Anti-GST tag antibody [Agarose]

BACKGROUND Non-target-site resistance (NTSR) to herbicides is a serious threat to global agriculture. Although metabolic resistance is the dominant mechanism of NTSR, the molecular mechanisms are not yet well-characterized. This study aimed to uncover the likely metabolism-related genes inBeckmannia syzigachne(American sloughgrass) resistant to fenoxaprop-p-ethyl. RESULTS Ultra-performance liquid chromatography - tandem mass spectrometry experiments showed that the resistant American sloughgrass biotype (R, SD-04-SS) showed enhanced degradation of this herbicide compared to the susceptible biotype (S, SD-12). R and S biotype were harvested at 24 h after fenoxaprop-p-ethyl treatment to conduct RNA sequencing (RNA-Seq) analysis to investigate the likely fenoxaprop-p-ethyl metabolic genes. The RNA-Seq libraries yield 417 969 980 clean reads. Thede novoassembly generated 115 112 unigenes, of which 57 906 unigenes were annotated. Finally, we identified 273 cytochrome P450s, 178 oxidases, 47 glutathione S-transferases (GSTs), 166 glucosyltransferases (GTs) and 180 ABC transporter genes to determine the likely fenoxaprop-p-ethyl metabolism-related genes in R biotype. Twelve overlapping up-regulated genes in the R biotype (fenoxaprop-p-ethyl-treated R/non-treated R, fenoxaprop-p-ethyl-treated R/fenoxaprop-p-ethyl-treated S) were identified by RNA-Seq and the results were validated using qRT-PCR. Ten were identified as fenoxaprop-p-ethyl metabolism-related genes, including three P450s (homologous toCYP71D7,CYP99A2andCYP71D10), one GST (homologous toGSTF1), two GTs (homologous toUGT90A1andUGT83A1) and four oxidase genes. CONCLUSION This work demonstrates that the NTSR mechanism by means of enhanced detoxification of fenoxaprop-p-ethyl in American sloughgrass is very likely driven by herbicide metabolism related genes. The RNA-Seq data presented here provide a valuable resource for understanding the molecular mechanism of NTSR in American sloughgrass. (c) 2020 Society of Chemical Industry

The development of new models to study diabetes in invertebrates is important to ensure adherence to the 3R's principle and to expedite knowledge of the complex molecular events underlying glucose toxicity. Streptozotocin (STZ)-an alkylating and highly toxic agent that has tropism to mammalian beta cells-is used as a model of type 1 diabetes in rodents, but little is known about STZ effects in insects. Here, the cockroach; Nauphoeta cinerea was used to determine the acute toxicity of 74 and 740 nmol of STZ injection per cockroach. STZ increased the glucose content, mRNA expression of glucose transporter 1 (GLUT1) and markers of oxidative stress in the head. Fat body glycogen, insect survival, acetylcholinesterase activity, triglyceride content and viable cells in head homogenate were reduced, which may indicate a disruption in glucose utilization by the head and fat body of insects after injection of 74 and 740 nmol STZ per nymph. The glutathione S-transferase (GST) activity and reduced glutathione levels (GSH) were increased, possibly via activation of nuclear factor erythroid 2 related factor as a compensatory response against the increase in reactive oxygen species. Our data present the potential for metabolic disruption in N. cinerea by glucose analogues and opens paths for the study of brain energy metabolism in insects. We further phylogenetically demonstrated conservation between N. cinerea glucose transporter 1 and the GLUT of other insects in the Neoptera infra-class.