In-vitro toxicity assessment of Eucalyptus robusta Smith extracts via whole-cell bioreporter

Eucalyptus is widely planted in China for wood industries, and there are increasing concerns about its ecotoxicity in the environment. This study explored the in-vitro toxicity of Eucalyptus extracts by assessing the impacts of water-soluble and dimethylsulfoxide (DMSO)-soluble fractions via a whole-cell bioreporter, Acinetobacter baylyi ADPWH_recA. Compounds identified in Eucalyptus extracts included one tannin, two phenolic acids, four terpenoids, four glycosides, and five flavonoids. The leaf extracts contained more biological-active components than barks and roots. Genotoxicity induced by Eucalyptus extracts was mainly associated with water extracts (e.g., flavonoids, phenolic acids) instead of DMSO extracts. The significant cytotoxicity was explained by programmed cell death (PCD), suggested by the results of propidium iodide (PI) and 2',7'-dichlorofluorescein-diacetate (DCFH-DA) assays. Generally, water-soluble fractions contributed more toxicities than DMSO-soluble fractions, particularly at high concentrations. A robust linear regression was built between the compromised toxicity and PCD index (Compromised toxicity = -2.192 × PCD index + 2.219; R² = 0.8886), suggesting a PCD-dependent compromised toxicity which was greatly underestimated. Our results implied non-neglectable ecotoxicological risks of Eucalyptus extracts, hinting at the possible magnified ecological impacts of its large-scale plantation and the potential adverse outcomes to the surrounding ecosystems.