Chlorophyll Fluorescence Analysis Research Articles

Promotion of Vegetative Growth in Force-ventilated Protea cynaroides L. Explants Cultured in Modified Temporary Immersion Culture Vessels

Temporary immersion culture vessels were modified to culture Protea cynaroides L. microshoots on semisolid growth medium. The effects of different ventilation treatments and sucrose concentrations on the vegetative growth and physiological characteristics of P. cynaroides L. microshoots were investigated. Three ventilation treatments were used: microshoots were either ventilated naturally or forced ventilated for 2 minutes/2 hours or 2 minutes/4 hours. In addition, two sucrose concentrations were used in the growth medium: 30 and 10 g·L−1. Significant interaction effects were found between ventilation and sucrose in the number of shoots formed. When cultured on growth medium with 10 g·L−1 sucrose, microshoots force-ventilated for 2 minutes/2 hours produced significantly higher number of shoots than those naturally ventilated or force-ventilated for 2 minutes/4 hours. In the 30 g·L−1 sucrose treatment, no significant differences in shoot numbers were observed among all ventilation treatments. The highest leaf areas were found in microshoots cultured in the 2 minutes/4 hours forced ventilation treatment, which were significantly higher than microshoots in the other ventilation treatments, irrespective of the sucrose concentration. Chlorophyll content was significantly higher in leaves of microshoots that were cultured in 30 g·L−1 sucrose compared with those grown in 10 g·L−1 sucrose in all ventilation treatments. Analysis of chlorophyll fluorescence of the leaves revealed that the Fv/Fm value of microshoots grown on 30 g·L−1 sucrose and force-ventilated for 2 minutes/4 hours was significantly higher than those naturally ventilated in the same sucrose treatment. Overall, the use of 30 g·L−1 sucrose in combination with 2 minutes/4 hours ventilation provided the best conditions for culturing P. cynaroides microshoots. This study demonstrated that these modified temporary immersion culture vessels can be used as a forced ventilation system to culture P. cynaroides microshoots and promote vegetative growth as well as improve their photosynthetic characteristics. The system described here introduces a simple and novel method of converting commercially available temporary immersion systems into force ventilation systems.

Chlorophyll fluorescence analysis revealed essential roles of FtsH11 protease in regulation of the adaptive responses of photosynthetic systems to high temperature

BackgroundPhotosynthetic systems are known to be sensitive to high temperature stress. To maintain a relatively “normal” level of photosynthetic activities, plants employ a variety of adaptive mechanisms in response to environmental temperature fluctuations. Previously, we reported that the chloroplast-targeted AtFtsH11 protease played an essential role for Arabidopsis plants to survive at high temperatures and to maintain normal photosynthetic efficiency at moderately elevated temperature. To investigate the factors contributing to the photosynthetic changes in FtsH11 mutant, we performed detailed chlorophyll fluorescence analyses of dark-adapted mutant plants and compared them to Col-0 WT plants under normal, two moderate high temperatures, and a high light conditions.ResultsWe found that mutation of FtsH11 gene caused significant decreases in photosynthetic efficiency of photosystems when environmental temperature raised above optimal. Under moderately high temperatures, the FtsH11 mutant showed significant 1) decreases in electron transfer rates of photosystem II (PSII) and photosystem I (PSI), 2) decreases in photosynthetic capabilities of PSII and PSI, 3) increases in non-photochemical quenching, and a host of other chlorophyll fluorescence parameter changes. We also found that the degrees of these negative changes for utilizing the absorbed light energy for photosynthesis in FtsH11 mutant were correlated with the level and duration of the heat treatments. For plants grown under normal temperature and subjected to the high light treatment, no significant difference in chlorophyll fluorescence parameters was found between the FtsH11 mutant and Col-0 WT plants.ConclusionsThe results of this study show that AtFtsH11 is essential for normal photosynthetic function under moderately elevated temperatures. The results also suggest that the network mediated by AtFtsH11 protease plays critical roles for maintaining the thermostability and possibly structural integrity of both photosystems under elevated temperatures. Elucidating the underlying mechanisms of FtsH11 protease in photosystems may lead to improvement of photosynthetic efficiency under heat stress conditions, hence, plant productivity.

Heat and water stressed field-grown soybean: A multivariate study on the relationship between physiological-biochemical traits and yield

Heat stress (HS) combined with water stress (WS) negatively impact soybean [Glycine max (L.) Merr.] production. The aims of this research were to quantify the combined effect of HS and WS during grain filling (GF) on grain yield, and to identify physiological-biochemical traits strongly associated to crop performance and yield determination processes under these stressful field environments. Two soybean cultivars were exposed during GF to ambient temperature or to HS (>32 °C for 6 h per d) during 21 d; and to field capacity or to WS (20% of available soil water content) during 35 d. Yield, grain number and weight (GN and GW) were significantly reduced under HS combined with WS. In irrigated HS yield and GN were also reduced, albeit final GW was similar to that of control, highlighting compensation mechanisms. Chlorophyll fluorescence analysis indicated that detrimental effects of HS and WS on photosynthesis were via structural capacity aspects but also through damage to photosystem II. The HS × WS decreased the availability of assimilate to grains, but also impaired the sink-grain metabolization of the remobilized products from leaves. A multivariate analysis highlighted strong correlations between quantum efficiency of photosystem II photochemistry, chlorophyll content (SPAD value), and the antioxidant state of leaves, which were all positively correlated with yield. Canopy temperature throughout grain filling (CTGF) was negatively associated with GN and GW, the latter also positively associated with SPAD. Explaining these physiological traits 81, 50, and 82% of the total variability of yield, GN and GW, respectively. To our knowledge this is the first report that integrating different scales of study, demonstrates the potential of physiological-biochemical parameters to explain yield variations in field-grown soybean under HS and WS conditions.

Taxonomic classification of algae by the use of chlorophyll a fluorescence

Natural water reservoirs are very important ecosystems thus they should be under continuous monitoring and protection. In water of low quality, the algal blooms develop develops vastly. The knowledge of algal species composition is necessary for understanding this process. There are a few traditional group-specific methods of microalgae classification, but they are often labour-intense and time-consuming. Moreover, the samples must be prepared and/or collected before getting any results. Non-invasive chlorophyll fluorescence analysis offers an alternative approach and potentially allows in situ estimation of algal concentration. In this work the fluorometric methods to estimate algae content in water and to differentiate algal populations is presented.

A Proteomic Approach to Investigate the Drought Response in the Orphan Crop Eragrostis tef.

The orphan crop, Eragrostis tef, was subjected to controlled drought conditions to observe the physiological parameters and proteins changing in response to dehydration stress. Physiological measurements involving electrolyte leakage, chlorophyll fluorescence and ultra-structural analysis showed tef plants tolerated water loss to 50% relative water content (RWC) before adverse effects in leaf tissues were observed. Proteomic analysis using isobaric tag for relative and absolute quantification (iTRAQ) mass spectrometry and appropriate database searching enabled the detection of 5727 proteins, of which 211 proteins, including a number of spliced variants, were found to be differentially regulated with the imposed stress conditions. Validation of the iTRAQ dataset was done with selected stress-related proteins, fructose-bisphosphate aldolase (FBA) and the protective antioxidant proteins, monodehydroascorbate reductase (MDHAR) and peroxidase (POX). Western blot analyses confirmed protein presence and showed increased protein abundance levels during water deficit while enzymatic activity for FBA, MDHAR and POX increased at selected RWC points. Gene ontology (GO)-term enrichment and analysis revealed terms involved in biotic and abiotic stress response, signaling, transport, cellular homeostasis and pentose metabolic processes, to be enriched in tef upregulated proteins, while terms linked to reactive oxygen species (ROS)-producing processes under water-deficit, such as photosynthesis and associated light harvesting reactions, manganese transport and homeostasis, the synthesis of sugars and cell wall catabolism and modification, to be enriched in tef downregulated proteins.

Continuous high and low temperature induced a decrease of photosynthetic activity and changes in the diurnal fluctuations of organic acids in Opuntia streptacantha.

Opuntia plants grow naturally in areas where temperatures are extreme and highly variable in the day during the entire year. These plants survive through different adaptations to respond to adverse environmental conditions. Despite this capability, it is unknown how CAM photosynthetic activity and growth in Opuntia plantlets is affected by constant heat or cold. Therefore, the main objective of this research was to evaluate the short-term effect of high (40°C) and low (4°C) continuous temperatures on the photosynthetic efficiency, the organic acid content (malic acid) and the relative growth rate (RGR) in seven-month-old Opuntia streptacantha plantlets during 5, 10, and 15 days. Chlorophyll fluorescence analysis allowed us to determine that high temperatures negatively impact the photosynthetic efficiency of O. streptacantha plantlets, which exhibited the lowest values of maximum quantum efficiency of the photosystem II (Fv/Fm = 52%, Fv/F0 = 85%), operational quantum yield of PS (ΦPSII = 65%) and relative electron transport rate (rETR = 65%), as well as highest values of basal fluorescence (F0 = 226%) during 15 days of treatment. Similarly, low temperatures decreased Fv/Fm (16%), Fv/F0 (50%), ΦPSII and rETR (16%). High temperatures also decreased nocturnal acidification in approximately 34–50%, whereas low temperatures increased it by 30–36%. Additionally, both continuous temperatures affected drastically diurnal consumption of malic acid, which was related to a significant RGR inhibition, where the specific photosynthetic structure area component was the most affected. Our results allowed determining that, despite the high tolerance to extreme temperatures described for Opuntia plants, young individuals of O. streptacantha suffered photosynthetic impairment that led to the inhibition of their growth. Thus, the main findings reported in this study can help to predict the potential impact of climatic change on the establishment and survival of succulent species of arid and semiarid regions of Mexico.

The Effect of Sugar Beet Broadleaf Herbicides on Fluorescence Induction Curves in Amaranthus retroflexus L. and Portulaca oleracea L.

Chlorophyll fluorescence analysis is a simple and rapid method for detecting herbicide effects after a short time following their application in photosynthetic apparatus in plants. Chlorophyll fluorescence measurements were carried out against two broad of weeds to describe how the Kautsky curve and its parameters were affected by herbicides. Desmedipham + phenmedipham + ethofumesate changed the chlorophyll fluorescence induction curve at all time intervals except four hours after spring (HAS) in Amaranthus retroflexus L. and at all doses of Portulaca oleracea L. 4 HAS. In contrast, chlorophyll fluorescence inhibition was evident by chloridazon at doses of 650 and 325 g a.i. ha-1 in P. oleracea and A. retroflexus respectively, for all time intervals. Furthermore, chlorophyll fluorescence decays only occurred by clopyralid in A. retroflexus at the highest dose. A biomass effective dose (ED50 and/or ED90) based on log-logistic dose-response curves for A. retroflexus were considerably higher than that of P. oleracea. The maximum quantum efficiency (FV/Fm) was stable, whereas the relative changes at the J step (Fvj) and area (the area between the Kautsky curve and the maximum fluorescence (Fm)) was more sensitive to all three herbicides. There was a relatively good correlation between fluorescence parameters taken 24 hours after the spraying and the dry matter taken three weeks later, for both species under study.

Loss of LHCI system affects LHCII re-distribution between thylakoid domains upon state transitions.

LHCI, the peripheral antenna system of Photosystem I, includes four light-harvesting proteins (Lhca1-Lhca4) in higher plants, all of which are devoid in the Arabidopsis thaliana knock-out mutant ΔLhca. PSI absorption cross-section was reduced in the mutant, thus affecting the redox balance of the photosynthetic electron chain and resulting in a more reduced PQ with respect to the wild type. ΔLhca plants developed compensatory response by enhancing LHCII binding to PSI. However, the amplitude of state transitions, as measured from changes of chlorophyll fluorescence in vivo, was unexpectedly low than the high level of PSI-LHCII supercomplex established. In order to elucidate the reasons for discrepancy, we further analyzed state transition in ΔLhca plants. The STN7 kinase was fully active in the mutant as judged from up-regulation of LHCII phosphorylation in state II. Instead, the lateral heterogeneity of thylakoids was affected by lack of LHCI, with LHCII being enriched in stroma membranes with respect to the wild type. Re-distribution of this complex affected the overall fluorescence yield of thylakoids already in state I and minimized changes in RT fluorescence yield when LHCII did connect to PSI reaction center. We conclude that interpretation of chlorophyll fluorescence analysis of state transitions becomes problematic when applied to mutants whose thylakoid architecture is significantly modified with respect to the wild type.

Non-intrusive Assessment of Photosystem II and Photosystem I in Whole Coral Tissues

Reef building corals (phylum Cnidaria) harbour endosymbiotic dinoflagellate algae (genus Symbiodinium) that generate photosynthetic products to fuel their host’s metabolism. Non-invasive techniques such as chlorophyll (Chl) fluorescence analyses of Photosystem II (PSII) have been widely used to estimate the photosynthetic performance of Symbiodinium in hospite. However, since the spatial origin of PSII chlorophyll fluorescence in coral tissues is uncertain, such signals give limited information on depth-integrated photosynthetic performance of the whole tissue. In contrast, detection of absorbance changes in the near infrared (NIR) region integrates signals from deeper tissue layers due to weak absorption and multiple scattering of NIR light. While extensively utilised in higher plants, NIR bio-optical techniques are seldom applied to corals. We have developed a non-intrusive measurement method to examine photochemistry of intact corals, based on redox kinetics of the primary electron donor in Photosystem I (P700) and chlorophyll fluorescence kinetics (Fast-Repetition Rate fluorometry, FRRf). Since the redox state of P700 depends on the operation of both PSI and PSII, important information can be obtained on the PSII-PSI intersystem electron transfer kinetics. Under moderate, sub-lethal heat stress treatments (33 ˚C for ~20 min), the coral Pavona decussata exhibited down-regulation of PSII electron transfer kinetics, indicated by slower rates of electron transport from QA to plastoquinone (PQ) pool, and smaller relative size of oxidised PQ with concomitant decrease of a specifically-defined P700 kinetics area, which represents the active pool of PSII. The maximum quantum efficiency of PSII (Fv/Fm) and functional absorption cross-section of PSII (σPSII) remained unchanged. Based on the coordinated response of P700 parameters and PSII-PSI electron transport properties, we propose that simple P700 kinetics parameters as employed here serve as indicators of the integrity of PSII-PSI electron transfer dynamics in corals.

Photosynthetic properties of spring geophytes assessed by chlorophyll fluorescence analysis

Since spring ephemerals are credited to be all “sun” species with unusually elevate photosynthesis, in contrast to shade-tolerant trees and understory geophytes with a long aboveground cycle, we examined the photosynthetic efficiency of 6 woody species, 9 long-cycle geophytes, and 8 spring ephemeral geophytes using blue flashes of increasing energy with the Imaging PAM fluorometer. Several parameters were obtained: quantum yield of electron transport (ΦETR) or of PSII (ΦPSII), maximum measured photosynthesis rate (ETRhv), maximum extrapolated rate of photosynthesis (ETRem), half-saturating photon flux density (KPAR), and in some cases photochemical (qP) and non-photochemical quenching (NPQ). Results confirm the ecological consistency of the three plant groups, with internal differences. Woody species have low ETRem and KPAR values with good ΦETR; long-cycle herbs have low ETRem and ΦETR and moderate KPAR values; spring ephemerals have elevate ΦETR, ETRem and KPAR values. The mean ETRem of ephemerals of 91 μmol m−2 s−1 exceeds that of long-cycle herbs 2.9-fold and woody species 4.8-fold, and corresponds to 19 μmol CO2 m−2 s−1 by assuming an ETR/ΦCO2 ratio of 4.7. Highest photosynthesis rates and KPAR were exhibited by five ephemerals (Eranthis, Erythronium, Narcissus, Scilla, Tulipa) with peak ETRem values equivalent to ∼40 μmol CO2 m−2 s−1 or ∼60 μmol CO2 (g Chl)−1 s−1 (“sun” species). According to a new, fluorescence based heliophily index, all trees and five long-cycle herbs were definitely “shade” species, while four long-cycle herbs and three ephemerals were intermediate shade-tolerant.

Acclimation responses to high light by Guazuma ulmifolia Lam. (Malvaceae) leaves at different stages of development.

The re-composition of deforested environments requires the prior acclimation of seedlings to full sun in nurseries. Seedlings can overcome excess light either through the acclimation of pre-existing fully expanded leaves or through the development of new leaves that are acclimated to the new light environment. Here, we compared the acclimation capacity of mature (MatL, fully expanded at the time of transfer) and newly expanded (NewL, expanded after the light shift) leaves of Guazuma ulmifolia Lam. (Malvaceae) seedlings to high light. The seedlings were initially grown under shade and then transferred to full sunlight. MatL and NewL were used for chlorophyll fluorescence and gas exchange analyses, pigment extraction and morpho-anatomical measurements. After the transfer of seedlings to full sun, the MatL persisted and acclimated to some extent to the new light condition, since they underwent alterations in some morpho-physiological traits and maintained a functional electron transport chain and positive net photosynthesis rate. However, long-term exposure to high light led to chronic photoinhibition in MatL, which could be related to the limited plasticity of leaf morpho-anatomical attributes. However, the NewL showed a high capacity to use the absorbed energy in photochemistry and dissipate excess energy harmlessly, attributes that were favoured by the high structural plasticity exhibited by these leaves. Both the maintenance of mature, photosynthetically active leaves and the production of new leaves with a high capacity to cope with excess energy were important for acclimation of G.ulmifolia seedlings.

Influence of different degrees of N limitation on photosystem II performance and heterogeneity of Chlorella vulgaris

Nitrogen (N) limitation is considered as the most efficient strategy to induce the accumulation of lipids, carbohydrates or other target compounds in microalgal biomass. However, along with biomass biochemical composition, alterations in N limitation affect the photosynthetic apparatus and result in decreased growth. In this study, Chlorella vulgaris was cultivated in semi-continuous mode with different degrees of N limitation and chlorophyll (Chl) fluorescence analyses were used to investigate the effect of N limitation on photosystem II (PSII) performance, in terms of structural and functional heterogeneity. As expected, N limitation resulted in the decrease of quantum yield and calculated OJIP parameters related to PSII performance. N limitation resulted in a significant increase of trapped energy per reaction center (RC) and subsequently to higher dissipation of excess energy. However, despite the negative effect of N limitation on the number of RCs, the electron transport beyond QA as well the capacity of reducing/re-oxidation of plastiquinone were not negatively affected, implying that performance of RCs was not affected by N limitation. Photochemical quenching (qp) increased of as N limitation increased while the curve of non-photochemical quenching (NPQ) was unimodal, i.e. increased up to a level of N limitation and then decreased as N limitation degree increased further. The overall results of the present study suggest that the decrease of PSII performance was due to a reduction of the number of RCs accompanied with higher energy dissipation a probable outcome of the decreased need for reductant by cells due to lower metabolic activity under N limitation.

Establishment of integrated protocols for automated high throughput kinetic chlorophyll fluorescence analyses

BackgroundAutomated plant phenotyping has been established as a powerful new tool in studying plant growth, development and response to various types of biotic or abiotic stressors. Respective facilities mainly apply non-invasive imaging based methods, which enable the continuous quantification of the dynamics of plant growth and physiology during developmental progression. However, especially for plants of larger size, integrative, automated and high throughput measurements of complex physiological parameters such as photosystem II efficiency determined through kinetic chlorophyll fluorescence analysis remain a challenge.ResultsWe present the technical installations and the establishment of experimental procedures that allow the integrated high throughput imaging of all commonly determined PSII parameters for small and large plants using kinetic chlorophyll fluorescence imaging systems (FluorCam, PSI) integrated into automated phenotyping facilities (Scanalyzer, LemnaTec). Besides determination of the maximum PSII efficiency, we focused on implementation of high throughput amenable protocols recording PSII operating efficiency (ΦPSII). Using the presented setup, this parameter is shown to be reproducibly measured in differently sized plants despite the corresponding variation in distance between plants and light source that caused small differences in incident light intensity. Values of ΦPSII obtained with the automated chlorophyll fluorescence imaging setup correlated very well with conventionally determined data using a spot-measuring chlorophyll fluorometer. The established high throughput operating protocols enable the screening of up to 1080 small and 184 large plants per hour, respectively. The application of the implemented high throughput protocols is demonstrated in screening experiments performed with large Arabidopsis and maize populations assessing natural variation in PSII efficiency.ConclusionsThe incorporation of imaging systems suitable for kinetic chlorophyll fluorescence analysis leads to a substantial extension of the feature spectrum to be assessed in the presented high throughput automated plant phenotyping platforms, thus enabling the simultaneous assessment of plant architectural and biomass-related traits and their relations to physiological features such as PSII operating efficiency. The implemented high throughput protocols are applicable to a broad spectrum of model and crop plants of different sizes (up to 1.80 m height) and architectures. The deeper understanding of the relation of plant architecture, biomass formation and photosynthetic efficiency has a great potential with respect to crop and yield improvement strategies.

Copper bioaccumulation, photosystem II functioning, and oxidative stress in the seagrass Cymodocea nodosa exposed to copper oxide nanoparticles.

Photosynthetic activity, oxidative stress, and Cu bioaccumulation in the seagrass Cymodocea nodosa were assessed 4, 12, 24, 48, and 72h after exposure to two copper oxide nanoparticle (CuO NP) concentrations (5 and 10mgL-1). CuO NPs were characterized by scanning electron microscopy (SEM) and dynamic light scattering measurements (DLS). Chlorophyll fluorescence analysis was applied to detect photosystem II (PSII) functionality, while the Cu accumulation kinetics into the leaf blades was fitted to the Michaelis-Menten equation. The uptake kinetics was rapid during the first 4h of exposure and reached an equilibrium state after 10h exposure to 10mgL-1 and after 27h to 5mgL-1 CuO NPs. As a result, 4-h treatment with 5mgL-1 CuO NPs, decreased the quantum yield of PS II photochemistry (Φ PSΙΙ ) with a parallel increase in the regulated non-photochemical energy loss in PSII (Φ NPQ ). However, the photoprotective dissipation of excess absorbed light energy as heat, through the process of non-photochemical quenching (NPQ), did not maintain the same fraction of open reaction centers (q p ) as in control plants. This reduced number of open reaction centers resulted in a significant increase of H2O2 production in the leaf veins serving possibly as an antioxidant defense signal. Twenty-four-hour treatment had no significant effect on Φ PSΙΙ and q p compared to controls. However, 24h exposure to 5mgL-1 CuO NPs increased the quantum yield of non-regulated energy loss in PSII (Φ NO ), and thus the formation of singlet oxygen (1O2) via the triplet state of chlorophyll, possible because the uptake kinetics had not yet reached the equilibrium state as did 10mgL-1. Longer-duration treatment (48 and 72h) had less effect on the allocation of absorbed light energy at PSII and the fraction of open reaction centers, compared to 4-h treatment, suggesting the function of a stress defense mechanism. The response of C. nodosa leaves to CuO NPs fits the "Threshold for Tolerance Model" with a threshold time (more than 4h) required for induction of a stress defense mechanism, through H2O2 production.

Effects of CO2 enrichment on primary photochemistry, growth and astaxanthin accumulation in the chlorophyte Haematococcus pluvialis

The atmospheric CO2 level is limiting for growth of phototrophic organisms such as microalgae, so CO2 enrichment boosts the growth and photosynthesis of microalgal cultures. Still, excessive CO2 injection might inhibit photosynthesis of microalgae. We investigated the effect of continuous sparging of the cultures of Haematococcus pluvialis BM 1 (IPPAS H-2018) (Chlorophyceae), the richest natural source of the value-added pigment astaxanthin. H. pluvialis cultures with CO2-enriched air-gas mixtures (with CO2 level from the atmospheric to 20%) on growth and astaxanthin accumulation in the microalga. Special attention was paid to photosynthetic activity and non-photochemical excited chlorophyll states quenching in the microalgal cells, which was monitored via chlorophyll fluorescence analysis. We also report on the capability of CO2 capture by H. pluvialis derived from direct measurements of its elemental carbon content. The beneficial effect of the moderately high (5%) CO2 levels on the culture growth and astaxanthin accumulation under stress results in a higher overall astaxanthin productivity. However, increase of the CO2 level to 10% or 20% was deteriorative for growth, photosynthesis and carbon assimilation. The results support the possibility of combining a traditional two-stage H. pluvialis cultivation with CO2 bio-capture although a dilution of the flue gas before its injection is required.

Difference in oxidative stress tolerance between rice cultivars estimated with chlorophyll fluorescence analysis

BackgroundOxidative stress is considered to be involved in growth retardation of plants when they are exposed to a variety of biotic and abiotic stresses. Despite its potential importance in improving crop production, comparative studies on oxidative stress tolerance between rice (Oryza sativa L.) cultivars are limited. This work describes the difference in term of oxidative stress tolerance between 72 rice cultivars.Methods72 rice cultivars grown under naturally lit greenhouse were used in this study. Excised leaf discs were subjected to a low concentration of methyl viologen (paraquat), a chemical reagent known to generate reactive oxygen species in chloroplast. Chlorophyll fluorescence analysis using a two-dimensional fluorescence meter, ion leakage analysis as well as the measurement of chlorophyll contents were used to evaluate the oxidative stress tolerance of leaf discs. Furthermore, fluorescence intensities were finely analyzed based on new fluorescence theories that we have optimized.ResultsTreatment of leaf discs with methyl viologen caused differential decrease of maximum quantum yield of photosystem II (Fv/Fm) between cultivars. Decrease of Fv/Fm was also closely correlated with increase of ion leakage and decrease of chlorophyll a/b ratio. Fv/Fm was factorized into photochemical and non-photochemical parameters to classify rice cultivars into sensitive and tolerant ones. Among the 72 compared rice cultivars, the traditional cultivar Co13 was identified as the most tolerant to oxidative stress. Koshihikari, a dominant modern Japonica cultivar in Japan as well as IR58, one of the modern Indica breeding lines exhibited a strong tolerance to oxidative stress.ConclusionsClose correlation between Fv/Fm and chlorophyll a/b ratio provides a simple method to estimate oxidative stress tolerance, without measurement of chlorophyll fluorescence with special equipment. The fact that modern cultivars, especially major cultivars possessed tolerance to oxidative stress suggests that oxidative stress tolerance is one of the agricultural traits prerequisite for improvement of modern rice cultivars. Data presented in this study would enable breeding of rice cultivars having strong tolerance to oxidative stress.

Morphological and physiological differences between two morphotypes of Durvillaea antarctica (Phaeophyceae) from the sub-Antarctic ecoregion of Magallanes, Chile

The coastal marine ecosystems of the sub-Antarctic ecoregion of Magallanes in southern Chile are strongly characterized by environmental heterogeneity with a unique and high biodiversity of benthic macroalgae. The kelp Durvillaea antarctica (Phaeophyceae) is abundant in the intertidal in this region. Two morphotypes, one with elongated-cylindrical (“morphotype EC”) and one with laminar (“morphotype L”) blades, have been collected from a wave-protected site at the Strait of Magellan and the wave-exposed shores of the isolated Otway Sound. Morphometric and physiological (by chlorophyll fluorescence) analyses showed significant differences between the two morphotypes regarding the morphology, the maximum electron transport rates (ETRmax), and the chlorophyll c concentration. These morphological and physiological differences can be linked to their hydrodynamic environment. The presence of two different species in the region of Magallanes based on the measured parameters is discussed. This study may be useful for future genetic and biotechnological investigations and draws attention to D. antarctica as a species of commercial value among the marine resources of Chile.

Investigation on metal tolerance and phytoremoval activity in the poplar hybrid clone “Monviso” under Cu-spiked water: Potential use for wastewater treatment

A serious concern for the environmental and human health is represented by the increasing copper (Cu) occurrence in agricultural soils and waters, because of the possible food contamination and bioaugmentation along the trophic chain. The request for the decontamination of different matrices with an environmentally sustainable technology as the phytoremediation should be addressed by selecting plant materials with improved pollutant tolerance and removal capability. With this purpose, plants of the hybrid poplar clone “Monviso” (Populus×generosa A. Henry×P. nigra L.) were grown in growth chamber under hydroponics and exposed to excess Cu concentrations (T1, 75μM Cu; T2, 150μM Cu), selected as about 5 and 10 times higher than those allowed by the Italian regulation on water use. Results evidenced a notable Cu tolerance by poplar plants, particularly at the lowest Cu concentration. At organ level, the root system was the most affected by Cu treatment, especially in T2-exposed plants. Copper determinations revealed that the metal was mostly bioaccumulated in the roots, with a limited amount reaching the shoots. Chlorophyll content and fluorescence analyses confirmed the visible symptoms in leaves, highlighting a good physiological status in T1-exposed plants. Contrarily, an impairment of the main processes associated to photosynthesis was observed in T2-exposed plants also by gas exchange measurements. Remarkably, the Cu content analysis of the spiked water solutions revealed that poplar plants succeeded in removing almost the 50% of the total Cu amount added. These results strengthen the evidence that poplar plants represent a useful eco-friendly bio-tool for the decontamination of metal polluted waters.

Effects of phosphorus stress on the photosynthetic and physiological characteristics of Chlorella vulgaris based on chlorophyll fluorescence and flow cytometric analysis

This paper discusses the effects of phosphorus stress on the photosynthetic and physiological characteristics of Chlorella vulgaris. The results indicate that the growth and photosynthesis of C. vulgaris were affected under non-phosphorus control and non-orthophosphate treatments (pyrophosphate, tripolyphosphate and glycerophosphate). Orthophosphate promoted the growth of C. vulgaris most significantly, while the growth of C. vulgaris under non-phosphorus control was slow and soon entered a stable period. The growth of C. vulgaris was also promoted under non-orthophosphate treatments, but slower than orthophosphate treatment, with no significant differences among the three. A reversible inactivation occurred in the PSII system of C. vulgaris under non-phosphorus control after 24h, while the same phenomenon occurred after exposure for 7 days under non-orthophosphate treatments. The decrease in actual photochemical efficiency was related to the fall in the ratios of the opening reaction centers in the PSII system. For non-orthophosphate treatments, it was also related to other factors, as not all opening reaction centers transferred electrons. Phosphorus stress did not affect the integrity of the cell membranes, demonstrating that phosphorus stress could result in the inhibition of cell yield, rather than cell death. Phosphorus stress could also decrease the esterase activities and increase the mean size of algal cells.

Effects of a ladle furnace slag added to soil on morpho-physiological and biochemical parameters of Amaranthus paniculatus L. plants

Industrial slag from steelwork activities is considered a by-product by the EU legislation and it can be used for civil construction. In this work, an experiment in a greenhouse was conducted over a 6-week period to investigate the effect of soil enrichment with ladle furnace slag on morpho-physiological parameters of Amaranthus paniculatus L. plants. Results showed that the addition of 5% (w/w) slag to soil did not alter the plant growth, highlighting a high tolerance to this slag concentration. Contrarily, plants cultivated in a soil with 10% (w/w) slag showed a marked reduction both in growth and biometric parameters. Moreover, plants grown on a slag-rich soil (20% w/w) highlighted a very low survival rate. This behaviour was confirmed by the biochemical and physiological investigations on chlorophyll a and b content, gas exchange and chlorophyll fluorescence analyses. Metal(loid)s determination showed the accumulation of Ni, Se, Sn, As, Sb and Cd in 10% slag-treated plants, while revealed an increase in Ni, Cd, As and Pb in 5% slag-treated plants. Results are discussed highlighting the profitability of the cultivation of Amaranthus plants on slag enriched soil, as this plant species is largely used both as feedstock for energy production and for environmental restoration.