Rare earth elements,aluminium and silicon distribution in the fern Dicranopteris linearis revealed by μPIXE Maia analysis

BackgroundThe fern Dicranopteris linearis is a hyperaccumulator of rare earth elements (REEs), aluminium (Al) and silicon (Si). However, the physiological mechanisms of tissue-level tolerance of high concentrations of REE and Al, and possible interactions with Si, are currently incompletely known.MethodsA particle-induced X-ray emission (μPIXE) microprobe with the Maia detector, scanning electron microscopy with energy-dispersive spectroscopy and chemical speciation modelling were used to decipher the localization and biochemistry of REEs, Al and Si in D. linearis during uptake, translocation and sequestration processes.ResultsIn the roots >80 % of REEs and Al were in apoplastic fractions, among which the REEs were most significantly co-localized with Si and phosphorus (P) in the epidermis. In the xylem sap, REEs were nearly 100 % present as REEH₃SiO₄²⁺, without significant differences between the REEs, while 24–45 % of Al was present as Al-citrate and only 1.7–16 % Al was present as AlH₃SiO₄²⁺. In the pinnules, REEs were mainly concentrated in necrotic lesions and in the epidermis, and REEs and Al were possibly co-deposited within phytoliths (SiO₂). Different REEs had similar spatial localizations in the epidermis and exodermis of roots, the necrosis, veins and epidermis of pinnae of D. linearis.ConclusionsWe posit that Si plays a critical role in REE and Al tolerance within the root apoplast, transport within the vascular bundle and sequestration within the blade of D. linearis.     

Optical spectroscopic studies of light-harvesting by pigment-reconstituted peridinin-chlorophyll-proteins at cryogenic temperatures

Low temperature, steady-state, optical spectroscopic methods were used to study the spectral features of peridinin-chlorophyll-protein (PCP) complexes in which recombinant apoprotein has been refolded in the presence of peridinin and either chlorophyll a (Chl a), chlorophyll b (Chl b), chlorophyll d (Chl d), 3-acetyl-chlorophyll a (3-acetyl-Chl a) or bacteriochlorophyll a (BChl a). Absorption spectra taken at 10 K provide better resolution of the spectroscopic bands than seen at room temperature and reveal specific pigment–protein interactions responsible for the positions of the Q_y bands of the chlorophylls. The study reveals that the functional groups attached to Ring I of the two protein-bound chlorophylls modulate the Q_y and Soret transition energies. Fluorescence excitation spectra were used to compute energy transfer efficiencies of the various complexes at room temperature and these were correlated with previously reported ultrafast, time-resolved optical spectroscopic dynamics data. The results illustrate the robust nature and value of the PCP complex, which maintains a high efficiency of antenna function even in the presence of non-native chlorophyll species, as an effective tool for elucidating the molecular details of photosynthetic light-harvesting.     

Rare earth elements in forest-floor herbs as related to soil conditions and mineral nutrition

Mixtures of rare earth elements (REEs) in fertilizers are widely used in Chinese agriculture to improve crop nutrition. REE concentrations in wild-growing plants, especially herbs, are little known. This study describes differences in the concentrations and proportions of REEs in eight forest-floor herbaceous plants and relates these differences to soil and mineral nutrient conditions. REEs studied were yttrium (Y), lanthanum (La), cerium (Ce), praseodymium (Pr), neodymium (Nd), samarium (Sm), europium (Eu), gadolinium (Gd), terbium (Tb), dysprosium (Dy), holmium (Ho), erbium (Er), thulium (Tm), ytterbium (Yb), and lutetium (Lu). Leaf concentrations of sum REEs differed more than one order of magnitude between species, being highest in Anemone nemorosa (10.1 nmol/g dry mass) and lowest in Convallaria majalis (0.66 nmol/g) from the same site. Leaf concentrations of all REEs correlated positively (p<0.001), as did sum REE with calcium (Ca) and strontium (Sr) concentrations (p<0.001). A negative relationship (r=−0.83, (p<0.001) was measured between phosphorus (P) concentrations and sum REE concentrations in leaves. However, the proportions of the single REEs in the REE sum differed among species. In A. nemorosa, 57% of the molar REE sum was taken by Y+La, and only 21% by Ce. The other extreme was Maianthemum bifolium, with 37% La+Y and 41% Ce. These two species had 2.7–3.0% of the REE sum as heavier lanthanides, compared to 4.1–5.2% in the six other species. No clear relationship between soil properties or REE contents and leaf REE concentrations was detected. For La, however, an overrepresentation in leaves prevailed throughout all species compared to soils, whereas particularly Nd, Sm, and Tb had a lower proportion in the leaves of all species than in their soils. Possible uptake mechanisms of REEs in plants are discussed.     

Extraction of chlorophylls a and b from phytoplankton using standard extraction techniques *

SUMMARY. Pigments extracted in methanol, acetone and ethanol from three cultures of green algae and one blue-green alga revealed different extraction efficiencies depending on the species, the extraction solvent used and the extraction time. Chromatographic identification and quantitative measurements of chlorophylls a and b were made from six green algae. When extraction of pigments was incomplete, chlorophyll-b was extracted faster than chlorophyll-a. This effect was more pronounced for acetone extractions, whereas methanol extractions gave the stable ratios of chlorophyll b/a after about 6–10 h. When green algae are frequent, a 6–10 h methanol extraction, without any extra manipulations, is sufficient to ensure reliable ratios of chlorophyll b/a and extraction of the major proportion of the chlorophylls without risk of induced destruction of the chlorophylls.     

Light-harvesting properties of zinc complex of chlorophyll-<Emphasis Type="Italic">a</Emphasis> from <Emphasis Type="Italic">spirulina</Emphasis> in surfactant micellar media

Zn chlorophyll-a was prepareted from Mg chlorophyll-a from spirulina and the optical properties of the ground state and the photoexited state of Zn chlorophyll-a in aqueous surfactant micellar media were studied using UV-vis absorption, fluorescence emission spectra, electrochemical and fluorescence lifetime measurements. In comparison of the UV-vis absorption and fluorescence emission spectra of Zn chlorophyll-a and Mg chlorophyll-a, the blue-shift in the absorption bands and emission peak of Zn chlorophyll-a was observed. The energies of the first excited singlet state of Zn chlorophyll-a was 1.87eV. The first oxidation and reduction potentials of the photoexcited singlet state of Zn chlorophyll-a were −0.67 and 0.60V, respectively. Fluorescence lifetime of Zn chlorophyll-a was 9.0 ns in CTAB micellar solution. The fluorescence lifetime of Zn chlorophyll-a is shorter than that of Mg chlorophyll-a (9.8 ns). The photositability of Zn chlorophyll-a was superior to that of Mg chlorophyll-a in various pH conditions. Published online December 2004     

The relation of biotic and abiotic interactions to eutrophication in Tjeukemeer,The Netherlands

During the summer months of 1974–1985 chlorophyll-a and total P concentration, biomass of Daphnia hyalina, smelt Osmerus eperlanus, bream Abramis brema and pikeperch Stizostedion lucioperca, water temperature and water intake from lake IJsselmeer were monitored in Tjeukemeer. During this period there were manipulations with the bream and pikeperch stocks as a consequence of the termination of a gill-net fishery in 1977, and larval smelt immigrated each year from the large lake IJsselmeer and contributed largely to the yearly smelt recruitment.The correlation matrix of the nine variables mentioned above showed a positive correlation between bream and chlorophyll-a, but surprisingly a negative one between smelt and chlorophyll-a. The latter can only be explained when smelt is the dependent variable. In a multi-linear regression there was a negative effect of temperature, chlorophyll a and pikeperch on smelt and a positive effect of water intake. Daphnia hyalina was negatively influenced by the biomass of smelt and the water intake of lake IJsselmeer. The positive relation of Daphnia hyalina and chlorophyll-a was probably related to better survival chances of D. hyalina in an Oscillatoria-rich environment when smelt is the most important predator. An increasing biomass of bream coincided with higher total-P levels and probably contributed to higher chlorophyll-a levels.     

Recovery and Separation of Rare Earth Elements Using Salmon Milt

Recycling rare earth elements (REEs) used in advanced materials such as Nd magnets is important for the efficient use of REE resources when the supply of several REEs is limited. In this work, the feasibility of using salmon milt for REE recovery and separation was examined, along with the identification of the binding site of REEs in salmon milt. Results showed that (i) salmon milt has a sufficiently high affinity to adsorb REEs and (ii) the adsorption capacity of the milt is 1.04 mEq/g, which is comparable with that of commercial cation exchange resin. Heavier REEs have higher affinity for milt. A comparison of stability constants and adsorption patterns of REEs discussed in the literature suggests that the phosphate is responsible for the adsorption of REE in milt. The results were supported by dysprosium (Dy) and lutetium (Lu) L_III-edge extended x-ray absorption fine structure (EXAFS) spectroscopy. The REE-P shell was identified for the second neighboring atom, which shows the importance of the phosphate site as REE binding sites. The comparison of REE adsorption pattern and EXAFS results between the milt system and other adsorbent systems (cellulose phosphate, Ln-resin, bacteria, and DNA-filter hybrid) revealed that the coordination number of phosphate is correlated with the slope of the REE pattern. The separation column loaded with milt was tested to separate REE for the practical use of salmon milt for the recovery and separation of REE. However, water did not flow through the column possibly because of the hydrophobicity of the milt. Thus, sequential adsorption–desorption approach using a batch-type method was applied for the separation of REE. As an example of the practical applications of REE separation, Nd and Fe(III) were successfully separated from a synthetic solution of Nd magnet waste by a batch-type method using salmon milt.     

Use of rare‐earth elements in the phyllosphere colonizer Methylobacterium extorquens PA1

Until recently, rare‐earth elements (REEs) had been thought to be biologically inactive. This view changed with the discovery of the methanol dehydrogenase XoxF that strictly relies on REEs for its activity. Some methylotrophs only contain xoxF, while others, including the model phyllosphere colonizer Methylobacterium extorquens PA1, harbor this gene in addition to mxaFI encoding a Ca²⁺‐dependent enzyme. Here we found that REEs induce the expression of xoxF in M. extorquens PA1, while repressing mxaFI, suggesting that XoxF is the preferred methanol dehydrogenase in the presence of sufficient amounts of REE. Using reporter assays and a suppressor screen, we found that lanthanum (La³⁺) is sensed both in a XoxF‐dependent and independent manner. Furthermore, we investigated the role of REEs during Arabidopsis thaliana colonization. Element analysis of the phyllosphere revealed the presence of several REEs at concentrations up to 10 μg per g dry weight. Complementary proteome analyses of M. extorquens PA1 identified XoxF as a top induced protein in planta and a core set of La³⁺‐regulated proteins under defined artificial media conditions. Among these was a REE‐binding protein that is encoded next to a gene for a TonB‐dependent transporter. The latter was essential for REE‐dependent growth on methanol indicating chelator‐assisted uptake of REEs.     

Dynamics and distribution of microphytobenthic chlorophyll-a in the Western Scheldt estuary (SW Netherlands)

The temporal dynamics and spatial distribution of microphytobenthic chlorophyll-a in the layer 0–1 cm were determined in the Western Scheldt estuary over the period 1991–1992. Connections between the annually averaged benthic chlorophyll-a and station elevation and sediment composition (as a measure of the hydrodynamic energy caused by currents and waves) were also examined.Microphytobenthic chlorophyll-a showed one main peak in early summer and a smaller peak in autumn. The mean chlorophyll-a concentration of 113 mg Chl-a m^–2 in the upper centimeter is of the same order of magnitude as in other estuarine areas. The average annual primary production of the microphytobenthos has been estimated at 136 g C m^–2 y^–1 The primary production of sediment inhabiting microalgae is at least 17% of the total primary production in the estuary.Considerable differences in annually averaged chlorophyll-a emerges between the stations. These differences are related mainly to the interaction between station elevation and clay content of the sediment.     

Visible light induced hydrogen production with Mg chlorophyll-a from spirulina and colloidal platinum

Photoinduced hydrogen production with Mg chlorophyll-a from spirulina as a visible light photosensitizer by use of three component system consisting of nicotineamide adenine dinucleotide phosphate, reduced form (NADPH) as an electron donor, methylviologen as electron relay reagent and colloidal platinum as hydrogen evolution catalyst was investigated. By the addition of NADPH, the photostability of Mg chlorophyll-a was increased. The effective visible-light induced hydrogen production system with colloidal platinum was established using Mg chlorophyll-a.     

Population dynamics of zooxanthellae during a bacterial bleaching event

Each summer 80–90% of the colonies of Oculina patagonica undergo bleaching off the Mediterranean coast of Israel. To investigate fluctuations through a yearly bleaching cycle, monthly measurements of zooxanthella density, mitotic index and chlorophyll-a concentration were conducted. Results showed (1) a significant negative correlation between sea surface temperature (SST) and zooxanthella density; (2) both significantly lower zooxanthella mitotic index and higher chlorophyll-a per zooxanthella content during the bleaching season compared with the non-bleaching period; (3) prior to bleaching, a lag between the peak of zooxanthella density and chlorophyll-a concentration followed by a similar lag during recovery. Zooxanthella density declined significantly between March and May while chlorophyll-a concentration peaked in April, and then declined. Zooxanthella density increased significantly in November while chlorophyll-a concentration increased significantly in January. We conclude that during bacterial bleaching events, zooxanthellae are severely damaged. However, by the time of the following bleaching event the coral tissues regain their “normal” (pre-bleaching) zooxanthella population density.     

Dynamic change in photosynthetic pigments and chlorophyll degradation elicited by cereal aphid feeding

The concentration of photosynthetic pigments (i.e., chlorophylls a and b, and carotenoids) and chlorophyll degradation enzyme (i.e., chlorophyllase, oxidative bleaching, and Mg-dechelatase) activities on aphid-damaged and non-damaged regions of the infested leaves were determined with two infestation periods (6 and 12 days). Russian wheat aphid [Diuraphis noxia (Mordvilko) (Hemiptera: Aphididae)] feeding caused significant losses of chlorophylls a and b and carotenoids in the damaged regions. However, bird cherry-oat aphid [Rhopalosiphum padi (L.) (Hemiptera: Aphididae)] feeding did not, except a significantly lower level of carotenoids was observed in the damaged regions from the short-infestation (6-day) samples. Interestingly, the non-damaged regions of D. noxia-infested leaves on both sampling dates had a significant increase of chlorophylls a and b and carotenoid concentrations when compared with the uninfested leaves. Although D. noxia feeding did not cause any changes in either chlorophyll a/b or chlorophyll (a+b)/carotenoid ratio between the damaged and non-damaged leaf regions on short-infestation (6-day) samples, a significantly lower chlorophyll a/b ratio was detected in long-infestation (12-day) samples. The assays of chlorophyllase and oxidative bleaching activities showed no significant differences between the damaged and non-damaged regions of the infested leaves on either sampling date. Mg-dechelatase activity, however, was significantly higher in D. noxia-damaged than non-damaged leaf regions from the short-infestation samples, while no differences were detected from the long-infestation samples. Furthermore, the long-infestation samples showed that Mg-dechelatase activity from both D. noxia-damaged and non-damaged regions increased significantly in comparison with the respective regions of either uninfested or R. padi-infested leaves. We infer that non-damaged regions of D. noxia-infested leaves compensate for the pigment losses in the damaged regions, and that Mg-dechelatase activity changed dynamically from a localized response to a systemic response as infestation duration extends. The findings from this study on cereal aphid-elicited chlorosis (or desistance) would help us to elucidate plant resistance mechanisms, in particular plant tolerance to non-defoliating herbivory.     

Photo-oxidative Degradation of Chlorophyll-a and Unsaturated Lipids in Liposomal Dispersions at Low-Temperature

Liposomal dispersions in water were used as a tool to study photo-oxidation of chlorophyll-a and photo-oxidation of unsaturated lipids at 1 or 4°C. The presence of monogalactosyl diglyceride stimulated chlorophyll-a degradation. In addition the level of linolenic acid was decreased in liposomal dispersions containing chlorophyll-a, dipalmitoyl phosphatidyl choline, and monogalactosyl diglyceride, indicating that monogalactosyl diglyceride and chlorophyll-a were coupled in the preparations. In liposomal dispersions containing equal (molar) quantities of a-tocopherol, monogalactosyl diglyceride, and chlorophyll-a, a-tocopherol fully protected linolenic acid against photo-oxidative degradation, while chlorophyll-a degradation was only slightly reduced. In liposomal preparations containing a-tocopherol, chlorophyll-a and phosphatidyl choline, a-tocopherol catalyzed degradation of chlorophyll-a. Absorption spectra of the liposomal dispersions showed that the presence of a-tocopherol caused increased absorption in red light, which was attributed to structural changes in the liposomal preparations and thus could explain the noted effects. Tocopherol itself was rapidly degraded in chlorophyll-a containing liposomal preparations. Complex formation between chlorophyll-a and monogalactosyl diglyceride in chloroplasts is suggested and protection by a-tocopherol against photo-oxidation in chilling-sensitive plants; a suggestion which is founded on the similarities that exist between chloroplast preparations and liposomal preparations containing chlorophyll-a and monogalactosyl diglyceride as regards photo-oxidative degradation of chlorophyll-a, a-tocopherol and linolenic acid.     

Minor complexes at work: light-harvesting by carotenoids in the photosystem II antenna complexes CP24 and CP26

Plant photosynthesis relies on the capacity of chlorophylls and carotenoids to absorb light. One of the roles of carotenoids is to harvest green-blue light and transfer the excitation energy to the chlorophylls. The corresponding dynamics were investigated here for the first time, to our knowledge, in the CP26 and CP24 minor antenna complexes. The results for the two complexes differ substantially. In CP26 fast transfer (80 fs) occurs from the carotenoid S₂ state to chlorophylls a absorbing at 675 and 678 nm, whereas transfer from the hot S₁ state to the lowest energy chlorophylls is observed in <1 ps. In CP24, energy transfer from the S₂ state leads in 80 fs to the population of chlorophylls b and high-energy chlorophylls a absorbing at 670 nm, whereas the low-energy chlorophylls a are populated only in several picoseconds. The results suggest that CP26 has a structural and functional organization similar to that of LHCII, whereas CP24 differs substantially from the other Lhc complexes, especially regarding the lutein L1 binding domain. No energy transfer from the carotenoid S₁ state to chlorophylls was observed in either complex, suggesting that this state is energetically below the chlorophyll Qy state and therefore may play a role in the quenching of chlorophyll excitations.     

Visible and near-IR light induced biohydrogen production using the system containing Mg chlorophyll-a from Spirulina and colloidal platinum

Photoinduced hydrogen production with Mg chlorophyll-a from Spirulina as a visible and near-IR light photosensitizer by use of three component system consisting of nicotineamide adenine dinucleotide phosphate, reduced form (NADPH) as an electron donor, methylviologen as electron relay reagent and colloidal platinum as hydrogen production catalyst was investigated. After 4 h irradiation, the amount of hydrogen production with Mg chlorophyll-a and MgTPP, which was artificial model compound for chlorophyll, were c.a. 2.7 and 1.8 mol, respectively. When the near-IR light was irradiated, little change of hydrogen production was observed. Thus, the effective visible and near IR light induced hydrogen production system with colloidal platinum was established using Mg chlorophyll-a.     

Pigment alterations in the brown mussel Perna perna

Potential sex and/or gametogenic stage differences in the metabolism of chlorophyll-a and carotenoids in the brown mussel Perna perna of southern Brazil were studied using high performance liquid chromatography (HPLC). Carotenoids derived directly from diet (phytoplankton) were fucoxanthin plus diatoxanthin (diatoms), alloxanthin (cryptophytes) and zeaxanthin (mainly cyanobacteria). Females accumulated carotenoid-diols and epoxides (~ 3–4 mg/g-dry wt.) while males had much lower concentrations (~ 0.7 mg/g-dry wt.). An antioxidant/free radical scavenging role is proposed for carotenoids in females. Mean ratios of chlorophyll plus derivatives (Chlns-a) to carotenoids for male and female P. perna were 50:1 and 4:1, respectively. The higher ratio in males relates to both higher carotenoid contents in females plus higher total Chlns-a in males (~ 22 mg/g-dry wt.), relative to the females (~ 4 mg/g-dry wt.). Chlorophyll-a metabolism in both sexes followed two distinct pathways. First, cyclization of pyropheophorbide-a gave 13², 17³-cyclopheophorbide-a-enol (CPPaE) which was further oxidized to hydroxy-chlorophyllone. Second, chlorophyll-a derivatives retaining the 13²-carbomethoxy moiety were oxidized to purpurin-18 which was hydrolyzed to chlorin-p₆. In both cases, metabolism of dietary chlorophyll-a was oxidative and derivatives could either serve as antioxidants or merely be the results of non-specific digestive processes.     

Nomenclature for membrane-bound light-harvesting complexes of cyanobacteria

Accessory chlorophyll-binding proteins (CBP) in cyanobacteria have six transmembrane helices and about 11 conserved His residues that might participate in chlorophyll binding. In various species of cyanobacteria, the CBP proteins bind different types of chlorophylls, including chlorophylls a, b, d and divinyl-chlorophyll a, b. The CBP proteins do not belong to the light-harvesting complexes (LHC) superfamily of plant and algae. The proposed new name of CBP for this class of proteins, which is a unique accessory light-harvesting superfamily in cyanobacteria, clarifies the confusion of names of prochlorophytes chlorophyll binding protein (Pcb), PSII-like light-harvesting proteins and iron-stress-induced protein A (IsiA). The CBP complexes are a member of a larger family that includes the chlorophyll a-binding proteins CP43 and CP47 that function as core antennas of photosystem II.     

Effects of Aluminium on Pigments and Pigment-Protein Complexes of Soybean

The effect of aluminium (0.5 –1.0 mM) on contents of phosphorus, pigments, and pigment-protein complexes was studied in soybean (Glycine max Merril.) grown in different nutrient medium with and without P. Increased Al concentrations led to the decrease in the contents of chlorophylls (Chl) a and b, and carotenoids (Car) in soybean leaves, but Chl a/b ratio did not vary significantly. In long-term experiments, P ameliorates the negative effects of Al.     

The chequered history of the development and use of simultaneous equations for the accurate determination of chlorophylls a and b

Over the last half century, the most frequently used assay for chlorophylls in higher plants and green algae, the Arnon assay [Arnon DI (1949) Plant Physiol 24: 1–15], employed simultaneous equations for determining the concentrations of chlorophylls a and b in aqueous 80% acetone extracts of chlorophyllous plant and algal materials. These equations, however, were developed using extinction coefficients for chlorophylls a and b derived from early inaccurate spectrophotometric data. Thus, Arnon's equations give inaccurate chlorophyll a and b determinations and, therefore, inaccurate chlorophyll a/b ratios, which are always low. This paper describes how the ratios are increasingly and alarmingly low as the proportion of chlorophyll a increases. Accurate extinction coefficients for chlorophylls a and b, and the more reliable simultaneous equations derived from them, have been published subsequently by many research groups; these new post-Arnon equations, however, have been ignored by many researchers. This Minireview records the history of the development of accurate simultaneous equations and some difficulties and anomalies arising from the retention of Arnon's seriously flawed equations. This revised version was published online in June 2006 with corrections to the Cover Date.