Apical Region Of Root Research Articles

Distribution and Toxic Effects of Cadmium and Lead on Maize Roots

Two-day-old seedlings of maize ( Zea mays L.) were incubated on Cd and Pb nitrate solutions at the concentrations that inhibited root growth approximately by 50% after two-day-long incubation (LC 50 ; 10 -4 and 10 -3 M, respectively) or completely terminated growth of the primary root after one-day-long incubation (LC; 5 × 10 -4 and 10 -2 M, respectively). Cd and Pb contents were measured using an anodic inversion voltammetric technique in a flow injection system and a histochemical method. At LC 50 , Cd and Pb were discerned, by his- tochemical techniques, in all root apical tissues, whereas in the root hair zone, the heavy metals were primarily accumulated in the apoplast of the rhizodermis and cortex and to a lesser extent, in the vascular tissues and parenchyma cells surrounding the metaxylem vessels. Insignificant accumulation of Cd and Pb in the pericycle probably explains why root branching was tolerant to these agents. At LC, Cd and Pb were found in the apoplast of all root tissues, in accordance with the practically complete inhibition of root growth and branching. Irre- spectively of Cd and Pb concentrations in the external solution, the metal contents in the root apex exceeded those in the basal region. Procion dyes were used to assess cell death inflicted by Cd and Pb. At LC, the root cap and meristematic cells perished, together with the rhizodermal cells and the outer cortical cells of the root apex, whereas only the rhizodermal cells in the root apical region died at LC 50 . The evidence that Cd and Pb cross the endodermal barrier at LC presumes that, at lower metal concentrations, the Casparian strip and plas- malemma of the endodermis regulate the transport of these metals into the central cylinder. The authors con- clude that the identical barriers control Cd and Pb transport in root tissues.

Changes in Phytohormone Status in Stems and Roots after Decapitation of Pea Seedlings

Experiments were performed on the first and second internodes and 4-cm-long apical segments of main roots of pea (Pisum sativum L.) seedlings, grown in the light and decapitated above the second node on the seventh day after seed germination. Endogenous phytohormones were measured by the enzyme-linked immunosorbent assay during three days after decapitation of seedlings. The IAA level in the internodes decreased 2–3 times on the second day after decapitation of seedlings while the cytokinin level increased 5–6 times for zeatin and zeatin riboside (Z and ZR) and 1.5–2 times for isopentenyl adenine and isopentenyl adenosine (IP and IPA). In contrast to internodes, the IP and IPA contents in the roots of decapitated seedlings did not change, but the levels of Z and ZR increased 1.5–2 times compared to intact plant roots. The IAA level in the apical region of root remained almost unchanged after the removal of shoot apex. It was concluded that the apical meristem of the main root is not the site of the cytokinin response to the auxin signal coming from the stem apex and that a slight accumulation of Z and ZR after decapitation is due to upper zones of the root. There was no difference in the content of gibberellin-like substances between the internodes of intact and decapitated seedlings. However, the content of gibberellins (GA) in the root tip decreased after decapitation of seedling, which suggests an essential role of apical bud in supplying the root with GA and/or intermediates for their biosynthesis.

Expression pattern of diacylglycerol acyltransferase-1, an enzyme involved in triacylglycerol biosynthesis, in Arabidopsis thaliana.

Triacylglycerol (TAG) is the major carbon storage reserve in oilseeds such as Arabidopsis. Acyl-CoA:diacylglycerol acyltransferase (DGAT) catalyses the final step of the TAG synthesis pathway. Although TAG is mainly accumulated during seed development, and DGAT has presumably the highest activity in developing seeds, we show here that TAG synthesis is also actively taking place during germination and seedling development in Arabidopsis. The expression pattern of the DGAT1 gene was studied in transgenic plants containing the reporter gene beta-glucuronidase (GUS) fused with DNA sequences flanking the DGAT1 coding region. GUS activity was not only detected in developing seeds and pollen, which normally accumulate storage TAG, but also in germinating seeds and seedlings. Western blots showed that DGAT1 protein is present in several tissues, though is most abundant in developing seeds. In seedlings, DGAT1 is expressed in shoot and root apical regions, correlating with rapid cell division and growth. The expression of GUS in seedlings was consistent with the results of RNA gel blot analyses, precursor feeding and DGAT assay. In addition, DGAT1 gene expression is up-regulated by glucose and associated with glucose-induced changes in seedling development.

Compartmentation of Cadmium and Iron in Mesembryanthemum crystallinum Plants during the Adaptation to Cadmium Stress

The common ice plants (Mesembryanthemum crystallinum) at the stage of five leaf pairs were exposed to cadmium chloride solutions (1, 0.1, and 0.01 mM) under the conditions of water culture. After five days, the partition of cadmium and iron in the plant organs and in the cell structures of the apical root region were investigated. Plant adaptation to excess cadmium in the environment was assessed by an increase in the leaf and root weight, a change in peroxidase activity, and an accumulation of proline. The common ice plant accumulated cadmium mainly in the root system. At a high concentration of cadmium in the nutrient solution (1 mM), its content in the root exceeded 2 g/kg fr wt, while at a concentration of 0.01 mM, it was as low as 10 mg/kg. Dithizone staining of transverse sections of the root apical region showed that, after a 48-h-long exposure of plants to 0.1 mM cadmium chloride, cadmium was localized in the cell walls of endodermis and metaxylem. The level of cadmium in leaves varied from 0.5 to 18 mg/kg fr wt. However, there was only a weak correlation between cadmium accumulation and the extent of a biomass decrease in the leaves of various stories, when cadmium concentration in the medium (1 mM cadmium chloride) was toxic. This fact could be related to a marked efflux of endogenous iron from old leaves into the young ones and to a change in the cadmium/iron ratio in the tissues. Proline accumulation in the third leaf pair and in the roots occurred at a relatively low cadmium content (10–12 mg/kg fr wt) in these organs. Maxima of activity of all three forms of peroxidase, viz., soluble, ionically-bound, and covalently-bound peroxidases, in roots were found at a high accumulation of cadmium in these organs (45 mg/kg fr wt). These maxima exceeded 3–4-fold the activity in aging leaves containing 5 mg cadmium/kg fr wt. A decrease in peroxidase activity in leaves was accompanied by a 3.3-fold decrease in iron content; thus, it could be caused by a deficiency of available iron necessary for the enzyme functioning. It was concluded that the resistance of Mesembryanthemum crystallinum, a halophyte, to excess cadmium content in the medium was achieved by its predominant accumulation in roots, where excess cadmium is compartmentalized in the apoplast and seems to be subjected to detoxification through pectate formation. Moreover, the leaves and, particularly, the roots are characterized by a high activity of the antioxidant systems, such as guaiacol-dependent peroxidases, and an occurrence of proline at modest cadmium concentrations.

Modification of expansin transcript levels in the maize primary root at low water potentials.

We previously demonstrated that maintenance of cell elongation in the apical region of maize primary roots at low water potentials (psi(w)) was associated with an increase in expansin activity and extractable expansin protein. Here, we characterized the spatial pattern of expansin gene expression along the growing maize root and studied the effect of low psi(w) on expansin gene expression. Roots were divided into three segments: apical 0 to 5 mm, subapical 5 to 10 mm, and non-growing 10 to 20 mm. Of the five expansin genes expressed in control roots, two alpha-expansins (Exp1 and Exp5) and two beta-expansins (ExpB2 and ExpB8) are expressed specifically in the growing region, whereas expression of beta-expansin ExpB6 is shifted basipetally. After seedlings were transplanted to vermiculite with a psi(w) of -1.6 MPa, transcripts for Exp1, Exp5, and ExpB8 rapidly accumulated in the apical region of the root. These mRNA changes correlated with the maintenance of root elongation and increases in wall extensibility found previously. The beta-expansins ExpB2 and ExpB6 showed distinctive patterns of expression and responses to low psi(w,) indicative of distinctive functions. Inhibition of abscisic acid (ABA) accumulation at low psi(w) (by fluridone treatment) had no effect on expansin expression, except that ExpB2 transcript level showed a minor dependence on ABA. Gene-specific regulation of alpha- and beta-expansin mRNA pools likely contributes to growth alterations of the maize (Zea mays) root as it adapts to a low psi(w), but these changes do not appear to be mediated by changes in ABA content.

Rice and Phragmites: effects of organic acids on growth, root permeability, and radial oxygen loss to the rhizosphere

Young Phragmites plants were grown in two cocktails of monocarboxylic acids (C(1)-C(5)) at pH 6, where the concentration of each acid was innocuous and the total undissociated (potentially toxic) concentrations were 0.35 mmol/L and 0.42 mmol/L. Rice plants were subjected to 1.5 mmol/L acetic acid at pH 4.5 (undissociated concentration = 1.05 mmol/L). In Phragmites, each cocktail curtailed root growth especially and induced premature shoot senescence. In both species, after 3-5 d of treatment, radial oxygen loss (ROL) from apical regions of adventitious roots, and from Phragmites laterals, was reduced to very low values and associated with cell wall lignification and suberization in the surface cell layers. At later stages of treatment, rice responded to acetic acid in similar ways to Phragmites, with the development of intercellular and callus type occlusions in the gas space system, vascular blockages, and the failure of laterals to emerge. The results are relevant to the supply of oxygen from Phragmites roots to sediments for the phytopurification of waste waters, to the efflux of methane and carbon dioxide from wetlands, and to rice cultivation.

An overview of the effects of phytotoxins on Phragmites australis in relation to die-back

The accumulation in sediments of the low molecular weight, volatile, monocarboxylic acids and/or sulphide, generated by the decay of (i) the underground parts of the reed and/or (ii) organic deposits produced under eutrophic conditions, may play a crucial part in Phragmites die-back. In the field high levels of some of these phytotoxins have been detected at certain die-back sites and in sediments containing the rotting underground parts of the plant. Symptoms of die-back include a clumped habit, stunting and death of roots and shoots, bud death, premature senescence of shoots, weakened stems, impeded aeration of the underground parts of the plant due to callus development, blockages within the vascular systems, lignification and suberisation of laterals and apical regions of adventitious roots and lower levels of starch in rhizomes. Poor convective aeration of the rhizome system has also been associated with higher proportions of alanine, γ-amino butyric acid and serine in culm bases, indicating hypoxic metabolism in the underground system. In laboratory experiments plants developed almost all of the growth, morphological and anatomical symptoms of die-back when treated with various single phytotoxins, e.g. acetic, propionic, n- and iso-butyric and n-caproic acids and sulphide at concentrations reported for die-back sites and/or associated with the decaying underground parts of the plant. For each acid alone, a concentration of ca. 1 mM, was highly toxic at pH 4.5, but relatively non-toxic at pH 6.0. However, when a cocktail of the five acids was applied (where each acid was 1 mM) the mixture proved to be toxic even at pH 6.

A morphological study of the mycorrhiza of Entoloma clypeatum f. hybridum on Rosa multiflora

Mycorrhizas ofEntoloma clypeatum f.hybridum onRosa multiflora in the field in Japan were studied by stereo, light and electron microscopy. In most mycorrhizas, the root cap, meristem, and apical region of the cortex disappeared, but in a few mycorrhizas, these tissues remained. Fungal hyphae of the mycorrhizas invaded root tissues and branched palmately. Hyphae in contact with cortical cells were larger than those far from the root cells and contained many mitochondria, cisternae of endoplasmic reticulum and transitional vesicles. Invading hyphae were undulate in the apical part of the mycorrhiza, and some of them lacked distinct organelles. Electron-dense granules accumulated in the root cells adjacent to the fungal hyphae. Both the remnants of the plant cells and the fungal hyphae were included in the amorphous materials on the tip of the stele. These observations suggest the destructive infection by fungal hyphae of the root cells and their collapse near the tip of the stele.

Interdental acrylic stabilisation of canine tooth root and mandibular fractures in a dog

A two-year-old labrador had bilateral open fractures of the apical region of the mandibular canine tooth roots and the adjacent mandible. The fractures were reduced and held in place with...

Histological and histochemical quantification of root resorption incident to the application of intrusive force to rat molars.

This study was conducted to investigate the nature of root resorption resulting from intrusive forces applied to the rat lower molars, by means of histological and histochemical techniques with tartrate resistant acid phosphatase (TRAP). Thirty-eight 13-week-old Wistar strain male rats were used. Intrusive force was created by a fixed appliance which was adjusted to exert an initial force of 50 g for the duration of 1, 2, and 3 weeks. The degree of root resorption and distribution of TRAP positive cells were evaluated. On the root surface, the TRAP positive scores were low in the apical regions. Significant differences in the scores were found in the inter-radicular region of the roots between the experimental and control groups for the 2- and 3-week groups. More active resorption of bone occurred during the experimental period, as denoted by greater TRAP positive scores on the bone than on the root surface. Root resorption scores in the apical root region were larger in the 2- and 3-week groups than in the 1-week group. Significant differences in the root resorption scores were also found between the 1- and 3-week groups in the inter-radicular region, indicating that intrusive force application of a longer duration may lead to a higher frequency of root resorption. It is shown that, irrespective of the level of TRAP positive cells and root resorption scores, the degree of root resorption activity is higher in the apical root region than in the inter-radicular area. These results indicate that cellular cementum may be resorbed more easily because of its richer organic components and low mineralized structure.

Shoot regeneration from cultured root explants of spinach (Spinacia oleracea L.): a system for Agrobacterium transformation.

A reliable plant regeneration system is described for the production of adventitious shoots from root explants of spinach. Explants from roots of axenic shoots and roots induced on cultured hypocotyl explants were used for adventitious shoot induction. Explants from apical, middle and basal root regions were incubated on Nitsch and Nitsch medium supplemented with α-naphthaleneacetic acid, gibberellic acid and kinetin. Optimum shoot regeneration was from explants of apical and middle root regions on medium with 20 µM α-naphthaleneacetic acid and 5.0 µM gibberellic acid. Shoots originated directly from root tissues without an intervening callus phase. Adventitious shoots were rooted and were grown to maturity in the glasshouse. This plant regeneration procedure has been exploited in preliminary studies of Agrobacterium-mediated transformation.

Flow cytometry, sorting and immunocharacterization with proliferating cell nuclear antigen of cycling and non-cycling cells in synchronized pea root tips.

In the 3-d-old 2-mm root tip of Pisum sativum L. cv. Lincoln the percentage of actively proliferating cells is estimated to be 70%. The remaining cells are non-cycling and arrested with 2C and 4C DNA content in G0 and in G2Q, respectively. In this work we studied the kinetic significance of these quiescent cells, using the sorting capabilities of flow cytometry and immunofluorescence techniques to detect the proliferation marker PCNA (proliferating cell nuclear antigen) inside cells within the different cell-cycle compartments. While in animal cells, PCNA is present at a high level only in actively proliferating cells, in 3-d-old pea root tips 95% of the cells are PCNA-positive. After flow cytometry and sorting of pea non-cycling nuclear populations, all G2Q nuclei appeared strongly PCNA-positive, indicating that these cells had recently left the cell cycle. By contrast, most G0 nuclei showed a low level of PCNA immunofluorescence intensity, as measured by image analysis, with about 25% of the nuclei being PCNA-negative. This small percentage was found to correspond to root cap cells, as could be observed in the root tip section. These are the only cells in the root apical region which are fully differentiated and which, therefore, lack the competence to enter the cell cycle. In contrast, the more or less PCNA-positive G0 nuclei could represent a kinetically heterogeneous population of cells competent to proliferate, but which have either recently left the cell cycle or are progressing to the G0-G1 transition.

Complete mechanical impedance increases the turgor of cells in the apex of pea roots

ABSTRACTIn this paper we describe an experimental approach which allows turgor (p) in an impeded root to be measured without the need to remove the root from the impeding environment. The maximum axial growth pressure (σmax) generated by completely impeded pea (Pisum sativum L.) roots was measured using a novel apparatus incorporating a force transducer. The apparatus was designed so that it was possible to gain access to the impeded root with the microcapillary of a pressure probe and so obtain in situ measurements of P. Turgor in cells in the apical region of impeded roots was 0.78 MPa, compared with 0.55 MPa in unimpeded roots. In impeded roots, σmax was 0.52 MPa, showing that the pressure component resulting from cell wall tension (W, where W=P–σ) decreased from 0.55 to 0.26 MPa as the roots became impeded. When impeded roots were removed from the apparatus, there was no decrease in P over the following 90 min. Impedance did not cause P to change in the non‐elongating part of the roots further from the apex.

Changes in pH at the dentin surface in roots obturated with calcium hydroxide pastes

The purpose of this study was to determine the pH, after defined periods of time, in cavities prepared in the facial surface of the cervical, middle, and apical regions of roots obturated with calcium hydroxide pastes. Root canal instrumentation was performed on 40 recently extracted, single-rooted human teeth. Cavities 1.5 mm in diameter and 0.75 mm in depth were prepared in the cervical, middle, and apical regions of the facial surface of each root. Teeth were randomly divided into four groups. One group was left unobturated and served as a control. The three remaining groups were obturated with either aqueous calcium hydroxide, calcium hydroxide mixed with camphorated monochlorophenol. or Pulpdent pastes. Access cavities and apical foramina were closed with Cavit. Each tooth was stored individually in a vial containing unbuffered isotonic saline. pH at the surface was measured in the cervical, middle, and apical cavities at 0 and 3, 7, 14, 21, 28, 45, 60, 90, and 120 days. Results indicate that hydroxyl ions derived from calcium hydroxide pastes diffused through root dentin at all regions over the experimental period of 120 days. The pattern of pH change at the tooth surface was similar in all regions of the root, regardless of the type of calcium hydroxide paste used. This was a rapid rise in pH from a control value of pH 7.6, to greater than pH 9.5 by 3 days, followed by a small decline to pH 9.0 over the next 18 days, before finally rising and remaining at, or above pH 10.0 for the remainder of the experimental period. Pulpdent paste in the apical region was the only exception in this pattern, producing a pH rise nearly one full unit below the other pastes, pH 9.3. These results indicate that, for all pastes tested, a high pH is maintained at the root surface for at least 120 days.

Adventitious shoot regeneration from root tips of intact seedlings ofAegle marmelos

SummaryIntact seedlings of Aegle marmelos Corr. were tested for their ability to produce adventitious shoots by direct culture of mature seeds on Murashige and Skoog (MS) medium supplemented with different concentrations of benzyladenine (BA); 1–2 mg l–1 BA was found to be the optimum concentration. Addition of 0.1 mg l–1 indoleacetic acid (IAA) further increased shoot proliferation efficiency. Shoot buds originated from regions adjacent to the bases of cotyledons and cotyledonary axils and roots. In many cases, adventitious shoots were produced from the enlarged apical region of root. Shoots were rooted on MS medium supplemented with indolebutyric acid (IBA). Rooted plantlets were successfully established in soil.

Aluminum accumulation and associated effects on 15NO3 − influx in roots of two soybean genotypes differing in Al tolerance

A study was conducted to examine aluminum (Al) exclusion by roots of two differentially tolerant soybean (Glycine max L. Merr.) lines, Pl-416937 (Al-tolerant) and Essex (Al-sensitive). Following exposure to 80μM Al for up to 2 h, roots were rinsed with a 10 mM potassium citrate solution and rapidly dissected to allow estimation of intracellular Al accumulation in morphologically distinct root regions. Using 10 min exposures to 300μM 15NO3 − and dissection, accompanying effects on NO3 − uptake were measured. With Al exposures of 20 min or 2 h, there was greater Al accumulation in all root regions of Essex than in those of Pl-416937. The genotypic difference in Al accumulation was particularly apparent at the root apex, both in the tip and in the adjacent root cap and mucilage. Exposure of roots to Al inhibited the uptake of 15NO3 − to a similar extent in all root regions. The results are consistent with Al exclusion from cells in the root apical region being an important mechanism of Al tolerance.

Phloem transport and differential unloading in pea seedlings after source and sink manipulations

Phloem transport was investigated in pea seedlings after application of [14C]sucrose to the cotyledons. The accumulation of the label in segments of young seedlings shows a differential unloading along the plant axis. Shoot and root exhibit tip-to-base gradients of sink strength. In the primary root, the sink-strength profiles reflect not only the importance of the apical meristem, but show also the starting points of cambial activity and production of secondary vascular elements. Experiments including partial removal of the source and manipulations of the sink strength indicate that translocation of pea seedlings is sink-regulated and responds rapidly to changed apoplastic conditions in the apical root region. Here, a lowered water potential leads to an increase of phloem unloading that is suggested to supply the assimilate demand for the short-term osmoregulation of affected cells via the symplasmic pathway.

Phloem transport of carboxyfluorescein through tomato roots infected with Meloidogyne incognita

The fluorescent acidic dye, carboxyfluorescein (CF) was used to study phloem transport through M. incognita infected tomato roots. Following application to leaves, CF moved via the internal and external phloem of the stem towards the roots. In infected intact roots, dye accumulated inside galls. Movement to more apical root regions occurred via irregularly arranged phloem elements. At the infection sites, dye accumulated inside the nematode induced giant transfer cells. When CF was applied directly to the xylem vessels, the giant cells failed to accumulate dye. These observations suggest that a symplastic transport pathway exists between the phloem and giant cells. Continuous dye application over prolonged periods to triple-split, singly infected shoot-root systems, revealed a transport pathway for solutes from the giant cells, via the body of the nematode, out of the root. This pathway may contribute to the higher leakage of solutes known to occur from root-knot nematode infected roots.

Developmental regulation and phytochrome-mediated induction of mRNAs encoding a proline-rich protein, glycine-rich proteins, and hydroxyproline-rich glycoproteins in Phaseolus vulgaris L.

We have studied developmental and light regulation of mRNAs encoding a putative cell wall proline-rich protein (PvPRP1), cell wall glycine-rich proteins (GRPs), and cell wall hydroxyproline-rich glycoproteins (HRGPs) in bean (Phaseolus vulgaris). Light increases the levels of these mRNAs 2- to 150-fold in highly spatially regulated patterns during seedling development. These mRNA changes include differential regulation of transcripts derived from the GRP and HRGP multigene families. In 6-day-old light-grown seedlings, the PvPRP1 and GRP1.0 mRNAs were most abundant in the apical region of hypocotyls, epicotyls, and roots. In contrast, several HRGP transcripts were most abundant in the mature region of hypocotyls and roots in light-grown seedlings. When etiolated 6-day-old seedlings were illuminated with white light for 8 hr, maximal accumulation of PvPRP1 and GRP1.0 mRNAs occurred in the apical hook, whereas HRGP and GRP1.8 mRNAs accumulated in the mature region of hypocotyls. Etiolated seedlings subjected to a pulse of red light accumulated PvPRP1, GRP, and HRGP mRNAs in the hypocotyls. Far-red light inhibited red light induction of these mRNAs, indicating a phytochrome-mediated process. The possible roles of PRPs, GRPs, and HRGPs in cell differentiation and photomorphogenesis are discussed.

Xyloglucan Endotransglycosylase Activity, Microfibril Orientation and the Profiles of Cell Wall Properties Along Growing Regions of Maize Roots

A series of physical and chemical analyses were made on the expanding zone of maize seedling roots grown in hydroponics. Comparison of longitudinal profiles of local relative elemental growth rate and turgor pressure indicated that cell walls become looser in the apical 5 mm and then tighten 5-10 mm from the root tip. Immersion of roots in 200 mol m-3 mannitol (an osmotic stress of 0-48 MPa) rapidly and evenly reduced turgor pressure along the whole growing region. Growth was reduced to a greater extent in the region 5-10 mm from the root tip than in the apical region. This indicated rapid wall-loosening in the root tip, but not in the more basal regions. Following 24 h immersion in 400 mol m-3 mannitol (an osmotic stress of 0-96 MPa) turgor had recovered to pre-stressed values. Under this stress treatment, growth was reduced in the region 4-10 mm from the root tip, despite the recovery of turgor, indicating a tightening of the wall. In the root apex, local relative elemental growth rate was unchanged in comparison to control tissue, showing that wall properties here were similar to the control values. Cellulose microfibrils on the inner face of cortical cell walls became increasingly more parallel to the root axis along the growth profile of both unstressed and stressed roots. Orientation did not correlate with the wall loosening in the apical region of unstressed roots, or with the tightening in the region 5-10 mm from the root tip following 24 h of osmotic stress. Longitudinal profiles of the possible wall-loosening enzyme xyloglucan endotransglycosylase (XET) had good correspondence with an increase in wall loosening during development. In the zone of wall tightening following osmotic stress, XET activity was decreased per unit dry weight (compared with the unstressed control), but not per unit fresh weight.