Toxin-antitoxin (TA) systems are found in nearly all prokaryotic genomes and usually consist of a pair of co-transcribed genes, one of which encodes a stable toxin and the other, its cognate labile antitoxin. Certain environmental and physiological cues trigger the degradation of the antitoxin, causing activation of the toxin, leading either to the death or stasis of the host cell. TA systems have a variety of functions in the bacterial cell, including acting as mediators of programmed cell death, the induction of a dormant state known as persistence and the stable maintenance of plasmids and other mobile genetic elements. Some bacterial TA systems are functional when expressed in eukaryotic cells and this has led to several innovative applications, which are the subject of this review. Here, we look at how bacterial TA systems have been utilized for the genetic manipulation of yeasts and other eukaryotes, for the containment of genetically modified organisms, and for the engineering of high expression eukaryotic cell lines. We also examine how TA systems have been adopted as an important tool in developmental biology research for the ablation of specific cells and the potential for utility of TA systems in antiviral and anticancer gene therapies.

Bacterial toxin-antitoxin systems usually comprise of a pair of genes encoding a stable toxin and its cognate labile antitoxin and are located in the chromosome or in plasmids of several bacterial species. Chromosomally-encoded toxin-antitoxin systems are involved in bacterial stress responses and activation of the toxins usually leads to cell death or dormancy. Overexpression of the chromosomally-encoded YoeB toxin from the yefM-yoeB toxin-antitoxin locus of the Gram-positive bacterium Streptococcus pneumoniae has been shown to cause cell death in S. pneumoniae as well as E. coli. Induction of a YoeB-GFP fusion protein using a 17-β-estradiol-inducible plant expression system in Arabidopsis thaliana Col 0, was lethal in all T2 progeny. Examination of plants by fluorescent confocal microscopy showed GFP fluorescence in all parts of the leaves at 24 hours after 17-β-estradiol induction, continuing up to plant death. Quantitative RT-PCR analysis revealed that the expression of the yoeB toxin gene peaked at 3 days after induction with 17-β-estradiol, coinciding with the onset of visible effects on the plants. Moreover, we detected DNA laddering in the transgenic plants at 24 hours after yoeB induction, indicative of apoptosis. Expression of the YoeB toxin from Streptococcus pneumoniae is lethal in Arabidopsis. We believe this is the first report of a toxin from a bacterial toxin-antitoxin system functioning in plants. The results presented here mark an important milestone towards the development of a cell ablation based bio-containment strategy, which may be useful for functional studies and for the control of spread of transgenic plants.

Phytoremediation is a green technology; however, very few species of arid environments have been identified as hyperaccumulators and fast growers. Therefore, a greenhouse experiment was performed to evidence the phytoaccumulation potential of Conocarpus erectus, Syzygium cumini, Populus deltoides and Morus alba at the sapling stage. Six-month-old plant saplings were subjected to control (CK; 0 µM) and cadmium treatments (Cd; CdCl₂; 200 µM). The results depicted that plant growth, dry biomass production (leaf and stem) and chl a, b and carotenoid contents decreased significantly in all four species under Cd treatment; however, the lowest decrease was evidenced in Conocarpus erectus. The concentration of hydrogen peroxide and superoxide radical increased significantly in all four species, with the highest increase observed in Morus alba. Osmolytes production, antioxidant enzymes activity (superoxide dismutase, peroxidase, catalase and ascorbate peroxidase) and Cd accumulation in the leaves, stem and root increased significantly in all four species under Cd treatment, with the highest increase observed in Conocarpus erectus. The translocation factor was >1 in Conocarpus erectus, Syzyngoim cumini and Populus deltoides and was <1 in Morus alba. The study revealed a better Cd tolerance in Conocarpus erectus, which was driven by the effective osmolyte balance and antioxidant enzymes mechanism.

Bacterial toxin-antitoxin (TA) systems have various cellular functions, including as part of the general stress response. The genome of the Gram-positive human pathogen Streptococcus pneumoniae harbors several putative TA systems, including yefM-yoeB_Spn, which is one of four systems that had been demonstrated to be biologically functional. Overexpression of the yoeB_Spn toxin gene resulted in cell stasis and eventually cell death in its native host, as well as in Escherichia coli. Our previous work showed that induced expression of a yoeB_Spn toxin-Green Fluorescent Protein (GFP) fusion gene apparently triggered apoptosis and was lethal in the model plant, Arabidopsis thaliana. In this study, we investigated the effects of co-expression of the yefM_Spn antitoxin and yoeB_Spn toxin-GFP fusion in transgenic A. thaliana. When co-expressed in Arabidopsis, the YefM_Spn antitoxin was found to neutralize the toxicity of YoeB_Spn-GFP. Interestingly, the inducible expression of both yefM_Spn antitoxin and yoeB_Spn toxin-GFP fusion in transgenic hybrid Arabidopsis resulted in larger rosette leaves and taller plants with a higher number of inflorescence stems and increased silique production. To our knowledge, this is the first demonstration of a prokaryotic antitoxin neutralizing its cognate toxin in plant cells.