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Oscillations of axial plant organs.

The tips of roots and shoots commonly show lateral movements as they grow forwards. These occur as both circumnutations (with long periods and large amplitudes) and micronutations (with short periods and small amplitudes). Their properties are reviewed, with emphasis on roots, and possible ways in which they could be regulated are discussed. The mechanisms could include long-range controls (for circumnutations) that depend on transmissible signals using steps common to gravitropism, and short-range controls (for micronutations) that operate within the elongation zone. The former are a property of the apex as a whole, while the latter may be confined to localized groups of cells. Simulation of nutations is presented with a view to isolating key physiological processes. However, this approach is limited by the current inadequate understanding of the growth mechanisms involved.

Cell Division↗

In vitro propagation of Lagerstromia parviflora Roxb. from adult tree.

A micropropagation protocol based on axillary bud proliferation has been developed from mature Lagerstromia parviflora adult tree. Nodal segments cultured on woody plant medium supplemented with 5.0 microl. BAP and 0.25 microm IAA gave maximum (86.9%) morphogenetic response. Proliferated shoots (10.7 per explants) were elongated to 3.9 cm within 6 weeks. In vitro produced micro-shoots were subjected to an IBA treatment (500 ppm for 2 min. dip) and placed under misting conditions for rooting. Misting beds were prepared with sand: soil (3:1) for 80.6% rooting and was acclimatized. Shoot length seems to be important to induce adventitious roots. The highest (91.7%) rooting was recorded on shoots ranging a length between 3.1-4.0 cm. Rooted and hardened plants were later transferred to poly bags and maintained in shadenet house. The protocol has the realizes capacity to produce 260 plants from a single explants within 10 months multiplication cycle.

Botany↗

Hyperaccumulation of cadmium by hairy roots of Thlaspi caerulescens.

Hairy roots were used to investigate cadmium uptake by Thlaspi caerulescens, a metal hyperaccumulator plant with potential applications in phytoremediation and phytomining. Experiments were carried out in nutrient media under conditions supporting root growth. Accumulation of Cd in short-term (9-h) experiments varied with initial medium pH and increased after treating the roots with H(+)-ATPase inhibitor. The highest equilibrium Cd content measured in T. caerulescens roots was 62,800 microg g(-1) dry weight, or 6.3% dry weight, at a liquid Cd concentration of 3710 ppm. Cd levels in live T. caerulescens roots were 1.5- to 1.7-fold those in hairy roots of nonhyperaccumulator species exposed to the same Cd concentration, but similar to the Cd content of autoclaved T. caerulescens roots. The ability to grow at Cd concentrations of up to 100 ppm clearly distinguished T. caerulescens hairy roots from the nonhyperaccumulators. The specific growth rate of T. caerulescens roots was essentially unaffected by 20 to 50 ppm Cd in the culture medium; in contrast, N. tabacum roots turned dark brown at 20 ppm and growth was negligible. Up to 10,600 microg g(-1) dry weight Cd was accumulated by growing T. caerulescens hairy roots. Measurement of Cd levels in whole roots and in the cell wall fraction revealed significant differences in the responses of T. caerulescens and N. tabacum roots to 20 ppm Cd. Most metal was transported directly into the symplasm of N. tabacum roots within 3 days of exposure; in contrast, T. caerulescens roots stored virtually all of their Cd in the wall fraction for the first 7 to 10 days. This delay in transmembrane uptake may represent an important defensive strategy against Cd poisoning in T. caerulescens, allowing time for activation of intracellular mechanisms for heavy metal detoxification.

Cadmium↗

Quantification of neural tissue injury in a rat radiculopathy model: comparison of local deformation, behavioral outcomes, and spinal cytokine mRNA for two surgeons.

Clinical and experimental work indicate that a variety of factors contribute to radicular pain mechanisms, including mechanical injury. While it has been qualitatively suggested that the magnitude of nerve root mechanical injury affects the nature of the pain response, no study has quantified the local in vivo injury biomechanics in these models. Therefore, it was the purpose of this study to develop and implement an in vivo method to quantify compressive nerve root injury strain severity and characterize its effect on the resulting responses in an existing lumbar radiculopathy rat model. Male Holtzman rats were divided into a sham group with only nerve root exposure or a ligation group with the nerve root tightly ligated using silk suture. Using image analysis, nerve root radial strains were calculated at the time of injury for two surgeons. Mechanical allodynia was continuously assessed throughout the study and spinal cord cytokine mRNA levels were assayed on postoperative day 7. The degree of intersurgeon variability for imposing a ligation injury in this model was also assessed. Mean compressive injury strains in the nerve root were 32.8+/-14.2% and were not different for the two experimenters. Animals undergoing more severe ligation strains exhibited significantly heightened allodynia following injury and greater upregulation of the inflammatory cytokines IL-1alpha/beta, IL-6, and IL-10. Results indicate a direct correlation of local nerve root injury severity with the ensuing physiologic responses associated with nociception.

Animals↗

Short and long term effects of root and shoot chilling of ransom soybean.

The immediate short term effects on some physiological processes and the long term effects on morphology and reproductive development of root- and shoot-chilled soybeans (Glycine max L. cv Ransom) were studied. Roots or shoots of 16- or 17-day-old plants were chilled at 10 degrees C for one week, and then rewarmed to 25 degrees C. Leaf elongation rate, net CO(2) uptake rate, and stomatal conductance decreased during root or shoot chilling. Root chilling had only temporary effects on water relations, while shoot chilling caused large changes in potentials during chilling. Most processes measured returned to control levels after two days of rewarming. Root-chilled plants harvested 90 days after emergence were similar in morphology and seed weight to controls. Shoot-chilled plants showed a large increase over controls in axillary branch growth, but an early abortion of flowers and a delayed resumption of flowering caused a 78% reduction in seed weight. Root chilling in this study was found to have little or no long term effect on the plants, while shoot chilling caused significant changes in vegetative morphology, and a delay in flowering and subsequent pod filling.

Journal Article↗

Physiological basis of reduced AL tolerance in ditelosomic lines of Chinese Spring wheat.

Aluminum tolerance was assessed in the moderately Al-tolerant wheat (Triticum aestivum L.) cultivar Chinese Spring and a set of ditelosomic lines derived from Chinese Spring. Three ditelosomic lines lacking chromosome arms 4DL, 5AS and 7AS, respectively, exhibited decreased Al tolerance relative to the euploid parent Chinese Spring based on reduced root growth in Al-containing solutions. The physiological basis of the reduced Al tolerance was investigated. Measurements by inductively coupled argon plasma mass spectroscopy of root apical Al accumulation demonstrated that two of these three lines had a decreased ability to exclude Al from the root apex, the site of Al phytotoxicity. As Al-induced malate exudation has been suggested to be an important physiological mechanism of Al tolerance in wheat, this parameter was quantified and malate exudation was shown to be smaller in all three deletion lines compared with Chinese Spring. These results suggest that the decreased Al tolerance in at least two of the three ditelosomic lines is due to the loss of different genes independently influencing a single Al-tolerance mechanism, rather than to the loss of genes encoding alternative Al-tolerance mechanisms.

Adaptation, Physiological↗

Plant regeneration of Ilex paraguariensis (Aquifoliaceae) by in vitro culture of nodal segments.

In vitro regeneration of complete plants from nodal single bud segments of "yerba mate" (Ilex paraguariensis St. Hil.) was studied under defined nutritional and environmental conditions. Nodal segments harvested from actively growing shoots of conventionally raised plants were cultured on nutrient medium with the mineral salts and vitamins of Murashige and Skoog medium at 1/4 strength, supplemented with various concentrations of sucrose and 6-benzyladenine (BAP). Shoot regeneration from explants of both young (2 years old) and adult (20 years old) mother plants were readily achieved in the medium supplemented with 0.04-0.09 M sucrose with or without BAP. As many as 60-65% of the nodal segments cultured formed shoots. Rooting of regenerated shoots was observed in 50% of the explants harvested from young plants, whereas 25% of the explants rooted when the nodal explants were harvested from adult plants. The best rooting induction was achieved on 1/4 strength MS medium with vermiculite as the substrate and supplemented with 1-1.5% IBA (indolebutyric acid) and 1-2% PPZ (3-methyl-1-phenyl-2 pyrazolin-5-one). Plantlets were successfully transferred to soil.

Plant Development↗

In vitro mass multiplication of Ophiorrhiza mungo Linn.

A protocol for in vitro mass multiplication of plants through seedling (shoot) cultures was established for Ophiorrhiza mungo. Maximum number of adventitious shoots per shoot culture (10.4 +/- 1.72) was initiated on MS solid medium supplemented with BAP (2.22 microM) after 3 weeks. Shoots were further multiplied (12.8 +/- 2.8) through subculture of intact shoots and reculture of nodal segments of aseptic shoots (6.5 +/- 0.94) in MS solid medium containing BAP (0.89 microM). Shoot elongation (1.27 +/- 0.12 cm) was achieved in the medium containing GA3 (1.44 microM) in two weeks. Rooting was favoured in basal agar medium supplemented with IBA (12.3 microM) plus NAA (1.07 microM). The plants were successfully established (100%) in the pots containing sand and top soil (1:1) mixture in a period of two weeks.

Agar↗

Nitric oxide plays a central role in determining lateral root development in tomato.

Nitric oxide (NO) is a bioactive molecule that functions in numerous physiological processes in plants, most of them involving cross-talk with traditional phytohormones. Auxin is the main hormone that regulates root system architecture. In this communication we report that NO promotes lateral root (LR) development, an auxin-dependent process. Application of the NO donor sodium nitroprusside (SNP) to tomato ( Lycopersicon esculentum Mill.) seedlings induced LR emergence and elongation in a dose-dependent manner, while primary root (PR) growth was diminished. The effect is specific for NO since the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (CPTIO) blocked the action of SNP. Depletion of endogenous NO with CPTIO resulted in the complete abolition of LR emergence and a 40% increase in PR length, confirming a physiological role for NO in the regulation of root system growth and development. Detection of endogenous NO by the specific probe 4,5-diaminofluorescein diacetate (DAF-2 DA) revealed that the NO signal was specifically located in LR primordia during all stages of their development. In another set of experiments, SNP was able to promote LR development in auxin-depleted seedlings treated with the auxin transport inhibitor N-1-naphthylphthalamic acid (NPA). Moreover, it was found that LR formation induced by the synthetic auxin 1-naphthylacetic acid (NAA) was prevented by CPTIO in a dose-dependent manner. All together, these results suggest a novel role for NO in the regulation of LR development, probably operating in the auxin signaling transduction pathway.

Indoleacetic Acids↗

Signaling mechanisms integrating root and shoot responses to changes in the nitrogen supply.

During their life cycle, plants must be able to adapt to wide variations in the supply of soil nitrogen (N). Changes in N availability, and in the relative concentrations of NO(3) (-)and NH(4) (+), are known to have profound regulatory effects on the N uptake systems in the root, on C and N metabolism throughout the plant, and on root and shoot morphology. Optimising the plant's responses to fluctuations in the N supply requires co-ordination of the pathways of C and N assimilation, as well as establishment of the appropriate allocation of resources between root and shoot growth. Achieving this integration of responses at the whole plant level implies long-distance signaling mechanisms that can communicate information about the current availability of N from root-to-shoot, and information about the C/N status of the shoot in the reverse direction. In this review we will discuss recent advances which have contributed to our understanding of these long-range signaling pathways.

Biological Transport, Active↗

Ca(2+) dynamics in the lumen of the endoplasmic reticulum in sensory neurons: direct visualization of Ca(2+)-induced Ca(2+) release triggered by physiological Ca(2+) entry.

In cultured rat dorsal root ganglia neurons, we measured membrane currents, using the patch-clamp whole-cell technique, and the concentrations of free Ca(2+) in the cytosol ([Ca(2+)](i)) and in the lumen of the endoplasmic reticulum (ER) ([Ca(2+)](L)), using high- (Fluo-3) and low- (Mag-Fura-2) affinity Ca(2+)-sensitive fluorescent probes and video imaging. Resting [Ca(2+)](L) concentration varied between 60 and 270 microM. Activation of ryanodine receptors by caffeine triggered a rapid fall in [Ca(2+)](L) levels, which amounted to only 40--50% of the resting [Ca(2+)](L) value. Using electrophysiological depolarization, we directly demonstrate the process of Ca(2+)-induced Ca(2+) release triggered by Ca(2+) entry through voltage-gated Ca(2+) channels. The amplitude of Ca(2+) release from the ER lumen was linearly dependent on I(Ca).

Animals↗

Physiological ecology of Clostridium glycolicum RD-1, an aerotolerant acetogen isolated from sea grass roots.

An anaerobic, H(2)-utilizing bacterium, strain RD-1, was isolated from the highest growth-positive dilution series of a root homogenate prepared from the sea grass Halodule wrightii. Cells of RD-1 were gram-positive, spore-forming, motile rods that were linked by connecting filaments. Acetate was produced in stoichiometries indicative of an acetyl coenzyme A (acetyl-CoA) pathway-dependent metabolism when RD-1 utilized H(2)-CO(2), formate, lactate, or pyruvate. Growth on sugars or ethylene glycol yielded acetate and ethanol as end products. RD-1 grew at the expense of glucose in the presence of low initial concentrations (up to 6% [vol/vol]) of O(2) in the headspace of static, horizontally incubated culture tubes; the concentration of O(2) decreased during growth in such cultures. Peroxidase, NADH oxidase, and superoxide dismutase activities were detected in the cytoplasmic fraction of cells grown in the presence of O(2). In comparison to cultures incubated under strictly anoxic conditions, acetate production decreased, higher amounts of ethanol were produced, and lactate and H(2) became significant end products when RD-1 was grown on glucose in the presence of O(2). Similarly, when RD-1 was grown on fructose in the presence of elevated salt concentrations, lower amounts of acetate and higher amounts of ethanol and H(2) were produced. When the concentration of O(2) in the headspace exceeded 1% (vol/vol), supplemental H(2) was not utilized. The 16S rRNA gene of RD-1 had a 99.7% sequence similarity to that of Clostridium glycolicum DSM 1288(T), an organism characterized as a fermentative anaerobe. Comparative experiments with C. glycolicum DSM 1288(T) demonstrated that it had negligible H(2)- and formate-utilizing capacities. However, carbon monoxide dehydrogenase was detected in both RD-1 and C. glycolicum DSM 1288(T). A 91.4% DNA-DNA hybridization between the genomic DNA of RD-1 and that of C. glycolicum DSM 1288(T) confirmed that RD-1 was a strain of C. glycolicum. These results indicate that (i) RD-1 metabolizes certain substrates via the acetyl-CoA pathway, (ii) RD-1 can tolerate and consume limited amounts of O(2), (iii) oxic conditions favor the production of ethanol, lactate, and H(2) by RD-1, and (iv) the ability of RD-1 to cope with limited amounts of O(2) might contribute to its survival in a habitat subject to daily gradients of photosynthesis-derived O(2).

Acetates↗

The nucleus of differentiated root plant cells: modifications induced by arbuscular mycorrhizal fungi.

The nuclei of plant cells show marked differences in chromatin organisation, related to their DNA content, which ranges from the type with large strands of condensed chromatin (reticulate or chromonematic nuclei) to one with mostly decondensed chromatin (chromocentric or diffuse nuclei). A loosening of the chromatin structure generally occurs in actively metabolising cells, such as differentiating and secretory cells, in relation to their high transcriptional activity. Endoreduplication may occur, especially in plants with a small genome, which increases the availability of nuclear templates, the synthesis of DNA, and probably regulates gene expression. Here we describe structural and quantitative changes of the chromatin and their relationship with transcription that occur in differentiated cells following an increase of their metabolism. The nuclei of root cortical cells of three plants with different 2C DNA content (Allium porrum, Pisum sativum and Lycopersicon esculentum) and their modifications induced by arbuscular mycorrhization, which strongly increase the metabolic activity of colonised cells, are taken as examples.

Cell Nucleus↗

Ion transport in roots: measurement of fluxes using ion-selective microelectrodes to characterize transporter function.

The transport of mineral ions into and out of tissues and cells is central to the life of plants. Ion transport and the plasma membrane transporters themselves have been studied using a variety of techniques. In the last 15 years, measurement of specific ion fluxes has contributed to the characterization of transport systems. Progress in molecular genetics is allowing gene identification and controlled expression of transporter molecules. However the molecular expression of transporter gene products must be characterized at the functional level. The ion-selective microelectrode technique to measure specific ion fluxes non-invasively is ideally suited to this purpose. This technique, its theory, its links with others and its application and prospects in plant science, are discussed. Ions studied include hydrogen, potassium, sodium, ammonium, calcium, chloride and nitrate. Applications discussed include: solute ion uptake by roots; gravitropism and other processes in the root cap, meristematic and elongation zones; Nod factor effect on root hairs; osmotic and salt stresses; oscillations; the effects of light and temperature. Studies have included intact roots, leaf mesophyll and other tissues, protoplasts and bacterial biofilms. A multi-ion capability of the technique will greatly assist functional genomics, particularly when coupled with imaging techniques, patch clamping and the use of suitable mutants.

Cell Membrane↗

Ionic signaling in plant gravity and touch responses.

Plant roots are optimized to exploit resources from the soil and as each root explores this environment it will encounter a range of biotic and abiotic stimuli to which it must respond. Therefore, each root must possess a sensory array capable of monitoring and integrating these diverse stimuli to direct the appropriate growth response. Touch and gravity represent two of the biophysical stimuli that plants must integrate. As sensing both of these signals requires mechano-transduction of biophysical forces to biochemical signaling events, it is likely that they share signal transduction elements. These common signaling components may allow for cross-talk and so integration of thigmotropic and gravitropic responses. Indeed, signal transduction events in both plant touch and gravity sensing are thought to include Ca(2+)- and pH-dependent events. Additionally, it seems clear that the systems responsible for root touch and gravity response interact to generate an integrated growth response. Thus, primary and lateral roots of Arabidopsis respond to mechanical stimuli by eliciting tropic growth that is likely part of a growth strategy employed by the root to circumvent obstacles in the soil. Also, the mechano-signaling induced by encountering an obstacle apparently down-regulates the graviperception machinery to allow this kind of avoidance response. The challenge for future research will be to define how the cellular signaling events in the root cap facilitate this signal integration and growth regulation. In addition, whether other stimuli are likewise integrated with the graviresponse via signal transduction system cross-talk is an important question that remains to be answered.

Arabidopsis↗

The influence of ventricular input impedance on the hydrodynamic performance of bioprosthetic aortic roots in vitro.

BACKGROUND AND AIM OF THE STUDY: Hydrodynamic function testing using pulsatile flow simulators provides a valuable means of comparative assessment of heart valves in vitro. The majority of pulsatile flow simulators consist of modular rigid chambers and a positive displacement pump with an infinite input impedance, in which the inertia of the test fluid results in pressure oscillations when the valves under test are opening and closing. For mechanical and stented bioprosthetic valves these pressure oscillations decay quickly. However, due to the highly compliant nature of tissue roots, the resulting pressure and flow oscillations are extreme and extend throughout systole. With increasing interest in the use of free-sewn roots and valves it is most desirable to improve this hydrodynamic model. The aim of this study was to investigate the influence in changes in ventricular input impedance on the hydrodynamic characteristics of free-sewn aortic roots and stented valves. METHODS: The Leeds pulsatile flow simulator was modified to incorporate additional compliance chambers in the form of a viscoelastic impedance adaptor (VIA) at the pump/ventricular interface. Six 23 mm bioprosthetic aortic roots fixed with 0.5% buffered glutaraldehyde at zero pressure, and a size 23 mm stented porcine aortic bioprosthesis were tested in this modified simulator, at the conditions of maximum and minimum input compliance. RESULTS: The pressure and flow waveforms for the fixed aortic roots showed considerable differences at the conditions of maximum and minimum input compliance. Indeed, the extreme pressure oscillations observed at minimum compliance (infinite input impedance) were not present at maximum compliance, and the forward flow waveform was much smoother. In contrast, for the stented valve, the differences in the pressure and flow waveforms between maximum and minimum input compliance were minimal, but this was expected due to the lack of compliance in the stented valve itself. In addition, the flow and pressure waveforms at maximum compliance in the VIA were comparable for the fixed aortic roots and the stented bioprosthesis, thus allowing direct comparison of the characteristics of these two different devices. Using test conditions of maximum input compliance, effective orifice area for the roots was 1.69 cm2 compared with 1.47 cm2 for the stented valve. CONCLUSION: An appropriate physiological model for the hydrodynamic testing of compliant tissue roots has been established.

Aortic Valve↗

A conceptual framework for investigating plant growth movements, with special reference to root gravitropism, utilizing a microgravity environment.

Microgravity (micro-g) has become established as an important environment in which to conduct experiments that aid in the understanding of gravity-responsive processes in plant growth and development. The gravitropic movement of roots is one example of a gravity-driven differential growth process that can be so investigated, autonomous nutational movements may be another. Although comparison of root movements of agravitropic mutants and their isogenic graviresponsive wild-types in l-g can answer many questions about their gravity-driven processes, they often relate only to gravity perception. Micro-g, however, can help define other constitutive processes which are also influenced by gravity. Examples of the latter are the so-called "tonic" effects, where organ or protoplast weight influences root growth and cellular polarity. A flow-chart is presented which orders the processes of differential growth in roots for analysis using both agravitropic mutants insensitive to gravity and mutant and wild-type roots grown in the micro-g environment. This approach addresses the question whether gravitropism and nutation of root tips are independent or co-existent processes, and also whether gravity determines any features of the nutation, e.g. its amplitude and period. While agravitropic mutants at 1-g can partially answer these questions, micro-g can validate the conclusions and indicate requirements for postulating additional influences such as tonic effects on signal transmission. Gravitropism of roots normally depends on the presence of the root cap, but embryonic tissue and its connection with the seed or grain can also influence gravitropic growth. A hypothetical scheme, based on experimental observations, is proposed for this grain-root-cap interaction. Testing the scheme could be achieved by physically dissociating the compartments presumed to interact in bringing about the tropism. The use of micro-g and agravitropic mutants to compare results with those obtained in 1-g would be of great value in further defining this system.

Gravitation↗

Amino acids regulate salinity-induced potassium efflux in barley root epidermis.

The amino acid content increases substantially in salt-stressed plants. The physiological relevance of this phenomenon remains largely unknown. Using the MIFE ion flux measuring technique, we studied the effects of physiologically relevant concentrations of 26 amino acids on NaCl-induced K(+) flux from barley root epidermis. We show that 21 (of 26) amino acids caused a significant mitigation of the NaCl-induced K(+) efflux, while valine and ornithine substantially enhanced the detrimental effects of salinity on K(+) homeostasis. Our results suggest that physiologically relevant concentrations of free amino acids might contribute to plant adaptive responses to salinity by regulating K(+) transport across the plasma membrane, thus enabling maintenance of an optimal K(+)/Na(+) ratio as opposed to being merely a symptom of plant damage by stress. Investigating the specific mechanisms of such amelioration remains a key issue for future studies.

Amino Acids↗