New insights into N-utilization efficiency in tomato (Solanum lycopersicum L.) under N limiting condition.

Understanding Nitrogen Use Efficiency (NUE) physiological and molecular mechanisms in high N demanding crops has become decisive for improving NUE in sustainable cropping systems. How the Nitrogen Utilization Efficiency (NUtE) component contributes to the NUE enhancement under nitrate limiting conditions in tomato remains to be elucidated. This study deals with the changes in several important nitrate metabolism related gene expressions (nitrate assimilation, transport, remobilization and storage/sequestration) engendered by short and long-term limiting nitrate exposure in two selected NUE-contrasting genotypes, Regina Ostuni (RO) and UC82, efficient and inefficient, respectively. At short-term, nitrate limiting supply triggered higher SlCLCa and SlNRT1.7 expressions in RO root and shoot, respectively, suggesting a higher nitrate storage and remobilization compared to UC82, explaining how RO withstood the nitrate deficiency better than UC82. At long-term, nitrate reductase (SlNR) and nitrite reductase (SlNIR) expression were not significantly different between nitrate treatments in RO, while significantly down-regulated under nitrate limiting treatment in UC82. In addition, SlCLCa and SlNRT1.8 transcript levels were significantly lower in RO, while those of SlNRT1.5 and SlNR appeared significantly higher. This suggested that the efficient genotype stored less nitrate compared to UC82, which was allocated and assimilated to the shoot. More interestingly, the expression of SlNRT2.7 was significantly higher in RO shoot compared to UC82 and strongly correlated to RO higher growth as well as to NUE and NUtE component. Our findings underlined the differential regulation of N-metabolism genes that may confer to NUtE component a pivotal role in NUE enhancement in tomato.

Short-term effects of the allelochemical umbelliferone on Triticum durum L. metabolism through GC-MS based untargeted metabolomics.

The present study used untargeted metabolomics to investigate the short-term metabolic changes induced in wheat seedlings by the specialized metabolite umbelliferone, an allelochemical. We used 10 day-old wheat seedlings treated with 104 µM umbelliferone over a time course experiment covering 6 time points (0 h, 6 h, 12 h, 24 h, 48 h, and 96 h), and compared the metabolomic changes to control (mock-treated) plants. Using gas chromatography mass spectrometry (GCMS)-based metabolomics, we obtained quantitative data on 177 metabolites that were derivatized (either derivatized singly or multiple times) or not, representing 139 non-redundant (unique) metabolites. Of these 139 metabolites, 118 were associated with a unique Human Metabolome Database (HMDB) identifier, while 113 were associated with a Kyoto Encyclopedia of Genes and Genomes (KEGG) identifier. Relative quantification of these metabolites across the time-course of umbelliferone treatment revealed 22 compounds (sugars, fatty acids, secondary metabolites, organic acids, and amino acids) that changed significantly (repeated measures ANOVA, P-value < 0.05) over time. Using multivariate partial least squares discriminant analysis (PLS-DA), we showed the grouping of samples based on time-course across the control and umbelliferone-treated plants, whereas the metabolite-metabolite Pearson correlations revealed tightly formed clusters of umbelliferone-derived metabolites, fatty acids, amino acids, and carbohydrates. Also, the time-course umbelliferone treatment revealed that phospho-l-serine, maltose, and dehydroquinic acid were the top three metabolites showing highest importance in discrimination among the time-points. Overall, the biochemical changes converge towards a mechanistic explanation of the plant metabolic responses induced by umbelliferone. In particular, the perturbation of metabolites involved in tryptophan metabolism, as well as the imbalance of the shikimate pathways, which are strictly interconnected, were significantly altered by the treatment, suggesting a possible mechanism of action of this natural compound.

Origanum vulgare essential oils inhibit glutamate and aspartate metabolism altering the photorespiratory pathway in Arabidopsis thaliana seedlings.

Essential oils (EOs) have been extensively studied as valuable eco-friendly compounds with herbicidal activity for weed management. Phytotoxic potential of EOs, extracted from a wild population of Origanum vulgare ssp. hirtum (Link) Ietswaart, has been here evaluated on plant model Arabidopsis, through a physiological and metabolomic approach. The EOs composition was mainly characterized by monoterpenes and sesquiterpenes, with a strong abundance of two monoterpenic phenols, namely carvacrol and thymol, and the monoterpene o-cymene. The in vitro bioassay confirmed a strong phytotoxic effect of EOs on Arabidopsis rosettes, showing by both a strong growth reduction and highly chlorotic leaves. In well-developed seedlings, EOs firstly caused growth reduction and leaf chlorosis, together with a series of interconnected metabolic alterations: i) impairing the nitrogen assimilation into amino acids, which affects in particular the glutamine metabolism; and as consequence ii) excessive accumulation of toxic ammonia into the leaves, associated with oxidative stress and damage; iii) declining the efficiency of the photosynthetic apparatus, connected to the reduced CO2 fixation and photooxidation protection; iv) impairing the photorespiratory pathway. Overall, the results highlights that EOs alters principally the ability of Arabidopsis seedlings to incorporate inorganic nitrogen into amino acids, principally glutamine, leading to a dramatic accumulation of ammonia in leaf cells. This primary effect induces, in turn, a cascade of reactions that limits the efficiency of PSII, inducing oxidative stress and finally causing a strong plant growth reduction, leaf necrosis and eventually plant death. These findings suggest that O. vulgare EOs might be proficiently exploited as a potential bioherbicide in an ecofriendly agriculture. Moreover, its multitarget activity could be advantageous in limiting weed resistance phenomenon.

Coumarin enhances nitrate uptake in maize roots through modulation of plasma membrane H+ -ATPase activity.

Coumarin is one of the simplest plant secondary metabolites, widely distributed in the plant kingdom, affecting root form and function, including anatomy, morphology and nutrient uptake. Although, some plant responses to coumarin have been described, comprehensive knowledge of the physiological and molecular mechanisms is lacking. Maize seedlings exposed to different coumarin concentrations, alone or in combination with 200 µm nitrate (NO3- ), were analysed, through a physiological and molecular approach, to elucidate action of coumarin on net NO3- uptake rate (NNUR). In detail, the time course of NNUR, plasma membrane (PM) H+ -ATPase activity, proton pumping and related gene expression (ZmNPF6.3, ZmNRT2.1, ZmNAR2.1, ZmHA3 and ZmHA4) were evaluated. Coumarin alone did not affect nitrate uptake, PM H+ -ATPase activity or transcript levels of ZmNRT2.1 and ZmHA3. In contrast, coumarin alone increased ZmNPF6.3, ZmNAR2.1 and ZmHA4 expression in response to abiotic stress. When coumarin and NO3- were concurrently added to the nutrient solution, a significant increase in the NNUR, PM H+ -ATPase activity, together with ZmNAR2.1:ZmNRT2.1 and ZmHA4 expression was observed, suggesting that coumarin affected the inducible component of the high affinity transport system (iHATS), and this effect appeared to be mediated by nitrate. Moreover, results with vanadate, an inhibitor of the PM H+ -ATPase, suggested that this enzyme could be the main target of coumarin. Surprisingly, coumarin did not affect PM H+ -ATPase activity by direct contact with plasma membrane vesicles isolated from maize roots, indicating its possible elicitor role in gene transcription.

Terpenoid trans-caryophyllene inhibits weed germination and induces plant water status alteration and oxidative damage in adult Arabidopsis.

trans-Caryophyllene (TC) is a sesquiterpene commonly found as volatile component in many different aromatic plants. Although the phytotoxic effects of trans-caryophyllene on seedling growth are relatively explored, not many information is available regarding the phytotoxicity of this sesquiterpenes on weed germination and on adult plants. The phytotoxic potential of TC was assayed in vitro on weed germination and seedling growth to validate its phytotoxic potential on weed species. Moreover, it was assayed on the metabolism of Arabidopsis thaliana adult plants, through two different application ways, spraying and watering, in order to establish the primary affected organ and to deal with the unknown mobility of the compound. The results clearly indicated that TC inhibited both seed germination and root growth, as demonstrated by comparison of the ED50 values. Moreover, although trans-caryophyllene-sprayed adult Arabidopsis plants did not show any effect, trans-caryophyllene-watered plants became strongly affected. The results suggested that root uptake was a key step for the effectiveness of this natural compound and its phytotoxicity on adult plants was mainly due to the alteration of plant water status accompanied by oxidative damage.

Genetic diversity and population structure of an Italian landrace of runner bean (Phaseolus coccineus L.): inferences for its safeguard and on-farm conservation.

The landraces are considered important sources of valuable germplasm for breeding activities to face climatic changes as well as to satisfy the requirement of new varieties for marginal areas. Runner bean (Phaseolus coccineus L.) is one of the most cultivated Phaseolus species worldwide, but few studies have been addressed to assess the genetic diversity and structure within and among landrace populations. In the present study, 20 different populations of a runner bean landrace from Central Italy named "Fagiolone," together with 41 accessions from Italy and Mesoamerica, were evaluated by using 14 nuclear SSRs to establish its genetic structure and distinctiveness. Results indicated that "Fagiolone" landrace can be considered as a dynamic evolving open-pollinated population that shows a significant level of genetic variation, mostly detected within populations, and the presence of two main genetic groups, of which one distinguished from other Italian runner bean landraces. Results highlighted also a relevant importance of farmers' management practices able to influence the genetic structure of this landrace, in particular the seed exchanges and selection, and the past introduction in cultivation of landraces/cultivars similar to seed morphology, but genetically rather far from "Fagiolone." The most suitable on-farm strategies for seed collection, conservation and multiplication will be defined based on our results, as a model for threatened populations of other allogamous crop species. STRUCTURE and phylogenetic analyses indicated that Mesoamerican accessions and Italian landraces belong to two distinct gene pools confirming the hypothesis that Europe could be considered a secondary diversification center for P. coccineus.

The inhibitory effects of coumarin on the germination of durum wheat (Triticum turgidum ssp. durum, cv. Simeto) seeds.

The event chain leading to germination blockage in durum wheat (Triticum turgidum ssp. durum Desf.) seeds exposed to the allelochemical coumarin (2H-chromen-2-one) was studied. The physiological and biochemical aspects thought to be critical for a successful seed germination were measured. At concentrations above 200 microM: , coumarin inhibited seed germination in a concentration-dependent manner. Inhibition occurred early during seed imbibition (phase I), was rapid, and irreversible. During phase I, coumarin inhibited water uptake, electrolyte retention capacity, and O(2) consumption. Later on, coumarin delayed the reactivation of peroxidases, enhanced the activity of superoxide dismutase, decreased the activities of selected marker enzymes for metabolic resumption, and repressed the transcription of molecular chaperones involved in secretory pathways. Insufficient and/or late seed rehydration caused by coumarin could have delayed membrane stabilization or decreased respiratory O(2) consumption, both of which are conducive to an overproduction of reactive O(2) species. Being unbalanced by an adequate upsurge of antioxidant defense systems, the resulting oxidative stress might have ultimately interfered with the germination program.

Acute leukemia with lymphoid/myeloid cell populations (hybrid leukemia). Case reports.

Based on conventional and immunological cell markers the great majority of acute leukemias can be classified as of lymphoid or myeloid origin. However, in some instances, leukemic cells with both lymphoid and myeloid features are found. For these cases the term of hybrid leukemia has been suggested; moreover, it has been shown that dual markers can be expressed by the same cell (biphenotypic leukemia) or myeloid and lymphoid cell populations coexist (biclonal leukemia). The accurate typing of acute leukemia is crucial in order to entail treatment and predict survival; therefore, the identification of acute hybrid leukemias may be important not only in terms of biological significance but also of management and outcome. We report here two further cases of acute hybrid leukemia with poor response to chemotherapy and very short survival.