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Transcriptomic Responses of Mycoplasma bovis upon Treatments of trans-Cinnamaldehyde, Carvacrol, and Eugenol

ABSTRACT: Through RNA sequencing, we report here that Carvacrol (CAR) affected the expression of 153 genes which included the down-regulation of energy generation-related proteins, pentose phosphate pathway, and up-regulation of ribosomes and translation-related proteins. Few differentially expressed genes were found when M. bovis were treated with Trans-cinnamaldehyde (TC), Eugenol (EU), or Carvacrol (CAR) when the three PDAs were double or triple combined. Our results suggest that, as opposed to the effect of CAR, the growth inhibitory effects of TC and EU at levels tested may be exerted through mechanisms other than gene expression regulations.

ORGANISM(S): Mycoplasmopsis bovis PG45

PROVIDER: GSE198086 | GEO | 2022/06/09

REPOSITORIES: GEO

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Project description:Antimicrobial resistance pose a global thread nowadays. Compounds of natural origin are an important source of drugs used in clinical practice. However, it is important to understand both their principles of efficacy and their molecular mechanism of action. In this study we evaluated antimicrobial potential of t-cinnamaldehyde which is an organic compound found in many plant species, especially in the Cinnamomum genus, such as Cinnamomum zeylanicum and cassia. Cinnamon oil extracted from the bark of these plants contains up to 80% trans-cinnamaldehyde. Although CNMA has shown antimicrobial properties against numerous Gram+ and Gram- species, its mode of action against pathogens remains not fuly elucidated. Therefore, this project aims to determine CNMA activity at the level of gene expression. Total RNA was isolated and checked for quality using the Bioanalyzer 2100. The sequencing run was conducted on the Illumina NovaSeq6000 platform. 30 million pair-end reads per samples were assessed with 101 pb read length. Reference E. coli MG1655 genome sequence and annotations were downloaded from GenBank. Differentially expressed analysis of 0.25 x MIC CNMA was performed against untreated control in indicated time with p ≤ 0.001 and log2FC ≥ 1.5. We have discovered many changes bacterial transcriptome. For instance: following the treatment with 0.25×MIC of CNMA, we found 292 and 140 upregulated and 107 and 96 downregulated genes at time points 30 and 60 min, respectively. Among the most enriched genes, were those related to the tricarboxylic acid (TCA) cycle, flagellum synthesis, amino acid transport, and oxidoreductase activity. According to these findings we can conclude that observed transcriprional pattern indicates severe metabolic downshift in treated cells, and consequently activation of stress processes. These was in line with our secondary experiments which revealed drop in growth kinnetic, cytoplasm shrinkage, NAD/NADH level alteration and elevation of stringent response alarmones ((p)ppGpp). Taken together, this suggests that CNMA-treated E. coli bacteria undergo major metabolic changes that finally result in cell death.

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Transcriptomic Responses of Mycoplasma bovis upon Treatments of trans-Cinnamaldehyde, Carvacrol, and Eugenol

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