Molecular characterization of CeOLE6, a diverged SH oleosin gene, preferentially expressed in Cyperus esculentus tubers.

MAIN CONCLUSION: CeOLE6, a tuber-specific gene in tigernut, encodes a diverged SH oleosin that functions in oil accumulation via homo and heteromultimerization. Tigernut (Cyperus esculentus L.) is a rare example accumulating high levels of triacylglycerols (TAGs) in underground tubers; however, the mechanism underlying is poorly understood. Given essential roles of oleosins (OLEs) in oil accumulation, in this study, structural and functional analyses were conducted for CeOLE6, an oleosin gene preferentially expressed in tigernut tubers. Phylogenetic analysis revealed that CeOLE6 encodes a diverged oleosin in Clade SH, which also includes CeOLE4 and -5. Further synteny analysis and sequence comparison indicated that CeOLE6 is more likely to be a whole-genome duplication (WGD) repeat of CeOLE4, which underwent rapid evolution and deletion of the typical C-terminal insertion for SHs. Nevertheless, CeOLE6 retains the capacity of oligomerization and oil accumulation, because (i) CeOLE6 could not only interact with itself but also with CeOLE2 and -5, two tuber-dominant members belonging to Clades SL and SH, respectively, and (ii) overexpressing CeOLE6 in tobacco leaves could significantly enhance the TAG content. Though CeWRI1 exhibits a similar expression pattern as CeOLE6 during tuber development, both CeWRI1 and -3 could not activate the CeOLE6 promoter, implying that they are not transcription factors contributing tuber-specific activation of CeOLE6. These findings not only provide insights into CeOLE genes in tuber oil accumulation, but also lay a foundation for further genetic improvement in tigernut and other species.

Comparative transcriptome analysis of Cyperus esculentus and C. rotundus with contrasting oil contents in tubers defines genes and regulatory networks involved in oil accumulation.

Plant vegetative organs present great potential for lipid storage, with tubers of Cyperus esculentus as a unique example. To investigate the genome and transcriptomic features of C. esculentus and related species, we sequenced and assembled the C. esculentus genome at the contig level. Through a comparative study of high-quality transcriptomes across 36 tissues from high-oil and intermediate-oil C. esculentus and low-oil Cyperus rotundus, we identified potential genes and regulatory networks related to tuber oil accumulation. First, we identified tuber-specific genes in two C. esculentus cultivars. Second, genes involved in fatty acid (FA) biosynthesis, triacylglycerol synthesis, and TAG packaging presented increased activity in the later stages of tuber development. Notably, tubers with high oil contents presented higher levels of these genes than those with intermediate oil contents did, whereas tubers with low oil contents presented minimal gene expression. Notably, a large fragment of the FA biosynthesis rate-limiting enzyme-encoding gene BCCP1 was missing from the C. rotundus transcript, which might be responsible for blocking FA biosynthesis in its tubers. WGCNA pinpointed a gene module linked to tuber oil accumulation, with a coexpression network involving the transcription factors WRI1, MYB4, and bHLH68. The ethylene-related genes in this module suggest a role for ethylene signaling in oil accumulation, which is supported by the finding that ethylene (ETH) treatment increases the oil content in C. esculentus tubers. This study identified potential genes and networks associated with tuber oil accumulation in C. esculentus, highlighting the role of specific genes, transcription factors, and ethylene signaling in this process.

Physiological adaptation of Cyperus esculentus L. seedlings to varying concentrations of saline-alkaline stress: Insights from photosynthetic performance.

Soil salinization effects plant photosynthesis in a number of global ecosystems. In this study, photosynthetic and physiological parameters were used to elucidate the impacts of saline-alkaline stress on Cyperus esculentus L. (C. esculentus) seedling photosynthesis. The results demonstrate that salt stress, alkali stress and mixed salt and alkali stress treatments all have similar bell-shaped influences on photosynthesis. At low concentrations (0-100 mmol L-1), saline-alkaline stress promoted net photosynthetic rate, transpiration rate and water use efficiency in C. esculentus. However, as the treatments increased in intensity (100-200 mmol L-1), plant photosynthetic parameters began to decline. We interpreted this as the capacity of C. esculentus to improve osmoregulatory capacity in low saline-alkaline stress treatments by accumulating photosynthetic pigment, proline and malondialdehyde to counterbalance the induced stress - an adaptive mechanism that failed once concentrations reached a critical threshold (100 mmol L-1). Stomatal conductance, maximum photosynthetic rate and actual photosynthetic rate all decreased with increasing concentration of the stress treatments, and intercellular carbon dioxide showed a decreasing and then increasing trend. These results indicated that when the saline-alkaline stress concentrations were low, C. esculentus seedlings showed obvious adaptive ability, but when the concentration increased further, the physiological processes of C. esculentus seedlings were significantly affected, with an obvious decrease in photosynthetic efficiency. This study provides a new understanding of the photosynthetic adaptation strategies of C. esculentus seedlings to varying concentrations of saline-alkaline stress.

Genome-wide identification and molecular evolution of Dof transcription factors in Cyperus esculentus.

Dof transcription factor family in Cyperus esculentus genome was identified and analyzed using bioinformatics. The analysis results revealed that C.esculentus genome contains 29 Dof genes (CesDof), all of which are located in the nucleus according to subcellular localization prediction. CesDof proteinrs have a range of 124 to 512 amino acids, with most being basic proteins. Their secondary structure was mainly irregular curl. The promoter sequence of CesDof genes contains cis-acting elements that respond to light, drought, hormones, low temperature, and circadian rhythm. Codon preference analysis showed that CesDof genes' codon preference ends in T/A. Collinearity analysis revealed that C.esculentus had three pairs of collinear CesDof genes. Additionally, there were 15 pairs of collinear genes between C.esculentus and Arabidopsis thaliana. The genetic relationship between C.esculentus and Rhynchospora pubera was found to be the closest. Phylogenetic tree analysis revealed that 29 CesDof genes of C.esculentus can be classified into 4 subgroups. Additionally, 144 miRNAs were predicted to target these CesDof genes. Furthermore, protein interaction analysis indicated that 15 Dof proteins in C.esculentus had interactions. The qRT-PCR verification results of drought stress and salt stress treatment experiments showed that most CesDof genes were involved in drought stress and salt stress responses, and the gene expression trends under drought stress and salt stress conditions were consistent. These results lay a theoretical foundation for further studying the molecular functions of Dof gene family in C.esculentus and its molecular mechanisms in regulating the life activities of C.esculentus.

Enhancing biodegradation of aged hydrocarbon-contaminated soils through toluene addition: assessing effects on solid and slurry phase treatments.

The main challenge in treating aged soils highly contaminated with total petroleum hydrocarbons (TPH) is to enhance their bioavailability for microbial degradation. Hydrocarbons in soils undergo chemical changes that make them more resistant to biodegradation. This study investigates toluene's efficacy in enhancing the biodegradation of aged hydrocarbon-contaminated soil containing 292,000 mg TPH kg-1 dry soil. Toluene's effect was compared between solid phase (SOP) and slurry phase (SLP) treatments using a microbial consortium isolated from Cyperus laxus rhizosphere. TPH biodegradation and microbial respiration were measured, the latter to estimate the respiratory quotient (RQ, the ratio between moles of carbon dioxide released and moles of oxygen absorbed during respiration). Toluene significantly accelerated TPH biodegradation in both treatments, achieving ~ 30% higher removal than in a non-solvent control, possibly through improved bioavailability of aromatic compounds and other low molecular weight compounds. According to the RQ analysis, toluene enhanced microbial respiratory processes and hydrocarbon catabolism with higher hydrocarbon mineralization (RQ = ~ 0.5) in both SOP and SLP assays. Our results reveal toluene's potential to increase hydrocarbon availability and microbial degradation efficiency in aged contaminated soils; its use in various bioremediation techniques could be of broad applicability across diverse soil types and pollutants.

RNA-Seq Reveals That Multiple Pathways Are Involved in Tuber Expansion in Tiger Nuts (Cyperus esculentus L.).

The tiger nut (Cyperus esculentus L.) is a usable tuber and edible oil plant. The size of the tubers is a key trait that determines the yield and the mechanical harvesting of tiger nut tubers. However, little is known about the anatomical and molecular mechanisms of tuber expansion in tiger nut plants. This study conducted anatomical and comprehensive transcriptomics analyses of tiger nut tubers at the following days after sowing: 40 d (S1); 50 d (S2); 60 d (S3); 70 d (S4); 90 d (S5); and 110 d (S6). The results showed that, at the initiation stage of a tiger nut tuber (S1), the primary thickening meristem (PTM) surrounded the periphery of the stele and was initially responsible for the proliferation of parenchyma cells of the cortex (before S1) and then the stele (S2-S3). The increase in cell size of the parenchyma cells occurred mainly from S1 to S3 in the cortex and from S3 to S4 in the stele. A total of 12,472 differentially expressed genes (DEGs) were expressed to a greater extent in the S1-S3 phase than in S4-S6 phase. DEGs related to tuber expansion were involved in cell wall modification, vesicle transport, cell membrane components, cell division, the regulation of plant hormone levels, signal transduction, and metabolism. DEGs involved in the biosynthesis and the signaling of indole-3-acetic acid (IAA) and jasmonic acid (JA) were expressed highly in S1-S3. The endogenous changes in IAA and JAs during tuber development showed that the highest concentrations were found at S1 and S1-S3, respectively. In addition, several DEGs were related to brassinosteroid (BR) signaling and the G-protein, MAPK, and ubiquitin-proteasome pathways, suggesting that these signaling pathways have roles in the tuber expansion of tiger nut. Finally, we come to the conclusion that the cortex development preceding stele development in tiger nut tubers. The auxin signaling pathway promotes the division of cortical cells, while the jasmonic acid pathway, brassinosteroid signaling, G-protein pathway, MAPK pathway, and ubiquitin protein pathway regulate cell division and the expansion of the tuber cortex and stele. This finding will facilitate searches for genes that influence tuber expansion and the regulatory networks in developing tubers.

Greenhouse gases emissions and carbon budget estimation in horizontal subsurface flow constructed wetlands with different plant species.

Constructed wetlands (CWs) are pivotal for wastewater treatment due to their high efficiency and numerous advantages. The impact of plant species and diversity on greenhouse gas (GHG) emissions from CWs requires a more comprehensive evaluation. Moreover, controversial perspectives persist about whether CWs function as carbon sinks or sources. In this study, horizontal subsurface flow (HSSF) CWs vegetated with Cyperus alternifolius, Typhae latifolia, Acorus calamus, and the mixture of these three species were constructed to evaluate pollutant removal efficiencies and GHG emissions, and estimate carbon budgets. Polyculture CWs can stably remove COD (86.79 %), NH4+-N (97.41 %), NO3--N (98.55 %), and TP (98.48 %). They also mitigated global warming potential (GWP) by suppressing N2O emissions compared with monoculture CWs. The highest abundance of the Pseudogulbenkiania genus, crucial for denitrification, was observed in polyculture CWs, indicating that denitrification dominated in nitrogen removal. While the highest nosZ copy numbers were observed in CWs vegetated with Cyperus alternifolius, suggesting its facilitation of denitrification-related microbes. Selecting Cyperus alternifolius to increase species diversity is proposed for simultaneously maintaining the water purification capacity and reducing GHG emissions. Carbon budget estimations revealed that all four types of HSSF CWs were carbon sinks after six months of operation, with carbon accumulation capacity of 4.90 ± 1.50 (Cyperus alternifolius), 3.31 ± 2.01 (Typhae latifola), 1.78 ± 1.30 (Acorus calamus), and 2.12 ± 0.88 (polyculture) kg C/m2/yr. This study implies that under these operation conditions, CWs function as carbon sinks rather than sources, aligning with carbon peak and neutrality objectives and presenting significant potential for carbon reduction efforts.

Assessment of sulfamethoxazole removal by three wetland plant species under hydroponic conditions: uptake, accumulation, and physiological responses.

Plants play a crucial role as a removal pathway in constructed wetlands, demonstrating the ability to absorb and tolerate antibiotics from wastewater. However, the specific contribution of plants in this regard has not yet to be sufficiently established. To gain a more comprehensive insight into the associated processes, we selected three common wetland plant species, Canna indica L. (C. indica), Cyperus alternifolius L. (C. alternifolius), and Thalia dealbata Fraser (T. dealbata), to evaluate their capacity for uptake, accumulation, and physiological response in the removal of sulfamethoxazole (SMX) at varying initial concentrations (10, 30, 100, and 300 µg/L) under hydroponic conditions. The results showed that SMX removal was more efficient at lower concentrations (10 and 30 µg/L) than at higher concentrations (100 and 300 µg/L). Moreover, plant systems were found to consistently outperform unplanted systems in SMX removal. Among the assessed species, C. indica was identified as being relatively effective in the removal of SMX, whereas the performance of C. alternifolius was notably less pronounced. A positive correlation was observed between the concentration of SMX in the plant tissues and that in the external aqueous medium. However, plant tissue residues contributed only a minor fraction to the overall removal of SMX. Wetland plants absorb SMX through their roots, and we accordingly detected significantly higher concentrations in submerged plant tissues. Furthermore, we also detected reductions in net photosynthetic rates indicative of potential phytotoxicity, which is associated with the accumulation of antibiotic in the shoot tissues. Accumulation of SMX in the roots and rhizomes was also found to be associated with the development of shorter roots, with this effect becoming more pronounced with an increase in the concentration of exogenous SMX. However, despite these adverse effects, plants can detoxify antibiotics via the glutathione pathway. Of the assessed plant species, C. indica was identified as the most SMX tolerant, as indicated by Km and Vmax values, with C. alternifolius being the least tolerant. Our findings in this study reveal the potential value of wetland plants in the sequestration of antibiotics and provide evidence for the underlying mechanisms of action. These findings could make an important contribution to the implementation of phytoremediation in antibiotic-contaminated water.

Three wetland plants with fibrous root systems, namely Canna indica, Cyperus alterniflius, and Thalia dealbata, were selected to investigate the removal efficiencies of sulfamethoxazole in the hydroponic system by different emergent plants, quantify the contribution of uptake and accumulation for sulfamethoxazole in plant tissues, and assess the physiological responses of plants and their effect on the removal of sulfamethoxazole. The knowledge obtained from this study shows the potential use of wetland plants for removing antibiotics and the inherent mechanisms, which will be useful for the application of phytoremediation in antibiotic contaminated water.

Impact of nanoplastics on the biodegradation, ecotoxicity, and key genes involved in imidacloprid metabolic pathways in papyrus (Cyperus papyrus L.).

Both nanoplastics (NPs) and imidacloprid (IMI) are widely distributed in the environment and have attracted significant attention due to their adverse effects on ecosystems. Constructed wetlands have the potential to remove IMI, but there is still limited understanding of how wetland plants interact with IMI, especially when influenced by different charged NPs. This study assessed their ecotoxicological effects, as well as the fate and transformation of IMI in papyrus (Cyperus papyrus L.) under the influence of different charged NPs and identified key driving genes in the plant. Results show that simultaneous exposure to positively charged PS-NH2 and IMI inhibited plant growth. The combined action of NPs and IMI intensified their toxicity, enhancing lipid peroxidation and altering antioxidant enzyme activities. The IMI removal efficiency, which was primarily driven by biodegradation, was 80.61%, 88.91%, and 74.71% in the IMI-alone, co-IMI/PS_COOH, and co-IMI/PS_NH2 systems, respectively. PS-NH2 restricted the roots-to-shoots translocation ability of IMI. PS-COOH enhanced IMI oxidation and nitro reduction, while PS-NH2 inhibited 2-OH-IMI dehydrogenation to IMI-olefin in papyrus. Transcriptomics and gene network analysis identified the genes encoding CYP450 enzymes, reductases, hydrolases, dehydrogenases, and peroxidases as those influencing IMI biodegradation. These enzymes play a crucial role in the hydroxylation, dehydrogenation, reduction, and oxidation processes during biodegradation of IMI in the presence of NPs. This study expands the understanding of the impact of differently charged NPs on the IMI remediation efficacy of papyrus, thus providing new insights into the phytoremediation of organic contaminants in constructed wetlands.

CePP2C19 confers tolerance to drought by regulating the ABA sensitivity in Cyperus esculentus.

BACKGROUND: Tiger nut (Cyperus esculentus) is widely known as an additional source of food, oil and feed worldwide. The agricultural production of tiger nut has been greatly hindered by drought stress, reducing both yield and quality. Protein phosphatase 2 C (PP2Cs) plays an important role in plant responses to drought stress however, the molecular mechanism of PP2Cs in tiger nuts still unclear. RESULTS: In this study, we identified a putative group A PP2C-encoding gene (CePP2C19) from tiger nut using transcriptome analysis, which is highly induced by drought stress. The transient expression assay suggested that CePP2C19 was localized to nucleus. Furthermore, the interaction between CePP2C19 and CePYR1, a coreceptor for ABA signaling, was first detected using a yeast two-hybrid assay and then verified using a bimolecular fluorescence complementation (BiFC) analysis. In addition, the transgenic Arabidopsis lines overexpressing CePP2C19 exhibited extreme tolerance to ABA and mannitol stresses during seed germination and root growth. At the mature stage, overexpression of CePP2C19 resulted in a higher tolerance to drought stress in transgenic Arabidopsis, as confirmed by a visible phenotype and several physiological parameters. Noticeably, the silencing of CePP2C19 by virus-induced gene silencing (VIGS) showed obvious reduction in drought tolerance in tiger nut plants. CONCLUSIONS: The CePP2C19 emerges as a pivotal gene involved in the ABA signaling pathway, which likely reduce ABA sensitivity and thus enhances drought tolerance in Cyperus esculentus.

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.

Target and nontarget mechanisms of AHAS inhibitor cross-resistance patterns in Cyperus difformis.

Weed resistance to acetohydroxyacid synthase (AHAS) inhibiting herbicides has been a critical issue for rice growers worldwide since the early 1990's. In California, resistance to bensulfuron-methyl was first detected in Cyperus difformis in 1993. Since then, populations of most major weeds of rice in California have been reported to show resistance to at least one AHAS inhibitor. We sought to describe the magnitude and mechanisms of AHAS inhibitor cross-resistance in California populations of C. difformis. Sixty-two populations were collected and screened for cross-resistance to bensulfuron-methyl (BEN), halosulfuron-methyl (HAL), bispyribac­sodium (BIS), and penoxsulam (PEN), revealing six major patterns of cross-resistance. Representative C. difformis populations from each cross-resistance pattern were then subjected to dose-response, cytochrome P450 inhibition, AHAS gene sequencing, and metabolic studies with the same herbicides as in the screening. Dose-response confirmed the detected resistances in the representative populations, and suggested that the majority of observed resistance was dose-dependent. Cytochrome P450 inhibition via malathion revealed evidence of increased metabolic activity in resistant populations to BEN, BIS, and PEN. AHAS gene sequencing revealed amino acid substitutions in five of six populations: R3 (Pro197-Ser), R4 (Pro97-His), R10 (Asp376), R41 (Ala122-Asn), and R18 (Trp574-Leu). Metabolic studies confirmed evidence of increased activity of cytochrome P450s in all populations. Metabolic BEN and HAL analysis did not yield similar results to malathion inhibition, suggesting different P450's or other pathways. Taken together, the results of the studies confirm the complexity of AHAS inhibitor cross-resistance in C. difformis, and the presence of both target-site and metabolic resistance in most of the representative populations underscores the importance of proper herbicide selection, rotation, and scouting in fields.

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.

Chromosome-scale Genome Assembly of the Yellow Nutsedge (Cyperus esculentus).

The yellow nutsedge (Cyperus esculentus L. 1753) is an unconventional oil plant with oil-rich tubers, and a potential alternative for traditional oil crops. Here, we reported the first high-quality and chromosome-level genome assembly of the yellow nutsedge generated by combining PacBio HiFi long reads, Novaseq short reads, and Hi-C data. The final genome size is 225.6 Mb with an N50 of 4.3 Mb. More than 222.9 Mb scaffolds were anchored to 54 pseudochromosomes with a BUSCO score of 96.0%. We identified 76.5 Mb (33.9%) repetitive sequences across the genome. A total of 23,613 protein-coding genes were predicted in this genome, of which 22,847 (96.8%) were functionally annotated. A whole-genome duplication event was found after the divergence of Carex littledalei and Rhynchospora breviuscula, indicating the rich genetic resources of this species for adaptive evolution. Several significantly enriched GO terms were related to invasiveness of the yellow nutsedge, which may explain its plastic adaptability. In addition, several enriched Kyoto Encyclopedia of Genes and Genomes pathways and expanded gene families were closely related with substances in tubers, partially explaining the genomic basis of characteristics of this oil-rich tuber.

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.

A seed-like proteome in oil-rich tubers.

There are numerous examples of plant organs or developmental stages that are desiccation-tolerant and can withstand extended periods of severe water loss. One prime example are seeds and pollen of many spermatophytes. However, in some plants, also vegetative organs can be desiccation-tolerant. One example are the tubers of yellow nutsedge (Cyperus esculentus), which also store large amounts of lipids similar to seeds. Interestingly, the closest known relative, purple nutsedge (Cyperus rotundus), generates tubers that do not accumulate oil and are not desiccation-tolerant. We generated nanoLC-MS/MS-based proteomes of yellow nutsedge in five replicates of four stages of tuber development and compared them to the proteomes of roots and leaves, yielding 2257 distinct protein groups. Our data reveal a striking upregulation of hallmark proteins of seeds in the tubers. A deeper comparison to the tuber proteome of the close relative purple nutsedge (C. rotundus) and a previously published proteome of Arabidopsis seeds and seedlings indicates that indeed a seed-like proteome was found in yellow but not purple nutsedge. This was further supported by an analysis of the proteome of a lipid droplet-enriched fraction of yellow nutsedge, which also displayed seed-like characteristics. One reason for the differences between the two nutsedge species might be the expression of certain transcription factors homologous to ABSCISIC ACID INSENSITIVE3, WRINKLED1, and LEAFY COTYLEDON1 that drive gene expression in Arabidopsis seed embryos.

Characterisation of two novel genes encoding Δ9 fatty acid desaturases (CeSADs) for oleic acid accumulation in the oil-rich tuber of Cyperus esculentus.

Cyperus esculentus is considered one of the most promising oil crops due to its oil-rich tuber, wide adaptability and large biomass production. Preferable triacylglycerol (TAG) composition, especially high oleic acid content, makes tuber oil suitable for human consumption and biodiesel production. However, the mechanism underlying oleic acid enrichment in the tuber remains unknown. Plastidial stearoyl-ACP desaturase (SAD) catalyses the formation of monounsaturated fatty acids (MUFAs), which may function crucially for high accumulation of oleic acid in C. esculentus tubers. In this study, two full-length cDNAs encoding SAD were isolated from the developing tubers of C. esculentus, namely, CeSAD1 and CeSAD2, with ORFs of 1194 bp and 1161 bp, respectively. Quantitative RT-PCR analysis showed that CeSAD genes were highly expressed in tubers. The expression pattern during tuber formation was also significantly correlated with fatty acid and oil accumulation dynamics. Overexpression of each CeSAD gene could restore the normal growth of the defective yeast BY4389, indicating that both CeSADs had fatty acid desaturase activity to catalyse MUFA biosynthesis. A tobacco genetic transformation assay demonstrated that both CeSAD enzymes had high enzyme activity. Exogenous addition of exogenous fatty acids to feed yeast revealed that CeSAD1 has a more substantial substrate preference ratio for C18:0 than CeSAD2 did. Moreover, the overexpression of CeSAD1 significantly increased host tolerance against low-temperature stress. Our data add new insights into the deep elucidation of oleic acid-enriched oils in Cyperus esculentus tubers, showing CeSAD, especially CeSAD1, as the target gene in genetic modification to increase oil and oleic yields in oil crops as well as stress tolerance.

Flavonoids naringenin chalcone, naringenin, dihydrotricin, and tricin are lignin monomers in papyrus.

Recent studies demonstrate that several polyphenolic compounds produced from beyond the canonical monolignol biosynthetic pathways can behave as lignin monomers, participating in radical coupling reactions and being incorporated into lignin polymers. Here, we show various classes of flavonoids, the chalconoid naringenin chalcone, the flavanones naringenin and dihydrotricin, and the flavone tricin, incorporated into the lignin polymer of papyrus (Cyperus papyrus L.) rind. These flavonoids were released from the rind lignin by Derivatization Followed by Reductive Cleavage (DFRC), a chemical degradative method that cleaves the ß-ether linkages, indicating that at least a fraction of each was integrated into the lignin as ß-ether-linked structures. Due to the particular structure of tricin and dihydrotricin, whose C-3' and C-5' positions at their B-rings are occupied by methoxy groups, these compounds can only be incorporated into the lignin through 4'-O-ß bonds. However, naringenin chalcone and naringenin have no substituents at these positions and can therefore form additional carbon-carbon linkages, including 3'- or 5'-ß linkages that form phenylcoumaran structures not susceptible to cleavage by DFRC. Furthermore, Nuclear Magnetic Resonance analysis indicated that naringenin chalcone can also form additional linkages through its conjugated double bond. The discovery expands the range of flavonoids incorporated into natural lignins, further broadens the traditional definition of lignin, and enhances the premise that any phenolic compound present at the cell wall during lignification could be oxidized and potentially integrated into the lignin structure, depending only on its chemical compatibility. This study indicates that papyrus lignin has a unique structure, as it is the only lignin known to date that integrates such a diversity of phenolic compounds from different classes of flavonoids. This discovery will open up new ways to engineer and design lignins with specific properties and for enhanced value.

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.

Chemical composition, energy content, and amino acid digestibility in Cyperus esculentus co-products fed to growing pigs.

This study was conducted to determine the chemical composition, DE, ME, and NE contents, and the apparent and standardized ileal digestibility (AID and SID) of AA in Cyperus esculentus co-products fed to growing pigs. The five C. esculentus co-products included expeller-pressed C. esculentus cake (EPCC), cold-pressed C. esculentus cake (CPCC), solvent-extracted C. esculentus meal (SECM), C. esculentus distillers's dried grains with solubles (CE DDGS), and C. esculentus meal (CEM). In Exp. 1, a total of 36 crossbred growing pigs (Duroc × Landrace × Yorkshire; BW: 50.12 ± 2.91 kg) were fed one of six diets in a completely randomized design. The diets included a corn-soybean meal basal diet and five experimental diets containing 24.31% C. esculentus co-products. In Exp. 2, 12 same breed of growing pigs (BW: 47.12 ± 3.2 kg), surgically fitted with a T-cannula in the distal ileum, were allotted to one of four experimental diets in a 2-period Youden Square design. The diets included one N-free diet and three experimental diets containing 50% C. esculentus co-products (including EPCC, SECM, and CE DDGS). Results indicated that the SECM and CE DDGS had the greatest contents of starch and CP, respectively. The contents of CF, NDF, and ADF were the greatest in CEM and the lowest in SECM. On a DM basis, the DE, ME, predicted NE, and apparent total tract digestibility (ATTD) of GE values of the 5 C. esculentus co-products ranged from 1,203 to 3,897 kcal/kg, 1,127 to 3,621 kcal/kg, 536 to 2,871 kcal/kg, and 28% to 79%, respectively. The EPCC and CPCC had the greatest DE, ME, and predicted NE values, and CPCC, EPCC, and SECM had the greatest ATTD of GE, whereas CEM had the lowest DE, ME, NE, and ATTD of GE (P < 0.001). The NDF and ADF were negatively correlated with DE, ME, and NE (P < 0.05). The AID and SID of CP varied from 53.57 % to 57.86% and from 69.99% to 87.85%, respectively. The EPCC and SECM had greater SID of CP, Ile, Met, Val, Asp, Cys, and Tyr compared to those of CE DDGS (P < 0.05). These results indicated that the chemical composition, DE, ME, and NE as well as the most AA digestibility of C. esculentus co-products obtained from different processing techniques varied greatly. Based on the energy contents and AA digestibility, the EPCC is a better feedstuff for growing pigs compared with the other 4 C. esculentus co-products.