Molecular characterization of oleosin genes in Cyperus esculentus, a Cyperaceae plant producing oil in underground tubers.

KEY MESSAGE: CeOLE genes exhibit a tuber-predominant expression pattern and their mRNA/protein abundances are positively correlated with oil accumulation during tuber development. Overexpression could significantly increase the oil content of tobacco leaves. Oleosins (OLEs) are abundant structural proteins of lipid droplets (LDs) that function in LD formation and stabilization in seeds of oil crops. However, little information is available on their roles in vegetative tissues. In this study, we present the first genome-wide characterization of the oleosin family in tigernut (Cyperus esculentus L., Cyperaceae), a rare example accumulating high amounts of oil in underground tubers. Six members identified represent three previously defined clades (i.e. U, SL and SH) or six out of seven orthogroups (i.e. U, SL1, SL2, and SH1-3) proposed in this study. Comparative genomics analysis reveals that lineage-specific expansion of Clades SL and SH was contributed by whole-genome duplication and dispersed duplication, respectively. Moreover, presence of SL2 and SH3 in Juncus effuses implies their appearance sometime before Cyperaceae-Juncaceae divergence, whereas SH2 appears to be Cyperaceae specific. Expression analysis showed that CeOLE genes exhibit a tuber-predominant expression pattern and transcript levels are considerably more abundant than homologs in the close relative Cyperus rotundus. Moreover, CeOLE mRNA and protein abundances were shown to positively correlate with oil accumulation during tuber development. Additionally, two dominant isoforms (i.e. CeOLE2 and -5) were shown to locate in LDs as well as the endoplasmic reticulum of tobacco (Nicotiana benthamiana) leaves, and are more likely to function in homo and heteromultimers. Furthermore, overexpression of CeOLE2 and -5 in tobacco leaves could significantly increase the oil content, supporting their roles in oil accumulation. These findings provide insights into lineage-specific family evolution and putative roles of CeOLE genes in oil accumulation of vegetative tissues, which facilitate further genetic improvement for tigernut.

Differential distribution of amino acids in plants.

Plants are a rich source of amino acids and their individual abundance in plants is of great significance especially in terms of food. Therefore, it is of utmost necessity to create a database of the relative amino acid contents in plants as reported in literature. Since in most of the cases complete analysis of profiles of amino acids in plants was not reported, the units used and the methods applied and the plant parts used were different, amino acid contents were converted into relative units with respect to lysine for statistical analysis. The most abundant amino acids in plants are glutamic acid and aspartic acid. Pearson's correlation analysis among different amino acids showed that there were no negative correlations between the amino acids. Cluster analysis (CA) applied to relative amino acid contents of different families. Alismataceae, Cyperaceae, Capparaceae and Cactaceae families had close proximity with each other on the basis of their relative amino acid contents. First three components of principal component analysis (PCA) explained 79.5% of the total variance. Factor analysis (FA) explained four main underlying factors for amino acid analysis. Factor-1 accounted for 29.4% of the total variance and had maximum loadings on glycine, isoleucine, leucine, threonine and valine. Factor-2 explained 25.8% of the total variance and had maximum loadings on alanine, aspartic acid, serine and tyrosine. 14.2% of the total variance was explained by factor-3 and had maximum loadings on arginine and histidine. Factor-4 accounted 8.3% of the total variance and had maximum loading on the proline amino acid. The relative content of different amino acids presented in this paper is alanine (1.4), arginine (1.8), asparagine (0.7), aspartic acid (2.4), cysteine (0.5), glutamic acid (2.8), glutamine (0.6), glycine (1.0), histidine (0.5), isoleucine (0.9), leucine (1.7), lysine (1.0), methionine (0.4), phenylalanine (0.9), proline (1.1), serine (1.0), threonine (1.0), tryptophan (0.3), tyrosine (0.7) and valine (1.2).

Effect of dissimilatory iron and sulfate reduction on arsenic dynamics in the wetland rhizosphere and its bioaccumulation in wetland plants (Scirpus actus).

Microbial redox transformations of arsenic (As) are coupled to dissimilatory iron and sulfate reduction in the wetlands, however, the processes involved are complex and poorly defined. In this study, we investigated the effect of dissimilatory iron and sulfate reduction on As dynamics in the wetland rhizosphere and its bioaccumulation in plants using greenhouse mesocosms. Results show that high Fe (50µM ferrihydrite/g solid medium) and SO42- (5mM) treatments are most favorable for As sequestration in the presence of wetland plants (Scirpus actus), probably because root exudates facilitate the microbial reduction of Fe(III), SO42-, and As(V) to sequester As(III) by incorporation into iron sulfides and/or plant uptake. As retention in the solid medium and accumulation in plants were mainly controlled by SO42- rather than Fe levels. Compared to the low SO42- (0.1mM) treatment, high SO42- resulted in 2 times more As sequestered in the solid medium, 30 times more As in roots, and 49% less As in leaves. An As speciation analysis in pore water indicated that 19% more dissolved As was reduced under high SO42- than low SO42- levels, which is consistent with the fact that more dissimilatory arsenate-respiring bacteria were found under high SO42- levels.

Phytoremediation of cadmium by the facultative halophyte plant Bolboschoenus maritimus (L.) Palla, at different salinities.

The cadmium phytoremediation capacity of the halophyte plant Bolboschoenus maritimus (L.) Palla and the influence of water salinity were assessed in a greenhouse experiment, in order to better understand the bioremediation capacity of this plant. Three concentrations of cadmium (0, 50 and 100 µg l(-1)) and four salinity conditions (0, 5, 10 and 20) were chosen to evaluate the cadmium accumulation, in order to test these plants as a potential phytoremediation tool in brackish environments. The cadmium content in water and plants (underground organs, stems and leaves) was analysed with graphite furnace atomic absorption spectrometry. All the plants submitted to salinity 20 and in the three cadmium treatments died. The plants' survival was highest in the lowest salinities, where highest growth and biomasses were also obtained. The plants presented more cadmium content in the rhizomes, followed by stems and even less in leaves. The salt stress of the plants interfered with their cadmium accumulation capacity. The highest cadmium accumulation in the rhizomes occurred at salinity 0, while the salinities 0 and 5 were the most adequate for stems and leaves. The experiment pointed out that B. maritimus represents a good possible intervenient for cadmium bioremediation in freshwater and low salinity brackish water environments, but its use is limited in the habitats of higher salinity.

Differences in phytotoxicity and dissipation between ionized and nonionized oil sands naphthenic acids in wetland plants.

Naphthenic acids (NAs) are composed of alkyl-substituted acyclic and cycloaliphatic carboxylic acids and, because they are acutely toxic to fish, are of toxicological concern. During the caustic hot-water extraction of oil from the bitumen in oil sands deposits, NAs become concentrated in the resulting tailings pond water. The present study investigated if dissipation of NAs occurs in the presence of hydroponically grown emergent macrophytes (Typha latifolia, Phragmites australis, and Scirpus acutus) to determine the potential for phytoremediation of these compounds. Plants were grown with oil sands NAs (pKa approximately 5-6) in medium at pH 7.8 (predominantly ionized NAs) and pH 5.0 (predominantly nonionized NAs) to determine if, by altering their chemical form, NAs may be more accessible to plants and, thus, undergo increased dissipation. Whereas the oil sands NA mixture in its nonionized form was more toxic to wetland plants than its ionized form, neither form appeared to be sequestered by wetland plants. The present study demonstrated that plants may selectively enhance the dissipation of individual nonionized NA compounds, which contributes to toxicity reduction but does not translate into detectable total NA dissipation within experimental error and natural variation. Plants were able to reduce the toxicity of a NA system over 30 d, increasing the median lethal concentration (LC50; % of hydroponic solution) of the medium for Daphnia magna by 23.3% +/- 8.1% (mean +/- standard error; nonionized NAs) and 37.0% +/- 2.7% (ionized NAs) as determined by acute toxicity bioassays. This reduction in toxicity was 7.3% +/- 2.6% (nonionized NAs) and 45.0% +/- 6.8% (ionized NAs) greater than that in unplanted systems.

Isoprene emission from a subarctic peatland under enhanced UV-B radiation.

Isoprene is a reactive hydrocarbon with an important role in atmospheric chemistry, and emissions from vegetation contribute to atmospheric carbon fluxes. The magnitude of isoprene emissions from arctic peatlands is not known, and it may be altered by increasing UV-B radiation. Isoprene emission was measured with the dynamic chamber method from a subarctic peatland under long-term enhancement of UV-B radiation targeted to correspond to a 20% loss in the stratospheric ozone layer. The site type of the peatland was a flark fen dominated by the moss Warnstorfia exannulata and sedges Eriophorum russeolum and Carex limosa. The relationship between species densities and the emission was also assessed. Isoprene emissions were significantly increased by enhanced UV-B radiation during the second (2004) and the fourth (2006) growing seasons under the UV-B exposure. Emissions were related to the density of E. russeolum. The dominant moss, W. exannulata, proved to emit small amounts of isoprene in a laboratory trial. Subarctic fens, even without Sphagnum moss, are a significant source of isoprene to the atmosphere, especially under periods of warm weather. Warming of the Arctic together with enhanced UV-B radiation may substantially increase the emissions.

The effect of the size of particles on mineralization of Oxycaryum cubense (Poepp. & Kunth) Lye

Ensaios foram realizados para avaliar os efeitos de tamanho dos detritos na mineralização de uma macrófita aquática, a Oxycaryum cubense. Foram coletadas amostras de planta e de água de uma lagoa marginal, a lagoa do Infernão (21° 35' S e 47° 51' W), localizada na planície de inundação do rio Mogi Guaçu. As plantas foram levadas ao laboratório, lavadas em água corrente, secas (50 °C) e fracionadas em seis grupos de acordo com o tamanho: 100, 10, 1,13, 0,78, 0,61 e 0,25 mm. Foram preparadas câmaras de decomposição adicionando aproximadamente 1,0 g de fragmentos de planta em 4,1 L de água da lagoa. Na seqüência, as incubações foram aeradas e as concentrações de oxigênio dissolvido, o pH, a condutividade elétrica e a temperatura foram monitorados durante 120 dias. A ocorrência de processos de anaeróbios foi evitada por reoxigenação das soluções. Os resultados experimentais foram ajustados a um modelo cinético de primeira ordem e o consumo de oxigênio dissolvido decorrente da mineralização foi obtido. De maneira geral, o processo físico de fragmentação não tende a envolver maiores quantidades de oxigênio durante a mineralização aeróbia dos detritos, significando que a fragmentação não interfere no balanço de oxigênio dissolvido deste sistema aquático; entretanto favoreceu a acidificação e também a liberação de íons dissolvidos na lagoa do Infernão.

Stilbenoids of Kobresia nepalensis (Cyperaceae) exhibiting DNA topoisomerase II inhibition.

Resveratrol oligomers, nepalensinol A, B and C, were isolated from the stem of Kobresia nepalensis (Cyperaceae). The structures were established on the basis of chemical properties and spectroscopic evidence including 2D NMR spectroscopic analysis. Nepalensinol A, B and C showed a potent inhibitory effect on topoisomerase II -- stronger than etoposide (VP-16), a topoisomerase II inhibitor used as an anti-cancer drug. Nepalensinol B, in particular, exhibited the most potent activity with an IC(50) of 0.02 microg/ml.

Hazard assessment of a simulated oil spill on intertidal areas of the St. Lawrence River with SPMD-TOX.

Phytoremediation in a simulated crude oil spill was studied with a "minimalistic" approach. The SPMD-TOX paradigm-a miniature passive sorptive device to collect and concentrate chemicals and microscale tests to detect toxicity-was used to monitor over time the bioavailability and potential toxicity of an oil spill. A simulated crude oil spill was initiated on an intertidal freshwater grass-wetland along the St. Lawrence River southwest of Quebec City, Quebec, Canada. Several phytoremediation treatments were investigated; to dissipate and ameliorate the spill, treatments included nutrient amendments with inorganic nitrogen sources (ammonium nitrate and sodium nitrate) and phosphate (super triple phosphate) with and without cut plants, with natural attenuation (no phytoremedial treatment) as a control. Sequestered oil residues were bioavailable in all oil-treated plots in Weeks 1 and 2. Interestingly, the samples were colored and fluoresced under ultraviolet light. In addition, microscale tests showed that sequestered residues were acutely toxic and genotoxic, as well as that they induced hepatic P(450) enzymes. Analysis of these data suggested that polycyclic aromatic hydrocarbons were among the bioavailable residues sequestered. In addition, these findings suggested that the toxic bioavailable fractions of the oil spill and degradation products dissipated rapidly over time because after the second week the water column contained no oil or detectable degradation products in this riverine intertidal wetland. SPMD-TOX revealed no evidence of bioavailable oil products in Weeks 4, 6, 8, and 12. All phytoremediation efforts appeared to be ineffective in changing either the dissipation rate or the ability to ameliorate the oil toxicity. SPMD-TOX analysis of the water columns from these riverine experimental plots profiled the occurrence, dissipation, and influence of phytoremediation on the bioavailability and toxicity of oil products (parent or degradation products).