Willow and poplar for the phyto-treatment of landfill leachate in Mediterranean climate.

Phytotechnological approaches using living plants are currently being proposed to address a wide range of environmental purposes including the treatment of landfill leachate (LL). Despite their popularity, few studies have investigated this possibility under actual Mediterranean conditions using fast-growing trees. This research reports the results of a two-year project where poplar and willow grown in mesocosm were tested for their ability to withstand and remove specific pollutants from different [Low: 7% (1st year) and 15% (2nd year); High: 15% (1st year) and 30% (2nd year)] amounts of LL. Results indicate that both species were able to treat 340 (Low) and 680 (High) m3 ha-1 in the establishment year (70 days) and 2470 (Low) and 4950 (High) m3 ha-1 in the second year (150 days). Both species yielded the same aboveground biomass, but under high LL treatment, poplar performed better than willow. Poplar showed on average significantly higher extraction rates for Cd, Cu, P, and N than willow. Moreover, under high LL treatment, poplar also seemed more efficient than willow in decreasing the concentration of specific pollutants (BOD5, COD and As) in output effluent. However, with low LL loads both species were able to significantly reduce other compounds (i.e. NH4-N, Cu and Ni). By contrast, Cl, surfactants, and NO3-N, had a tendency to accumulate over time in the effluent and could still represent an actual constraint to large-scale application of the technique. The fate of such pollutants should be investigated with further research to better inform strategies used to manage low amounts of high-concentrated effluent.

Na+ extrusion from the cytosol and tissue-specific Na+ sequestration in roots confer differential salt stress tolerance between durum and bread wheat.

The progress in plant breeding for salinity stress tolerance is handicapped by the lack of understanding of the specificity of salt stress signalling and adaptation at the cellular and tissue levels. In this study, we used electrophysiological, fluorescence imaging, and real-time quantitative PCR tools to elucidate the essentiality of the cytosolic Na+ extrusion in functionally different root zones (elongation, meristem, and mature) in a large number of bread and durum wheat accessions. We show that the difference in the root's ability for vacuolar Na+ sequestration in the mature zone may explain differential salinity stress tolerance between salt-sensitive durum and salt-tolerant bread wheat species. Bread wheat genotypes also had on average 30% higher capacity for net Na+ efflux from the root elongation zone, providing the first direct evidence for the essentiality of the root salt exclusion trait at the cellular level. At the same time, cytosolic Na+ accumulation in the root meristem was significantly higher in bread wheat, leading to the suggestion that this tissue may harbour a putative salt sensor. This hypothesis was then tested by investigating patterns of Na+ distribution and the relative expression level of several key genes related to Na+ transport in leaves in plants with intact roots and in those in which the root meristems were removed. We show that tampering with this sensing mechanism has resulted in a salt-sensitive phenotype, largely due to compromising the plant's ability to sequester Na+ in mesophyll cell vacuoles. The implications of these findings for plant breeding for salinity stress tolerance are discussed.

Cell-Type-Specific H+-ATPase Activity in Root Tissues Enables K+ Retention and Mediates Acclimation of Barley (Hordeum vulgare) to Salinity Stress.

While the importance of cell type specificity in plant adaptive responses is widely accepted, only a limited number of studies have addressed this issue at the functional level. We have combined electrophysiological, imaging, and biochemical techniques to reveal the physiological mechanisms conferring higher sensitivity of apical root cells to salinity in barley (Hordeum vulgare). We show that salinity application to the root apex arrests root growth in a highly tissue- and treatment-specific manner. Although salinity-induced transient net Na+ uptake was about 4-fold higher in the root apex compared with the mature zone, mature root cells accumulated more cytosolic and vacuolar Na+, suggesting that the higher sensitivity of apical cells to salt is not related to either enhanced Na+ exclusion or sequestration inside the root. Rather, the above differential sensitivity between the two zones originates from a 10-fold difference in K+ efflux between the mature zone and the apical region (much poorer in the root apex) of the root. Major factors contributing to this poor K+ retention ability are (1) an intrinsically lower H+-ATPase activity in the root apex, (2) greater salt-induced membrane depolarization, and (3) a higher reactive oxygen species production under NaCl and a larger density of reactive oxygen species-activated cation currents in the apex. Salinity treatment increased (2- to 5-fold) the content of 10 (out of 25 detected) amino acids in the root apex but not in the mature zone and changed the organic acid and sugar contents. The causal link between the observed changes in the root metabolic profile and the regulation of transporter activity is discussed.

Sensory, spectrometric (PTR-ToF-MS) and chemometric analyses to distinguish extra virgin from virgin olive oils.

Olive oil samples were obtained from six cultivars grown in different environments, and graded by chemical analyses as extra virgin (EVOOs). These were evaluated for flavors and off-flavors, and relative VOCs spectrum as determined by PTR-ToF-MS. A hierarchical clustering of Panel test data separated olive oil in three groups, one including the samples with perceived off-flavor (VOOs), regardless of cultivar and environment. The Pearson's correlation coefficients between the mass data from PTR-ToF-MS and the sensory characteristics perceived by the Panel test were determined. A mass-to-sensory attributes correlation index was calculated. A color-coded card was built up based on the intensities (ncps) of five selected protonated mass data that was able to distinguish EVOOs from VOOs olive oil samples.

Developing and validating a high-throughput assay for salinity tissue tolerance in wheat and barley.

MAIN CONCLUSION: Leaf tissue tolerance was strongly and positively correlated with overall salt tolerance in barley, but not in wheat where the inability of sensitive varieties to exclude Na(+) is compensated by their better ability to handle Na(+) accumulated in the shoot via tissue tolerance mechanisms. A new high-throughput assay was developed to use the excised leaves to eliminate the confounding contribution of sodium exclusion mechanisms and evaluate genetic variability in salinity tissue tolerance in a large number of wheat (Triticum aestivum and Triticum turgidum ssp. durum) and barley (Hordeum vulgare) accessions. The changes in relative chlorophyll content (measured as chlorophyll content index, CCI) in excised leaves exposed to 50 mM NaCl for 48 h were found to be a reliable indicator of leaf tissue tolerance. In both wheat and barley, relative CCI correlated strongly with the overall plant salinity tolerance (evaluated in glasshouse experiments). To a large extent, this tissue tolerance was related to more efficient vacuolar Na(+) sequestration in leaf mesophyll, as revealed by fluorescent Na(+) dye imaging experiments. However, while in barley this correlation was positive, tissue tolerance in wheat correlated negatively with overall salinity tolerance. As a result, more salt-sensitive durum wheat genotypes possessed higher tissue tolerance than bread wheat plants, and this negative correlation was present within each of bread and durum wheat clusters as well. Overall, these results indicate that the lack of effective Na(+) exclusion ability in sensitive wheat varieties is compensated by their better ability to handle Na(+) accumulated in the shoot via tissue tolerance mechanisms. Implications of these findings for plant breeding for salinity tolerance are discussed.

PTR-TOF-MS analysis of volatile compounds in olive fruits.

BACKGROUND: Volatile compounds of Cellina di Nardò and Ogliarola Barese, two typical Italian olive varieties, have been characterised at different ripening stages. Proton transfer reaction-time-of-flight-mass spectrometry (PTR-TOF-MS) was used for the first time on these fruits with the aim of characterising the volatile profile and, in the case of Ogliarola, the changes which may occur during the maturation process. RESULTS: PTR-TOF-MS does not involve any sample pre-treatment, and allows high-resolution measurements, large spectra and small fragmentation of the volatiles. Therefore it allows both compound identification and data statistical treatments. In the present work, about 40 compounds that contribute to the discrimination between samples of the two varieties have been identified. CONCLUSIONS: Three groups of compounds were identified: (1) compounds that are typical of mature fruits of Ogliarola, (2) compounds that tend to decrease during the change from green to mature fruits, and (3) compounds that increase during the maturation process.

Understanding interdisciplinary perspectives of plant intelligence: Is it a matter of science, language, or subjectivity?

BACKGROUND: Evidence suggests that plants can behave intelligently by exhibiting the ability to learn, make associations between environmental cues, engage in complex decisions about resource acquisition, memorize, and adapt in flexible ways. However, plant intelligence is a disputed concept in the scientific community. Reasons for lack of consensus can be traced back to the history of Western philosophy, interpretation of terminology, and due to plants lacking neurons and a central nervous system. Plant intelligence thus constitutes a novel paradigm in the plant sciences. Therefore, the perspectives of scientists in plant-related disciplines need to be investigated in order to gain insight into the current state and future development of this concept. METHODS: This study analyzed opinions of plant intelligence held by scientists from different plant-related disciplines, including ethnobiology and other biological sciences, through an online questionnaire. RESULTS: Our findings show that respondents' personal belief systems and the frequency of taking into account other types of knowledge, such as traditional knowledge, in their own field(s) of study, were associated with their opinions of plant intelligence. Meanwhile, respondents' professional expertise, background (discipline), or familiarity with evidence provided on plant intelligence did not affect their opinions. CONCLUSIONS: This study emphasizes the influential role of scientists' own subjective beliefs. In response, two approaches could facilitate transdisciplinary understanding among scientists: (1) effective communication designed to foster change in agreement based on presented information; and (2) holding space for an interdisciplinary dialogue where scientists can express their own subjectivities and open new opportunities for collaboration.

Modelling botanical biofiltration of indoor air streams contaminated by volatile organic compounds.

Botanical filtration is a biological-based treatment method suitable for removing hazardous volatile organic compounds (VOCs) from air streams, based on forcing an air flow through a porous substrate and foliage of a living botanical compartment. The pathways and removal mechanisms during VOC bioremediation have been largely investigated; however, their mathematical representation is well established only for the non-botanical components of the system. In this study, we evaluated the applicability of such a modelling scheme to systems which include a botanical compartment. We implemented a one-dimensional numerical model and performed a global sensitivity analysis to measure the input parameters influence on the transient and steady biofilter responses. We found that the most sensitive parameters on the transient-state behaviour were the mass transfer coefficient between gas and solid surfaces, and the fraction of solid surfaces covered by the biofilm; the steady-state response was primarily influenced by the biofilm specific surface area and the fraction of surfaces covered by the biofilm. We calibrated the identified set of parameters and successfully validated the model against data from a pilot-scale installation. The results showed that the application of the model to systems with a botanical compartment is feasible, although under a strict set of assumptions.

On the mechanism underlying photosynthetic limitation upon trigger hair irritation in the carnivorous plant Venus flytrap (Dionaea muscipula Ellis).

Mechanical stimulation of trigger hairs on the adaxial surface of the trap of Dionaea muscipula leads to the generation of action potentials and to rapid leaf movement. After rapid closure secures the prey, the struggle against the trigger hairs results in generation of further action potentials which inhibit photosynthesis. A detailed analysis of chlorophyll a fluorescence kinetics and gas exchange measurements in response to generation of action potentials in irritated D. muscipula traps was used to determine the 'site effect' of the electrical signal-induced inhibition of photosynthesis. Irritation of trigger hairs and subsequent generation of action potentials resulted in a decrease in the effective photochemical quantum yield of photosystem II (Φ(PSII)) and the rate of net photosynthesis (A(N)). During the first seconds of irritation, increased excitation pressure in photosystem II (PSII) was the major contributor to the decreased Φ(PSII). Within ∼1 min, non-photochemical quenching (NPQ) released the excitation pressure at PSII. Measurements of the fast chlorophyll a fluorescence transient (O-J-I-P) revealed a direct impact of action potentials on the charge separation-recombination reactions in PSII, although the effect seems to be small rather than substantial. All the data presented here indicate that the main primary target of the electrical signal-induced inhibition of photosynthesis is the dark reaction, whereas the inhibition of electron transport is only a consequence of reduced carboxylation efficiency. In addition, the study also provides valuable data confirming the hypothesis that chlorophyll a fluorescence is under electrochemical control.

Relationship between Leachate Pollution Index and growth response of two willow and poplar hybrids: Implications for phyto-treatment applications.

Phytoremediation is a potentially suitable technique for the reclamation of toxic landfill leachate (LL) by decreasing its volume through water uptake and improving its composition by uptake, accumulation and amelioration of pollutants. We investigated the use of two parameters, the LL concentration and the Leachate Pollution Index (LPI), a method used to determine the phytotoxicity potential of a leachate source based on a weighted sum of its components, to set the best LL dilution to apply when poplar clone 'Orion' and willow clone 'Levante' are selected for phytoremediation. Cuttings were watered with five LL concentrations ranging from 0 to 100%. The poplar clone showed significantly higher values than the willow clone for lowest effective concentration index (LOEC) for leaf (i.e. 11.3% vs 10.5%; p = 0.0284) and total biomass (i.e. 10.9% vs 10.6%; p = 0.0402) and for lowest effective LPI for leaf (i.e. 12.3 vs 12.1; p = 0.0359) and total biomass (i.e. 12.8 versus 12.2; p = 0.0365), respectively, with effectiveness demonstrating the LOEC or LPI value at which the parameter is negatively affected. Photosynthetic rates were higher in poplar than willow in both control and the lowest LL dilution, but rapidly declined in both at higher LL dilutions. Although a direct translation of data from bench trials to field conditions should be investigated, we concluded that in the establishment phase, the poplar hybrid is more tolerant than the willow hybrid to LL. We also provide evidence for LPI as a potential predictor for setting LL irrigation levels in the initial phase of a phyto-treatment approach.

Specificity of polyamine effects on NaCl-induced ion flux kinetics and salt stress amelioration in plants.

Polyamine (PA) levels in plants increase considerably under saline conditions. Because such an increase is believed to be beneficial for stress resistance, exogenous application of PAs has often been advocated as a means of ameliorating the detrimental effects of salinity. Results, however, are rather controversial, ranging from a significant amelioration to being ineffective or even toxic. The reasons for this controversy remain elusive. The ability of a root to retain K(+) in the presence of NaCl was used as a physiological indicator to evaluate the ameliorative effects of PA. Pre-treatment with 1 mM Spm(4+) (spermine), Spd(3+) (spermidine) or Put(2+) (putrescine) prevented salt-induced K(+) leak only in the mature root zone of hydroponically grown maize and Arabidopsis. In contrast, in the distal elongation root zone, PA pre-treatment resulted in an even larger NaCl-induced K(+) efflux, with the effect ranging from Spm(4+) >Spd(3+ )= Put(2+). A similar sequence has been also reported for H(+) pump inhibition, measured for both root zones. It appears that PAs affect cell membrane transporters in a highly specific way, with a relatively narrow 'window' in which amelioration is observed. We suggest that the ameliorative affect of PAs is the result of a complex combination of factors which might potentially include PA transport and accumulation in the cell cytosol, their metabolization and the functional expression of the specific target proteins or signaling elements.

Salt acclimation process: a comparison between a sensitive and a tolerant Olea europaea cultivar.

Saline soils are highly heterogeneous in time and space, and this is a critical factor influencing plant physiology and productivity. Temporal changes in soil salinity can alter plant responses to salinity, and pre-treating plants with low NaCl concentrations has been found to substantially increase salt tolerance in different species in a process called acclimation. However, it still remains unclear whether this process is common to all plants or is only expressed in certain genotypes. We addressed this question by assessing the physiological changes to 100 mM NaCl in two contrasting olive cultivars (the salt-sensitive Leccino and the salt-tolerant Frantoio), following a 1-month acclimation period with 5 or 25 mM NaCl. The acclimation improved salt tolerance in both cultivars, but activated substantially different physiological adjustments in the tolerant and the sensitive cultivars. In the tolerant Frantoio the acclimation with 5 mM NaCl was more effective in increasing plant salt tolerance, with a 47% increase in total plant dry mass compared with non-acclimated saline plants. This enhanced biomass accumulation was associated with a 50% increase in K+ retention ability in roots. On the other hand, in the sensitive Leccino, although the acclimation process did not improve performance in terms of plant growth, pre-treatment with 5 and 25 mM NaCl substantially decreased salt-induced leaf cell ultrastructural changes, with leaf cell relatively similar to those of control plants. Taken together these results suggest that in the tolerant cultivar the acclimation took place primarily in the root tissues, while in the sensitive they occurred mainly at the shoot level.

Acclimation improves salt stress tolerance in Zea mays plants.

Plants exposure to low level salinity activates an array of processes leading to an improvement of plant stress tolerance. Although the beneficial effect of acclimation was demonstrated in many herbaceous species, underlying mechanisms behind this phenomenon remain poorly understood. In the present study we have addressed this issue by investigating ionic mechanisms underlying the process of plant acclimation to salinity stress in Zea mays. Effect of acclimation were examined in two parallel sets of experiments: a growth experiment for agronomic assessments, sap analysis, stomatal conductance, chlorophyll content, and confocal laser scanning imaging; and a lab experiment for in vivo ion flux measurements from root tissues. Being exposed to salinity, acclimated plants (1) retain more K(+) but accumulate less Na(+) in roots; (2) have better vacuolar Na(+) sequestration ability in leaves and thus are capable of accumulating larger amounts of Na(+) in the shoot without having any detrimental effect on leaf photochemistry; and (3) rely more on Na(+) for osmotic adjustment in the shoot. At the same time, acclimation affect was not related in increased root Na(+) exclusion ability. It appears that even in a such salt-sensitive species as maize, Na(+) exclusion from uptake is of a much less importance compared with the efficient vacuolar Na(+) sequestration in the shoot.

Networks of plants: how to measure similarity in vegetable species.

Despite the common misconception of nearly static organisms, plants do interact continuously with the environment and with each other. It is fair to assume that during their evolution they developed particular features to overcome similar problems and to exploit possibilities from environment. In this paper we introduce various quantitative measures based on recent advancements in complex network theory that allow to measure the effective similarities of various species. By using this approach on the similarity in fruit-typology ecological traits we obtain a clear plant classification in a way similar to traditional taxonomic classification. This result is not trivial, since a similar analysis done on the basis of diaspore morphological properties do not provide any clear parameter to classify plants species. Complex network theory can then be used in order to determine which feature amongst many can be used to distinguish scope and possibly evolution of plants. Future uses of this approach range from functional classification to quantitative determination of plant communities in nature.

Morphologic and Aerodynamic Considerations Regarding the Plumed Seeds of Tragopogon pratensis and Their Implications for Seed Dispersal.

Tragopogon pratensis is a small herbaceous plant that uses wind as the dispersal vector for its seeds. The seeds are attached to parachutes that increase the aerodynamic drag force and increase the total distance travelled. Our hypothesis is that evolution has carefully tuned the air permeability of the seeds to operate in the most convenient fluid dynamic regime. To achieve final permeability, the primary and secondary fibres of the pappus have evolved with complex weaving; this maximises the drag force (i.e., the drag coefficient), and the pappus operates in an "optimal" state. We used computational fluid dynamics (CFD) simulations to compute the seed drag coefficient and compare it with data obtained from drop experiments. The permeability of the parachute was estimated from microscope images. Our simulations reveal three flow regimes in which the parachute can operate according to its permeability. These flow regimes impact the stability of the parachute and its drag coefficient. From the permeability measurements and drop experiments, we show how the seeds operate very close to the optimal case. The porosity of the textile appears to be an appropriate solution to achieve a lightweight structure that allows a low terminal velocity, a stable flight and a very efficient parachute for the velocity at which it operates.

The electrical network of maize root apex is gravity dependent.

Investigations carried out on maize roots under microgravity and hypergravity revealed that gravity conditions have strong effects on the network of plant electrical activity. Both the duration of action potentials (APs) and their propagation velocities were significantly affected by gravity. Similarly to what was reported for animals, increased gravity forces speed-up APs and enhance synchronized electrical events also in plants. The root apex transition zone emerges as the most active, as well as the most sensitive, root region in this respect.

Linking salinity stress tolerance with tissue-specific Na(+) sequestration in wheat roots.

Salinity stress tolerance is a physiologically complex trait that is conferred by the large array of interacting mechanisms. Among these, vacuolar Na(+) sequestration has always been considered as one of the key components differentiating between sensitive and tolerant species and genotypes. However, vacuolar Na(+) sequestration has been rarely considered in the context of the tissue-specific expression and regulation of appropriate transporters contributing to Na(+) removal from the cytosol. In this work, six bread wheat varieties contrasting in their salinity tolerance (three tolerant and three sensitive) were used to understand the essentiality of vacuolar Na(+) sequestration between functionally different root tissues, and link it with the overall salinity stress tolerance in this species. Roots of 4-day old wheat seedlings were treated with 100 mM NaCl for 3 days, and then Na(+) distribution between cytosol and vacuole was quantified by CoroNa Green fluorescent dye imaging. Our major observations were as follows: (1) salinity stress tolerance correlated positively with vacuolar Na(+) sequestration ability in the mature root zone but not in the root apex; (2) contrary to expectations, cytosolic Na(+) levels in root meristem were significantly higher in salt tolerant than sensitive group, while vacuolar Na(+) levels showed an opposite trend. These results are interpreted as meristem cells playing a role of the "salt sensor;" (3) no significant difference in the vacuolar Na(+) sequestration ability was found between sensitive and tolerant groups in either transition or elongation zones; (4) the overall Na(+) accumulation was highest in the elongation zone, suggesting its role in osmotic adjustment and turgor maintenance required to drive root expansion growth. Overall, the reported results suggest high tissue-specificity of Na(+) uptake, signaling, and sequestration in wheat roots. The implications of these findings for plant breeding for salinity stress tolerance are discussed.

Gravity affects the closure of the traps in Dionaea muscipula.

Venus flytrap (Dionaea muscipula Ellis) is a carnivorous plant known for its ability to capture insects thanks to the fast snapping of its traps. This fast movement has been long studied and it is triggered by the mechanical stimulation of hairs, located in the middle of the leaves. Here we present detailed experiments on the effect of microgravity on trap closure recorded for the first time during a parabolic flight campaign. Our results suggest that gravity has an impact on trap responsiveness and on the kinetics of trap closure. The possible role of the alterations of membrane permeability induced by microgravity on trap movement is discussed. Finally we show how the Venus flytrap could be an easy and effective model plant to perform studies on ion channels and aquaporin activities, as well as on electrical activity in vivo on board of parabolic flights and large diameter centrifuges.

Oxidative stress and NO signalling in the root apex as an early response to changes in gravity conditions.

Oxygen influx showed an asymmetry in the transition zone of the root apex when roots were placed horizontally on ground. The influx increased only in the upper side, while no changes were detected in the division and in the elongation zone. Nitric oxide (NO) was also monitored after gravistimulation, revealing a sudden burst only in the transition zone. In order to confirm these results in real microgravity conditions, experiments have been set up by using parabolic flights and drop tower. The production of reactive oxygen species (ROS) was also monitored. Oxygen, NO, and ROS were continuously monitored during normal and hyper- and microgravity conditions in roots of maize seedlings. A distinct signal in oxygen and NO fluxes was clearly detected only in the apex zone during microgravity, with no significant changes in normal and in hypergravity conditions. The same results were obtained by ROS measurement. The detrimental effect of D'orenone, disrupting the polarised auxin transport, on the onset of the oxygen peaks during the microgravity period was also evaluated. Results indicates an active role of NO and ROS as messengers during the gravitropic response, with probable implications in the auxin redistribution.

Biomimetics on seed dispersal: survey and insights for space exploration.

Seeds provide the vital genetic link and dispersal agent between successive generations of plants. Without seed dispersal as a means of reproduction, many plants would quickly die out. Because plants lack any sort of mobility and remain in the same spot for their entire lives, they rely on seed dispersal to transport their offspring throughout the environment. This can be accomplished either collectively or individually; in any case as seeds ultimately abdicate their movement, they are at the mercy of environmental factors. Thus, seed dispersal strategies are characterized by robustness, adaptability, intelligence (both behavioral and morphological), and mass and energy efficiency (including the ability to utilize environmental sources of energy available): all qualities that advanced engineering systems aim at in general, and in particular those that need to enable complex endeavors such as space exploration. Plants evolved and adapted their strategy according to their environment, and taken together, they enclose many desirable characteristics that a space mission needs to have. Understanding in detail how plants control the development of seeds, fabricate structural components for their dispersal, build molecular machineries to keep seeds dormant up to the right moment and monitor the environment to release them at the right time could provide several solutions impacting current space mission design practices. It can lead to miniaturization, higher integration and packing efficiency, energy efficiency and higher autonomy and robustness. Consequently, there would appear to be good reasons for considering biomimetic solutions from plant kingdom when designing space missions, especially to other celestial bodies, where solid and liquid surfaces, atmosphere, etc constitute and are obviously parallel with the terrestrial environment where plants evolved. In this paper, we review the current state of biomimetics on seed dispersal to improve space mission design.