Microplastics increase cadmium absorption and impair nutrient uptake and growth in red amaranth (Amaranthus tricolor L.) in the presence of cadmium and biochar.

Microplastic (MP) pollution in terrestrial ecosystems is gaining attention, but there is limited research on its effects on leafy vegetables when combined with heavy metals. This study examines the impact of three MP types-polyethylene (PE), polyethylene terephthalate (PET), and polystyrene (PS)-at concentrations of 0.02, 0.05, and 0.1% w/w, along with cadmium (Cd) and biochar (B), on germination, growth, nutrient absorption, and heavy metal uptake in red amaranth (Amaranthus tricolor L.). We found that different MP types and concentrations did not negatively affect germination parameters like germination rate, relative germination rate, germination vigor, relative germination vigor, and germination speed. However, they increased phytotoxicity and decreased stress tolerance compared to an untreated control (CK1). The presence of MPs, particularly the PS type, reduced phosphorus and potassium uptake while enhancing Cd uptake. For example, treatments PS0.02CdB, PS0.05CdB, and PS0.1CdB increased Cd content in A. tricolor seedlings by 158%, 126%, and 44%, respectively, compared to the treatment CdB (CK2). Additionally, MP contamination led to reduced plant height, leaf dry matter content, and fresh and dry weights, indicating adverse effects on plant growth. Moreover, the presence of MPs increased bioconcentration factors and translocation factors for Cd, suggesting that MPs might act as carriers for heavy metal absorption in plants. On the positive side, the addition of biochar improved several root parameters, including root length, volume, surface area, and the number of root tips in the presence of MPs, indicating potential benefits for plant growth. Our study shows that the combination of MPs and Cd reduces plant growth and increases the risk of heavy metal contamination in food crops. Further research is needed to understand how different MP types and concentrations affect various plant species, which will aid in developing targeted mitigation strategies and in exploring the mechanisms through which MPs impact plant growth and heavy metal uptake. Finally, investigating the potential of biochar application in conjunction with other amendments in mitigating these effects could be key to addressing MP and heavy metal contamination in agricultural systems.

Role of glutathione S-transferases in the mode of action of herbicides that inhibit amino acid synthesis in Amaranthus palmeri.

Acetolactate synthase inhibitors (ALS inhibitors) and glyphosate are two classes of herbicides that act by inhibiting an enzyme in the biosynthetic pathway of branched-chain or aromatic amino acids, respectively. Besides amino acid synthesis inhibition, both herbicides trigger similar physiological effects in plants. The main aim of this study was to evaluate the role of glutathione metabolism, with special emphasis on glutathione S-transferases (GSTs), in the mode of action of glyphosate and ALS inhibitors in Amaranthus palmeri. For that purpose, plants belonging to a glyphosate-sensitive (GLS) and a glyphosate-resistant (GLR) population were treated with different doses of glyphosate, and plants belonging to an ALS-inhibitor sensitive (AIS) and an ALS-inhibitor resistant (AIR) population were treated with different doses of the ALS inhibitor nicosulfuron. Glutathione-related contents, GST activity, and related gene expressions (glutamate-cysteine ligase, glutathione reductase, Phi GST and Tau GST) were analysed in leaves. According to the results of the analytical determinations, there were virtually no basal differences between GLS and GLR plants or between AIS and AIR plants. Glutathione synthesis and turnover did not follow a clear pattern in response to herbicides, but GST activity and gene expression (especially Phi GSTs) increased with both herbicides in treated sensitive plants, possibly related to the rocketing H2O2 accumulation. As GSTs offered the clearest results, these were further investigated with a multiple resistant (MR) population, compressing target-site resistance to both glyphosate and the ALS inhibitor pyrithiobac. As in single-resistant plants, measured parameters in the MR population were unaffected by herbicides, meaning that the increase in GST activity and expression occurs due to herbicide interactions with the target enzymes.

Assessment of health risks associated with prediction of vegetable inorganic arsenic concentrations given different soil properties.

Inorganic arsenic (iAs) is a carcinogen. Vegetables such as water spinach (Ipomoea aquatica Forssk.) and amaranth (Amaranthus mangostanus L.) are recognized as high-risk sources of iAs exposure because they can accumulate significant amounts of iAs and are widely consumed. To ensure safe cultivation conditions, this study aimed to establish prediction models for iAs concentration in the edible parts of water spinach and amaranth based on soil properties. Subsequently, health risk assessments associated with iAs exposure through the consumption of these vegetables were conducted using prediction models. Soil samples were collected from agricultural fields in Taiwan and used in the pot experiments. Pearson correlation and partial correlation analyses were used to explore the relationship between soil properties, including total As, clay, organic matter, iron oxides and available phosphates, and iAs concentration in edible parts of water spinach and amaranth. Prediction models based on soil properties were developed by stepwise multiple linear regression. Health risk assessments were conducted using the Monte Carlo algorithm. The results indicate that total As and organic matter contents in soil were major predictors of iAs concentration in water spinach, whereas those in amaranth were total As and clay contents. Therefore, higher health risks for consuming water spinach and amaranth are associated with higher levels of organic matter and clay contents in soil, respectively, and these are crucial factors to consider to ensure food safety. This study suggested that As-elevated soils enriched with organic matter and clay contents should be avoided when growing water spinach and amaranth, respectively.

Effect of inoculation with arbuscular mycorrhizal fungi on the energy metabolism of selenite-rich amaranth.

A series of pot trials were undertaken to examine the impact of four arbuscular mycorrhizal fungi (AMF), namely Glomus mosseae (G.m), Glomus etunicatum (G.e), Corymbiglomus tortuosum (C.t), and the combined application of Glomus etunicatum and Corymbiglomus tortuosum (G.e + C.t), on the energy metabolism of amaranth plants grown in soil enriched with selenite at a concentration of 0.5 mg kg-1 . The inoculation of four AMFs resulted in an increase in both amaranth biomass and selenium (Se) content in leaves. The activities of phosphoglucose isomerase (PGI) and glucose-6-phosphate dehydrogenase + 6-phosphogluconate dehydrogenase were observed to decrease when AMFs were inoculated, as compared with the absence of AMF inoculation. The inoculation with G.m, C.t, and G.e + C.t resulted in an increase in succinate dehydrogenase activity; however, the inoculation with G.m, G.e, and G.e + C.t led to an increase in ascorbate oxidase activity. Furthermore, the inoculation of all four AMFs resulted in an increase in cytochrome c oxidase activity and the concentrations of oxidized coenzyme I (NAD) and reduced coenzyme I (NADH). The polyphenol oxidase activity of amaranth plants increased when inoculated with G.m and G.e, whereas it decreased when inoculated with C.t and G.e + C.t. Furthermore, the application of all four AMF treatments resulted in a reduction in adenosine triphosphate (ATP) levels and energy charge. It was worth mentioning that there was a clear inverse relationship between the energy charge and the biomass, Se concentration in the leaves. The findings presented in this research indicated that AMF may have an impact on energy metabolism and ultimately the biomass of amaranth by influencing the uptake of Se.

Amaranthus cruentus L. Seed Oil Counteracts UVA-Radiation-Induced Inhibition of Collagen Biosynthesis and Wound Healing in Human Skin Fibroblasts.

The effect of Amaranthus cruentus L. seed oil (AmO) on collagen biosynthesis and wound healing was studied in cultured human dermal fibroblasts exposed to UVA radiation. It was found that UVA radiation inhibited collagen biosynthesis, prolidase activity, and expression of the ß1-integrin receptor, and phosphorylated ERK1/2 and TGF-ß, while increasing the expression of p38 kinase. The AmO at 0.05-0.15% counteracted the above effects induced by UVA radiation in fibroblasts. UVA radiation also induced the expression and nuclear translocation of the pro-inflammatory NF-κB factor and enhanced the COX-2 expression. AmO effectively suppressed the expression of these pro-inflammatory factors induced by UVA radiation. Expressions of ß1 integrin and IGF-I receptors were decreased in the fibroblasts exposed to UVA radiation, while AmO counteracted the effects. Furthermore, AmO stimulated the fibroblast's migration in a wound healing model, thus facilitating the repair process following exposure of fibroblasts to UVA radiation. These data suggest the potential of AmO to counteract UVA-induced skin damage.

Microplastics could alter invasive plant community performance and the dominance of Amaranthus palmeri.

The increase in alien plant invasions poses a major threat to global biodiversity and ecosystem stability. However, the presence of microplastics (MPs) as an environmental stressor could impact the interactions between invasive and native species in an invasive plant community. Nevertheless, the community alterations and underlying mechanisms resulting from these interactions remain unclear. Herein, we systematically investigated the impacts of polyethylene (PE) and polypropylene (PP) on invasive plant communities invaded by Amaranthus palmeri through soil seed bank. The results illustrated that MPs markedly declined community height and biomass, and altered community structure, low-dose MPs could prominently increase community invasion resistance, but reduced community stability. The niche width and niche overlap of A. palmeri and S. viridis declined when exposed to high-dose MPs, but MPs elicited a significant rise in the niche width of S. salsa. PP had the potential to reduce the diversity of invasive plant community. Structural equation model revealed that PP addition could change soil total phosphorus content, thereby leading to a reduction of the community stability. Our study helps to fill the knowledge gap regarding the effects of MPs on invasive plant communities and provide new perspectives for invasive plant management.

Citronella essential oil-based nanoemulsion as a post-emergence natural herbicide.

A natural herbicide nanoemulsion was fabricated from citronella (Cymbopogon nardus L.) essential oil (CEO) and a nonionic surfactant Tween 60 mixed with Span 60 at hydrophilic-lipophilic balance 14 using a microfluidization method. The main constituents of CEO were citronellol (35.244%), geraniol (21.906%), and citronellal (13.632%). CEO nanoemulsion droplet size and polydispersity index (PI) were evaluated by dynamic light scattering (DLS). The smallest droplet size (33.2 nm, PI 0.135) was obtained from a microfluidizer at 20,000 psi, 7 cycles. Nanoemulsion droplet in transmission electron microscopy correlated with DLS confirmed CEO to successfully produce nanoemulsion. The herbicidal activity of the nanoemulsion as a foliar spray was evaluated against Echinochloa cruss-galli and Amaranthus tricolor as representative narrow- and broadleaf weed plants, both of which presented visual toxicity symptoms. The modes of action of the nanoemulsion were then determined in terms of membrane integrity (relative electrolyte leakage; REL), malondialdehyde (MDA), and photosynthetic pigment contents. The results showed increase in REL and MDA which indicated the destruction of the treated plants; additionally, chlorophylls and carotenoid contents were decreased. Consequently, CEO nanoemulsion may have the possibility to act as a natural herbicide resource, and natural herbicides from citronella nanoemulsions could be good alternatives for use in sustainable agriculture.

Cysteine proteases are activated in sensitive Amaranthus palmeri populations upon treatment with herbicides inhibiting amino acid biosynthesis.

The herbicides glyphosate and pyrithiobac inhibit the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) in the aromatic amino acid biosynthetic pathway and acetolactate synthase (ALS) in the branched-chain amino acid biosynthetic pathway, respectively. Here we characterise the protease activity profiles of a sensitive (S), a glyphosate-resistant (GR) and a multiple-resistant (MR) population of Amaranthus palmeri in response to glyphosate and pyrithiobac. Amino acid accumulation and cysteine protease activities were induced with both herbicides in the S population and with pyrithiobac in the GR population, suggesting that the increase in cysteine proteases is responsible for the increased degradation of the available proteins and the observed increase in free amino acids. Herbicides did not induce any changes in the proteolytic activities in the populations with target-site resistance, indicating that this effect was only induced in sensitive plants.

Chemical and cell antioxidant activity of amaranth flour and beverage after simulated gastrointestinal digestion. Role of peptides.

The potential of peptides generated by simulated gastrointestinal digestion (SGID) of two products derived from Amaranthus manteggazianus seeds, flour (F) and beverage (B), to exert peroxyl scavenging activity (ORAC) and antioxidant action on intestinal cells was studied. B was prepared by solubilisation of seed proteins, with the addition of gums and the application of a pasteurization treatment. The gastrointestinal digests FD and BD showed some differences in the peptide/polypeptide composition. The SGID produced increased ORAC activity for both samples, with some differences in the ORAC of the whole digests BD and FD and of some gel filtration fractions. Bioaccessible fractions (FDdbs and BDdbs) were obtained after treatment with cholestyramine resin to remove bile salts due to their cytotoxicity and oxidative effect. BDdbs presented a greater ORAC potency (IC50: 0.05 ± 0.01 and 0.008 ± 0.004 mg protein/ml for FDdbs and BDdbs, respectively). These fractions showed low cytotoxicity values (measured by LDH release) and produced high intracellular ROS inhibition (around 80 %), increased the SOD activity and the GSH content, with no effect on GPx activity in Caco2-TC7 cells exposed to H2O2. Several fractions with MM < 2.2 kDa presented also these cellular actions; fractions from FD induced higher increases in GSH concentration. Amaranth flour and a processed matrix like the beverage are shown as sources of bioactive peptides with potential cell antioxidant activity.

Transcriptomic analysis of Amaranthus retroflex resistant to PPO-inhibitory herbicides.

Amaranthus retroflexus L. is one of the malignant weeds which can cause a reduction in the soybean yield. We found a population of A. retroflexus (R-Q) resistant to fomesafen through the initial screening of whole-plant dose response bioassay in the research. The resistance index of the population (R-Q) was 183 times of the sensitive population (S-N). The resistant and sensitive populations were used as experimental materials in the paper. Strand-specific RNA-Seq analyses of R‒Q and S‒N populations obtained from herbicide-treated and mock-treated leaf samples after treatment were conducted to generate a full-length transcriptome database. We analyzed differentially expressed genes (DEGs) among the R-Q and S‒N A. retroflexus populations treated with recommended dose and mock-treated on the 1st (24 h) and 3rd (72 h) days to identify genes involved in fomesafen resistance. All 82,287 unigenes were annotated by Blastx search with E-value < 0.00001 from 7 databases. A total of 94,815 DEGs among the three group comparisons were identified. Two nuclear genes encoding PPO (PPX1 and PPX2) and five unigenes belonging to the AP2-EREBP, GRAS, NAC, bHLH and bZIP families exhibited different expression patterns between individuals of S‒N and R-Q populations. The A. retroflexus transcriptome and specific transcription factor families which can respond to fomesafen in resistant and susceptible genotypes were reported in this paper. The PPX1 and PPX2 genes of the target enzyme were identified. The study establishes the foundation for future research and provides opportunities to manage resistant weeds better.

Protective Effect of Amaranthus cruentus L. Seed Oil on UVA-Radiation-Induced Apoptosis in Human Skin Fibroblasts.

Since the exposure of fibroblasts to prolonged UVA radiation induces oxidative stress and apoptosis, there is a need for effective skin protection compounds with cytoprotective and antioxidant properties. One of their sources is Amaranthus cruentus L. seed oil (AmO), which is rich in unsaturated fatty acids, squalene, vitamin E derivatives and phytosterols. The aim of this study was to evaluate whether AmO evokes a protective effect on the apoptosis induced by UVA radiation in human skin fibroblasts. UVA radiation at an applied dose of 10 J/cm2 caused a significant reduction in the survival of human skin fibroblasts and directed them into the apoptosis pathway. Increased expression of p53, caspase-3, caspase-9 and PARP proteins in UVA-treated fibroblasts suggests the intrinsic mechanism of apoptosis. Application of the oil at 0.1% and 0.15% concentrations to UVA-treated cells decreased the expression of these proteins, which was accompanied by increased cell survival. Similarly, the UVA-dependent decrease in the expression of p-Akt and mTOR proteins was restored under the effect of the studied oil. The molecular mechanism of this phenomenon was related to the stimulation of antioxidant processes through the activation of Nrf2. This suggests that AmO stimulated the antioxidant system in fibroblasts, preventing the effects of UVA-induced oxidative stress, which may lead to pharmaceutical and cosmetological applications as a sun-protective substance.

Changes in bioactive compounds and antioxidant activity of three Amaranthus L. genotypes from a model to household processing.

Amaranthus L. leaves are consumed as vegetables and are a rich source of secondary plant metabolites. The phenolic profiles of the three analyzed genotypes by LC-Q-TOF-MS/MS and HPLC-DAD were characterized by high amounts of hydroxycinnamic glucaric and -isocitric acids. 'Gartenfuchsschwanz' (A. hybridus L.) and 'Red Callaloo' (A. tricolor L.) had similar profiles. 'Gemüse-Amaranth' (A. tricolor L.) had a high amount of caffeoylglucaric acid 4, which was isolated, and afterward identified by NMR. Its antioxidant activity, measured by TEAC, DPPH, and TPC, was similar to 5-caffeoylquinic acid, common in many plant species. The antioxidant activity of Amaranthus L. can be explained rather by their different phenolic- and ascorbic acid concentrations than by their species. Household cooking reduces antioxidant activity due to oxidation processes while leaching into cooking water could be neglected. Amaranthus L. baked into a wheat-dough-matrix showed lower phenolic concentrations, presumably due to the formation of phenol-protein-bounds and thermal degradation.

Cadmium uptake and membrane transport in roots of hyperaccumulator Amaranthus hypochondriacus L.

Hyperaccumulator Amaranthus hypochondriacus L. has huge potential in the remediation of cadmium (Cd)-contaminated soils and is necessary to understand the mechanism of Cd uptake by the roots. In this study, the mechanism of Cd uptake into the root of A. hypochondriacus was investigated using the non-invasive micro-test technology (NMT) by analyzing the rate of Cd2+ fluxes at different regions of the root tip; also we assessed the impact of different channel blockers and inhibitors on the Cd accumulation in the roots, the real-time Cd2+ fluxes, and the distribution of Cd along the roots. The results showed that the Cd2+ influx was greater near the root tip (within 100 µm of the tip). All the inhibitors, ion-channel blockers, and metal cations had different degrees of inhibition on the absorption of Cd in the roots of A. hypochondriacus. The net Cd2+ flux in the roots was significantly decreased by the Ca2+ channel blockers lanthanum chloride (LaCl3) by up to 96% and verapamil by up to 93%; as for the K+ channel blocker tetraethylammonium (TEA), it also caused a 68%-reduction on the net Cd2+ flux in the roots. Therefore, we infer that the uptake by A. hypochondriacus roots is mainly through the Ca2+ channels. The Cd absorption mechanism appears to be related to the synthesis of plasma membrane P-type ATPase and phytochelatin (PC), which is reflected by the inhibition of Ca2+ upon addition of inorganic metal cations. In conclusion, access of Cd ions into the roots of A. hypochondriacus is achieved through various ion channels, with the most important being the Ca2+ channel. This study will further enhance the literature regarding Cd uptake and pathways of membrane transport in roots of Cd hyperaccumulators.

Extrachromosomal circular DNA-mediated spread of herbicide resistance in interspecific hybrids of pigweed.

Extrachromosomal circular DNAs (eccDNAs) are found in many eukaryotic organisms. EccDNA-powered copy number variation plays diverse roles, from oncogenesis in humans to herbicide resistance in crop weeds. Here, we report interspecific eccDNA flow and its dynamic behavior in soma cells of natural populations and F1 hybrids of Amaranthus sp. The glyphosate-resistance (GR) trait is controlled by eccDNA-based amplification harboring the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene (eccDNA replicon), the molecular target of glyphosate. We documented pollen-mediated transfer of eccDNA in experimental hybrids between glyphosate-susceptible Amaranthus tuberculatus and GR Amaranthus palmeri. Experimental hybridization and fluorescence in situ hybridization (FISH) analysis revealed that the eccDNA replicon in Amaranthus spinosus derived from GR A. palmeri by natural hybridization. FISH analysis also revealed random chromosome anchoring and massive eccDNA replicon copy number variation in soma cells of weedy hybrids. The results suggest that eccDNAs are inheritable across compatible species, contributing to genome plasticity and rapid adaptive evolution.

Effects of exogenous calcium on cadmium accumulation in amaranth.

Calcium oxalate (CaOx) crystals in plants act as a sink for excess Ca and play an essential role in detoxifying heavy metals (HMs). However, the mechanism and related influencing factors remain unclear. Amaranth (Amaranthus tricolor L.) is a common edible vegetable rich in CaOx and a potential Cd hyperaccumulation species. In this study, the hydroponic experiment was carried out to investigate the effect of exogenous Ca concentrations on Cd uptake by amaranth. The results showed that either insufficient or excess Ca supply inhibited amaranth growth, while the Cd bioconcentration factor (BCF) increased with Ca concentration. Meanwhile, the sequence extraction results demonstrated that Cd mainly accumulated as pectate and protein-bound species (NaCl extracted) in the root and stem, compared to pectate, protein, and phosphate-bound (acetic acid extractable) species in the leaf. Correlation analysis showed that the concentration of exogenous Ca was positively correlated with amaranth-produced CaOx crystals but negatively correlated with insoluble oxalate-bound Cd in the leaf. However, since the accumulated insoluble oxalate-bound Cd was relatively low, Cd detoxification via the CaOx pathway in amaranth is limited.

Amaranth proteins and peptides: Biological properties and food uses.

Amaranthus grains have attracted great attention due to its attractive health benefits. The grains have processing properties (e.g., starch related properties) similar to those of common cereals. Amaranth grains are gluten free and protein is a significant component of these grains. Proteins of the grains have been used in various food applications such as formulations of edible films and emulsions for controlled release of bioactive compounds. The proteins have been hydrolyzed using different enzymes to produce peptides and hydrolysates, which showed a range of biological functions including anti-hypertensive and antioxidant activities among others. They have been formulated into staple foods including breads and pastas for improved nutritional quality. This review summarizes the recent advances of the last 5 years in understanding the biological functions and food applications of proteins, protein hydrolysates and peptides from the grains of different Amaranthus species. Limitations in the studies summarized are critically discussed with an aim to improve the efficiency in amaranth grain protein and peptide research.

Three typical microplastics affect the germination and growth of amaranth (Amaranthus mangostanus L.) seedlings.

Microplastics (MPs) have been a global emerging contaminant and have aroused wide public concern. Currently, it is still unknown the phytotoxicity effect of MPs on amaranth (Amaranthus mangostanus L.). This study investigated the early responses of amaranth by exposing its seeds to suspensions of polystyrene (PS), polyethylene (PE), and polypropylene (PP) MPs. We observed the effects of MPs on seed germination and growth of amaranth, especially on the oxidative damage in amaranth roots. Impacts of MPs on the germination and growth of amaranth varied with the type, concentration, and particle size of MPs. PE MPs and PP MPs inhibited the shoot extension of amaranth, while the root length under PP MPs treatment was generally shorter than that under PS MPs and PE MPs. The accumulation of H2O2 in amaranth roots increased with the rising of MPs concentration. Compared with the control, a little number of dead cells were found in the roots of amaranth under high MPs treatment. It is noteworthy that only under 100 mg/L PP treatment, the amaranthus seedlings root cells were disorganized, due to the reactive oxygen species (ROS) damage in the roots. These findings provide essential information to assess the phytotoxicity of MPs in agricultural products, and provide insights into the underlying mechanisms of the observed phytotoxicity.

Development of a Functional Cookie Formulated with Chaya (Cnidoscolus aconitifolius (Mill.) I.M. Johnst) and Amaranth (Amaranthus cruentus).

Chaya and amaranth are Mexican traditional foods with a high nutritional value. Many studies have demonstrated the individual beneficial effect of each. However, there is no evidence of the use of these foods on the formulation of functional foods. This study evaluated the effect of replacing 5-20% of wheat flour with chaya and amaranth flours to generate four different formulations of cookies. Proximal analysis, total polyphenols and oxalate content, antioxidant activity, fatty acid profile, and sensory analysis were performed on the cookies. The results of the chemical composition showed that all cookies have a high protein content (9.21-10.10%), an adequate amount of fiber (5.34-6.63%), and a balanced ratio of unsaturated-saturated fatty acids (70:20), and they contain PUFAs (50.4-53.2 g/100 g of fatty acids), especially α-linolenic and oleic acids. All formulations presented antioxidant activity (2540 ± 65.9 to 4867 ± 61.7 Trolox equivalents (µmoles/100 g)) and polyphenols (328-790 mg/100 g); in particular, quercetin was identified in their composition. Results of the sensory analysis indicated that incorporation of chaya and amaranth flour in cookies does not affect the acceptability of the products. The inclusion of traditional foods, such as chaya and amaranth, in cookies enhances their nutritional value and increases the content of bioactive compounds associated with health effects.

Synthesis of Novel α-Trifluoroanisole Derivatives Containing Phenylpyridine Moieties with Herbicidal Activity.

To find novel herbicidal compounds with high activity and broad spectrum, a series of phenylpyridine moiety-containing α-trifluoroanisole derivatives were designed, synthesized, and identified via nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS). Greenhouse-based herbicidal activity assays revealed that compound 7a exhibited > 80% inhibitory activity against Abutilon theophrasti, Amaranthus retroflexus, Eclipta prostrate, Digitaria sanguinalis, and Setaria viridis at a dose of 37.5 g a.i./hm2, which was better than fomesafen. Compound 7a further exhibited excellent herbicidal activity against Abutilon theophrasti and Amaranthus retroflexus in this greenhouse setting, with respective median effective dose (ED50) values of 13.32 and 5.48 g a.i./hm2, both of which were slightly superior to fomesafen (ED50 = 36.39, 10.09 g a.i./hm2). The respective half-maximal inhibitory concentration (IC50) for compound 7a and fomesafen when used to inhibit the Nicotiana tabacum protoporphyrinogen oxidase (NtPPO) enzyme, were 9.4 and 110.5 nM. The docking result of compound 7a indicated that the introduction of 3-chloro-5-trifluoromethylpyridine and the trifluoromethoxy group was beneficial to the formation of stable interactions between these compounds and NtPPO. This work demonstrated that compound 7a could be further optimized as a PPO herbicide candidate to control various weeds.

Involvement of glutamine synthetase 2 (GS2) amplification and overexpression in Amaranthus palmeri resistance to glufosinate.

MAIN CONCLUSION: Amplification and overexpression of the target site glutamine synthetase, specifically the plastid-located isoform, confers resistance to glufosinate in Amaranthus palmeri. This mechanism is novel among glufosinate-resistant weeds. Amaranthus palmeri has recently evolved resistance to glufosinate herbicide. Several A. palmeri populations from Missouri and Mississippi, U.S.A. had survivors when sprayed with glufosinate-ammonium (GFA, 657 g ha-1). One population, MO#2 (fourfold resistant) and its progeny (sixfold resistant), were used to study the resistance mechanism, focusing on the herbicide target glutamine synthetase (GS). We identified four GS genes in A. palmeri; three were transcribed: one coding for the plastidic protein (GS2) and two coding for cytoplasmic isoforms (GS1.1 and GS1.2). These isoforms did not contain mutations associated with resistance. The 17 glufosinate survivors studied showed up to 21-fold increase in GS2 copies. GS2 was expressed up to 190-fold among glufosinate survivors. GS1.1 was overexpressed > twofold in only 3 of 17, and GS1.2 in 2 of 17 survivors. GS inhibition by GFA causes ammonia accumulation in susceptible plants. Ammonia level was analyzed in 12 F1 plants. GS2 expression was negatively correlated with ammonia level (r = - 0.712); therefore, plants with higher GS2 expression are less sensitive to GFA. The operating efficiency of photosystem II (ϕPSII) of Nicotiana benthamiana overexpressing GS2 was four times less inhibited by GFA compared to control plants. Therefore, increased copy and overexpression of GS2 confer resistance to GFA in A. palmeri (or other plants). We present novel understanding of the role of GS2 in resistance evolution to glufosinate.