Combined pesticides in field doses weaken honey bee (Apis cerana F.) flight ability and analyses of transcriptomics and metabolomics.

Imidacloprid, chlorpyrifos, and glyphosate rank among the most extensively employed pesticides worldwide. The effects of these pesticides and their combined on the flight capability of Apis cerana, and the potential underlying mechanisms remain uncertain. To investigate these effects, we carried out flight mill, transcriptome, and metabolome experiments. Our findings reveal that individual acute oral treatments with pesticides, specifically 20 µL of 10 ng/g imidacloprid (0.2 ng per bee), 30 ng/g chlorpyrifos (0.6 ng per bee), and 60 ng/g glyphosate (1.2 ng per bee), did not impact the flight capability of the bees. However, when bees were exposed to a combination of two or three pesticides, a notable reduction in flight duration and distance was observed. In the transcriptomic and metabolomic analyses, we identified 307 transcripts and 17 metabolites that exhibited differential expression following exposure to combined pesticides, primarily associated with metabolic pathways involved in energy regulation. Our results illuminate the intricate effects and potential hazards posed by combined pesticide exposures on bee behavior. These findings offer valuable insights into the synergistic potential of pesticide combinations and their capacity to impair bee behavior. Understanding these complex interactions is essential for comprehending the broader consequences of pesticide formulations on honey bee populations.

Multi-omics analysis identifies EcCS4 is negatively regulated in response to phytotoxin isovaleric acid stress in Echinochloa crus-galli.

BACKGROUND: Knowledge of herbicidal targets is critical for weed management and food safety. The phytotoxin isovaleric acid (ISA) is effective against weeds with a broad spectrum, carries low environmental risks, and is thus an excellent herbicide lead. However, the biochemical and molecular mechanisms underlying the action of ISA remain unclear. RESULTS: Multi-omics data showed that acetyl coenzyme A (acetyl-CoA) was the key affected metabolite, and that citrate synthase (CS) 4 was substantially down-regulated under ISA treatment in Echinochloa crus-galli leaves. In particular, the transcript level of EcCS4 was the most significantly regulated among the six genes involved in the top 10 different pathways. The EcCS4 encodes a protein of 472 amino acids and is localized to the cell membrane and mitochondria, similar to the CS4s of other plants. The protein content of EcCS4 was down-regulated after ISA treatment at 0.5 h. ISA markedly inhibited the CS4 activity in vitro in a concentration-dependent manner (IC50 = 41.35 µM). In addition, the transgenic rice plants overexpressing EcCS4 (IC50 = 111.8 mM for OECS4-8 line) were more sensitive, whereas loss-of-function rice mutant lines (IC50 = 746.5 mM for oscs4-19) were more resistant to ISA, compared to wild type (WT) plants (IC50 = 355.6 mM). CONCLUSION: CS4 was first reported as a negative regulator of plant responses to ISA. These results highlight that CS4 is a candidate target gene for the development of novel herbicides and for breeding herbicide-resistant crops. © 2023 Society of Chemical Industry.

Inhibitory activities of essential oils from Syzygium aromaticum inhibition of Echinochloa crus-galli.

Echinochloa crus-galli is a serious weed species in rice paddies. To obtain a new potential bioherbicide, we evaluated the inhibitory activities of 13 essential oils and their active substances against E. crus-galli. Essential oil from Syzygium aromaticum (L.) Merr. & L. M. Perry (SAEO) exhibited the highest herbicidal activity (EC50 = 3.87 mg mL-1) among the 13 essential oils evaluated. The SAEO was isolated at six different temperatures by vacuum fractional distillation, including 164°C, 165°C (SAEO-165), 169°C, 170°C 175°C and 180°C. The SAEO-165 had the highest inhibitory rate against E. crus-galli. Gas chromatography-mass spectrometry and high phase liquid chromatography identified eugenol (EC50 = 4.07 mg mL-1), α-caryophyllene (EC50 = 17.34 mg mL-1) and ß-caryophyllene (EC50 = 96.66 mg mL-1) as the three compounds in SAEO. Results from a safety bioassay showed that the tolerance of rice seedling (~ 20% inhibition) was higher than that of E. crus-galli (~ 70% inhibition) under SAEO stress. SAEO induced excessive generation of reactive oxygen species leading to oxidative stress and ultimately tissue damage in E. crus-galli. Our results indicate that SAEO has a potential for development into a new selective bio-herbicide. They also provide an example of a sustainable management strategy for E. crus-galli in rice paddies.

Design, Synthesis, and Herbicidal Activities of N-(5-(3,5-Methoxyphenyl)-(thiazole-2-yl))phenoxyacetamide Derivatives.

Thiazole and phenoxyacetic acid are key moieties in many natural and synthetic biologically active agents. A series of N-(5-(3,5-methoxyphenyl)-(thiazole-2-yl))phenoxyacetamide derivatives 6an-6bd were designed and synthesized, and their structures were confirmed by NMR and HRMS. Most of derivatives exhibited superior inhibition of Echinochloa crusgalli (E.c.) and Lactuca sativa (L.s.) seed germination by the Petri dish bioassay. Indeed, herbicidal bioassays indicated that 6an (2-(2,4-dichlorophenoxy)-N-(5-(3,5-dimethoxyphenyl)-1,3,4-thiadiazol-2-yl)acetamide) had the best inhibition against L.s. (IC50 = 42.7 g/ha, 375 g/ha at field experiments). 6an also had no harmful effect on Zea mays at 2- to 4-fold field usage. Moreover, transcriptomics and metabolomics analysis showed that 6an significantly influenced cell metabolism, including galactose metabolism and ascorbate and aldarate metabolism. These discoveries highlight that 6an shows promise to be developed as a potential herbicide.

Phytochemical profiling and allelopathic effect of garlic essential oil on barnyard grass (Echinochloa crusgalli L.).

In agriculture, barnyard grass (Echinochloa crusgalli L.) is one of the most harmful weeds in rice fields now. In order to identify active ingredients which had inhibiting effect on barnyard grass (Echinochloa crusgalli L.), we evaluated several possible natural plant essential oils. Essential oils from twelve plant species showed inhibitory activity against barnyard grass seedlings and root length. The garlic essential oil (GEO) had the most significant allelopathic effect (EC50 = 0.0126 g mL-1). Additionally, the enzyme activities of catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD) increased during the first 8 hours of treatment at a concentration of 0.1 g mL-1 and then declined. The activities of CAT, SOD and POD increased by 121%, 137% and 110% (0-8h, compared to control), and decreased (8-72h, compared to the maximum value) by 100%, 185% and 183%, respectively. The total chlorophyll content of barnyard grass seedlings decreased by 51% (0-72h) continuously with the same dosage treatment. Twenty constituents of GEO were identified by gas chromatography-mass spectrometry, and the herbicidal activity of two main components (diallyl sulfide and diallyl disulfide) was evaluated. Results showed that both components had herbicidal activity against barnyard grass. GEO had a strong inhibitory effect (~88.34% inhibition) on barnyard grass growth, but safety studies on rice showed it did not have much inhibitory effect on rice seed germination. Allelopathy of GEO provide ideas for the development of new plant-derived herbicides.

A first-in-class dimethyl 2-acetamido terephthalate inhibitor targeting Conyza canadensis SHMT1 with a novel herbicidal mode-of-action.

INTRODUCTION: Herbicide application is a highly efficiency method of weed control that boots agricultural output and assures food security. The development of novel herbicides focuses on improved bioactivity and new modes of action. The amino acid biosynthesis was validated as a promising novel mode of action for herbicidal compounds. However, the amino acid biosynthesis enzyme remains largely unexplored for herbicidal targets. OBJECTIVES: Serine hydroxymethyl transferase (SHMT) is an essentialenzyme in the photorespiratory cycle. The study aims to explore Conyza canadensis SHMT1 (CcSHMT1) as a promising target for herbicide discovery. METHODS: Structure determination of CcSHMT1 was resolved by X-ray crystallography. Virtual screening docking experiments were performed with Glide version 5.5. Novel derivatives of dimethyl 2-acetamido terephthalate were further designed, synthesized, and bioassay. The druggability of the inhibitor was evidenced by ultrastructural changes in mitochondria, in vivo and vitro enzyme activity assays, and genetics analysis. RESULTS: CcSHMT1 has a typical PLP-dependent enzyme 3D structure. The dimethyl 2-acetamido terephthalate-containing compounds had herbicidal activity. Dimethyl 2-(2-(4-(2-(4-bromo-2-chlorophenoxy) acetyl)piperazin-1-yl)acetamido) terephthalate (Compound 9ay, EC50 = 193.8 g a.i./ ha) exhibited the highest herbicidal activity on tested weed among the synthesized compounds. Compound 9ay had no obvious adverse effect on the growth of maize and honeybees. Compound 9ay was verified to target CcSHMT1 as an herbicide candidate. CONCLUSION: A first-in-class CcSHMT1 inhibitor that could be developed as a potent herbicide with a new mode of action and provide an avenue for discovering novel inhibitors of pyridoxal-5-phosphate-dependent enzymes.

Effects of novel bioorganic fertilizer application on soil enzymes and bacterial community in multi-site rice paddies in China.

Application of a novel bioorganic fertilizer (BIO) has been effectively used to inhibit weeds in rice paddies. To identify changes in soil bacterial community and enzymes in response to BIO treatments, field experiments were carried out in five major rice-growing areas in China. The dominant phylogenetic groups recorded included Proteobacteria, Actinobacteria, Bacteroidetes, Firmicutes and Acidobacteria. Anaeromyxobacter, Bacteroides, Bifidobacterium, Escherichia- Shigella, Geobacter and Haliangium were significantly different between BIO-treatment and untreated control and aided in general function (R), amino acid transport, metabolism (E) and transcription (K) clusters. The soil chemical properties and enzyme activities were less affected by BIO at these study sites. RDA analysis showed that soil bacterial community had a significant positive correlations among northern latitude, eastern longitude, exchangeable K, total K, total P, soil pH, and total N, except for organic matter, hydrolytic N and extractable P. Overall, our work showed that application of BIO does not alter the main community structure and functional diversity of soil bacteria in rice paddies and should be encouraged for use as a sustainable weed management strategy.