Drought stress modulates secondary metabolites in Brassica oleracea L. convar. acephala (DC) Alef, var. sabellica L.

BACKGROUND: Consumer preference today is for the consumption of functional food and the reduction of chemical preservatives. Moreover, the antimicrobial properties and health-promoting qualities of plant secondary metabolites are well known. Due to forecasted climate changes and increasing human population, agricultural practices for saving water have become a concern. In the present study, the physiological responses of curly kale Brassica oleracea L. convar. Acephala (DC) var. sabellica to drought stress and the impact of water limitation on the concentration of selected secondary metabolites were investigated under laboratory-controlled conditions. RESULTS: Results indicated that drought stress increased the content of trans-2-hexenal, phytol and δ-tocopherol, and decreased chlorophyll content. Moreover, drought stress increased antioxidant capacity and the expression of AOP2, a gene associated with the biosynthesis of aliphatic alkenyl glucosinolates, and of three genes - TGG1, TGGE and PEN2 - encoding for myrosinases, the enzymes involved in glucosinolate breakdown. CONCLUSION: The present study shows that water limitation during the growing phase might be exploited as a sustainable practice for producing curly kale with a high concentration of nutritionally important health-promoting bioactive metabolites. © 2019 Society of Chemical Industry.

Cross-talk between physiological and biochemical adjustments by Punica granatum cv. Dente di cavallo mitigates the effects of salinity and ozone stress.

Plants are exposed to a broad range of environmental stresses, such as salinity and ozone (O3), and survive due to their ability to adjust their metabolism. The aim of this study was to evaluate the physiological and biochemical adjustments adopted by pomegranate (Punica granatum L. cv. Dente di cavallo) under realistic field conditions. One-year-old saplings were exposed to O3 [two levels denoted as ambient (AO) and elevated (EO) O3 concentrations] and salinity [S (salt, 50 mM NaCl)] for three consecutive months. No salt (NS) plants received distilled water. Despite the accumulation of Na+ and Cl- in the aboveground biomass, no evidence of visible injury due to salt (e.g. tip yellow-brown lesions) was found. The maintenance of leaf water status (i.e. unchanged values of electrolytic leakage and relative water content), the significant increase of abscisic acid, proline and starch content (+98, +65 and +59% compared to AO_NS) and stomatal closure (-24%) are suggested to act as adaptive mechanisms against salt stress in AO_S plants. By contrast, EO_NS plants were unable to protect cells against the negative impact of O3, as confirmed by the reduction of the CO2 assimilation rate (-21%), accumulation of reactive oxygen species (+10 and +225% of superoxide anion and hydrogen peroxide) and malondialdehyde by-product (about 2-fold higher than AO_NS). Plants tried to preserve themselves from further oxidative damage by adopting some biochemical adjustments [i.e. increase in proline content (+41%) and induction of catalase activity (8-fold higher than in AO_NS)]. The interaction of the two stressors induced responses considerably different to those observed when each stressor was applied independently. An analysis of the antioxidant pool revealed that the biochemical adjustments adopted by P. granatum under EO_S conditions (e.g. reduction of total ascorbate; increased activities of superoxide dismutase and catalase) were not sufficient to ameliorate the O3-induced oxidative stress.

Root colonization by Pseudomonas chlororaphis primes tomato (Lycopersicum esculentum) plants for enhanced tolerance to water stress.

Previous research demonstrated that Pseudomonas chlororaphis subsp. aureofaciens strain M71, a plant growth promoting bacterium (PGPB), exerts beneficial effects on plant metabolism and primes tolerance mechanisms against biotic stresses in tomatoes. We designed an experiment to assess whether root colonization with P. chlororaphis is also able to improve tolerance to water stress in tomatoes. Our results show that inoculation with P. chlororaphis stimulates the antioxidant activity of well-watered tomatoes while maintaining a steady-state level of reactive oxygen species (ROS), increases the expression of genes encoding for the biosynthesis of leaf terpenes, stimulates the production of both the phytohormones ABA and IAA, in turn affecting plant shape (number of leaves) and height (length of internodes), without altering photosynthesis. Upon exposure to mild water stress conditions, an improved antioxidant activity in tomatoes 'primed' by P. chlororaphis inoculation limited the accumulation of reactive oxygen species (ROS) in leaves and thus enhanced tolerance, also through increase of the (osmolyte) proline content. Moreover, P. chlororaphis inoculation further enhanced the ABA level in leaves of water-stressed tomatoes allowing a more efficient modulation of stomatal closure that resulted in an improved water use efficiency (WUE) and biomass accumulation.

Can nutrient fertilization mitigate the effects of ozone exposure on an ozone-sensitive poplar clone?

We tested the independent and interactive effects of nitrogen (N; 0 and 80 kg ha-1), phosphorus (P; 0, 40 and 80 kg ha-1), and ozone (O3) application/exposure [ambient concentration (AA), 1.5 × AA and 2.0 × AA] for five consecutive months on biochemical traits of the O3-sensitive Oxford poplar clone. Plants exposed to O3 showed visible injury and an alteration of membrane integrity, as confirmed by the malondialdehyde by-product accumulation (+3 and +17% under 1.5 × AA and 2.0 × AA conditions, in comparison to AA). This was probably due to O3-induced oxidative damage, as documented by the production of superoxide anion radical (O2-, +27 and +63%, respectively). Ozone per se, independently from the concentrations, induced multiple signals (e.g., alteration of cellular redox state, increase of abscisic acid/indole-3-acetic acid ratio and reduction of proline content) that might be part of premature leaf senescence processes. By contrast, nutrient fertilization (both N and P) reduced reactive oxygen species accumulation (as confirmed by the decreased O2- and hydrogen peroxide content), resulting in enhanced membrane stability. This was probably due to the simultaneous involvement of antioxidant compounds (e.g., carotenoids, ascorbate and glutathione) and osmoprotectants (e.g., proline) that regulate the detoxification processes of coping with oxidative stress by reducing the O3 sensitivity of Oxford clone. These mitigation effects were effective only under AA and 1.5 × AA conditions. Nitrogen and P supply activated a free radical scavenging system that was not able to delay leaf senescence and mitigate the adverse effects of a general peroxidation due to the highest O3 concentrations.

A L-type lectin gene is involved in the response to hormonal treatment and water deficit in Volkamer lemon.

Combination of biotic and abiotic stress is a major challenge for crop and fruit production. Thus, identification of genes involved in cross-response to abiotic and biotic stress is of great importance for breeding superior genotypes. Lectins are glycan-binding proteins with a functions in the developmental processes as well as in the response to biotic and abiotic stress. In this work, a lectin like gene, namely ClLectin1, was characterized in Volkamer lemon and its expression was studied in plants exposed to either water stress, hormonal elicitors (JA, SA, ABA) or wounding to understand whether this gene may have a function in the response to multiple stress combination. Results showed that ClLectin1 has 100% homology with a L-type lectin gene from C. sinensis and the in silico study of the 5'UTR region showed the presence of cis-responsive elements to SA, DRE2 and ABA. ClLectin1 was rapidly induced by hormonal treatments and wounding, at local and systemic levels, suggesting an involvement in defence signalling pathways and a possible role as fast detection biomarker of biotic stress. On the other hand, the induction of ClLectin1 by water stress pointed out a role of the gene in the response to drought. The simultaneous response of ClLectin1 expression to water stress and SA treatment could be further investigated to assess whether a moderate drought stress may be useful to improve citrus performance by stimulating the SA-dependent response to biotic stress.

Proteome changes during yeast-like and pseudohyphal growth in the biofilm-forming yeast Pichia fermentans.

The Pichia fermentans strain DISAABA 726 is a biofilm-forming yeast that has been proposed as biocontrol agent to control brown rot on apple. How ever, when inoculated on peach, strain 726 shows yeast-like to pseudohyphal transition coupled to a pathogenic behaviour. To identify the proteins potentially involved in such transition process, a comparative proteome analysis of P. fermentans 726 developed on peach (filamentous growth) vs apple (yeast-like growth) was carried out using two-dimensional gel electrophoresis coupled with mass spectrometry analysis. The proteome comparison was also performed between the two different cell morphologies induced in a liquid medium amended with urea (yeast-like cells) or methionine (filamentous cells) to exclude fruit tissue impact on the transition. Seventy-three protein spots showed significant variations in abundance (±twofold, p < 0.01, confidence intervals 99 %) between pseudohyphal vs yeast-like morphology produced on fruits. Among them, 30 proteins changed their levels when the two morphologies were developed in liquid medium. The identified proteins belong to several pathways and functions, such as glycolysis, amino acid synthesis, chaperones, and signalling transduction. The possible role of a group of proteins belonging to the carbohydrate pathway in the metabolic re-organisation during P. fermentans dimorphic transition is discussed.

Salt-stress induced changes in the leaf proteome of diploid and tetraploid mandarins with contrasting Na+ and Cl- accumulation behaviour.

To understand the genotypic variation of citrus to mild salt stress, a proteomic approach has been carried out in parallel on two citrus genotypes ('Cleopatra' and 'Willow leaf' mandarins), which differ for Na(+) and Cl(-) accumulation, and their cognate autotetraploids (4×). Using two-dimensional electrophoresis approximately 910 protein spots were reproducibly detected in control and salt-stressed leaves of all genotypes. Among them, 44 protein spots showing significant variations at least in one genotype were subjected to mass spectrometry analysis for identification. Salt-responsive proteins were involved in several functions, including photosynthetic processes, ROS scavenging, stress defence, and signalling. Genotype factors affect the salt-responsive pattern, especially that of carbon metabolism. The no ion accumulator 'Cleopatra' mandarin genotype showed the highest number of salt-responsive proteins, and up-regulation of Calvin cycle-related proteins. Conversely the ion accumulator 'Willow leaf' mandarin showed high levels of several photorespiration-related enzymes. A common set of proteins (twelve spots) displayed higher levels in salt-stressed leaves of 2× and 4× 'Cleopatra' and 4× 'Willow leaf' mandarin. Interestingly, antioxidant enzymes and heat shock proteins showed higher constitutive levels in 4× 'Cleopatra' mandarin and 4× 'Willow leaf' mandarin compared with the cognate 2× genotype. This work provides for the first time information on the effect of 8 weeks of salt stress on citrus genotypes contrasting for ion accumulation and their cognate autotetraploids. Results underline that genetic factors have a predominant effect on the salt response, although a common stress response independent from genotype was also found.

A stress responsive alternative splicing mechanism in Citrus clementina leaves.

Chitinases are often considered pathogenesis-related proteins since their activity can be induced by viral infections, fungal and bacterial cell wall components, and also by more general sources of stress such as wounding, salicylic acid, ethylene, auxins and cytokinins. In the present study, comparative proteomic analysis showed the defense-related acidic chitinase II to be specifically induced in Citrus clementina leaves infested by the two-spotted spider mite Tetranychus urticae or treated with MeJA. In parallel, changes in the mRNA profiles of two partially homologous chitinase forms were shown by RT-PCR. In particular, the appearance of an additional cDNA chitinase fragment in T. urticae-infested and MeJA-treated leaves was observed. This finding may indicate a specific regulatory mechanism of chitinase expression. We report evidence for alternative splicing in T. urticae-infested C. clementina, where a premature stop codon after the first 135 amino acids was introduced. We observed inducible chitinase activity after MeJA treatment, indicative of a rapid plant response to infestation. This work provides the first evidence of chitinase alternative splicing in C. clementina. In addition, the presence of the dual-band pattern for chitinase cDNA by RT-PCR may represent a suitable predictive marker for early diagnosis of plant biotic stress.