Volatolomics to Decrypt the Monophagous Nature of a Rice Pest, Scirpophaga Incertulas (Walker).

Scirpophaga incertulas Walker (Lepidoptera: Crambidae, yellow stem borer, YSB) is a monophagous insect pest that causes significant yield loss in rice (Oryza staiva L.). Semiochemical based pest management is being sought as an alternate to chemical pesticides to reduce pesticide footprints. We hypothesized differential release of volatiles from host rice and two companion non-host weeds, Echinochloa colona and Echinochloa crus-galli could be responsible for oviposition and biology of YSB and these chemicals could be used for YSB management. Number of eggs laid, and number of larvae hatched were significantly higher in rice plant as compared to weeds. YSB could only form dead hearts in rice plants. YSB significantly preferred host-plant volatiles compared to the non-host plants both in choice and no-choice tests in an Y-tube olfactometer. 2-Hexenal, hexanal, 2,4-hexadienal, benzaldehyde, nonanal, methyl salicylate and decanal were found in the leaf volatolomes of both the host and non-host plants in HS-SPME-GC-MS (Headspace-Solid phase micro extraction-Gas chromatography-Mass spectrometer). Pentene-3-one, 2-pentyl furan, 2,4-heptadienal, 2-octenal, 2-octenol and menthol were present only in the non-host plants. Fourteen rice unique compounds were also detected. The built-in PCA (Principal Component Analysis) and PLS-DA (Partial least squares-discriminant analysis) analysis in the MS-DIAL tool showed that the volatiles emitted from TN1 formed a cluster distinct from Echinochloa spp. and 2-octenal was identified as a unique compound. Olfactometer bioassays using synthetic compounds showed that rice unique compounds, like xylene, hexanal served as attractants whereas non-host unique compounds, like 2-pentylfuran, 2-octenal acted as repellent. The results indicate that the rice unique compounds xylene, hexanal along with other volatile compounds could be responsible for higher preference of YSB towards rice plants. Similarly, the non-host unique compounds 2-pentylfuran, 2-octenal could possibly be responsible for lower preference and defence against YSB. These compounds could be utilised for devising traps for YSB monitoring and management.

Barnyard grass [Echinochloa crus-galli (L.) Beauv] leaves extract against tomato pests.

BACKGROUND: Tomato is one of the widely cultivated crops worldwide that is affected by several pests, such as fungi (Fusarium oxysoporum, Alternaria solani), bacteria (Pectobacterium carotovorum) and weeds (Cyperus iria L., Amaranthus spinosus). A growing interest has emerged for developing plant-derived pesticidal compounds to counteract these pests. One attractive alternative is to use barnyard grass (Echinochloa crus-galli), known to be widely resistant to synthetic herbicides, as a potential biopesticide compound source. RESULTS: Phytochemical screening of the crude extract showed that phenolic compounds were the most abundant component present in barnyard grass. The crude extract was evaluated for antifungal, antibacterial and herbicidal activities. Bioassays showed inhibition against F. oxysporum (10.73 ± 1.30%) and A. solani (20.47 ± 3.51%), the causative agent of Fusarium rot and early blight disease in tomato, respectively. Antibacterial activity against P. carotovorum gave a mean zone of inhibition (paper disc diffusion assay) of 17.00 ± 1.00 mm and an IC50 (dose-response assay) of 2.26 mg mL-1 was observed. Dose-responsive herbicidal activity on the lettuce seed germination bioassay produced an IC50 of 459.30 ppm. Selectivity studies showed inhibition towards C. iria and A. spinosus with no effect on tomato. Lastly, bioassay-guided fractionation coupled with untargeted metabolomics studies using ultra-performance liquid chromatography with diode array detection-tandem mass spectrometry mass analyses revealed loliolide and tricin as the putative metabolites present in barnyard grass. CONCLUSION: To date, this is the first reported study on using barnyard grass as a potential alternative biopesticide against tomato pests such as fungi, bacteria and weeds. © 2021 Society of Chemical Industry.

Vanillin extracted from Proso and Barnyard millets induce apoptotic cell death in HT-29 human colon cancer cell line.

In the present study, we hypothesized that the active compound extracted from Proso and Barnyard millets inhibits cell proliferation and apoptosis induction in colon cancer cell line. The bioactive compounds from these millets were purified by supercritical fluid extraction and their structure was elucidated using spectroscopic methods. Extracted bioactive components from these millets were similar in chemical structure to the phenolic aldehyde-Vanillin [4-Hydroxy-3-methoxybenzaldehyde]. Cell proliferative effect was assessed by MTT assay using HT-29 cell line. Compound 1 significantly inhibited the proliferation of HT-29 cells when treated with concentrations of 250 µg/ml and 1,000 µg/ml for 48 h, while compound 2 moderately inhibited the proliferation of the HT-29 cell line at the same concentration and time period. Cytotoxic activity of extracted compounds by the release of lactate dehydrogenase confirms that these compounds were not toxic to the cells at 250 µg/ml of compounds 1 and 2. In addition, flow cytometry results show a significant cell arrest in the G0/G1 phase and increase in the apoptotic cells in sub G0 phase, in a dose-dependent manner when compared with the control. The conclusion of this study suggests that the anticancer property of these millets is mediated through the presence of vanillin.

Diversity of Harpin-Like Defense Peptides from Barnyard Grass (Echinochloa crusgalli L.) Seeds.

The results of the study of the structure and function of harpin-like peptides (alpha-harpinins) of the EcAMP group from the barnyard grass (E. crusgalli) seeds and the possibility of their involvement in the innate immunity to biotic stresses are presented.

Hepatoprotective studies on methanolic extract of caryopses of Echinochloa colona Link.

The caryopses (seeds) of Echinochloa colona Link of family Poaceae are traditionally used for the treatment of jaundice. The methanolic extract of caryopses of Echinochloa colona (ECME) was evaluated for its hepatoprotective activity in paracetamol (3g/kg per oral) and ethanol (5g/kg per oral) intoxicated rats while its antihepatotoxic activity against D-galactosamine (400mg/kg body weight intra peritoneal). The activity of the extract was assessed on the basis of improvement in the altered level of various serum biochemical parameters and in the changes occurred in the histology of liver of the rats. The extract was also investigated for its antioxidant potential by employing different in vitro methods. The extract exhibited ferrous ion reducing power, 1,1 Diphenyl-1-picryl hydrazyl (DPPH), superoxide, nitric oxide and hydroxyl radical scavenging activities. The significant (p<0.001) hepatoprotective and antioxidant activities exhibited by the extract ECME, in different in vivo models and in vitro studies respectively may be attributed to the flavonoids and phenolic compounds present in the extract.

Characterization of Hydroxyproline-Containing Hairpin-Like Antimicrobial Peptide EcAMP1-Hyp from Barnyard Grass (Echinochloa crusgalli L.) Seeds: Structural Identification and Comparative Analysis of Antifungal Activity.

Herein, we describe a modified form of the antimicrobial hairpin-like peptide EcAMP1, isolated from barnyard grass (E. crusgalli) seeds, which is structurally characterized by a combination of high-pressure liquid chromatography, mass spectrometry, and automated Edman sequencing. This derivate has a single amino acid substitution (Pro19Hyp) in the second α-helical region of the molecule, which is critical for the formation of the hydrophobic core and the secondary structure elements. Comparing the antifungal activity of these two peptides, we found that the modified EcAMP1-Hyp had a significantly weaker activity towards the most-sensitive plant pathogenic fungus Fusarium solani. Molecular dynamics simulations and in vitro binding to the commercial polysaccharides allowed us to conclude that the Pro-19 residue is important for binding to carbohydrates located in the spore cell wall and it chiefly exhibits a fungistatic action representing the hyphal growth inhibition. These data are novel and significant for understanding a role of α-hairpinins in plant immunity.

The role of grass volatiles on oviposition site selection by Anopheles arabiensis and Anopheles coluzzii.

BACKGROUND: The reproductive success and population dynamics, of Anopheles malaria mosquitoes is strongly influenced by the oviposition site selection of gravid females. Mosquitoes select oviposition sites at different spatial scales, starting with selecting a habitat in which to search. This study utilizes the association of larval abundance in the field with natural breeding habitats, dominated by various types of wild grasses, as a proxy for oviposition site selection by gravid mosquitoes. Moreover, the role of olfactory cues emanating from these habitats in the attraction and oviposition stimulation of females was analysed. METHODS: The density of Anopheles larvae in breeding sites associated with Echinochloa pyramidalis, Echinochloa stagnina, Typha latifolia and Cyperus papyrus, was sampled and the larvae identified to species level. Headspace volatile extracts of the grasses were collected and used to assess behavioural attraction and oviposition stimulation of gravid Anopheles arabiensis and Anopheles coluzzii mosquitoes in wind tunnel and two-choice oviposition assays, respectively. The ability of the mosquitoes to differentiate among the grass volatile extracts was tested in multi-choice tent assays. RESULTS: Anopheles arabiensis larvae were the most abundant species found in the various grass-associated habitats. The larval densities described a hierarchical distribution, with Poaceae (Echinochloa pyramidalis and Echinochloa stagnina)-associated habitat sites demonstrating higher densities than that of Typha-associated sites, and where larvae were absent from Cyperus-associated sites. This hierarchy was maintained by gravid An. arabiensis and An. coluzzii mosquitoes in attraction, oviposition and multi-choice assays to grass volatile extracts. CONCLUSIONS: The demonstrated hierarchical preference of gravid An. coluzzii and An. arabiensis for grass volatiles indicates that vegetation cues associated with larval habitats are instrumental in the oviposition site choice of the malaria mosquitoes. Identifying volatile cues from grasses that modulate gravid malaria mosquito behaviours has distinct potential for the development of tools to be used in future monitoring and control methods.

A new lignan and a new alkaloid, and α-glucosidase inhibitory compounds from the grains of Echinochloa utilis Ohwi & Yabuno.

A new lignan, utilisin (1), and a new alkaloid, echinoutilin (2), together with eleven known compounds 3-13 were isolated from the grains of Echinochloa utilis Ohwi & Yabuno. Their structures were identified through the analysis of spectroscopic data. The absolute configuration of 2 was determined by Mosher's method. These compounds were evaluated for α-glucosidase inhibitory activity. Among them, compounds 2, 3 and 6 exhibited considerable α-glucosidase inhibitory activity with IC50 values of 42.1±1.3, 58.9±3.7, and 40.9±1.1µM, respectively. The results indicate that the grains of E. utilis will be useful in the treatment of diabetes control agents.

Isolation of Functional Components ß-Glucan and γ-Amino Butyric Acid from Raw and Germinated Barnyard Millet (Echinochloa frumentaceae) and their Characterization.

The study was carried out to analyze the characteristics of two functional constituents' viz. γ-amino butyric acid (GABA) and ß-glucan extracted from raw and germination barnyard millet (var. PRJ-1). A significant (P ≤ 0.05) effect of germination (sprouting) was observed in yield, chemical composition, functional, rheological and antioxidant properties of ß-glucan and GABA. The yield of GABA extract was 12.34 % and the content increased from 6.37 mg/100 g in raw to 35.70 mg/100 g in germinated sample. The DPPH, total antioxidant and hydrogen peroxide scavenging activities of GABA extract increased after germination from 45.34 to 65.34 %, 15.3 to 33.3 millimole/g and 38.4 to 64.7 millimole/g, respectively. The yield of ß-glucan extract of raw and germinated flour was 6.05 and 5.01 % whereas the ß-glucan contents were 83.30 and 79.64 %, respectively. The functional properties of ß-glucan i.e., swelling power, water binding capacity and DPPH scavenging activity increased from 1.45 to 1.76 g/g, 2.13 to 2.32 g/g and 44.39 to 57.42 %, respectively, after germination. Similarly there was an increase in the storage modulus after germination process which attributes a better viscoelastic capacity of ß-glucan at low frequencies. The results exploit that the ß-glucan and GABA might promise a polymeric incipient to be implemented as food additives with variable functional and structural characteristics.

Comparison of arsenic uptake ability of barnyard grass and rice species for arsenic phytoremediation.

In this research, the relative performance in arsenic (As) remediation was evaluated among some barnyard grass and rice species under hydroponic conditions. To this end, four barnyard grass varieties and two rice species were selected and tested for their remediation potential of arsenic. The plants were grown for 2 weeks in As-rich solutions up to 10 mg As L(-1) to measure their tolerance to As and their uptake capabilities. Among the varieties of plants tested in all treatment types, BR-29 rice absorbed the highest amount of As in the root, while Nipponbare translocated the maximum amount of As in the shoot. Himetainubie barnyard grass produced the highest biomass, irrespective of the quantity of As in the solution. In all As-treated solutions, the maximum uptake of As was found in BR-29 followed by Choto shama and Himetainubie. In contrast, while the bioaccumulation factor was found to be the highest in Nipponbare followed by BR-29 and Himetainubie. The results suggest that both Choto shama and Himetainubie barnyard grass varieties should exhibit a great potential for As removal, while BR-29 and Nipponbare rice species are the best option for arsenic phytoremediation.

Elicitation of nutritional, antioxidant, and antidiabetic potential of barnyard millet (Echinochloa esculenta (A. Braun) H. Scholz) sprouts and microgreens through in vitro bio-accessibility assessment.

This study has redirected focus towards the untapped potential of millets, exploring their utilization as small-scale vegetables like sprouts and microgreens. This study assessed the metabolite profiles and therapeutic efficacy of barnyard millets as sprouts and microgreens for antioxidant, anti-diabetic, and bioaccessibility properties. Based on the study, sprouts contained 456.52 mg GE/g of starch and microgreens contained 470.04 mg GE/g of carbohydrates, whereas the gastric phase of microgreens showed 426.85 mg BSAE/g, 397.6 mg LE/g, 348.19 g RE/g, and 307.40 g AAE/g of proteins, amino acids, vitamin A and vitamin C respectively. Secondary metabolites were significantly concentrated in the microgreen stage which is responsible for their increased antioxidant and antidiabetic potential than sprouts. This study validated the therapeutic and nutritional value of millet sprouts and microgreens by demonstrating their significant nutritional composition.

Physiological Basis for the Mechanism of Selectivity of Tripyrasulfone between Rice (Oryza sativa) and Barnyard Grass (Echinochloa crus-galli).

Tripyrasulfone is currently the only HPPD-inhibiting herbicide that possesses outstanding selectivity even for direct-seeded rice (Oryza sativa) when applied POST to control grass weeds; however, the underlying mechanisms remain unclear. In this study, the inhibitory effects of the real active HDT of tripyrasulfone on recombinant 4-hydroxyphenylpyruvate dioxygenase (HPPDs) from rice and barnyard grass (Echinochloa crus-galli) were similar, with consistent structural interactions and similar binding energies predicted by molecular docking. However, the HPPD expression level in rice was significantly greater than that in barnyard grass after tripyrasulfone treatment. Tripyrasulfone was rapidly taken up and hydrolyzed into HDT, which was similarly distributed within the whole plants of rice and barnyard grass at 24 h after treatment. Compared with barnyard grass, rice has more uniform epicuticular wax in the cuticle of its leaves, absorbing less tripyrasulfone and metabolizing much more tripyrasulfone. Overall, to a greater extent, the different sensitivities to tripyrasulfone between barnyard grass and rice resulted from metabolic variations.

Disulfide-stabilized helical hairpin structure and activity of a novel antifungal peptide EcAMP1 from seeds of barnyard grass (Echinochloa crus-galli).

This study presents purification, activity characterization, and (1)H NMR study of the novel antifungal peptide EcAMP1 from kernels of barnyard grass Echinochloa crus-galli. The peptide adopts a disulfide-stabilized α-helical hairpin structure in aqueous solution and thus represents a novel fold among naturally occurring antimicrobial peptides. Micromolar concentrations of EcAMP1 were shown to inhibit growth of several fungal phytopathogens. Confocal microscopy revealed intensive EcAMP1 binding to the surface of fungal conidia followed by internalization and accumulation in the cytoplasm without disturbance of membrane integrity. Close spatial structure similarity between EcAMP1, the trypsin inhibitor VhTI from seeds of Veronica hederifolia, and some scorpion and cone snail toxins suggests natural elaboration of different functions on a common fold.

Vanillin extracted from proso and barnyard millets induces cell cycle inhibition and apoptotic cell death in MCF-7 cell line.

CONTEXT: Consuming whole grain food has been motivated due to numerous health benefits arising from their bioactive components. AIMS: This study aims to study whether the active compound extracted from Proso and Barnyard millets inhibits cell proliferation and induces apoptotic cell death in MCF-7 cell line. MATERIALS AND METHODS: Cell proliferative effect was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay using MCF-7 cell line. Cytotoxicity was determined by release of lactate dehydrogenase (LDH) enzyme from cells. Apoptotic morphological changes in MCF-7 cells were observe under fluorescence microscope using double staining of Hoeschst 33342/propidium iodide (PI). Induction of apoptosis was analyzed using Annexin V-fluorescein isothiocyanate/PI through flow cytometry. RESULTS: In this study, cell proliferative effect of the bioactive compounds from proso millet (Compound 1) and barnyard millet (Compound 2) was evaluated using MCF-7 cell line. Both the compounds significantly inhibited the proliferation of MCF-7 cells after treated with 250 µg/ml and 1000 µg/ml concentration for 48 h. Cytotoxic activity of compounds was assessed by the release of LDH showed that these extracted compounds were not toxic to the cells. Apoptosis was confirmed by Hoechst 33,342/PI dual-staining, Annexin V-FTIC/PI staining, and flow cytometry results of cell cycle analysis shows that there was a significant cell arrest in the G0/G1 phase and increased the apoptotic cells in sub-G0 phase in a dose-dependent manner. CONCLUSIONS: This study suggests that the extracted vanillin compound from these millets have effectively induced apoptotic cell death in breast cancer cell line.

Comparison of the ability of organic acids and EDTA to enhance the phytoextraction of metals from a multi-metal contaminated soil.

Chelates have been shown to enhance the phytoextraction of metal from contaminated soil. In this study, we evaluated the ability of chelates to enhance the phytoextraction of metals by barnyard grass (Echinochloa crus-galli) from soils contaminated with multiple metals. The results revealed that EDTA increased the ability of barnyard grass to take up Cd, Cu and Pb, but that it resulted in increased soil leaching. Conversely, citric acid induced the removal of Cd, Cu and Pb from soil without increasing the risk of leaching. Furthermore, E.crus-galli showed no signs of phytotoxicity in response to treatment with citric acid, whereas its shoot growth decreased in response to treatment with EDTA (p < 0.05). Taken together, these results demonstrate that citric acid is a good agent for the enhancement of the phytoextraction of metals.

The Genomes of the Allohexaploid Echinochloa crus-galli and Its Progenitors Provide Insights into Polyploidization-Driven Adaptation.

The hexaploid species Echinochloa crus-galli is one of the most detrimental weeds in crop fields, especially in rice paddies. Its evolutionary history is similar to that of bread wheat, arising through polyploidization after hybridization between a tetraploid and a diploid species. In this study, we generated and analyzed high-quality genome sequences of diploid (E. haploclada), tetraploid (E. oryzicola), and hexaploid (E. crus-galli) Echinochloa species. Gene family analysis showed a significant loss of disease-resistance genes such as those encoding NB-ARC domain-containing proteins during Echinochloa polyploidization, contrary to their significant expansionduring wheat polyploidization, suggesting that natural selection might favor reduced investment in resistance in this weed to maximize its growth and reproduction. In contrast to the asymmetric patterns of genome evolution observed in wheat and other crops, no significant differences in selection pressure were detected between the subgenomes in E. oryzicola and E. crus-galli. In addition, distinctive differences in subgenome transcriptome dynamics during hexaploidization were observed between E. crus-galli and bread wheat. Collectively, our study documents genomic mechanisms underlying the adaptation of a major agricultural weed during polyploidization. The genomic and transcriptomic resources of three Echinochloa species and new insights into the polyploidization-driven adaptive evolution would be useful for future breeding cereal crops.

Inhibition of protein glycoxidation and advanced glycation end-product formation by barnyard millet (Echinochloa frumentacea) phenolics.

Protein glycation plays a vital role in the progression of various diabetes complications. Therefore, inhibition of protein glycation could be a key strategy to prevent these diabetic abnormalities. Evaluation of phenolic compositions and their antiglycation activity revealed that p-coumaric and chlorogenic acids were major phenolic acids in barnyard millet. These phenolics exhibited multiple antioxidant activities in various mechanisms and protected the oxidative DNA damage and hydroxyl radical-induced protein fragmentation. Millet phenolics were very effective in scavenging >78% reactive carbonyl intermediates in the reaction and protected protein thiol group oxidation. Furthermore, 68.3% inhibition of protein glycation and reduced formation of protein aggregates were also observed with millet phenolics. Besides, fluorescence intensity measurements indicated a significant decrease in advance glycated end products and protection against glycoxidation-induced protein conformational changes at 100 µg/ml phenolics. These results suggest the potential utility of barnyard millet as an ingredient in functional foods for controlling protein glycation associated diabetic complications.

Dietary Japanese millet protein ameliorates plasma levels of adiponectin, glucose, and lipids in type 2 diabetic mice.

Millet is an important food crop in Asia and Africa, but the health benefits of dietary millet are little known. This study defined the effects of dietary Japanese millet on diabetic mice. Feeding of a high-fat diet containing Japanese millet protein concentrate (JMP, 20% protein) to type 2 diabetic mice for 3 weeks significantly increased plasma levels of adiponectin and high-density lipoprotein cholesterol (HDL cholesterol) and decreased the levels of glucose and triglyceride as compared to control. The starch fraction of Japanese millet had no effect on glucose or adiponectin levels, but the prolamin fraction beneficially modulated plasma glucose and insulin concentrations as well as adiponectin and tumor necrosis factor-alpha gene expression. Considering the physiological significance of adiponectin and HDL cholesterol levels in type 2 diabetes, insulin resistance, and cardiovascular disease, our findings imply that dietary JMP has the potential to ameliorate these diseases.

Short-term accumulation of atrazine by three plants from a wetland model system.

This work describes the accumulation and distribution of the herbicide atrazine in soil, water, and roots from three wetland model systems using the monocots Typha domingensis, Sagittaria lancifolia, and Echinochloa pyramidalis. Results were analyzed from a 3(3) full factorial experimental design, in order to describe the effect of accumulation of atrazine and times of exposure in the species evaluated. We found that accumulation depends on the species and the herbicide concentration; about 30% was accumulated in soil, 40% in roots, and 10-20% in water. By the end of the experiment, E. pyramidalis accumulated 8.47 mg/l of atrazine and 14.39 mg/l T. domingensis; in all cases, adsorption accounted for 1.4%, fitting a Langmuir model with a k(d) of 14.47.

[The purification and characterization of a novel lipid transfer protein from caryopsis of barnyard grass (Echinochloa crusgalli)].

A novel lipid transfer protein called Ec-LTP was isolated from resting caryopsis of weed barnyard grass Echinochloa crusgalli (L.) Beauv.; its molecular weight, amino acid content and N-terminal amino acid sequence were determined. Ec-LTP was a 9150 Da protein, containing eight cysteine residues, which formed four disulfide bonds. The isolated protein could significantly inhibit the development of pathogenic fungi Phytophthora infestans and Helminthosporium sativum, causing the late blight of potato and tomato and the root rot of herbs, respectively.