Chloroplasts are organelles subjected to extreme oxidative stress conditions. Biomolecules produced in the chloroplasts act as signals guiding plant metabolism toward stress tolerance and play a major role in regulating gene expression in the nucleus. Herein, we used transplastomic plants as an alternative approach to expression of transgenes in the nucleus for conferring stress tolerance to abiotic stresses and herbicides. To investigate the morphophysiological and molecular mechanisms and the role of plastid expressed GSTs in tobacco stress detoxification and stress tolerance, we used transplastomic tobacco lines overexpressing a theta class glutathione transferase (GST) in chloroplasts. The transplastomic plants were tested under drought (0, 100, and 200 mM mannitol) and salinity (0, 150, and 300 mM NaCl) in vitro, and under herbicide stress (Diquat). Our results suggest that ptAtGSTT lines were tolerant to herbicide-induced oxidative and salinity stresses and showed enhanced response tolerance to mannitol-induced osmotic stress compared to WT plants. Overexpression of the Arabidopsis thaliana AtGSTT in the chloroplasts resulted in enhanced photo-tolerance and turgor maintenance under stress. Whole-genome transcriptome analysis revealed that genes related to stress tolerance, were upregulated in ptAtGSTT2a line under both control and high mannitol stress conditions. Transplastomic plants overexpressing the ptAtGSTT2a in the chloroplast showed a state of acclimation to stress, as only limited number of genes were upregulated in the ptAtGSTT2a transplastomic line compared to WT under stress conditions while at the same time genes related to stress tolerance were upregulated in ptAtGSTT2a plants compared to WT in stress-free conditions. In parallel, the metabolic profile indicated limited perturbations of the metabolic homeostasis in the transplastomic lines and greater accumulation of mannitol, and soluble sugars under high mannitol stress. Therefore, transplastomic lines seem to be in a state of acclimation to stress under stress-free conditions, which was maintained even under high mannitol stress. The results help to elucidate the role of GSTs in plant abiotic stress tolerance and the underlying mechanisms of the GSTs expressed in the chloroplast, toward environmental resilience of cultivated crops.

Under the era of climate change, plants are forced to survive under increasingly adverse conditions. Application of biostimulants in plants is shown to mitigate the deleterious effects of abiotic stresses including salinity, enhancing plant tolerance and performance. The present study focuses on the effects of five biostimulants based on biocompost and biofertilizer compounds that have been applied to tomato plants grown in the presence (salt-stressed plants) or absence of salt stress (control plants). To study the beneficial effects of the biostimulants in tomato plants, a series of analyses were performed, including phenotypic and agronomic observations, physiological, biochemical and enzymatic activity measurements, as well as gene expression analysis (RT-qPCR) including genes involved in antioxidant defense (SlCu/ZnSOD, SlFeSOD, SlCAT1, SlcAPX), nitrogen (SlNR, SlNiR, SlGTS1) and proline metabolism (p5CS), potassium transporters (HKT1.1, HKT1.2), and stress-inducible TFs (SlWRKY8, SlWRKY31). Among all the biostimulant solutions applied to the plants, the composition of 70% biofertilizer and 30% biocompost (Bf70/Bc30) as well as 70% biocompost and 30% biofertilizer (Bc70/Bf30) formulations garnered interest, since the former showed growth promoting features while the latter displayed better defense responses at the time of harvesting compared with the other treatments and controls. Taken together, current findings provide new insight into the beneficial effects of biostimulants, encouraging future field studies to further evaluate the biostimulant effects in plants under a real environment which is compromised by a combination of abiotic and biotic stresses.

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