Fenbendazole, a broad spectrum anthelmintic used especially in veterinary medicine, may impact non-target organisms in the environment. Nevertheless, information about the effects of fenbendazole in plants is limited. We investigated the biotransformation of fenbendazole and the effect of fenbendazole and its metabolites on gene expression in the model plant Arabidopsis thaliana. High-sensitive UHPLC coupled with tandem mass spectrometry, RNA-microarray analysis together with qPCR verification and nanoLC-MS proteome analysis were used in this study. Twelve fenbendazole metabolites were identified in the roots and leaves of A. thaliana plants. Hydroxylation, S-oxidation and glycosylation represent the main fenbendazole biotransformation pathways. Exposure of A. thaliana plants to 5 μM fenbendazole for 24 and 72 h significantly affected gene and protein expression. The changes in transcriptome were more pronounced in the leaves than in roots, protein expression was more greatly affected in the roots at a shorter period of exposure (24 h) and in leaf rosettes over a longer period (72 h). Up-regulated (>2-fold change, p < 0.1) proteins are involved in various biological processes (electron transport, energy generating pathways, signal transduction, transport), and in response to stresses (e.g. catalase, superoxide dismutase, cytochromes P450, UDP-glycosyltransferases). Some of the proteins which were up-regulated after fenbendazole-exposure probably participate in fenbendazole biotransformation (e.g. cytochromes P450, UDP-glucosyltransferases). Finally, fenbendazole in plants significantly affects many physiological and metabolic processes and thus the contamination of ecosystems by manure containing this anthelmintic should be restricted.

• Klíčová slova
• Arabidopsis thaliana, Fenbendazole, Genomics, Metabolism, Proteomics, Veterinary drugs,
• MeSH
• anthelmintika metabolismus   MeSH
• Arabidopsis účinky léků   metabolismus   MeSH
• fenbendazol metabolismus   farmakokinetika   MeSH
• kořeny rostlin metabolismus   MeSH
• listy rostlin metabolismus   MeSH
• proteiny huseníčku účinky léků   metabolismus   MeSH
• proteom účinky léků   metabolismus   MeSH
• proteomika metody   MeSH
• regulace genové exprese u rostlin MeSH
• transkriptom účinky léků   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• anthelmintika MeSH
• fenbendazol MeSH
• proteiny huseníčku MeSH
• proteom MeSH

Arabidopsis (Arabidopsis thaliana) SPINDLY (SPY) is a putative serine and threonine O-linked N-acetylglucosamine transferase (OGT). While SPY has been shown to suppress gibberellin signaling and to promote cytokinin (CK) responses, its catalytic OGT activity was never demonstrated and its effect on protein fate is not known. We previously showed that SPY interacts physically and functionally with TCP14 and TCP15 to promote CK responses. Here, we aimed to identify how SPY regulates TCP14/15 activities and how these TCPs promote CK responses. We show that SPY activity is required for TCP14 stability. Mutation in the putative OGT domain of SPY (spy-3) stimulated TCP14 proteolysis by the 26S proteasome, which was reversed by mutation in CULLIN1 (CUL1), suggesting a role for SKP, CUL1, F-box E3 ubiquitin ligase in TCP14 proteolysis. TCP14 proteolysis in spy-3 suppressed all TCP14 misexpression phenotypes, including the enhanced CK responses. The increased CK activity in TCP14/15-overexpressing flowers resulted from increased sensitivity to the hormone and not from higher CK levels. TCP15 overexpression enhanced the response of the CK-induced synthetic promoter pTCS to CK, suggesting that TCP14/15 affect early steps in CK signaling. We propose that posttranslational modification of TCP14/15 by SPY inhibits their proteolysis and that the accumulated proteins promote the activity of the CK phosphorelay cascade in developing Arabidopsis leaves and flowers.

• MeSH
• cytokininy farmakologie   MeSH
• katalytická doména MeSH
• proteasomový endopeptidasový komplex metabolismus   MeSH
• proteiny huseníčku chemie   účinky léků   metabolismus   MeSH
• proteolýza účinky léků   MeSH
• represorové proteiny chemie   metabolismus   MeSH
• stabilita proteinů MeSH
• transkripční faktory metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytokininy MeSH
• proteasomový endopeptidasový komplex MeSH
• proteiny huseníčku MeSH
• represorové proteiny MeSH
• SPY protein, Arabidopsis MeSH Prohlížeč
• TCP14 protein, Arabidopsis MeSH Prohlížeč
• TCP15 protein, Arabidopsis MeSH Prohlížeč
• transkripční faktory MeSH