Genetic Differentiation of Reed Canarygrass (Phalaris arundinacea L.) within Eastern Europe and Eurasia.

Worldwide molecular research of economically important Phalaris arundinacea (Poaceae) is mainly focused on the invasions of this species from Europe to North America. Until the present study, the genetic diversity of the P. arundinacea had not been studied across the Baltic countries. The objective of this research is to evaluate the diversity of Lithuanian populations of P. arundinacea at simple sequence repeat (SSR) loci comparatively among populations of the Baltic countries, Luxembourg, and the Russian Far East (Eurasian), evaluating differentiation between Lithuanian populations and ornamental accessions, and relating these with environmental features. For six selected Lithuanian river basin populations, GBS low density SNPs were used to determine genetic diversity. Bayesian analysis showed that Eurasian populations of Phalaris arundinacea consist of two gene clusters. Statistically significant genetic differentiation among European and Eurasian populations was documented. Lithuanian genotypes growing naturally along rivers are genetically distinct from cultivated ornamentals. GBS-SNPs divided the six selected Nemunas river basins into three distinct groups with one, two, or three rivers in separate groupings for genetic diversity. Genetic diversity is primarily within, rather than among, Lithuanian, eastern European, and Eurasian populations of P. arundinacea across the continent. Thus, restoration efforts would benefit from local population seed origination.

Development of Low Glycemic Index Pancakes Formulated with Canary Seed (Phalaris Canariensis) Flour.

Consumers prefer food products that, in addition to nutritional properties, also have effects beneficial to health. Non-conventional food plants such as canary seed (Phalaris canariensis) constitute an alternative in the food industry due to nutritional properties, chemical, and bioactive potential. The aim of this research was to develop pancake formulations with the inclusion of canary seed flour to evaluate their proximate composition, overall score, taste and texture sensory acceptability, and glycemic index. Pancakes based on whole-wheat flour mixed with canary seed flour were developed at four substitution levels (control 0, 10, 30, and 50%). The formulations exhibited attractive nutritional properties, mainly due to the levels of protein (~ 8.7%), minerals (~ 5.1%), and total dietary fiber (4.7-5.9%). The overall score and taste showed a statistical difference (p < 0.05) between the formulations. The flours with 10 and 30% showed high sensory acceptability with scores of 6.22 and 6.67 respect to 7-point hedonic scale, between the categories "I like it" and "I like it a lot". All formulations presented a low glycemic index (34-39%) that was significantly influenced (p < 0.05) as the level of substitution increased. The findings represent a new approach to the use of canary seed in the development of healthy food products.

Application of Molecular Dynamics Simulations to Determine Interactions between Canary Seed (Phalaris canariensis L.) Bioactive Peptides and Skin-Aging Enzymes.

Food bioactive peptides are well recognized for their health benefits such as antimicrobial, antioxidant, and antihypertensive benefits, among others. Their drug-like behavior has led to their potential use in targeting skin-related aging factors like the inhibition of enzymes related with the skin-aging process. In this study, canary seed peptides (CSP) after simulated gastrointestinal digestion (<3 kDa) were fractioned by RP-HPLC and their enzyme-inhibition activity towards elastase and tyrosinase was evaluated in vitro. CSP inhibited elastase (IC50 = 6.2 mg/mL) and tyrosinase (IC50 = 6.1 mg/mL), while the hydrophobic fraction-VI (0.2 mg/mL) showed the highest inhibition towards elastase (93%) and tyrosinase (67%). The peptide fraction with the highest inhibition was further characterized by a multilevel in silico workflow, including physicochemical descriptor calculations, antioxidant activity predictions, and molecular dynamics-ensemble docking towards elastase and tyrosinase. To gain insights into the skin permeation process during molecular dynamics simulations, based on their docking scores, five peptides (GGWH, VPPH, EGLEPNHRVE, FLPH, and RPVNKYTPPQ) were identified to have favorable intermolecular interactions, such as hydrogen bonding of polar residues (W, H, and K) to lipid polar groups and 2-3 Å van der Waals close contact of hydrophobic aliphatic residues (P, V, and L). These interactions can play a critical role for the passive insertion of peptides into stratum corneum model skin-membranes, suggesting a promising application of CSP for skin-aging treatments.

Identification and Validation of Reference Genes for RT-qPCR Analysis in Reed Canary Grass during Abiotic Stress.

Reed canary grass (Phalaris arundinacea L.) is known for its tolerance to drought, heavy metals, and waterlogging, making it a popular choice for forage production and wetland restoration in the Qinghai-Tibet Plateau (QTP). To accurately assess gene expression in reed canary grass under different abiotic stresses, suitable reference genes need to be identified and validated. Thirteen candidate reference gene sequences were selected and screened using RT-qPCR to detect their expression levels in reed canary grass leaves under drought, salt, cadmium, and waterlogging stresses. Four algorithms were used to assess the stability of the expression levels of the candidate reference genes. The most stably expressed genes were UBC and H3 under drought Cd, ETF and CYT under salt stress, and ETF and TUB under waterlogging stress. GAPDH was found to be less stable under abiotic stresses. PIP-1, PAL, NAC 90, and WRKY 72A were selected as response genes for quantitative expression assessment under drought, salt, Cd, and waterlogging stresses to confirm the accuracy of the selected stable reference genes. These results provide a theoretical reference for assessing gene expression in reed canary grass under abiotic stresses.

Production, optimization, and purification of alkaline thermotolerant protease from newly isolated Phalaris minor seeds.

The present work aims to purify and perform a preliminary analysis on a thermostable serine alkaline protease from a recently identified P. minor. The enzyme was purified 2.7-fold with a 12.4 % recovery using Sephadex G-100 chromatography, DEAE-cellulose, and ammonium sulphate precipitation. The isolated enzyme has a specific activity of 473 U/mg. The purified protease had a molecular mass of 29 kDa, and just one band was seen, which matched the band obtained using SDS-PAGE. High thermostability was demonstrated by the enzymes, which had half-lives of 31.79 and 6.0 min (a 5.3-fold improvement), enthalpies of denaturation (ΔH°) of 119.53 and 119.35 KJ mol-1, entropies of denaturation (ΔS°) of 32.96 and 41.11 J/mol·K, and free energies of denaturation (ΔG°) of 108.87 and 105.58 KJ mol-1 for the protease enzyme. Studies on the folding and stability of alkaline proteases are important since their use in biotechnology requires that they operate in settings of extreme pH and temperature. According to the kinetic and thermodynamic properties, the protease produced by P. minor is superior to that produced by other sources and previously described plants, and it might find utility in a variety of industrial fields.

Recovery of microelements from municipal sewage sludge by reed canary grass and giant miscanthus.

The aim of the study was to evaluate the content, uptake, and utilization of Al, Mn, Fe, Co, and Mo from municipal sewage sludge by the reed canary grass (RCG) and the giant miscanthus (MG). The impact of sewage sludge on the mass ratios between microelements contained in the plant biomass was analyzed as well. The long-term field experiment consisted in application of four levels of fertilization with sewage sludge: 10, 20, 40, and 60 Mg DM·ha-1. A non-fertilized control object (0 Mg DM·ha-1) was used as well. The miscanthus biomass was characterized by higher content of Al and Mn, whereas higher levels of Fe, Co, and Mo were determined in the reed canary grass biomass. As in the case of the element content, the giant miscanthus was characterized by highest level of Al and Mn uptake, while the highest uptake of Fe, Co, and Mo was exhibited by the reed canary grass. In the group of the analyzed microelements, the giant miscanthus showed the highest the recovery of Mo (47%) followed by Mn (39%), Fe (35%), and Co (15%), and the lowest uptake was determined in the case of Al (5%).

In addition to the use of their biomass for the production of renewable energy, energy crops (reed canary grass, giant miscanthus) can be used to recover microelements from municipal sewage sludge. Energy crop biomass can therefore be a potential source of microelements in animal feed. Attention was paid to the mass ratios of Fe:Mn, Fe:Al, Fe:Co, Fe:Mo, Mn:Al, Mn:Co, Mn:Mo, and Co:Mo in the biomass yield of the species used.

Canary Seed (Phalaris canariensis L.) Peptides Prevent Obesity and Glucose Intolerance in Mice Fed a Western Diet.

Previous research showed that canary seed (Phalaris canariensis L.) peptides (CSP) possess robust in vitro antiobesity properties via inhibition of pancreatic lipase (PL). Nevertheless, no studies have yet explored their antiobesity properties in vivo. Consequently, we investigated the effects of CSP in C57BL/6J mice under a Western diet (WD). Mice were assigned into groups and fed a normal diet (ND) or a WD accompanied by an oral dose of CSP (250 or 500 mg/kg/day), orlistat (40 mg/kg/day), or distilled water. The results showed that consuming CSP can provide metabolic benefits, including preventing weight gain by up to 20%, increasing glucose tolerance, and reducing insulin, leptin, and LDL/VLDL levels in plasma. Conversely, total ghrelin was unaffected by CSP-500, but decreased by CSP-250, and amplified by orlistat. Surprisingly, CSP-250 was more effective in preventing weight gain and promoting satiety than CSP-500. Parallel to this, protein absorption in CSP-500 was decreased, supported by a rise in fecal crude protein (+3.5%). Similarly, fecal fat was increased by orlistat (38%) and was unaffected by CSP-250 (3.0%) and CSP (3.0%), comparatively to WD (2.5%). Despite this, both CSP treatments were equally effective in decreasing hepatic steatosis and avoiding hyperlipidemia. Furthermore, the enzymatic analysis showed that CSP-PL complexes dissociated faster (15 min) than orlistat-PL complexes (41 min). Lastly, CSP did not affect expression of hepatic lipid oxidation genes ACO and PPAR-α, but reduced the expression of the hydrolase gene LPL, and lipogenesis related genes FAS and ACC. Taken together, these results suggest that CSP antiobesity mechanism relies on lipid metabolism retardation to increase fat transit time and subsequently suppress hunger.

Enhanced biosorption of europium and cesium ions from aqueous solution onto phalaris seed peel as environmental friendly biosorbent: Equilibrium and kinetic studies.

The sorption characteristics of Eu(III) and Cs(I) removal from aqueous solution were estimated using phalaris seeds peel powder (PSP), a novel biosorbent that is economical and low-cost. Batch equilibrium experiments were carried out to investigate the effects of contact time, initial metal concentration, media pH, and interference ions on the sorption of Eu(III) and Cs(I). PSP powder characterization via Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TG/DSC), X-ray diffraction (XRD), and scanning electron microscope (SEM) revealed some changes before and after the adsorption process. This indicates that most likely, adsorption has taken place between the metal ions and the adsorbents in the aqueous solution. The obtained results show that Eu(III) has a higher selectivity than Cs(I). The kinetics of the adsorption of Eu3+ and Cs+ have been discussed. It was shown that the second-order kinetic equation could describe the sorption and was found to be the best fitted model (R2 = 0.999) for two metal ions. Langmuir and, Dubinin-Radushkevich (D-R), isotherm were found to best fit (R2 = 0.99) in this study. The separation factor (RL) value of less than 1.0 indicates that the biosorption of both metal ions on PSP is favorable. Thermodynamic parameters, such as ΔHo, ΔGo, and ΔSo, have been calculated by using the thermodynamic equilibrium coefficient obtained at different temperatures. The obtained results indicated the endothermic nature of the sorption process for both metal ions onto PSP. PSP powder has the potential to be used as a low-cost biosorbent for the removal of Eu(III) and Cs(I) from wastewater, according to the findings.

Seed germination ecology of hood canarygrass (Phalaris paradoxa L.) and herbicide options for its control.

Hood canarygrass (Phalaris paradoxa L.) is a problematic weed in winter crops of Australia. Experiments were conducted to determine the effects of environmental factors on seed germination of P. paradoxa and wheat (Triticum aestivum L.) and herbicide options for P. paradoxa control. Results revealed that P. paradoxa had higher germination (> 89%) at a temperature range from 15/5 ℃ to 25/15 ℃ [day/night (12 h/12 h] compared with 30/20 ℃ and 35/25 ℃. At a temperature regime of 30/20 ℃, P. paradoxa had 1% germination; however, wheat at this temperature range resulted in 79% germination. Exposure of seeds of P. paradoxa to > 150 ℃ pretreatment (radiant heat for 5 min) resulted in no germination. These results suggest that infestation of P. paradoxa could be reduced by residue burning or by planting wheat crops early in the season when the temperature is relatively high. At a water potential of -0.8 MPa, seed germination of P. paradoxa and wheat was 75 and 96%, respectively. Similarly, at the highest salt concentration (200 mM sodium chloride), seed germination of P. paradoxa and wheat was 73 and 79%, respectively. These observations suggest that like wheat, P. paradoxa is also highly tolerant to water and salt stress conditions, therefore, it could invade the agro-ecosystem under water and salt stress situations. Germination of P. paradoxa was found to be low (10%) on the soil surface, suggesting that no-till systems could inhibit the germination of P. paradoxa. Pre-emergent (PRE) herbicides, namely cinmethylin, pyroxasulfone, and trifluralin, provided 100% control of P. paradoxa; however, in the presence of 2 t ha-1 of residue cover, pyroxasulfone provided better control of P. paradoxa compared with other herbicides. Post-emergent (POST) herbicides clethodim, haloxyfop, and paraquat provided excellent control of P. paradoxa, even if the plant size was large (10-leaf stage). Knowledge generated from this study will help in strengthening the integrated management of P. paradoxa.

Possibility of Metal Accumulation in Reed Canary Grass (Phalaris arundinacea L.) in the Aquatic Environment of South-Western Polish Rivers.

A four-year research study was conducted on aquatic plants (reed canary grass) growing in the beds of three rivers and their tributaries in Lower Silesia, Poland. Metal contents (Cu, Cd, Ni, Pb, Zn, Fe, Mn) were determined in plant samples, metal accumulation in water (BCFw) and sediment (BCFB), Metal Pollution Index (MPI) and Enrichment Factor (EF) were calculated. The highest contents of copper, lead, nickel and cadmium were found in reed canary grass sampled from the Nysa Szalona River. The highest values were recorded for zinc in the Bystrzyca River, and for iron and manganese in the Strzegomka River. The series of metals were as follows: Nysa Szalona and Strzegomka: Cd < Ni < Pb < Cu < Zn < Mn < Fe, Bystrzyca: Cd < Ni < Cu < Pb < Zn < Mn < Fe. Throughout the study period, the lowest values of metals in plants were recorded in 2015 and 2018, and the highest in 2017. The general picture of MPI in aquatic plants is arranged in the series Bystrzyca < Strzegomka < Nysa Szalona. These values classify the studied material at a high level of pollution in all rivers. In the comparison of the two extreme sites, i.e., source−mouth, higher values were found at the mouth of the reservoir, which suggests that metals move with the water current and accumulate more with the direction of the river flow, which is most likely a consequence of the influence of the catchment area as the source of metals. The series of EF enrichment factor values were as follows: Bystrzyca­Ni < Cd < Fe < Cu < Zn < Mn < Pb, Nysa Szalona­Ni < Fe < Zn < Cd < Mn < Cu < Pb, Strzegomka­Ni < Cd < Fe < Zn < Cu < Pb < Mn. For all the samples studied, the values found in spring were much higher than in autumn, which indicates the great importance for research in that area. The levels of copper and iron were within the range of moderate values, lead and manganese reached very high and exceptionally high values, and the remaining metals were within the values described as significant. Bioaccumulation of metals determined relative to bottom sediments was highest in 2017 and lowest in 2018, while bioaccumulation relative to water was highest in 2018 and lowest in 2016. The four-year study found that the metal content in reed canary grass was mostly within the range of mean values presented in the literature from moderately polluted areas. Also, no significant deviation was found from levels that have been recorded for the same rivers for more than two decades.

Oxidative Stress Protection by Canary Seed (Phalaris canariensis L.) Peptides in Caco-2 Cells and Caenorhabditis elegans.

During oxidative stress, degenerative diseases such as atherosclerosis, Alzheimer's, and certain cancers are likely to develop. Recent research on canary seed (Phalaris canariensis) peptides has demonstrated the high in vitro antioxidant potential. Thus, this study aimed to assess the cellular and in vivo antioxidant capacity of a low-molecular-weight (<3 kDa) canary seed peptide fraction (CSPF) using Caco-2 cells and the Caenorhabditis elegans model. The results show that the CSPF had no cytotoxicity effect on Caco-2 cells at any tested concentration (0.3−2.5 mg/mL). Additionally, the cellular antioxidant activity (CAA) of the CSPF was concentration-dependent, and the highest activity achieved was 80% by the CSPF at 2.5 mg/mL. Similarly, incubation with the CSPF significantly mitigated the acute and chronic oxidative damage, extending the lifespan of the nematodes by 88 and 61%, respectively. Furthermore, it was demonstrated that the CSPF reduced the accumulation of reactive oxygen species (ROS) to safe levels after sub-lethal doses of pro-oxidant paraquat. Quantitative real-time PCR revealed that the CSPF increased the expression of oxidative-stress-response-related gene GST-4. Overall, these results show that the CSPFs relied on GST-4 upregulation and scavenging of free radicals to confer oxidative stress protection and suggest that a CSPF can be used as a natural antioxidant in foods for health applications.

Physiological and molecular responses of Phalaris arundinacea under salt stress on the Tibet plateau.

Phalaris arundinacea, with its characteristics of rapid growth and high biological yield, is regarded as an excellent forage grass in the Qinghai-Tibetan Plateau region of China. To explore the physiological and molecular response mechanism of Phalaris arundinacea under salt stress, we monitored the biomass and physiological indexes of two locally grown strains under conditions of exposure to 150 and 300 mM NaCl solution. Z0611 exhibited better salt stress tolerance than YS. Transcriptome sequencing analysis showed that YS and Z0611 had 1713 and 4290 differentially expressed genes (DEGs), respectively, including on metabolic processes, single-organism process, catalytic activity, and plant hormone signal transduction in the GO and KEGG databases. We also identified a large number of genes involved in hormone signaling, antioxidant systems, ion homeostasis, and photosynthetic systems. Our study provides physiological and molecular insight for establishing a salt resistance database and mining salt tolerance genes in Phalaris arundinacea, and also provides theoretical guidance for the restoration of saline-alkali land on the Qinghai-Tibet Plateau.

Aluminum Bioaccumulation in Reed Canary Grass (Phalaris arundinacea L.) from Rivers in Southwestern Poland.

This study aimed to determine aluminum levels in reed canary grass Phalaris arundinacea L.) in rivers in southwestern Poland­Bystrzyca, Strzegomka, and Nysa Szalona, together with their tributaries. The samples were collected in spring and autumn 2015−2018. The highest amounts of aluminum were recorded in the Nysa Szalona, and the lowest in the Bystrzyca. During the four-year cycle of studies, the highest values were recorded in the last year, and the lowest in the first year. The highest amounts of aluminum were found in all three rivers in the lowland tributaries. In the main rivers, higher amounts of aluminum were found at the mouth of the Nysa Szalona and Strzegomka reservoirs, while the opposite situation was found for the Bystrzyca. Higher aluminum contents were recorded in autumn than in spring, and the values of BCFW (aluminum bioaccumulation factor in relation to water) and BCFB (aluminum bioaccumulation factor in relation to bottom sediments) coefficients were also higher. The MPI (metal pollution index) was arranged in a series: Bystrzyca < Strzegomka < Nysa Szalona, while the degree of pollution was high for Bystrzyca and very high for the other two rivers. The variability in Al levels may be attributed to pollution level in the catchments, but also to successive modernization works carried out in the beds of the main rivers and their tributaries. All these works were carried out in a variable way and often covered only a fragment of the riverbed; therefore, the consequences of activity may have been visible in the catchment but not necessarily in the same vegetation cycles.

6-Ethoxy-4- N-(2-morpholin-4-ylethyl) -2-N-propan-2-yl-1,3, 5-triazine-2, 4-diamine endows herbicidal activity against Phalaris minor a weed of wheat crop field: An in -silico and experimental approaches of herbicide discovery.

Phalaris minor is a major weed of wheat crop which has evolved resistance against herbicides. Isoproturon is the most accepted herbicide developed resistance in 1992. Later, introduced herbicides also developed resistance and cross-resistance to their respective binding sites. Isoproturon binds at the QB binding site of the D1 protein of photosystem-II (PS-II), which blocks the electron transfer in photosynthesis. In this work, we have carried out a series of computational studies to prioritize the promising herbicides against D1 protein of P. minor. Through the computational studies, twenty-four lead molecules are prioritized which have shown a higher binding affinity and inhibition constant than the reference ligand molecule. The binding and conformational stability of docked complexes was evaluated by molecular dynamics simulations and binding free energy calculations i.e., MM/PBSA. A list of amino acids such as Ala225, Ser226, Phe227, and Asn229 present in the binding site of protein is obtained to be playing an important role in the stability of the protein-lead complex via hydrogen bond and π-π interactions. Binding free energy calculation revealed that the selected lead molecule binding is energetically favorable and driven by electrostatic interactions. Among 24 leads, computational results have uncovered eight promising compounds as potential herbicides which have shown comparable physiochemical profile, better docking scores, system stability, H-bond occupancy, and binding free energy than terbutryn, a reference molecule. These prioritized molecules were custom synthesized and evaluated for their herbicidal activity and specificity through whole plant assay under laboratory-controlled conditions. The lead molecule ELC5 (6-ethoxy-4-N-(2-morpholin-4-ylethyl)-2-N-propan-2-yl-1,3,5-triazine-2,4-diamine) has shown comparable activity to the reference herbicide(isoproturon) against P. minor.

Effect of phenyllactic acid on silage fermentation and bacterial community of reed canary grass on the Qinghai Tibetan Plateau.

BACKGROUND: This study aimed to investigate the effect of phenyllactic acid as an additive on silage fermentation and bacterial community of reed canary grass (RCG, Phalaris arundinacea L.) on the Qinghai Tibetan Plateau. At the heading stage, RCG was harvested, chopped and ensiled in small bag silos. The silage was treated without (control, 1.0 g/mL sterile water, on a fresh matter basis (FM)) or with phenyllactic acid (PLA, 3 mg/mL, FM), antimicrobial additive (PSB, a mixture of potassium sorbate and sodium benzoate, 2%, FM), lactic acid bacteria inoculant (LABi, L. plantarum + L. curvatus, 1 × 106 cfu/g, FM) and PLA + LABi, and then stored in a dark room at the ambient temperature (5 ~ 15 °C) for 60 days. RESULTS: Compared with control, PLA decreased lactic acid, acetic acid and ammonia-N contents, and subsequently increased CP content of RCG silage. PLA enhanced the growth of lactic acid bacteria and reduced the count of yeasts (P < 0.05) in RCG silage, with reduced bacterial richness index (Chao1), observed operational taxonomic units and diversity index (Simpson). In relative to control, moreover, PLA and PLA + LABi increased the relative abundance of Lactococcus in RCG silage by 27.73 and 16.93%, respectively. CONCLUSIONS: Therefore, phenyllactic acid at ensiling improved nutritional quality of RCG silage by advancing the disappearance of yeasts and the dominance of Lactococcus.

Variability in ITS1 and ITS2 sequences of historic herbaria and extant (fresh) Phalaris species (Poaceae).

BACKGROUND: Phalaris species (Poaceae) occupy diverse environments throughout all continents except Antarctica. Phalaris arundinacea is an important forage, ornamental, wetland restoration and biofuel crop grown globally as well as being a wetland invasive. The nuclear ribosomal internal transcribed spacer (ITS) region has been used for Phalaris barcoding as a DNA region with high nucleotide diversity for Phalaris species identification. Recent findings that P. arundinacea populations in Minnesota USA are most likely native and not European prompted this analysis to determine whether Eurasian vs. native North American P. arundinacea differed in ITS regions. Our objectives were to amplify and compare ITS regions (ITS1 and ITS2) of historic herbaria (1882-2001) and extant (fresh) Phalaris specimens; analyze ITS regions for species-specific polymorphisms (diagnostic SNPs) and compare ITS regions of historic Phalaris specimens with known, extant Phalaris species. RESULTS: We obtained complete ITS1 and ITS2 sequences from 31 Phalaris historic (herbaria samples, 1908 to 2001) and five extant (fresh) specimens. Herbaria Phalaris specimens did not produce new SNPs (single nucleotide polymorphisms) not present in extant specimens. Diagnostic SNPs were identified in 8/12 (66.6%) Phalaris species. This study demonstrates the use of herbaria tissue for barcoding as a means for improved species identification of Phalaris herbaria specimens. No significant correlation between specimen age and genomic DNA concentration was found. Phalaris arundinacea showed high SNP variation within its clade, with the North American being distinctly different than other USA and most Eurasian types, potentially allowing for future identification of specific SNPs to geographic origin. CONCLUSIONS: While not as efficient as extant specimens to obtain DNA, Phalaris herbaria specimens can produce high quality ITS sequences to evaluate historic genetic resources and facilitate identification of new species-specific barcodes. No correlation between DNA concentration and age of historic samples (119 year range) occurred. Considerable polymorphism was exhibited in the P. arundinacea clade with several N. American accessions being distinct from Eurasian types. Further development of within species- and genus-specific barcodes could contribute to designing PCR primers for efficient and accurate identification of N. American P. arundinacea. Our finding of misidentified Phalaris species indicates the need to exercise stringent quality control measures on newly generated sequence data and to approach public sequence databases in a critical way.

Physiological and transcriptional responses of Phalaris arundinacea under waterlogging conditions.

As a high-yielding forage grass, Phalaris arundinacea widely distributed in the Qinghai-Tibet Plateau region of China. To explore physiological and molecular response mechanism of Phalaris arundinacea under waterlogging, we analyzed the biomass and physiological indexes of three locally grown strains under the submerged condition of 10 cm. The material Z0611 showed the strongest waterlogging resistance while the YS showed the weakest performance. Transcriptome sequencing analysis demonstrated that the YS and Z0611 had 17010 and 7566 differently expression genes (DEGs), respectively, which were mainly concentrated in the metabolic process, cell, ribosome, phenylpropanoid biosynthesis pathway in GO and KEGG databases. We also identified a large number of genes involved in carbohydrate metabolism, hormone signaling regulation, transcription factors, antioxidant system, and ethylene signaling. Our research may provide a scientific basis for the restoration of wetland environment on the Qinghai-Tibet Plateau, and lay a foundation for further exploration of the waterlogging resistance genes of Phalaris arundinacea and breeding of new strains resistant with waterlogging stress.

Field application of allelopathic bacteria to control invasion of little seed canary grass in wheat.

Development of successful biological weed control can help to resolve various environmental challenges created by the chemical and mechanical weed control. The current study is aimed at investigating the potential of allelopathic bacteria (hereinafter as AB) for biological weed control in wheat rather than the traditional areas of plant allelopathy, phyto-pathology, and insect biocontrol agents. Eleven strains of AB were obtained that were inhibitory to little seed canary grass of which 5 also inhibited wheat in our previous studies. The remaining strains indicated the potential for biological control of this weed in wheat. Five efficient strains were selected for this purpose to conduct pot and field trials. Seeds of little seed canary grass were sown together in potted soils with wheat seeds inoculated with AB strains. A subsequent field trial was conducted at a site selected based on chronic infestations of canary grass. Seeds of the weed and inoculated wheat were co-seeded directly in field soil. For inoculation, these strains were formulated in sterilized peat and applied to seeds. In pot trials, the invasion of little seed canary grass in wheat reduced wheat grain yield up to 59.9%. Four strains suppressed the weed which resulted in recovery wheat grain yield losses from 20.1 to 66.9%. The field trial showed that the invasion of little seed canary grass reduced wheat grain yields up to 53.9%. Four strains suppressed the weed which resulted into recovery of grain wheat grain yield losses 34.3 to 64.3%. These findings were consistent with improvement of other agronomic, physiological, and chemical parameters of the crop where the four strains of AB (L9, T42, 7O0, and O010) were applied.

Scale up fractionation of components from novel glabrous brown and yellow canary seeds (Phalaris canariensis L.) and techno-functional properties of the resulting protein isolates.

Glabrous canary seed (Phalaris canariensis L.) is a novel true cereal grain produced primarily in Western Canada which has been approved for human consumption by the U.S. Food and Drug Administration and Health Canada in 2016. Due to its high protein content (22%), this new edible grain is emerging as an alternative source of plant proteins. In the present work, protein extractability from four novel glabrous (2 yellow and 2 brown) canary seeds varieties was improved based on the selection of optimal pH of protein solubilisation and precipitation. Solubilisation at pH 12 followed by acid precipitation at pH 5 were retained as optimal conditions. Scale up of the protein optimized wet fractionation process resulted in highly purified canary seed protein isolates (purity of 91 to 93%) with protein recovery yield of 65 to 69%. In parallel, for the others canary seed components, a good recovery yields were obtained for the oil fraction (6.1-6.7 g/100 g flour), starch fraction (48.1-54 g/100 g flour), and crude fiber fraction (15.1-19.7 g/100 g flour). The study of the functional properties of the obtained canary protein isolates revealed, higher solubility at acidic than alkaline region; enhanced fat and water holding capacities and notably higher foaming and emulsifying capacities than control soy protein isolate. With growing global demand for protein, glabrous canary seed has high potential in the food industry, particularly as a good source of functional gluten free cereal proteins.

A wetland plant, Phalaris arundinacea, accumulates nitrogen and phosphorus during senescence.

Secondary pollution resulting from shoot death is a difficult problem that complicated the application of wetland plants for water purification in northern wetlands. Phalaris arundinacea, a perennial herb with an obviously declining stage, or senescence, is a species that is often selected for water purification in Northern China; however, whether it reduces the secondary pollution risk via nitrogen (N) and phosphorus (P) accumulation during senescence or not remains unclear. To investigate this question, an experiment was conducted with containerized plants during the winter of 2016, after roughly half the leaves on the plants had withered. The experimental observations and analyses were conducted within 0, 2, 4, 6, and 8 weeks of the initiation of senescence. Results revealed that leaves continued to wither and shoot death occurred during weeks 4 to 6 and 8 to 10, respectively. However, no significant differences occurred in fresh biomass or in N and P accumulations of a single plant during senescence. The root biomass, root weight per volume, and total N content increased significantly, while total P content remained stable when leaves withered, respectively. H+-ATPase, a key enzyme for ion transportation, decreased after the leaves withered. However, root activity, evaluated by absorption surface per root volume, remained stable, and percentage of fine root length (diameter < 1 mm) increased significantly during senescence. In conclusion, the root activity and morphology enables P. arundinacea to accumulate N and P during senescence, which makes it a good choice for water purification in northern wetlands.