In this work, the involvement of heat shock proteins (HSP70) in barley (Hordeum vulgare) has been studied in response to drought and salinity. Thus, 3 barley genotypes usually cultivated and/or selected in Italy, 3 Middle East/North Africa landraces and genotypes and 1 improved genotype from ICARDA have been studied to identify those varieties showing the best stress response. Preliminarily, a bioinformatic characterization of the HSP70s protein family in barley has been made by using annotated Arabidopsis protein sequences. This study identified 20 putative HSP70s orthologs in the barley genome. The construction of un-rooted phylogenetic trees showed the partition into four main branches, and multiple subcellular localizations. The enhanced HSP70s presence upon salt and drought stress was investigated by both immunoblotting and expression analyses. It is worth noting the Northern Africa landraces showed peculiar tolerance behavior versus drought and salt stresses. The drought and salinity conditions indicated the involvement of specific HSP70s to counteract abiotic stress. Particularly, the expression of cytosolic MLOC_67581, mitochondrial MLOC_50972, and encoding for HSP70 isoforms showed different expressions and occurrence upon stress. Therefore, genotypes originated in the semi-arid area of the Mediterranean area can represent an important genetic source for the improvement of commonly cultivated high-yielding varieties.

The interaction between salinity and nitrogen metabolism has been investigated in two barley landraces, one tolerant (“100/1B”) and one susceptible to salinity (“Barley medenine”) from the Middle East and North Africa (MENA) region. Barley plants were exposed to 50 mM NaCl for 7 days; then, salinity was increased to 150 mM NaCl in the presence (10 mM) or limitation (1 mM) of ammonium as a nitrogen source. Upon salinity, “100/1B” was shown to support N assimilation by enhancing the glutamine synthetase (GS) and glutamine oxoglutarate aminotransferase (GOGAT) cycle under high N, and the stimulation of the glutamate dehydrogenase (GDH) pathway under low N treatment. In “Barley medenine”, salinity reduced the GS/GOGAT cycle, and increased GDH activity. Upon salinity, Heat Shock Proteins 70 and PEPC remained unchanged in “100/1B”, while they decreased in “Barley medenine”. The tolerance degree is a determining factor in enzymes’ occurrence and regulation: exposed to salinity, “100/1B” rapidly increased APX and PEPC activities, while this was delayed in “Barley medenine”. Salinity increased cyt-G6PDH levels in “100/1B”, while “Barley medenine” showed a decrease in G6PDH isoforms. Correlation analyses confirm GOGAT was related to G6PDH; GDH and APX with PEPC in “100/1B” under moderate salinity; severe salinity correlated GDH with G6PDH and PEPC. In “Barley medenine” under salinity, GOGAT was correlated with G6PDH, while APX showed a relation with PEPC. Therefore, specific enzymatic activities and occurrence can be used to determine stress responsiveness of different landraces. We suggest that the rapid increase in G6PDH, APX, and nitrogen assimilation enzymes activities represents an index of tolerance in “100/1B” and a stress symptom in “Barley medenine”.

This study aims to investigate the activities and expression of enzymes of primary metabolism and relate these data with the growth performance of three different durum wheat genotypes (Maali; YT13; and ON66) under osmotic stress. Growth traits—including plant height, dry weight (DW) and relative water content (RWC)—were measured to classify genotypes depending on their tolerance to stress. Several enzymes were investigated: Ascorbate peroxidase (APX), Glutamine Synthetase (GS), Glutamine dehydrogenase (GDH), Glutamate synthase (GOGAT), Glucose 6-phosphate dehydrogenase (G6PDH), and Phosphoenolpyruvate Carboxylase (PEPC). The expression of the cytosolic and plastidic glutamine synthetase (TaGS1 and TaGS2), high affinity nitrate transporters (TaNRT2.3) and Glutamate dehydrogenase (TaGDH) were also detected by qRT-PCR. The results indicated different growth performances among genotypes, indicating Maali and YT13 as tolerant genotypes and ON66 as a drought-susceptible variety. Data showed a decrease in PEPC and increase in APX activities under osmotic stress; a slight decrease in GS activity was observed, together with an increase in G6PDH in all genotypes; GS and NRT2 expressions changed in a similar pattern in the different genotypes. Interestingly, Maali and YT13 showed higher transcript abundance for GDH under stress compared to ON66, suggesting the implication of GDH in protective phenomena upon osmotic stress.

Nitrogen (N) is a crucial nutrient for plant growth and development. To optimize agricultural environments, N fertilizers represent a critical tool to regulate crop productivity. The improvement of nitrogen use efficiency (NUE) represents a promising tool that may enable cereal production to meet future food demand. Wheat reported contrasting behaviors in N utilization showing specific abilities depending on genotype. This study selected two landraces and two improved genotypes from Northern Africa to investigate grain yield (GY), grain protein content (GPC) and NUE. Plants were grown under three levels of N supply: 0, 75, 150 kg N ha⁻¹ and for two consecutive years. Results reported a better NUE (0.40 kg kg N⁻¹) obtained under 150 kg N ha⁻¹, while N utilization efficiency (NUtE) showed a 13% increase using 75 kg N ha⁻¹ compared with 150 kg N ha⁻¹. Under low nitrogen rate (0 N), crop N supply (CNS) and N uptake efficiency (NUpE) were shown as determinant factors for improved genotypes GY (R² = 0.72), while NUtE represented the most determinant component for GPC in landraces (R² = 0.92). Multivariate regression models explained the dependence in GPC on NUE, NUpE, and NUtE. In conclusion, our results recognize GPC and NUtE as suitable selection traits to identify durum wheat with higher NUE.