and the dibenzofurans eriobofuran and noreriobofuran. In pear, aucuparin, 2′-hydroxyaucuparin, noreriobofuran and in addition 3,4,5-trimethoxybiphenyl were detected. The total phytoalexin content in the transition zone of pear was 25 times lower than that in apple. Leaves and stems of mock-inoculated apple and pear shoots lacked phytoalexins. A number of biphenyls and dibenzofurans were tested for their in vitro antibacterial activity against some Erwinia amylovora strains. The most efficient compound was 3,5-dihydroxybiphenyl (MIC = 115 μg/ml), the immediate product of biphenyl synthase which initiates phytoalexin biosynthesis.     

The role of sucrose and different cytokinins in the in vitro floral morphogenesis of rose (hybrid tea) cv. “First Prize”

Shoots of rose (hybrid tea) cv. “First Prize” were induced to flower in vitro on Murashige and Skoog (MS) medium containing various sucrose concentrations (15, 30 or 45 g l⁻¹) and different phytohormone combinations of different cytokinins [N⁶-benzyladenine (BA); thidiazuron (TDZ) and zeatin] with α-naphthaleneacetic acid (NAA). Results indicate that sucrose is the key factor in floral morphogenesis while cytokinin increases the flowering percentage and helps the normal development of floral buds. From the three cytokinins that were used, BA and zeatin were considered to be more suitable as inductive flowering agents than TDZ. Reduced inorganic and organic salt concentration in MS media had a positive effect on in vitro flowering. The morphology of shoots bearing floral buds varied with different cytokinin treatments. The highest percentage (45%) of flowering was obtained on MS medium supplemented with 3.0 mg l⁻¹ BA, 0.1 mg l⁻¹ NAA and 30 g l⁻¹ sucrose.     

Effects of aluminium and micro-nutrients on the sorption of phosphorus byTrifolium repens L. cv. ‘Grasslands Huia’ from nutrient solutions during plant induced pH changes

Summary The sorption of phosphorus from nutrient solution and the pH change in the nutrient solution were monitored over a 24 hour period forTrifolium repens L. cv. ‘Grasslands Huia’ plants. Two different concentration levels of micro-nutrients (B, Cu, Fe, Mn and Zn) and Al formed the factors of a fractional replicate of a 2⁶ factorial design. Measurements were made at four time intervals (30 minutes after the plants were placed on the pots, 3 hours, 6 hours and 24 hours later). In addition to phosphorus, fourteen other nutrients (including nitrate and ammonium) were monitored throughout the experiment. The sorption of phosphorus was significantly influenced by both aluminium and iron. The effect of aluminium and iron on phosphorus sorption is attributed to physico-chemical sorption processes involving the root surface. However the effect on the removal of phosphorus by boron, copper, manganese and zinc was observed only as first order interaction effects —B−Zn, Cu−Zn, Mn−Zn. Thus these three elements (B, Cu and Mn) only affect phosphorus removal in conjunction with zinc. Aluminium and iron together had a separate but very significant effect on the removal of phosphorus at most periods throughout the experiment. In contrast, pH was affected only by aluminium, iron (the pH drop was enhanced) and manganese (the pH drop was decreased) as main effects independent of the other treatment elements.     

Cloning and sequence of the cDNA encoding the β4 integrin subunit in rat peripheral nerve.

β4 and α6 integrin subunits dimerize to form an adhesion receptor that is necessary to nucleate hemidesmosomes and to anchor epithelial cells to their basal laminae. β4 is also expressed in Schwann cells (which do not contain hemidesmosomes) in peripheral nerve, where it may function in the formation or maintenance of myelin. The cDNA for β4 integrin has been cloned from epithelia-derived human and mouse tissues. We cloned cDNAs encoding β4 integrin from libraries derived from rat peripheral nerve, and determined the complete nucleotide sequence encoding the signal peptide and mature protein. Comparison of the deduced amino acid (aa) sequence revealed 95.1% and 87.5% identity with the mouse and human epithelia-derived sequences, respectively. The amino acid sequence of postulated signal transduction domains in β4 was 100% identical among rat, mouse, and human. Our cDNA clones included two of the four postulated alternatively spliced variants previously described in epithelial clones. Despite the potentially diverse functions of β4 integrin in Schwann cells and keratinocytes, the cDNAs for nerve-derived β4 integrin are highly similar to those cloned from epithelia.     

Cytological investigations of the effect of colchicine on meiosis in Lilium hybrid cv. “Black Beauty” microsporocytes

Using colchicine, two methods have been successfully applied for the induction of tetraploid meiocytes at premeiotic mitosis in the near achiasmate diploid hybrid cultivar Black Beauty of Lilium. The experiments were aimed at understanding whether the achiasmate condition is attributable to insufficient homology for effective pairing or to some genie defect in order to interpret the data obtained from molecular studies. Cytological observations demonstrate a strong correlation between the induction of tetraploidy and subsequent chiasma formation. The chiasma frequency per cell in untreated diploid control meiocytes ranged from 0 to 8 (mean 2.25), while in colchicine treated undoubled cells sampled 14±2 days after the start of treatment it ranged from 0 to 2 (mean 0.148). By comparison, the chiasma frequency per cell in colchicine-doubled tetraploid cells ranged from 29 to 51 (mean 42.24), such cells showing complete or near complete bivalent pairing. These results, similar to those reported previously for other higher plant species, demonstrate that the achiasmatic condition in Black Beauty is not due to a genie defect. The methods developed have made possible hitherto inaccessible biochemical analyses of meiocytes that had been treated with colchicine or other chemicals at premeiotic stages.     

Absence of “void area” in freeze-etched vesicles of the Alnus crispa var. mollis Fern. root nodule endophyte

Nitrogen-fixing root nodules of Alnus crispa var. mollis Fern. were studied by transmission electron microscopy and by freeze-etching technique. Ultrathin sectioning of septate vesicles of the actinomycetal endophyte showed an electron transparent zone, the so-called void area, between the vesicle cell wall and its encapsulation material. This void area was not observed in the freeze-etching replicas of cryoprotected nodular tissue. It is suggested that the void area is the result of the coming-off of the vesicle cell wall from the capsule and that its formation reflects difficulty in fixing the voluminous mature vesicle of the root nodule endophyte.     

Histochemical and chemical studies on pre- and postnatal development of the different systems of “short” and “long” adrenergic neurons in peripheral organs of the rat

Summary The adrenergic innervation in the submaxillary gland, heart, kidney, small intestine, and accessory male genital organs and the development of the adrenal chromaffin cells and the sympathetic ganglia were studied in the rat from 15 days post coitum to 16 days post partum using the fluorescence histochemical method of Falck and Hillarp. The postnatal development of the noradrenaline concentrations in the heart and vas deferens was followed by fluorometric determinations.At about 15 days post coitum, the anlagen of the sympathetic chains were well visible in the form of two dorsal segmented columns of small branching sympathicoblasts exhibiting an intense catecholamine fluorescence. In the midline, ventrally to these two anlagen, another column of sympathicoblasts developed; this seemed to give rise to the prevertebral ganglia and to the short adrenergic neurons supplying the internal genital organs. At the level of the adrenal anlagen, small intensely fluorescent chromaffin cells were collected in two bilateral groups which became enclosed by adreno-cortical cells. This enclosure was, however, not complete even at two weeks post partum.Bundles of growing sympathetic nerves were visible in the periphery of the various organs studied at 19–21 days post coitum. A terminal innervation of the organs suggestive of a functional transmitter mechanism did not start to establish until at or immediately after birth. The final pattern of innervation was usually reached at about one week post partum, and the following development proceeded largely in the form of a quantitative increase in the number of nerves participating in the innervation apparatus. The adult level of noradrenaline in the heart and vas deferens was reached three to five weeks after birth. The small intestine was an exception in that the final pattern of innervation in the wall was attained immediately after birth.There was no overt difference in the rate of development of the terminal sympathetic innervation in organs supplied by short adrenergic neurons (accessory male genital organs) compared to the innervation of the submaxillary gland, heart and kidney, which receive classical long adrenergic neurons.The work was supported by a grant from the Association for the Aid of Crippled Children, New York, and was carried out within a research organization sponsored by the Swedish Medical Research Council (grants No. B71-14X-56-07A and B71-14X-712-06A).     

The effect of colchicine on meiosis in Lilium speciosum cv. “Rosemede”

Chromosome pairing and chiasma frequency were studied in meiocytes at diakinesis of Lilium speciosum cv. Rosemede fixed up to 21 days after the start of either continuous or 3 day pulse colchicine treatment. The two treatments gave similar results. In pulse treated pollen mother cells (PMCs) the mean chiasma frequency per cell fell from 26.4 in controls to 8.5 after fourteen days while the mean number of univalents per cell increased from 0.05 to 17.58. There was a negative correlation between mean chiasma frequency per bivalent and per PMC in colchicine treated buds; univalents were preferentially induced in bivalents with one chiasma, and preferentially excluded in bivalents with 4 chiasmata. Some chiasmata were redistributed to surviving bivalents despite the concurrent reduction in chiasma frequency per meiocyte. — Colchicine sensitivity began in premeiotic interphase and extended to mid or late zygotene in PMCs; ongoing synapsis was unaffected. However, susceptibility to univalency was asynchronous between bivalents occurring at zygotene in short chromosomes but at late premeiotic interphase in the longest chromosomes. The number of chiasmata per bivalent could be altered by colchicine without inducing univalents, but the ultimate effect was to reduce the number of chiasmata per bivalent (or per chromosome arm) directly to zero. The major factors determining the order and extent of reduced pairing and chiasma number were total chromosome length and arm length. Pairing and chiasma formation in embryo sac mother cells were less sensitive to colchicine than in PMCs, but their behavior was otherwise similar.     

The Sl TPL3–Sl WUS module regulates multi-locule formation in tomato by modulating auxin and gibberellin levels in the shoot apical meristem

Malformed fruits depreciate a plant’s market value.In tomato(Solanum lycopersicum),fruit malformation is associated with the multi-locule trait,which involves genes regulating shoot apical meristem(SAM) development.The expression pattern of TOPLESS3(SITPL3) throughout SAM development prompted us to investigate its functional significance via RNA interference(RNAi) and clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9(Cas9)-mediated gene editing.Lower SITPL3 t...     

Identification of C-banded chromosomes in meiosis and the analysis of nucleolar activity in Avena byzantina C. Koch cv ‘Kanota’

Summary The Giemsa C-banding technique was used to identify individual meiotic and somatic chromosomes in 21 monosomic lines of Avena byzantina C. Koch cv Kanota (genome designation AACCDD). The hexaploid complement is composed of three sets of seven chromosome pairs. The heterochromatin in the putative diploid progenitors is located at the telomeres (genome A), at the centromeric and interstitial regions (genome C), or more evenly spread throughout the set (genome D). Comparisons based on C-banding between A. byzantina and its diploid progenitor species allowed us to allocate individual chromosomes into specific genomes. The C-banding technique may be useful for interspecific chromosome pairing analyses. Nucleolar activity and competition were studied using a silver-staining procedure. Only three chromosome pairs showed nucleolar organizer regions, thus indicating that nucleolar competition occurs naturally in hexaploid oats.