Karel Klem

Re‐interpreting plant morphological responses to UV‐B radiation

There is a need to reappraise the effects of UV-B radiation on plant morphology in light of improved mechanistic understanding of UV-B effects, particularly elucidation of the UV RESISTANCE LOCUS 8 (UVR8) photoreceptor. We review responses at cell and organismal levels, and explore their underlying regulatory mechanisms, function in UV protection and consequences for plant fitness. UV-induced morphological changes include thicker leaves, shorter petioles, shorter stems, increased axillary branching and altered root:shoot ratios. At the cellular level, UV-B morphogenesis comprises changes in cell division, elongation and/or differentiation. However, notwithstanding substantial new knowledge of molecular, cellular and organismal UV-B responses, there remains a clear gap in our understanding of the interactions between these organizational levels, and how they control plant architecture. Furthermore, despite a broad consensus that UV-B induces relatively compact architecture, we note substantial diversity in reported phenotypes. This may relate to UV-induced morphological changes being underpinned by different mechanisms at high and low UV-B doses. It remains unproven whether UV-induced morphological changes have a protective function involving shading and decreased leaf penetration of UV-B, counterbalancing trade-offs such as decreased photosynthetic light capture and plant-competitive abilities. Future research will need to disentangle seemingly contradictory interactions occurring at the threshold UV dose where regulation and stress-induced morphogenesis overlap.

Detection of photosynthetic herbicides: Algal growth inhibition test vs. electrochemical photosystem II biosensor

We compared a novel PSII-biosensor assay with a standard algal growth inhibition test for detection of photosynthetic herbicides--diuron, atrazine and isoproturon in liquid samples. To evaluate the convenience and sensitivity, values of the parameters EC50 and LOD and the duration of assays were compared. The biosensor assay was made with an electrochemical biosensor toxicity analyser with immobilised Photosystem II (PSII) complex. Using the PSII-biosensor assay, higher sensitivity (LOD) to herbicides (10(-8)-10(-9)M) was achieved as compared to standard algal growth inhibition tests (about 10(-7)M). The results of both assays showed a good correlation as concerns their EC50 values while the interval of detectable concentrations is about twice wider for PSII-biosensor. A proposed measurement protocol includes the reference standard of phytotoxicity (RSP). The main advantage of the PSII-biosensor assay is that it can be completed in about 1h and is by 1-2 orders more sensitive than standard algal growth inhibition test, which takes 72 h.

Ultraviolet and photosynthetically active radiation can both induce photoprotective capacity allowing barley to overcome high radiation stress

The main objective of this study was to determine the effects of acclimation to ultraviolet (UV) and photosynthetically active radiation (PAR) on photoprotective mechanisms in barley leaves. Barley plants were acclimated for 7 days under three combinations of high or low UV and PAR treatments ([UV-PAR-], [UV-PAR+], [UV+PAR+]). Subsequently, plants were exposed to short-term high radiation stress (HRS; defined by high intensities of PAR - 1000 μmol m(-2) s(-1), UV-A - 10 W m(-2) and UV-B 2 W m(-2) for 4 h), to test their photoprotective capacity. The barley variety sensitive to photooxidative stress (Barke) had low constitutive flavonoid content compared to the resistant variety (Bonus) under low UV and PAR intensities. The accumulation of lutonarin and 3-feruloylquinic acid, but not of saponarin, was greatly enhanced by high PAR and further increased by UV exposure. Acclimation of plants to both high UV and PAR intensities also increased the total pool of xanthophyll-cycle pigments (VAZ). Subsequent exposure to HRS revealed that prior acclimation to UV and PAR was able to ameliorate the negative consequences of HRS on photosynthesis. Both total contents of epidermal flavonols and the total pool of VAZ were closely correlated with small reductions in light-saturated CO2 assimilation rate and maximum quantum yield of photosystem II photochemistry caused by HRS. Based on these results, we conclude that growth under high PAR can substantially increase the photoprotective capacity of barley plants compared with plants grown under low PAR. However, additional UV radiation is necessary to fully induce photoprotective mechanisms in the variety Barke. This study demonstrates that UV-exposure can lead to enhanced photoprotective capacity and can contribute to the induction of tolerance to high radiation stress in barley.

Impact of elevated CO2 concentration on dynamics of leaf photosynthesis in Fagus sylvatica is modulated by sky conditions.

It has been suggested that atmospheric CO2 concentration and frequency of cloud cover will increase in future. It remains unclear, however, how elevated CO2 influences photosynthesis under complex clear versus cloudy sky conditions. Accordingly, diurnal changes in photosynthetic responses among beech trees grown at ambient (AC) and doubled (EC) CO2 concentrations were studied under contrasting sky conditions. EC stimulated the daily sum of fixed CO2 and light use efficiency under clear sky. Meanwhile, both these parameters were reduced under cloudy sky as compared with AC treatment. Reduction in photosynthesis rate under cloudy sky was particularly associated with EC-stimulated, xanthophyll-dependent thermal dissipation of absorbed light energy. Under clear sky, a pronounced afternoon depression of CO2 assimilation rate was found in sun-adapted leaves under EC compared with AC conditions. This was caused in particular by stomata closure mediated by vapour pressure deficit.

Effect of Elevated Carbon Dioxide Concentration on Carbon Assimilation under Fluctuating Light

Natural fluctuations in light intensity may significantly affect the amount of CO assimilated by plants and ecosystems. Little is known, however, about the interactive effect of dynamic light conditions and atmospheric CO concentrations. The hypothesis that elevated CO concentration (EC; 700 μmol CO mol) increases photosynthetic efficiency in dynamic light environments as compared to ambient CO concentration (AC; 385 μmol CO mol) was tested. Sun leaves of European beech ( L.) and current-year shoots of Norway spruce [ (L). Karst.] were exposed to five dynamic light regimes (LRs) occurring within forest canopies due to variable cloud cover or self-shading of leaves and to a steady-state LR. The LRs differed in the time course of incident irradiance, whereas the overall duration (600 s) and total amount of radiation (35.88 mmol photons m) were the same in all LRs. The EC treatment enhanced the amount of CO assimilated under all LRs tested. While the stimulation was only 37 to 50% in beech, it was 52 to 85% in spruce. The hypothesis that photosynthetic efficiency is stimulated by EC was confirmed in LRs when the leaves were pre-exposed to low light intensity and photosynthetic induction was required. By contrast, only a minor effect of EC treatment was found on the rate of induction loss and postillumination CO fixation in both species studied.

Plant phenotyping: a perspective

Sustainable agriculture for feeding increasing population is a foremost global challenge. The “green revolution” based crop productivity has done wonders in the past, but it has limits, and, thus, we are compelled to look for new avenues to increase productivity of important crops. Plant phenomics is emerging as a promising area in which many imaging sensors developed in the past are being tested for mapping of genetic information expressed within plant phenotypes, and the integrated use of these sensors may help speed-up unraveling of underlying molecular, biochemical and physiological mechanisms. We provide here a review of methods used for phenotyping and understanding of abiotic stress (drought/cold) tolerance mechanisms in the context of dynamic challenges faced by plants during their life.

Morphological, biochemical and physiological traits of upper and lower canopy leaves of European beech tend to converge with increasing altitude

The present work has explored for the first time acclimation of upper versus lower canopy leaves along an altitudinal gradient. We tested the hypothesis that restrictive climatic conditions associated with high altitudes reduce within-canopy variations of leaf traits. The investigated beech (Fagus sylvatica L.) forest is located on the southern slope of the Hrubý Jeseník Mountains (Czech Republic). All measurements were taken on leaves from upper and lower parts of the canopy of mature trees (>85 years old) growing at low (400 m above sea level, a.s.l.), middle (720 m a.s.l.) and high (1100 m a.s.l.) altitudes. Compared with trees at higher altitudes, those growing at low altitudes had lower stomatal conductance, slightly lower CO(2) assimilation rate (A(max)) and leaf mass per area (LMA), and higher photochemical reflectance index, water-use efficiency and Rubisco content. Given similar stand densities at all altitudes, the different growth conditions result in a more open canopy and higher penetration of light into lower canopy with increasing altitude. Even though strong vertical gradients in light intensity occurred across the canopy at all altitudes, lower canopy leaves at high altitudes tended to acquire the same morphological, biochemical and physiological traits as did upper leaves. While elevation had no significant effect on nitrogen (N) and carbon (C) contents per unit leaf area, LMA, or total content of chlorophylls and epidermal flavonoids in upper leaves, these increased significantly in lower leaves at higher altitudes. The increases in N content of lower leaves were coupled with similar changes in A(max). Moreover, a high N content coincided with high Rubisco concentrations in lower but not in upper canopy leaves. Our results show that the limiting role of light in lower parts of the canopy is reduced at high altitudes. A great capacity of trees to adjust the entire canopy is thus demonstrated.

Does long-term cultivation of saplings under elevated CO2 concentration influence their photosynthetic response to temperature?

BACKGROUND AND AIMS Plants growing under elevated atmospheric CO2 concentrations often have reduced stomatal conductance and subsequently increased leaf temperature. This study therefore tested the hypothesis that under long-term elevated CO2 the temperature optima of photosynthetic processes will shift towards higher temperatures and the thermostability of the photosynthetic apparatus will increase. METHODS The hypothesis was tested for saplings of broadleaved Fagus sylvatica and coniferous Picea abies exposed for 4-5 years to either ambient (AC; 385 µmol mol(-1)) or elevated (EC; 700 µmol mol(-1)) CO2 concentrations. Temperature response curves of photosynthetic processes were determined by gas-exchange and chlorophyll fluorescence techniques. KEY RESULTS Initial assumptions of reduced light-saturated stomatal conductance and increased leaf temperatures for EC plants were confirmed. Temperature response curves revealed stimulation of light-saturated rates of CO2 assimilation (Amax) and a decline in photorespiration (RL) as a result of EC within a wide temperature range. However, these effects were negligible or reduced at low and high temperatures. Higher temperature optima (Topt) of Amax, Rubisco carboxylation rates (VCmax) and RL were found for EC saplings compared with AC saplings. However, the shifts in Topt of Amax were instantaneous, and disappeared when measured at identical CO2 concentrations. Higher values of Topt at elevated CO2 were attributed particularly to reduced photorespiration and prevailing limitation of photosynthesis by ribulose-1,5-bisphosphate (RuBP) regeneration. Temperature response curves of fluorescence parameters suggested a negligible effect of EC on enhancement of thermostability of photosystem II photochemistry. CONCLUSIONS Elevated CO2 instantaneously increases temperature optima of Amax due to reduced photorespiration and limitation of photosynthesis by RuBP regeneration. However, this increase disappears when plants are exposed to identical CO2 concentrations. In addition, increased heat-stress tolerance of primary photochemistry in plants grown at elevated CO2 is unlikely. The hypothesis that long-term cultivation at elevated CO2 leads to acclimation of photosynthesis to higher temperatures is therefore rejected. Nevertheless, incorporating acclimation mechanisms into models simulating carbon flux between the atmosphere and vegetation is necessary.

Changes in Vertical Distribution of Spectral Reflectance within Spring Barley Canopy as an Indicator of Nitrogen Nutrition, Canopy Structure and Yield Parameters

Abstract The main objective of this study was to evaluate the spectral reflectance in the vertical profile of spring barley canopy at the booting growth stage and to determine how the reflectance gradient changes in relation to crop density and nitrogen (N) nutrition. Vertical gradients of spectral reflectance were studied in field trials with three sowing densities (2, 4 and 6 million of germinating seeds/ha) and two levels of N nutrition (0 and 90 kg/ha). It was found that differences in vegetation indices caused by N nutrition are most pronounced in the second and third leaf from the top, and these increase with increasing sowing density. The vertical gradient of reflectance, specifically the ratio between the leaves F-3/F-1 for vegetation indices based on red-edge reflectance, represents a reliable indicator of number of ears per area unit (R = –0.87 for Normalised Red Edge-Red Index (NRERI) and –0.93 for Zarco-Teja-da and Miller Simple Ratio Index (ZM)). A close relationship to ear productivity was found almost for all observed vegetation indices and any leaf in vertical profile (R = 0.79–0.97). In contrast, the prediction of protein content in barley grain was the most reliable when the red-edge reflectance indices (ZM and NRERI) particularly from upper three leaves were used (R = 0.81–0.88). The results show that the knowledge of reflectance heterogeneity in the vertical profile of canopy can significantly contribute to the interpretation of the measured data, to the differentiation of the N nutrition effect from the response to canopy density, and finally to a more accurate estimation of yield parameters and protein content in grain.

Detection of herbicide effects on pigment composition and PSII photochemistry in Helianthus annuus by Raman spectroscopy and chlorophyll a fluorescence

The effects of herbicides from three mode-of-action groups - inhibitors of protoporphyrinogen oxidase (carfentrazone-ethyl), inhibitors of carotenoid biosynthesis (mesotrione, clomazone, and diflufenican), and inhibitors of acetolactate synthase (amidosulfuron) - were studied in sunflower plants (Helianthus annuus). Raman spectroscopy, chlorophyll fluorescence (ChlF) imaging, and UV screening of ChlF were combined to evaluate changes in pigment composition, photosystem II (PSII) photochemistry, and non-photochemical quenching in plant leaves 6d after herbicide application. The Raman signals of phenolic compounds, carotenoids, and chlorophyll were evaluated and differences in their intensity ratios were observed. Strongly augmented relative content of phenolic compounds was observed in the case of amidosulfuron-treated plants, with a simultaneous decrease in the chlorophyll/carotenoid intensity ratio. The results were confirmed by in vivo measurement of flavonols using UV screening of ChlF. Herbicides from the group of carotenoid biosynthesis inhibitors significantly decreased both the maximum quantum efficiency of PSII and non-photochemical quenching as determined by ChlF. Resonance Raman imaging (mapping) data with high resolution (150,000-200,000 spectra) are presented, showing the distribution of carotenoids in H. annuus leaves treated by two of the herbicides acting as inhibitors of carotenoid biosynthesis (clomazone or diflufenican). Clear signs were observed that the treatment induced carotenoid depletion within sunflower leaves. The depletion spatial pattern registered differed depending on the type of herbicide applied.