On the Mathematical Modelling of Competitive Invasive Weed Dynamics.

We explore the dynamics of invasive weeds by partial differential equation (PDE) modelling and applying dynamical system and phase portrait techniques. We begin by applying the method of characteristics to a preexisting PDE model of the spreading of T. fluminensis, an invasive weed which has been responsible for native forest depletion. We explore the system both at particular points in space and over all of space, in one dimension, as a function of time. Our model suggests that an increase in the rate of spread of the weed through space will increase the efficacy of control measures taken at the weed's spatial boundary. We then propose new competition models based on the previous model and explore the existence of travelling wave solutions. These models represent both the cases with (i) a competing native plant species which spreads through the forest and (ii) a non-mobile, established native plant species. In the former case, the model suggests that an increased mass-action coefficient between the competing species is sufficient and necessary for the transition of the forest into a state of coexistence. In the latter case, the result is not as strong: a sufficiently large rate of competition between the species excludes the possibility of native plant extinction and hence suggests that forest depletion will not occur, but does not imply coexistence. We perform some numerical simulations to support our analytic results. In all cases, we give a discussion on the physical and biological interpretations of our results. We conclude with some suggestions for future work and with a discussion of the advantages and disadvantages of the methods.

High resolution leaf spectral signature as a tool for foliar pigment estimation displaying potential for species differentiation.

Optical leaf profiles depend on foliar pigment type and content, as well as anatomical aspects and cellular ultrastructure, whose effects are shown in several species. Monocotyledon and Dicotyledon plants presenting natural pigment content variations and anatomical alterations were analyzed. Each plant species displays its own spectral signatures, which are, in turn, influenced by foliar pigment class (composition) and concentration, as well as anatomical and ultrastructural plant cell characteristics. Plants with no anthocyanin displayed increased reflectance and transmittance in the green spectral region (501-565 nm), while values decreased in the presence of anthocyanin. At wavelengths below 500 nm (350-500 nm), strong overlapping signatures of phenolics, carotenoids, chlorophylls, flavonoids and anthocyanins were observed. Using a partial least squares regression applied to 350-700 nm spectral data allowed for accurate estimations of different foliar pigment levels. In addition, a PCA and discriminant analysis were able to efficiently discriminate different species displaying spectra overlapping. The use of absorbance spectra only was able to discriminate species with 100 % confidence. Finally, a discussion on how different wavelengths are absorbed and on anatomical interference of light interaction in leaf profiles is presented.

Slow induction of chlorophyll a fluorescence excited by blue and red light in Tradescantia leaves acclimated to high and low light.

Tradescantia is a good model for assaying induction events in higher plant leaves. Chlorophyll (Chl) fluorescence serves as a sensitive reporter of the functional state of photosynthetic apparatus in chloroplasts. The fluorescence time-course depends on the leaf growth conditions and actinic light quality. In this work, we investigated slow induction of Chl a fluorescence (SIF) excited by blue light (BL, λmax = 455 nm) or red light (RL, λmax = 630 nm) in dark-adapted leaves of Tradescantia fluminensis acclimated to high light (~ 1000 µmol photons m-2 s-1; HL) or low light (~ 100 µmol photons m-2 s-1; LL). Our special interest was focused on the contribution of the avoidance response to SIF kinetics. Bearing in mind that BL and RL have different impacts on photoreceptors that initiate chloroplast movements within the cell (accumulation/avoidance responses), we have compared the SIF patterns during the action of BL and RL. The time-courses of SIF and kinetics of non-photochemical quenching (NPQ) of Chl a fluorescence revealed a certain difference when leaves were illuminated by BL or RL. In both cases, the yield of fluorescence rose to the maximal level P and then, after the lag-phase P-S-M1, the fluorescence level decreased toward the steady state T (via the intermediate phases M1-M2 and M2-T). In LL-acclimated leaves, the duration of the P-S-M1 phase was almost two times longer that in HL-grown plants. In the case of BL, the fluorescence decay included the transient phase M1-M2. This phase was obscure during the RL illumination. Non-photochemical quenching of Chl a fluorescence has been quantified as [Formula: see text], where [Formula: see text] and [Formula: see text] stand for the fluorescence response to saturating pulses of light applied to dark-adapted and illuminated samples, respectively. The time-courses of such a formally determined NPQ value were markedly different during the action of RL and BL. In LL-grown leaves, BL induced higher NPQ as compared to the action of RL. In HL-grown plants, the difference between the NPQ responses to BL and RL illumination was insignificant. Comparing the peculiarities of Chl a fluorescence induced by BL and RL, we conclude that the avoidance response can provide a marked contribution to SIF and NPQ generation. The dependence of NPQ on the quality of actinic light suggests that chloroplast movements within the cell have a noticeable impact on the formally determined NPQ value. Analyzing kinetics of post-illumination decay of NPQ in the context of solar stress resistance, we have found that LL-acclimated Tradescantia leaves are more vulnerable to strong light than the HL-grown leaves.

Tradescantia-based models: a powerful looking glass for investigation of photoacclimation and photoadaptation in plants.

Here, we summarize diverse evidence from species that belong to the genus Tradescantia, which we propose as handy and versatile models for studies of the ecology of photosynthesis and the mechanisms of photoacclimation in higher plants. A valuable feature of this genus is the amazingly broad range of ecological niches occupied by its species: from shady understory of tropical rainforest to deserts and semideserts. The former habitats demand shade tolerance (e.g. that featured by Tradescantia fluminensis), whereas the latter requires succulence and/or high light stress tolerance (evident in e.g. Tradescantia navicularis). At the same time, the acclimative traits of Tradescantia species seem quite moderate at first glance. Certainly, their basic principles of acclimation seem to differ in some aspects from the ones typical for most of other higher plants. This review presents a systematic analysis of irradiance responses of Tradescantia species studied on different timescales. The specifics of Tradescantia responses to irradiance make the plants of this genus a 'multitool' for studies in this field. Similarity of irradiance acclimation patterns is a characteristic feature in the ecologically contrasting Tradescantia species, which may inspire further insights into physiology and evolution of plants.

Evaluation of vehicular pollution using the TRAD-MCN mutagenic bioassay with Tradescantia pallida (Commelinaceae).

Biomonitoring is one of the tools used to assess the mutagenic potential of the atmosphere. In this study, the mutagenicity of Tradescantia pallida, a species of plant largely present in urban environments, was investigated. The objectives of this study was to estimate the mutagenic potential of vehicular flow through the TRAD-MCN bioassay in cities located at different altitudes in the southwest mesoregion of Mato Grosso do Sul, Brazil, to infer possible abiotic agents that may contribute to the effects of atmospheric pollutants, and finally to map the cities with greater risks to the health of the local population. To achieve these objectives, the Tradescantia-micronucleus test was performed on young buds of T. pallida collected between August 2015 and August 2016 in nine cities of Mato Grosso do Sul. These buds were exposed to traffic flows of various intensities. The data collected consisted of measurements of meteorological parameters and vehicular traffic counts for each city. The variables considered were: mean ambient temperature; micronuclei frequency; vehicular flow; altitude; relative humidity; pluviosity. The application of the Trad-MCN bioassay, with the consideration of environmental variables and altitudes, and the use of the Kernel interpolation technique, allowed us to map the areas with significant pollution risks to the population. The highest frequency of exposure to mutagens occurred in the cities with the highest vehicular traffic intensity. The average ambient temperature failed to show a linear association with the frequency of the micronuclei in the samples analyzed (r = 0.11ns). A positive correlation was observed between micronuclei frequency and vehicular flow, (r = 0.67; p ≤ 0.001%) and between micronuclei frequency and altitude (r = 0.24; p ≤ 0.05). A negative correlation was found between relative humidity and micronuclei frequency (r = -0.19; p ≤ 0.05%). Thus, higher micronuclei frequency tended to be present in locations with low relative humidity and high altitudes and vehicular flow.

Acclimation of shade-tolerant and light-resistant Tradescantia species to growth light: chlorophyll a fluorescence, electron transport, and xanthophyll content.

In this study, we have compared the photosynthetic characteristics of two contrasting species of Tradescantia plants, T. fluminensis (shade-tolerant species), and T. sillamontana (light-resistant species), grown under the low light (LL, 50-125 µmol photons m-2 s-1) or high light (HL, 875-1000 µmol photons m-2 s-1) conditions during their entire growth period. For monitoring the functional state of photosynthetic apparatus (PSA), we measured chlorophyll (Chl) a emission fluorescence spectra and kinetics of light-induced changes in the heights of fluorescence peaks at 685 and 740 nm (F 685 and F 740). We also compared the light-induced oxidation of P700 and assayed the composition of carotenoids in Tradescantia leaves grown under the LL and HL conditions. The analyses of slow induction of Chl a fluorescence (SIF) uncovered different traits in the LL- and HL-grown plants of ecologically contrasting Tradescantia species, which may have potential ecophysiological significance with respect to their tolerance to HL stress. The fluorometry and EPR studies of induction events in chloroplasts in situ demonstrated that acclimation of both Tradescantia species to HL conditions promoted faster responses of their PSA as compared to LL-grown plants. Acclimation of both species to HL also caused marked changes in the leaf anatomy and carotenoid composition (an increase in Violaxanthin + Antheraxantin + Zeaxanthin and Lutein pools), suggesting enhanced photoprotective capacity of the carotenoids in the plants grown in nature under high irradiance. Collectively, the results of the present work suggest that the mechanisms of long-term PSA photoprotection in Tradescantia are based predominantly on the light-induced remodeling of pigment-protein complexes in chloroplasts.

Light acclimation of shade-tolerant and light-resistant Tradescantia species: induction of chlorophyll a fluorescence and P700 photooxidation, expression of PsbS and Lhcb1 proteins.

In this work, we have compared photosynthetic performance and expression of the PsbS and Lhcb1 proteins in two contrast ecotypes of Tradescantia species, T. fluminensis (shade-tolerant) and T. sillamontana (light-resistant), grown at two intensities of light: 50-125 µmol photons m-2 s-1 (low light, LL) and 875-1000 µmol photons m-2 s-1 (high light, HL). Using the EPR method for measuring the P700 content, we have found that LL-grown plants of both species have higher (by a factor of ≈1.7-1.8) contents of PSI per fresh weight unit as compared to HL-grown plants. Acclimation of plants to LL or HL irradiation also influences the Chl(a + b) level and expression of the PsbS and Lhcb1 proteins. Immunoblotting analysis showed that acclimation to HL stimulates (by a factor of ≈1.7-1.8) the level of PsbS related to the total number of P700 centers. In light-resistant species T. sillamontana, the ratio PsbS/P700 is about 2-times higher than in shade-tolerant species T. fluminensis grown under the same conditions. This should enhance the capacity of their leaves for protection against the light stress. In agreement with these observations, the capacity of leaves for NPQ induction was enhanced during plant acclimation to HL. Kinetic studies of P700 photooxidation and light-induced changes in the yield of Chl a fluorescence also revealed that the short-term regulation of electron transport processes in chloroplasts, which manifested themselves in the kinetics of [Formula: see text] induction and the rate of Chl a fluorescence quenching, occurred more rapidly in HL-grown plants than in LL-grown plants. Thus, both factors, enhanced expression of PsbS and more rapid response of the photosynthetic electron transport chain to dark-to-light transitions should increase the capacity of HL-grown plants for their resistance to rapid fluctuations of solar light.

Probing plasmodesmata function with biochemical inhibitors.

To investigate plasmodesmata (PD) function, a useful technique is to monitor the effect on cell-to-cell transport of applying an inhibitor of a physiological process, protein, or other cell component of interest. Changes in PD transport can then be monitored in one of several ways, most commonly by measuring the cell-to-cell movement of fluorescent tracer dyes or of free fluorescent proteins. Effects on PD structure can be detected in thin sections of embedded tissue observed using an electron microscope, most commonly a Transmission Electron Microscope (TEM). This chapter outlines commonly used inhibitors, methods for treating different tissues, how to detect altered cell-to-cell transport and PD structure, and important caveats.

Assessment of genotoxicity and acute toxic effect of the imatinib mesylate in plant bioassays.

Imatinib mesylate (IM) is at present one of the most widely used cytostatic drugs in developed countries but information on its ecotoxicological activities is scarce. This article describes the results of the first investigation in which genotoxic and acute toxic properties of the drug were studied in higher plants. IM was tested in two widely used plant bioassays namely in micronucleus (MN) assays with meiotic tetrad cells of Tradescantia (clone #4430) and in mitotic root tip cells of Allium cepa. Additionally, acute toxic effects (inhibition of cell division and growth of roots) were monitored in the onions. Furthermore, we studied the impact of the drug on the fertility of higher plants in pollen abortion experiments with three wildlife species (Chelidonium majus, Tradescantia palludosa and Arabidopsis thaliana). In MN assays with Tradesacantia a significant effect was seen with doses ⩾10µM; the Allium MN assay was even more sensitive (LOEL⩾1.0µM). A significant decrease of the mitotic indices was detected at levels ⩾10µM in the onions and reduction of root growth with ⩾100µM. In the pollen fertility assays clear effects were observed at doses ⩾147.3mgkg(-1). Data concerning the annual use of the drug in European countries (France, Germany, Slovenia) enable the calculation of the predicted environmental concentration (PEC) values which are in the range between 3.3 and 5.0ngL(-1). Although comparisons with the genotoxic potencies of other commonly used cytostatic drugs and with highly active heavy metal compounds show that IM is an extremely potent genotoxin in higher plants, it is evident that the environmental concentrations are ⩾5 orders of magnitude lower as the levels which are required to cause adverse effects.

Is the signal from the mesophyll to the guard cells a vapour-phase ion?

Previous studies have suggested that the red light and CO2 responses of stomata are caused by a signal from the mesophyll to the guard cells. Experiments were conducted to test the idea that this signal is a vapour-phase ion. Stomata in isolated epidermes of Tradescantia pallida were found to respond to air ions created by an electrode that was positioned under the epidermes. Anthocyanins in the epidermes of this species were observed to change colour in response to these air ions, and this change in colour was attributed to changes in pH. A similar change in lower epidermal colour was observed in intact leaves upon illumination and with changes in CO2 concentration. Based on the change in epidermal colour, the pH of the epidermis was estimated to be approximately 7.0 in darkness and 6.5 in the light. Stomata in isolated epidermes responded to pH when suspended over (but not in contact with) solutions of different pH. We speculate that stomatal responses to CO2 and light are caused by vapour-phase ions, possibly hydronium ions that change the pH of the epidermis.

Effects of light environment on the induction of chlorophyll fluorescence in leaves: a comparative study of Tradescantia species of different ecotypes.

In this work, using a PAM-fluorimetry technique, we have compared effects of plant adaptation to the light or dark conditions on the kinetics of chlorophyll a fluorescence yield in Tradecantia leaves of several species (Tradescantia albiflora, Tradescantia fluminensis, Tradescantia navicularis, and Tradescantia sillamontana), which represent plants of different ecotypes. Two fluorescence parameters were used to assess photosynthetic performance in vivo: non-photochemical quenching (NPQ) of chlorophyll fluorescence (q(NPQ)) determined by energy losses in the light-harvesting antenna of photosystem 2 (PS2), and PS2 operating efficiency (Φ(PSII)). Comparative study of light-induced changes in q(NPQ) and Φ(PSII) has demonstrated that shade-tolerant Tradecantia species (T. albiflora Kunth, T. fluminensis Vell.) reveal higher capacities for NPQ and demonstrate slower transitions between the 'light-adapted' and 'dark-adapted' states than succulent species T. navicularis and T. sillamontana, which are typical habitats of semi-deserts. We analyze the photosynthetic performance of Tradescantia species in the context of their adaptabilities to variable environment conditions. The ability of shade-tolerant plants to retain a relatively long-term (∼40-60 min) 'memory' for illumination history may be associated with the regulatory mechanisms that provide the flexibility of photosynthetic apparatus in response to fluctuations of light intensity.

ESEM imaging of dynamic biological processes: the closure of stomatal pores.

Historically, electron microscopy of dynamic biological processes has been impossible to achieve in real time because conventional electron microscopy requires specimen fixation, dehydration and metallic coating. The advent of the environmental scanning electron microscope removes these restrictions, allowing fully hydrated samples to be imaged in their native state. We explore the possibility of secondary electron imaging of biological systems undergoing natural morphological changes in the microscope chamber and present a proof of principle study on the closure of stomatal pores in Tradescantia andersonia leaf tissue. An imaging protocol is developed and the advantages and limitations of this high-resolution imaging technique are considered, including a discussion of potential beam damage mechanisms.

Stomatal responses to humidity and temperature in darkness.

Stomatal responses to leaf temperature (T(l)) and to the mole fractions of water vapour in the ambient air (w(a)) and the leaf intercellular air spaces (w(i)) were determined in darkness to remove the potential effects of changes in photosynthesis and intercellular CO(2) concentration. Both the steady-state and kinetic responses of stomatal conductance (g(s)) to w(a) in darkness were found to be indistinguishable from those in the light. g(s) showed a steep response to the difference (Deltaw) between w(a) and w(i) when w(a) was varied. The response was much less steep when w(i) was varied. Although stomatal apertures responded steeply to T(l) when Deltaw was held constant at 17 mmol mol(-1), the response was much less steep when Deltaw was held constant at about zero. Similar results were obtained in the light for Deltaw = 15 mmol mol(-1) and Deltaw approximately 0 mmol mol(-1). These results are discussed in the context of mechanisms for the stomatal response to humidity.

Microinjecting FM4-64 validates it as a marker of the endocytic pathway in plants.

The amphiphilic dye FM4-64 is used to investigate endocytosis and vesicle trafficking in living eukaryotic cells. The standing hypothesis is that it is inserted into the outer leaflet of the plasma membrane and, from there, is passed on to intracellular membrane compartments by endocytosis. We tested this hypothesis by microinjecting FM4-64 into the cytoplasm and vacuole of Nicotiana tabacum BY-2 suspension culture cells and Tradescantia virginiana stamen hair cells. We found that the dye did not label any membranes when injected into the cytoplasm, but clearly labelled the tonoplast when injected directly into the vacuole. However, because the dye is pH-sensitive, the fluorescence intensity between the plasma membrane and tonoplast varied. We conclude that FM4-64 is a specific marker for the endocytic pathway. Nevertheless, little is known about the molecular interactions of FM4-64 with these particular phospholipid membrane leaflets. We, therefore, appeal for biochemical research to determine which membrane lipids FM4-64 interacts with.

The role of the mesophyll in stomatal responses to light and CO2.

Stomatal responses to light and CO(2) were investigated using isolated epidermes of Tradescantia pallida, Vicia faba and Pisum sativum. Stomata in leaves of T. pallida and P. sativum responded to light and CO(2), but those from V. faba did not. Stomata in isolated epidermes of all three species could be opened on KCl solutions, but they showed no response to light or CO(2). However, when isolated epidermes of T. pallida and P. sativum were placed on an exposed mesophyll from a leaf of the same species or a different species, they regained responsiveness to light and CO(2). Stomatal responses in these epidermes were similar to those in leaves in that they responded rapidly and reversibly to changes in light and CO(2). Epidermes from V. faba did not respond to light or CO(2) when placed on mesophyll from any of the three species. Experiments with single optic fibres suggest that stomata were being regulated via signals from the mesophyll produced in response to light and CO(2) rather than being sensitized to light and CO(2) by the mesophyll. The data suggest that most of the stomatal response to CO(2) and light occurs in response to a signal generated by the mesophyll.

Stomatal responses to humidity in isolated epidermes.

The ability of guard cells to hydrate and dehydrate from the surrounding air was investigated using isolated epidermes of Tradescantia pallida and Vicia faba. Stomata were found to respond to the water vapour pressure on the outside and inside of the epidermis, but the response was more sensitive to the inside vapour pressure, and occurred in the presence or absence of living, turgid epidermal cells. Experiments using helium-oxygen air showed that guard cells hydrated and dehydrated entirely from water vapour, suggesting that there was no significant transfer of water from the epidermal tissue to the guard cells. The stomatal aperture achieved at any given vapour pressure was shown to be consistent with water potential equilibrium between the guard cells and the air near the bottom of the stomatal pore, and water vapour exchange through the external cuticle appeared to be unimportant for the responses. Although stomatal responses to humidity in isolated epidermes are the result of water potential equilibrium between the guard cells and the air near the bottom of the stomatal pore, stomatal responses to humidity in leaves are unlikely to be the result of a similar equilibrium.

Cell-to-cell pathway dominates xylem-epidermis hydraulic connection in Tradescantia fluminensis (Vell. Conc.) leaves.

A steady supply of water is indispensable for leaves to fulfil their photosynthetic function. Understanding water movement in leaves, especially factors that regulate the movement of water flux from xylem to epidermis, requires that the nature of the transport pathway be elucidated. To determine the hydraulic linkage between xylem and epidermis, epidermal cell turgor pressure (P (t)) in leaves of Tradescantia fluminensis was monitored using a cell pressure probe in response to a 0.2 MPa step change in xylem pressure applied at the leaf petiole. Halftime of P (t) changes (T(x)(1/2)) were 10-30 times greater than that of water exchange across an individual cell membrane (T(m)(1/2)) suggesting that cell-to-cell water transport constitutes a significant part of the leaf hydraulic path from xylem to epidermis. Furthermore, perfusion of H(2)O(2) resulted in increases of both T(m)(1/2) and T(x)(1/2) by a factor of 2.5, indicating that aquaporins may play a role in the xylem to epidermis hydraulic link. The halftime for water exchange (T(m)(1/2)) did not differ significantly between cells located at the leaf base (2.5 s), middle (2.6 s) and tip (2.5 s), indicating that epidermal cell hydraulic properties are similar along the length of the leaf. Following the pressure application to the xylem (0.2 MPa), P (t) changed by 0.12, 0.06 and 0.04 MPa for epidermal cells at the base, middle and the tip of the leaf, respectively. This suggests that pressure dissipation between xylem and epidermis is significant, and that the pressure drop along the vein may be due to its structural similarities to a porous pipe, an idea which was further supported by measurements of xylem hydraulic resistance using a perfusion technique.

Dynamics of adaptation of stomatal behaviour to moderate or high relative air humidity in Tradescantia virginiana.

Chlorophyll fluorescence imaging was used to measure stomatal closure in response to desiccation of Tradescantia virginiana leaves grown under high (90%) and moderate (55%) relative humidities (RHs), or transferred between these humidities. Stomata in leaves grown at high RH were less responsive to desiccation than those of leaves grown at moderate RH. Stomata of plants transferred from moderate RH conditions to high RH showed the same diminished closure in response to desiccation as did stomata that developed at high RH. This response was found both when the leaves were fully expanded and when still actively expanding during the moderate RH pre-treatment. Four days of exposure to high RH was the minimal exposure time to induce the diminished closure response. When leaves were grown in high RH prior to a 10 d moderate RH treatment, the reduced stomatal closure response to desiccation was only reversed in leaves (regions) which were actively expanding during moderate RH treatment. This indicates that with respect to stomatal responses to desiccation, high RH leaf regions have a limited capacity to adapt to moderate RH conditions. The decrease in responsiveness to desiccation of the stomata, induced by long-term exposure to high RH, was not due to osmotic adjustment in the leaves. Within 1 d after transferring moderate RH-grown plants to a high RH, the abscisic acid (ABA) concentration of their leaves decreased to the low level of ABA found in high RH-grown leaves. The closure response in leaves exposed to high RH for 5 d, however, could not be fully restored by the application of ABA. Transferring plants from high to moderate RH resulted in increased ABA levels within 2 d without a recovery of the stomatal closing response. It is discussed that the diminished stomatal closure in plants exposed to high RH could be due to changes in the signalling pathway for ABA-related closure of stomata or to an increased sequestration of ABA by mesophyll tissue or the symplast in the epidermis, induced by a longer period (several days) of a low ABA level.

Potential of the Trad-MCN assay applied with inflorescences of Tradescantia pallida 'Purpurea' for evaluating air contamination by naphthalene.

The aims of this study were to determine clastogenic responses of Tradescantia pallida cv. Purpurea to naphthalene (NAPH) by means of the bioassay Trad-MCN with inflorescences of T. pallida cv. Purpurea and to verify if this assay might be an indicator of the potential risk imposed in a workplace, where solid insecticide containing NAPH is usually applied. The clastogenic potential of NAPH was assessed by using static and dynamic experimental systems. In both systems, increased micronucleus frequencies were observed in inflorescences submitted to increasing concentrations of solid or gaseous NAPH. The evident clastogenicity verified in inflorescences exposed experimentally to 25-50 mg m(-3) of NAPH during 6h points to a narrow threshold of plant sensitivity, indicating risks under lower NAPH levels than the standards established by OSHA and therefore revealing its suitability for biomonitoring purposes. However, the clastogenic risk should be carefully investigated by other monitoring methods if human health is taken into consideration.