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.

Analysis of oil synthesis pathway in Cyperus esculentus tubers and identification of oleosin and caleosin genes.

The tubers of the widely distributed Cyperus esculentus are rich in oil, and therefore, the plant is considered to have a high utilization value in the vegetable oil industry. Oleosins and caleosins are lipid-associated proteins found in oil bodies of seeds; however oleosins and caleosins genes have not been identified in C. esculentus. In this study, we performed transcriptome sequencing and lipid metabolome analysis of C. esculentus tubers at four developmental stages to obtain the information on their genetic profile, expression trends, and metabolites in oil accumulation pathways. Overall, 120,881 non-redundant unigenes and 255 lipids were detected; 18 genes belonged to the acetyl-CoA carboxylase (ACC), malonyl-CoA:ACP transacylase (MCAT), ß-ketoacyl-ACP synthase (KAS), and fatty acyl-ACP thioesterase (FAT) gene families involved in fatty acid biosynthesis, and 16 genes belonged to the glycerol-3-phosphate acyltransferase (GPAT), diacylglycerol acyltransferase 3 (DGAT3), phospholipid:diacylglycerol acyltransferase (PDAT), FAD2, and lysophosphatidic acid acyltransferase (LPAAT) gene families playing important roles in triacylglycerol synthesis. We also identified 9 oleosin- and 21 caleosin-encoding genes in C. esculentus tubers. These results provide detailed information on the C. esculentus transcriptional and metabolic profiles, which can be used as reference for the development of strategies to increase oil content in C. esculentus tubers.

Chemical Profile of Cyperus laevigatus and Its Protective Effects against Thioacetamide-Induced Hepatorenal Toxicity in Rats.

Cyperus species represent a group of cosmopolitan plants used in folk medicine to treat several diseases. In the current study, the phytochemical profile of Cyperus laevigatus ethanolic extract (CLEE) was assessed using UPLC-QTOF-MS/MS. The protective effect of CLEE at 50 and 100 mg /kg body weight (b.w.) was evaluated on hepatorenal injuries induced by thioacetamide (100 mg/kg) via investigation of the extract's effects on oxidative stress, inflammatory markers and histopathological changes in the liver and kidney. UPLC-QTOF-MS/MS analysis of CLEE resulted in the identification of 94 compounds, including organic and phenolic acids, flavones, aurones, and fatty acids. CLEE improved the antioxidant status in the liver and kidney, as manifested by enhancement of reduced glutathione (GSH) and coenzyme Q10 (CoQ10), in addition to the reduction in malondialdehyde (MDA), nitric oxide (NO), and 8-hydroxy-2'-deoxyguanosine (8OHdG). Moreover, CLEE positively affected oxidative stress parameters in plasma and thwarted the depletion of hepatorenal ATP content by thioacetamide (TAA). Furthermore, treatment of rats with CLEE alleviated the significant increase in plasma liver enzymes, kidney function parameters, and inflammatory markers. The protective effect of CLEE was confirmed by a histopathological study of the liver and kidney. Our results proposed that CLEE may reduce TAA-hepatorenal toxicity via its antioxidant and anti-inflammatory properties suppressing oxidative stress.

The antioxidant and antidiabetic potentials of polyphenolic-rich extracts of Cyperus rotundus (Linn.).

In this study, the rhizome of Cyperus rotundus L was investigated for its antioxidant and antidiabetic effects using in vitro and in silico experimental models. Its crude extracts (ethyl acetate, ethanol and aqueous) were screened in vitro for their antioxidant activity using ferric-reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picrylhydrazyl (DPPH), as well as their inhibitory effect on α-glucosidase enzyme. Subsequently, the extracts were subjected to Gas Chromatography-Mass Spectrometry (GC-MS) analysis to elucidate their possible bioactive compounds. Furthermore, computational molecular docking of selected phenolic compounds was conducted to determine their mode of α-glucosidase inhibitory activity. The aqueous extract displayed the highest level of total phenolic content and significantly higher scavenging activity in both FRAP and DPPH assays compared to ethyl acetate and ethanol extracts. In FRAP and DPPH assays, IC50 values of aqueous extract were 448.626 µg/mL and 418.74 µg/mL, respectively. Aqueous extract further presented higher α-glucosidase inhibitory activity with an IC50 value of 383.75 µg/mL. GC-MS analysis revealed the presence of the following phenolic compounds: 4-methyl-2-(2,4,4-trimethylpentan-2-yl) phenol, Phenol,2-methyl-4-(1,1,3,3-tetramethylbutyl)- and 1-ethoxy-2-isopropylbenzene. Molecular docking study revealed 1-ethoxy-2-isopropylbenzene formed two hydrogen bonds with the interacting residues in the active site of α-glucosidase enzyme. Furthermore, 4-methyl-2-(2,4,4-trimethylpentan-2-yl) phenol had the lowest binding energy inferring the best affinity for α-glucosidase active site. These results suggest the possible antioxidant and antidiabetic potential of Cyperus rotundus.Communicated by Ramaswamy H. Sarma.

Cyperus spp.: A Review on Phytochemical Composition, Biological Activity, and Health-Promoting Effects.

Cyperaceae are a plant family of grass-like monocots, comprising 5600 species with a cosmopolitan distribution in temperate and tropical regions. Phytochemically, Cyperus is one of the most promising health supplementing genera of the Cyperaceae family, housing ≈950 species, with Cyperus rotundus L. being the most reported species in pharmacological studies. The traditional uses of Cyperus spp. have been reported against various diseases, viz., gastrointestinal and respiratory affections, blood disorders, menstrual irregularities, and inflammatory diseases. Cyperus spp. are known to contain a plethora of bioactive compounds such as α-cyperone, α-corymbolol, α-pinene, caryophyllene oxide, cyperotundone, germacrene D, mustakone, and zierone, which impart pharmacological properties to its extract. Therefore, Cyperus sp. extracts were preclinically studied and reported to possess antioxidant, anti-inflammatory, antimicrobial, anticancer, neuroprotective, antidepressive, antiarthritic, antiobesity, vasodilator, spasmolytic, bronchodilator, and estrogenic biofunctionalities. Nonetheless, conclusive evidence is still sparse regarding its clinical applications on human diseases. Further studies focused on toxicity data and risk assessment are needed to elucidate its safe and effective application. Moreover, detailed structure-activity studies also need time to explore the candidature of Cyperus-derived phytochemicals as upcoming drugs in pharmaceuticals.

Selection and Validation of Reference Genes for qRT-PCR Analysis in the Oil-Rich Tuber Crop Tiger Nut (Cyperus esculentus) Based on Transcriptome Data.

Tiger nut (Cyperus esculentus), a perennial C4 plant of the Cyperaceae family, is an unconventional crop that is distinguished by its oil-rich tubers, which also possesses the advantages of strong resistance, wide adaptability, short life periods, and large biomass. To facilitate studies on gene expression in this species, we identified and validated a series of reference genes (RGs) based on transcriptome data, which can be employed as internal controls for qRT-PCR analysis in tiger nut. Fourteen putative candidate RGs were identified and evaluated across nine different tissues of two cultivars, and the RGs were analyzed using three different algorithms (geNorm, NormFinder, and BestKeeper). The stability rankings of the candidate RGs were merged into consensus lists with RankAggreg. For the below-ground storage organ of tiger nut, the optimal RGs were TUB4 and UCE2 in different developmental stages of tubers. UCE2 and UBL5 were the most stably expressed RGs among all tissues, while Rubisco and PGK exhibited the lowest expression stability. UCE2, UBL5 and Rubisco were compared to normalize the expression levels of the caleosin (CLO) and diacylglycerol acyltransferase 2-2 (DGAT2-2) genes across the same tissues. Our results showed that the RGs identified in this study, which exhibit more uniform expression patterns, may be utilized for the normalization of qRT-PCR results, promoting further research on gene expression in various tissues of tiger nut.

UPLC-qTOF-MS Phytochemical Profile and Antiulcer Potential of Cyperus conglomeratus Rottb. Alcoholic Extract.

Cyperus has been commonly used as a multi-use medicinal plant in folk medicine worldwide. The objectives of our study were to determine the different metabolites in the Cyperus conglomeratus Rottb. methanol extract, and to assess its in vivo gastroprotective effect in ethanol-induced gastric ulcer model in rats. Serum levels of galactin-3 and TNF-α were employed as biochemical markers. To pinpoint for active agents, comprehensive metabolites profiling of extract via UPLC-qTOF-MS/MS was employed. A total of 77 chromatographic peaks were detected, of which 70 were annotated. The detected metabolites were categorized into phenolic acids and their derivatives, flavonoids, stilbenes, aurones, quinones, terpenes, and steroids. Rats were divided into six groups; healthy control, ulcer control, standard drug group, and 25, 50, 100 mg/kg of C. conglomeratus treated rats. Pre-treatment with C. conglomeratus alcohol extract significantly reduced galactin-3, and TNF-α in ethanol-induced ulcer model at 25, 50, and 100 mg/kg. Further histopathological and histochemical studies revealed moderate erosion of superficial epithelium, few infiltrated inflammatory cells, and depletion of gastric tissue glycoprotein in the ulcer group. Treatment with the extract protected the gastric epithelial cells in a dose-dependent manner. It could be concluded that C. conglomeratus extract provides significant gastroprotective activity in ethanol-induced gastric ulcer and ought to be included in nutraceuticals in the future for ulcer treatment.

Functional Characterization of Three Novel Genes Encoding Diacylglycerol Acyltransferase (DGAT) from Oil-Rich Tubers of Cyperus esculentus.

Cyperus esculentus is probably the only plant that is known to accumulate large amounts of oil in its tubers. However, the underlying metabolic mechanism and regulatory factors involved in oil synthesis of tubers are still largely unclear. In this study, one gene encoding type I diacylglycerol acyltransferase (DGAT) (CeDGAT1) and two genes encoding type II DGAT (CeDGAT2a and CeDGAT2b) from C. esculentus were identified and functionally analyzed. All three DGAT genes were found to be expressed in tuber, root and leaf tissues. CeDGAT1 is highly expressed in roots and leaves, whereas CeDGAT2b is dominantly expressed in tubers. Furthermore, the temporal expression pattern of CeDGAT2b is well coordinated with the oil accumulation in developing tubers. When each CeDGAT was heterologously expressed in triacylglycerol (TAG)-deficient mutant of Saccharomyces cerevisiae, Arabidopsis thaliana wild type or its TAG1 mutant with AtDGAT1 disruption, only CeDGAT2b showed the ability to restore TAG biosynthesis with lipid body formation in yeast mutant, enhance seed oil production of Arabidopsis wild type and rescue multiple seed phenotypes of TAG1 mutant. In addition, CeDGAT2b was shown to have a substrate preference for unsaturated fatty acids toward TAG synthesis. Taken together, our results indicated that CeDGAT2b from C. esculentus is an actively functional protein and is most likely the major contributor to tuber oil biosynthesis containing common fatty acids, in contrast to oil-rich seeds and fruits where DGAT1 plays a more central role than DGAT2 in oil production accumulating normal fatty acids, whereas DGAT2 is a primary regulator for oil synthesis rich in unusual fatty acids.

Chemical composition of industrially and laboratory processed Cyperus esculentus rhizomes.

We report here the results of the study of the chemical composition of Cyperus esculentus rhizomes. Ethanolic extracts have been separated by column chromatography and analyzed by magnetic resonance spectroscopy and mass spectrometry. Quercetin, stigmasterol, and linoleic and oleic acid glycerol esters, together with 4-chlorobutyl oleate, oleamide, myricetin, tyramine and N-feruloyltyramine, found for the first time in Cyperus esculentus rhizomes, have been isolated and quantified in the extracts. Alkaloids have not been detected, and the presence of flavonoids and sterols is moderate.

Sucrose metabolism in developing oil-rich tubers of Cyperus esculentus: comparative transcriptome analysis.

BACKGROUND: Cyperus esculentus is unique in that it can accumulate significant amounts of oil, starch and sugar as major storage reserves in tubers with high tuber yield and therefore considered as a novel model to study carbon allocation into different storage reserves in underground sink tissues such as tubers and roots. Sucrose (Suc) plays a central role in control of carbon flux toward biosynthesis of different storage reserves; however, it remains unclear for the molecular mechanism underlying Suc metabolism in underground oil-rich storage tissues. In the present study, a comprehensive transcriptome analysis of C. esculentus oil tuber compared to other plant oil- or carbohydrate-rich storage tissues was made for the expression patterns of genes related to the Suc metabolism. RESULTS: The results revealed some species-specific features of gene transcripts in oil tuber of C. esculentus, indicating that: (i) the expressions of genes responsible for Suc metabolism are developmentally regulated and displayed a pattern dissimilar to other plant storage tissues; (ii) both of Suc breakdown and biosynthesis processes might be the major pathways associated with Suc metabolism; (iii) it was probably that Suc degradation could be primarily through the action of Suc synthase (SUS) other than invertase (INV) during tuber development. The orthologs of SUS1, SUS3 and SUS4 are the main SUS isoforms catalyzing Suc breakdown while the vacuolar INV (VIN) is the leading determinant controlling sugar composition; (iv) cytosolic hexose phosphorylation possibly relies more on fructose as substrate and uridine diphosphate glucose pyrophosphorylase (UGP) plays an important role in this pathway; (v) it is Suc-phosphate synthase (SPS) B- and C-family members rather than SPS A that are the principal contributors to SPS enzymes and play crucial roles in Suc biosynthesis pathway. CONCLUSIONS: We have successfully identified the Suc metabolic pathways in C. esculentus tubers, highlighting several conserved and distinct expressions that might contribute to sugar accumulation in this unique underground storage tissue. The specific and differential expression genes revealed in this study might indicate the special molecular mechanism and transcriptional regulation of Suc metabolism occurred in oil tubers of C. esculentus.

Untargeted metabolomics of fresh and heat treatment Tiger nut (Cyperus esculentus L.) milks reveals further insight into food quality and nutrition.

Tiger nut (Cyperus esculentus L.) is a crop traditionally grown in Valencia Region (Spain) and other temperate and tropical regions in the world, where its tubers are commonly consumed as tiger nut milk (horchata). Because of their nutritive potential and original taste, these products are beginning to spread internationally and, as consequence, analytical procedures to assess nutritional profiles, quality control issues are acquiring increasing relevance. The main objective of this study was to use an advance analytical method and chemometrics tools to determine if the ultra-high temperature (UHT) treatment necessary to extend the shelf life of tiger nut milk would affect the profile of nutrients when compared to fresh product. A cold solvent extraction followed by liquid chromatography coupled with high resolution mass spectrometry (UHPLC-HRMS) was used. Datasets obtained from UHT and fresh tiger nut milk data were analyzed through an untargeted metabolomics approach to compare chemical patterns, highlighting differences in citric acid esters of mono- diglycerides (CITREM) and monoacylglycerol (MAG) used as emulsifiers of UHT products, and a remarkably higher abundance of biotin, phosphatidic acid (PA) and L-arginine in fresh products. These results showed that untargeted metabolomics through high resolution tandem mass spectrometry allowed fine differences between food products to be found, therefore, the nutrient lost caused by UHT treatment was clearly discerned.

Effectiveness of Vegetated Drainage Ditches for Domestic Sewage Effluent Mitigation.

Plant species have an important role in eco-ditches; however, the Michaelis-Menten kinetic parameters of nutrient uptake, growth rate and purification efficiency of ditch plants and their influences on domestic sewage treatment efficiency are still unclear. Growth rates of all nine species, but especially Lemna gibba, Cladophora and Myriophyllum verticillatum were best in undiluted domestic sewage as opposed to a mixture of domestic sewage. Performance of species to accumulate nutrients was not only species-specific, but was also affected by both sewage treatments. Removal efficiency of nutrients was dependent on both plant species and treatment. Uptake kinetic parameters were significantly affected by both nutrient form and plant species. The maximum uptake rate (Vmax) of NH4-N was higher than NO3-N. Similarly, Km values for NH4-N were greater than NO3-N. These results could be used to identify plants for sewage treatment efficiency and enhance water quality in eco-ditch treatment systems.

Oil Biosynthesis in Underground Oil-Rich Storage Vegetative Tissue: Comparison of Cyperus esculentus Tuber with Oil Seeds and Fruits.

Cyperus esculentus is unique in that it can accumulate rich oil in its tubers. However, the underlying mechanism of tuber oil biosynthesis is still unclear. Our transcriptional analyses of the pathways from pyruvate production up to triacylglycerol (TAG) accumulation in tubers revealed many distinct species-specific lipid expression patterns from oil seeds and fruits, indicating that in C. esculentus tuber: (i) carbon flux from sucrose toward plastid pyruvate could be produced mostly through the cytosolic glycolytic pathway; (ii) acetyl-CoA synthetase might be an important contributor to acetyl-CoA formation for plastid fatty acid biosynthesis; (iii) the expression pattern for stearoyl-ACP desaturase was associated with high oleic acid composition; (iv) it was most likely that endoplasmic reticulum (ER)-associated acyl-CoA synthetase played a significant role in the export of fatty acids between the plastid and ER; (v) lipid phosphate phosphatase (LPP)-δ was most probably related to the formation of the diacylglycerol (DAG) pool in the Kennedy pathway; and (vi) diacylglyceroltransacylase 2 (DGAT2) and phospholipid:diacylglycerolacyltransferase 1 (PDAT1) might play crucial roles in tuber oil biosynthesis. In contrast to oil-rich fruits, there existed many oleosins, caleosins and steroleosins with very high transcripts in tubers. Surprisingly, only a single ortholog of WRINKLED1 (WRI1)-like transcription factor was identified and it was poorly expressed during tuber development. Our study not only provides insights into lipid metabolism in tuber tissues, but also broadens our understanding of TAG synthesis in oil plants. Such knowledge is of significance in exploiting this oil-rich species and manipulating other non-seed tissues to enhance storage oil production.

Natural attenuation of weathered oil using aquatic plants in a farm in Southeast Mexico.

An experiment was conducted in field for three years to assess the sustainability of aquatic plants Leersia hexandra, Cyperus articulatus, and Eleocharis palustris for use in the removal of total hydrocarbons of weathered oil in four areas contaminated with 60916-119373 mg/kg of hydrocarbons. The variables evaluated were coverage of plant, dry matter, density of plant growth-promoting rhizobacteria, and the removal of total weathered oil. The variables showed statistical differences (p = 0.05) due to the effects of time and the amount of oil in the soil. The three aquatic plants survived on the farm during the 36-month evaluation. The grass L. hexandra yielded the greatest coverage of plant but was inhibited by the toxicity of the oil, which, in contrast, stimulated the coverage of C. articulatus. The rhizosphere of L. hexandra in control soil was more densely colonized by N-fixing bacteria, while the density of phosphate and potassium solubilizing rhizobacteria was stimulated by exposure to oil. C. articulatus coverage showed positive relationship with the removal of weathered oil; positive effect between rhizosphere and L. hexandra grass coverage was also identified. These results contributed to the removal of weathered oil in Gleysols flooded and affected by chronic discharges of crude oil.

Purification of landscape water by using an innovative application of subsurface flow constructed wetland.

This study attempted to purify eutrophic landscape water under a low pollutant concentration and high hydraulic volume loading using an embedded subsurface flow (SSF) constructed wetland (CW). Three species of aquatic plants (i.e., Cyperus alternifolius subsp. flabelliformis, Canna indica, and Hydrocotyle verticillata) were found to be conducive to the requirements of purifying the low-polluted water. Field results of nearly 2 years of experiments showed that SSF CW purified the eutrophic water and maintained the landscape water in a visibly clear condition. In an environment approaching the SSF CW background concentration, pollutant removal processes were divided into modulation and optimum performance periods. Average concentrations of biochemical oxygen demand (BOD), ammonium-nitrogen (NH4 (+)-N), and total phosphorous (TP) in the optimum performance period were 0.69-1.00, 0.35-1.42, and 0.19-0.23 mg/L, respectively. Almost 500 days of BOD and NH4 (+)-N removals were necessary to perform optimally. A shorter period, 350 days, was required for TP optimum removal. This feature of two stage removals was not found in chlorophyll-a (Chl-a) and suspended solids (SS), whose averages were 11.86-17.98 and 13.30 µg/L, respectively. Filter cleaning and water replacement were unnecessary, while only water recharging was needed to compensate for the water lost by evapotranspiration. The field SSF CW has maintained its performance level for over 7 years.

The role of a hybrid phytosystem in landscape water purification and herbicides removal.

The performance of a hybrid phytosystem in landscape water purification and herbicides removal was investigated. The phytosystem operating in an arboretum is located in the Minhang Campus of Shanghai Jiao Tong University, China. The phytosystem is composed of two purification stages: sedimentation Stage 1 without external air supply; and Stage 2 with an external air supply. Stage 2 is also vegetated with three major kinds of plants, namely Pontederia cordata L., Typha latifolia L. and Cyperus alternifolius L. The system's hydraulic loading rate (HLR) was maintained at 1.632 m/day between December 2013 and November 2014. Sedimentation, filtration and adsorption by filter media, combined microbial processes in the rhizosphere (nitrification-denitrification) and plant uptake of the pollutants were all responsible for water purification in the phytosystem. The biological and physical parameters analyzed were total dissolved nitrogen (TDN), nitrate (NO3-N), nitrite (NO2-N), ammonia (NH3-N), total dissolved phosphorus (TDP), dissolved organic carbon (DOC), turbidity, chlorophyll-a and algal cells number. Highest removal efficiencies for TDN, TDP, turbidity, DOC, chlorophyll-a and algal cells were 56.9%, 73.3%, 92.4%, 29.9%, 94.3% and 91.0%, respectively. When the phytosystem was considered for herbicides removal, removal efficiencies of more than 25% were noted for all the herbicides.

Transcriptional transitions in Nicotiana benthamiana leaves upon induction of oil synthesis by WRINKLED1 homologs from diverse species and tissues.

BACKGROUND: Carbon accumulation and remobilization are essential mechanisms in plants to ensure energy transfer between plant tissues with different functions or metabolic needs and to support new generations. Knowledge about the regulation of carbon allocation into oil (triacylglycerol) in plant storage tissue can be of great economic and environmental importance for developing new high-yielding oil crops. Here, the effect on global gene expression as well as on physiological changes in leaves transiently expressing five homologs of the transcription factor WRINKLED1 (WRI1) originating from diverse species and tissues; Arabidopsis thaliana and potato (Solanum tuberosum) seed embryo, poplar (Populus trichocarpa) stem cambium, oat (Avena sativa) grain endosperm, and nutsedge (Cyperus esculentus) tuber parenchyma, were studied by agroinfiltration in Nicotiana benthamiana. RESULTS: All WRI1 homologs induced oil accumulation when expressed in leaf tissue. Transcriptome sequencing revealed that all homologs induced the same general patterns with a drastic shift in gene expression profiles of leaves from that of a typical source tissue to a source-limited sink-like tissue: Transcripts encoding enzymes for plastid uptake and metabolism of phosphoenolpyruvate, fatty acid and oil biosynthesis were up-regulated, as were also transcripts encoding starch degradation. Transcripts encoding enzymes in photosynthesis and starch synthesis were instead down-regulated. Moreover, transcripts representing fatty acid degradation were up-regulated indicating that fatty acids might be degraded to feed the increased need to channel carbons into fatty acid synthesis creating a futile cycle. RT-qPCR analysis of leaves expressing Arabidopsis WRI1 showed the temporal trends of transcripts selected as 'markers' for key metabolic pathways one to five days after agroinfiltration. Chlorophyll fluorescence measurements of leaves expressing Arabidopsis WRI1 showed a significant decrease in photosynthesis, even though effect on starch content could not be observed. CONCLUSIONS: This data gives for the first time a general view on the transcriptional transitions in leaf tissue upon induction of oil synthesis by WRI1. This yields important information about what effects WRI1 may exert on global gene expression during seed and embryo development. The results suggest why high oil content in leaf tissue cannot be achieved by solely transcriptional activation by WRI1, which can be essential knowledge in the development of new high-yielding oil crops.

Phosphorus removal from domestic wastewater by Nelumbo nucifera Gaertn. and Cyperus alternifolius L.

Domestic wastewater is a source of phosphorus contamination that causes eutrophication when it contaminates aquatic environments. Nelumbo nucifera Gaertn. and Cyperus alternifolius L. were applied for phosphorus removal from domestic wastewater. From the study, phosphorus in domestic wastewater was removed from the initial concentration of 1.038 ± 0.001 mgL(-1) to 0.094 ± 0.001 and 0.048 ± 0.004 mgL(-1) by N. nucifera and C. alternifolius, respectively, within 5 days. In addition, total Kjeldahl nitrogen (TKN) and chemical oxygen demand (COD) also decreased when wetland systems were applied the same as treatment with conventional method (chemical + activated sludge process). However, the plant removed TDS better than the conventional method. During 5 cycles of exposure, the two plants still survived and were healthy. The weight of plants increased after the experiment from 4060 ± 0.05 g to 4820 ± 0.17 g of N. nucifera, and from 4000 ± 0.00 g to 4600 ± 0.14 g of C. alternifolius. Phosphorus content also increased in both plants after the experiment. However, in the wetland system, phosphorus was removed mainly by the soil, followed by the plants, and then microorganisms. The domain group in the microbial community of both wetland systems was Pseudomas sp.

nrDNA ITS sequence based SCAR marker to authenticate Aconitum heterophyllum and Cyperus rotundus in Ayurvedic raw drug source and prepared herbal products.

To authenticate Ayurvedic medicinal plants Ativisha (Aconitum heterophyllum) and Musta (Cyperus rotundus) at the raw drug source and in prepared herbal products, nrDNA ITS sequence based SCAR markers were designed and validated spp.-specific SCAR primers gave amplicon of 415 bp and 134 bp, respectively, in authentic species. The SCAR primers (Cyr-FP and Cyr-RP) could identify tissue sample containing 750 µg to 4.76 mg/100mg of Musta in complex mixtures of DNA extracted from commercial herbal drugs. Ativisha could not be identified through SCAR markers suggesting that authentic species may not been used to prepare herbal drugs despite its being labelled as one of the ingredients in formulations. Analysis of individual tubers of Ativisha and Musta assures the presence of admixtures in raw drug trade of Ativisha, indicates the need to monitor the basic raw material supply and concludes, supplying plant materials through cultivation to manufacturing industries can minimize the risks of adulteration.

Efficiency of constructed wetland vegetated with Cyperus alternifolius applied for municipal wastewater treatment.

The treatment of municipal wastewater from Yazd city (center of Iran) by constructed wetland vegetated with Cyperus alternifolius was assessed. Two identical wetlands with a total working volume of 60 L and 10 cm sandy layer at the bottom were used. First wetland (W1) was control and had no Cyperus alternifolius plant. Second wetland (W2) had 100 Cyperus alternifolius shrubs with 40 cm height. Influent wastewater was provided from Yazd's septic tanks effluents and after a 4-day retention time in wetlands, reactors effluent was sampled for parameters analysis. Results show that chemical oxygen demand (COD), NO3 (-)-N, NH4 (+)-N, and PO4 (-3)-P in W1 were reduced to 72%, 88%, 32%, and 0.8%, and in W2, these parameters were removed in values of 83%, 81%, 47%, and 10%, respectively. In both wetlands, the highest and lowest removal efficiencies were related to COD and phosphorus, respectively. Also, the removed phosphorus can be released to stream when the soil saturated or influent phosphorus decreased and when the plant died. After a 4-day-retention time, the W2 wetland showed a statistically significantly lower COD and NH4 (+)-N in comparison with W2 wetland.