Seed priming protocols implement incomplete imbibition phases, as well as physical, chemical or biological treatments, to activate pre-germinative metabolism and stress response, thus improving germination performances, seedling establishment and stress tolerance according to agricultural productivity requirements. The dehydration phase following priming treatments represents a critical variable, since an excessively prolonged imbibition (overpriming) impairs desiccation tolerance, compromising seed viability and seedling establishment. Priming protocols generally optimize imbibition-dehydration timing empirically to avoid overpriming. Hence, a better understanding of the dynamics underlying the loss of desiccation tolerance represents a promising route to test and develop efficient and cost-effective priming techniques. In the present work, priming and overpriming conditions were defined to explore the role of desiccation tolerance in seed priming efficiency in the model legume Medicago truncatula. The positive effects of hydropriming and kinetin-mediated hormopriming on germination parameters were screened in combination with conditions of short/prolonged priming and mild/severe overpriming. Biometric analyses highlighted contrasting responses in terms of germination performances and seedling development, while ROS (reactive oxygen species) levels measured during dehydration positively correlate with the loss of desiccation tolerance in early seedlings, suggesting possible applications to monitor priming progression and predict overpriming occurrence.

Re-establishment of desiccation tolerance is essential for the survival of germinated seeds facing water deficit in the soil. The molecular and ultrastructural features of desiccation tolerance maintenance and loss within the nuclear compartment are not fully resolved. In the present study, the impact of desiccation-induced genotoxic stress on nucleolar ultrastructure and ribogenesis was explored along the rehydration−dehydration cycle applied in standard seed vigorization protocols. Primed and overprimed Medicago truncatula seeds, obtained through hydropriming followed by desiccation (dry-back), were analysed. In contrast to desiccation-tolerant primed seeds, overprimed seeds enter irreversible germination and do not survive dry-back.Reactive oxygen species, DNA damage and expression profiles of antioxidant/DNA Damage Response genes were measured, as main hallmarks of the seed response to desiccation stress. Nuclear ultrastructural features were also investigated. Overprimed seeds subjected to dry-back revealed altered rRNA accumulation profiles and up-regulation of genes involved in ribogenesis control. The signal molecule PAP (3′-phosphoadenosine 5′-phosphate) accumulated during dry-back only in primed seeds, as a distinctive feature of desiccation tolerance. The presented results show the molecular and ultrastructural landscapes of the seed desiccation response, including substantial changes in nuclear organization.

The tyrosyl-DNA phosphodiesterase 1 (TDP1) enzyme hydrolyzes the phosphodiester bond between a tyrosine residue and the 3′-phosphate of DNA in the DNA–topoisomerase I (TopI) complex, being involved in different DNA repair pathways. A small TDP1 gene subfamily is present in plants, where TDP1α has been linked to genome stability maintenance, while TDP1β has unknown functions. This work aimed to comparatively investigate the function of the TDP1 genes by taking advantage of the rich transcriptomics databases available for the Arabidopsis thaliana model plant. A data mining approach was carried out to collect information regarding gene expression in different tissues, genetic backgrounds, and stress conditions, using platforms where RNA-seq and microarray data are deposited. The gathered data allowed us to distinguish between common and divergent functions of the two genes. Namely, TDP1β seems to be involved in root development and associated with gibberellin and brassinosteroid phytohormones, whereas TDP1α is more responsive to light and abscisic acid. During stress conditions, both genes are highly responsive to biotic and abiotic treatments in a time- and stress-dependent manner. Data validation using gamma-ray treatments applied to Arabidopsis seedlings indicated the accumulation of DNA damage and extensive cell death associated with the observed changes in the TDP1 genes expression profiles.