MicroRNAs (miRNAs) have recently emerged as important regulators of gene expression in plants. Many miRNA families and their targets have been extensively studied in model species and major crops. We have characterized mature miRNAs along with their precursors and potential targets in Hypericum to generate a comprehensive list of conserved miRNA families and to investigate the regulatory role of selected miRNAs in biological processes that occur in the flower. St. John's wort (Hypericum perforatum L., 2n = 4x = 32), a medicinal plant that produces pharmaceutically important metabolites with therapeutic activities, was chosen because it is regarded as an attractive model system for the study of apomixis. A computational in silico prediction of structure, in combination with an in vitro validation, allowed us to identify 7 pre-miRNAs, including miR156, miR166, miR390, miR394, miR396, and miR414. We demonstrated that H. perforatum flowers share highly conserved miRNAs and that these miRNAs potentially target dozens of genes with a wide range of molecular functions, including metabolism, response to stress, flower development, and plant reproduction. Our analysis paves the way toward identifying flower-specific miRNAs that may differentiate the sexual and apomictic reproductive pathways.

Common wheat grains are characterised by low concentrations of Ca, K, and Mg, which can be partially removed with the bran during milling processes. This preliminary study investigated the effects of foliar fertilisation at the earing stage with nitrates of Ca, Mg, and K contemporarily, together with a small amount of urea and protein hydrolysate as potential carriers, in two contrasting common wheat varieties, i.e., Solehio (medium proteins content) and Vivendo (high proteins content). Based on the preliminary grain-to-straw concentration ratio of these minerals, two biofortification targets were applied in order to increase their grain contents by +20% and +40%, in comparison with untreated controls. Here, we demonstrate that the highest fertilisation dose was effective in increasing grain K by 13% and Mg by 16% in Vivendo, and Ca by 7% in Solehio, with no boosting effects of the co-formulants urea and protein hydrolysate. In addition to some qualitative benefits due to nitrates supply, negligible phytotoxicity symptoms were observed, as revealed by the NDVI vegetational index dynamics. Although the biofortification target was not fully achieved, this study firstly reports the possibility to increase at the same time Mg and K, and to a lower extent Ca in wheat grains. It is concluded that efficient multiple biofortification should consider a variety-depend response, while further studies are necessary to investigate the effects of different fertilisation timings and doses for improving the poor mineral translocation to the grains.

Micronutrient malnutrition affects more than half of the world population. Reduced bioavailability of microelements in the raw materials is considered one of the main causes of mineral deficiency in populations whose diet is largely based on the consumption of staple crops. In this context, the production of low phytic acid (lpa) cereals is a main goal of the breeding programs, as phytic acid (PA) binds essential mineral cations such as iron (Fe), zinc (Zn), manganese (Mn), potassium (K), calcium (Ca) and magnesium (Mg) precipitating in the form of phytate salts poorly digested by monogastric animals, including humans, due to the lack of phytases in the digestive tract. Since PA limits the bioavailability of microelements, it is widely recognized as an anti-nutritional compound. A Targeting Induced Local Lesions IN Genomes (TILLING) approach has been undertaken to silence the genes encoding the TdABCC13 proteins, known as Multidrug-Resistance associated Proteins 3 (TdMRP3), transporters involved in the accumulation of PA inside the vacuole in durum wheat. The TdMRP3 complete null genotypes showed a significant reduction in the content of PA and were able to accumulate a higher amount of essential micronutrients (Fe, Zn, Mn) compared to the control. The number of spikelets and seeds per spike, traits associated with the agronomic performances, were reduced compared to the control, but the negative effect was in part balanced by the increased grain weight. The TdMRP3 mutant lines showed morphological differences in the root apparatus such as a significant decrease in the number of root tips, root length, volume and surface area and an increase in root average diameter compared to the control plants. These materials represent a promising basis for obtaining new commercial durum wheats with higher nutritional value.