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Desiccation tolerance in vegetative tissues enables resurrection plants to remain quiescent under severe drought and rapidly recover full metabolism once water becomes available. Barbacenia graminifolia is a resurrection plant that occurs at high altitudes, typically growing on rock slits, exposed to high irradiance and limited water availability. We analyzed the levels of reactive oxygen species (ROS) and antioxidants, carotenoids and its cleavage products, and stress-related phytohormones in fully hydrated, dehydrated, and rehydrated leaves of B. graminifolia. This species exhibited a precise adjustment of its antioxidant metabolism to desiccation. Our results indicate that this adjustment is associated with enhanced carotenoid and apocarotenoids, α-tocopherol and compounds of ascorbate-glutathione cycle. While α-carotene and lutein increased in dried-leaves suggesting effective protection of the light-harvesting complexes, the decrease in β-carotene was accompanied of 10.2-fold increase in the content of β-cyclocitral, an apocarotenoid implicated in the regulation of abiotic stresses, compared to hydrated plants. The principal component analysis showed that dehydrated plants at 30 days formed a separate cluster from both hydrated and dehydrated plants for up to 15 days. This regulation might be part of the protective metabolic strategies employed by this resurrection plant to survive water scarcity in its inhospitable habitat.

Hairy roots are made after the integration of a small set of genes from Agrobacterium rhizogenes in the plant genome. Little is known about how this small set is linked to their hormone profile, which determines development, morphology, and levels of secondary metabolite production. We used C. asiatica hairy root line cultures to determine the putative links between the rol and aux gene expressions with morphological traits, a hormone profile, and centelloside production. The results obtained after 14 and 28 days of culture were processed via multivariate analysis and machine-learning processes such as random forest, supported vector machines, linear discriminant analysis, and neural networks. This allowed us to obtain models capable of discriminating highly productive root lines from their levels of genetic expression (rol and aux genes) or from their hormone profile. In total, 12 hormones were evaluated, resulting in 10 being satisfactorily detected. Within this set of hormones, abscisic acid (ABA) and cytokinin isopentenyl adenosine (IPA) were found to be critical in defining the morphological traits and centelloside content. The results showed that IPA brings more benefits to the biotechnological platform. Additionally, we determined the degree of influence of each of the evaluated genes on the individual hormone profile, finding that aux1 has a significant influence on the IPA profile, while the rol genes are closely linked to the ABA profile. Finally, we effectively verified the gene influence on these two specific hormones through feeding experiments that aimed to reverse the effect on root morphology and centelloside content.

Improved nutrient use efficiency together with the use of biostimulants have been little explored thus far to improve fruit yield and quality in economically relevant crops. The aim of this study was to determine the additive or synergistic effects, if any, of the application of an enzyme hydrolyzed animal protein biostimulant (Pepton) combined with priming with low nitrogen (N) in the production and quality of greenhouse tomatoes. Biostimulant treatment (Pepton at a dose equivalent of 4 kg/ha) was applied by ferti-irrigation for 2 months during the vegetative phase both in controls (watered with nutrient solution) and nutrient efficient crop (NEC), in which plants were primed with low N by exposing them to a 30% N deficiency for 2 months, and then recovered for 1 month before fruit production. Foliar water and N contents, pigments, maximum PSII efficiency (Fv/Fm ratio), and phytohormones [including abscisic acid (ABA), salicylic acid (SA), jasmonic acid (JA), and cytokinins] were measured prior and at 4 and 8 weeks after the first application. Fruit production and quality [as indicated by total soluble sugars (TSS) and acidity (TA), and the contents of lycopene, vitamin E, and vitamin C] were measured 1 month later at harvest. Priming with low N availability (NEC plants) doubled (p

Leaf senescence, the last stage of the developmental program of leaves, can be induced by both internal and external signals. Cold stress-induced leaf senescence is an efficient strategy to overcome winter temperatures. In this work, we studied leaf senescence in yellow flag (Iris pseudacorus L.) individuals growing in a natural wetland, not only considering its relationship with external and internal cues, but also the plant developmental program, and the biological significance of rhizomes, storage organs that remain viable through winter. Total chlorophyll contents and the maximum efficiency of PSII (Fv/Fm ratio) decreased in senescing leaves, which was associated with a sharp increase in abscisic acid (ABA) contents. Furthermore, total cytokinin and 2-isopentenyladenine contents decreased in December compared to November, as plants became more stressed due to a decline in air temperatures. ABA increases in senescing leaves increased in parallel to reductions in violaxanthin. Rhizomes also accumulated large amounts of ABA during winter, while roots did not, and neither roots nor rhizomes accumulated 9-cis-epoxycarotenoids, thus suggesting ABA, which might play a role in conferring cold tolerance to this subterranean organ, may result from phloem transport from senescing leaves. It is concluded that (i) leaf senescence is a highly regulated physiological process in yellow flag playing a key role in the modulation of the entire plant developmental program, and (ii) ABA plays a major role not only in the regulation of leaf senescence but also in the establishment of cold tolerance in rhizomes, two processes that appear to be intimately interconnected.

Agricultural practices in grapevines management include water restrictions due to its positive effect on wine quality, especially when applied at fruit ripening. Although the effects of water stress in some groups of phytohormones have already been described in leaves and whole grapes, information regarding tissue-specific variations in hormones during ripening in grapes is scarce. Field-grown grapevines from the cv. “Merlot” were subjected to two differential water supplies, including only rainfed, non-irrigated vines (T0) and vines additionally irrigated with 25Lweek−1 vine−1 (T1). Tissue-specific variations in the hormonal profiling of grapes [including changes in the contents of abscisic acid (ABA), jasmonic acid (JA), salicylic acid (SA), the ethylene precursor 1-amino-cyclopropane-1-carboxylic acid (ACC), the auxin indole-3-acetic acid, gibberellins 1, 3, 4, and 7 (GA1, GA3, GA4, and GA7), the cytokinins trans-zeatin, and 2-isopentenyl adenine, including as well their respective ribosylated forms] were periodically evaluated from veraison to harvest. The hormonal profiling in leaves was also measured at the beginning and end of the season for comparison. Results showed that grape growth dynamics were transiently affected by the differences in water regimes, the increased water supply leading to an accelerated growth, slightly reduced accumulation of sugars, and transiently lowered pH, although grape quality did not differ between treatments at harvest. Hormonal profiling of whole berries did not reveal any difference in the endogenous contents of phytohormones between treatments, except for a transient decrease in GA4 contents in T1 compared to T0 vines, which was not confirmed at the tissular level. Hormonal profiling at the tissue level highlighted a differential accumulation of phytohormones during ripening in berry tissues, with pulps being particularly poor in ABA, JA, and SA contents, seeds particularly accumulating ACC, gibberellins, and zeatin-type cytokinins, and the skin being particularly rich in auxin and active cytokinins. Changes in water supply led to very small and transient changes in the endogenous contents of phytohormones in the seeds, pulp, and skin of berries, the most remarkable variations being observed in cytokinin contents, which increased earlier [between 5 and 12days after veraison (DAV)] but later kept more constant in the skin from T1 compared to T0 vines and were also 3-fold higher at 40 DAV in seeds of T1 compared to T0 vines. It is concluded that small changes in water supply can trigger hormonal-driven physiological adjustments at the tissular level affecting the evolution of fruit growth and quality throughout grape berry ripening.

Sweet cherry is an important non-climacteric fruit with a high commercial interest, but exploitation of sweet cherry trees (Prunus avium L.) in orchards is usually subject to important economic losses due to fruit decay by pathogenic fungi and other microorganisms. Sweet cherries development and ripening are characterized by profound physiological changes in the fruit, among which the phytohormone abscisic acid (ABA) plays a pivotal role. In addition, sweet cherries are usually affected by fruit decay pathogens, and the role of other stress-related hormones such as jasmonic acid (JA) and salicylic acid (SA) may also be of paramount importance, not only from a developmental point of view, but also from a fruit-microbe interaction perspective. Here, a tissue-specific hormone quantification by LC-MS/MS, including the contents of JA, SA, and ABA, in the fruit exocarp and mesocarp of sweet cherries during fruit development from trees growing in a commercial orchard was carried out. Additionally, this study was complemented with the characterization of the culturable epiphytic and endophytic microbial communities of sweet cherries at various stages of fruit development and during cracking lesion formation. Our results revealed a completely differential behavior of phytohormones between both tissues (the exocarp and mesocarp), with a more dynamic exocarp in front of a more stable mesocarp, and with marked variations during fruit development. Microbial epiphytic community was mainly composed by yeasts, although rot-causing fungi like Alternaria spp. were always also present throughout fruit development. Endophytic colonization was poor, but it increased throughout fruit development. Furthermore, when the exocarp was naturally disrupted in sweet cherries suffering from cracking, the colonization by Alternaria spp. markedly increased. Altogether, results suggest that the fruit exocarp and mesocarp are very dynamic tissues in which endogenous phytohormones not only modulate fruit development and ripening but also fruit-microbe interactions.

The identification of drought-tolerant olive tree genotypes has become an urgent requirement to develop sustainable agriculture in dry lands. However, physiological markers linking drought tolerance with mechanistic effects operating at the cellular level are still lacking, in particular under severe stress, despite the urgent need to develop these tools in the current frame of global change. In this context, 1-year-old olive plants growing in the greenhouse and with a high intra-specific variability (using various genotypes obtained either from cuttings or seeds) were evaluated for drought tolerance under severe stress. Growth, plant water status, net photosynthesis rates, chlorophyll contents and the extent of photo- and antioxidant defenses (including the de-epoxidation state of the xanthophyll cycle, and the contents of carotenoids and vitamin E) were evaluated under well-watered conditions and severe stress (by withholding water for 60 days). Plants were able to continue photosynthesizing under severe stress, even at very low leaf water potential of −4 to −6 MPa. This ability was achieved, at least in part, by the activation of photo- and antioxidant mechanisms, including not only increased xanthophyll cycle de-epoxidation, but also enhanced α-tocopherol contents. “Zarrazi” (obtained from seeds) and “Chemlali” (obtained from cuttings) showed better performance under severe water stress compared to the other genotypes, which was associated to their ability to trigger a higher antioxidant protection. It is concluded that (i) drought tolerance among the various genotypes tested is associated with antioxidant protection in olive trees, (ii) the extent of xanthophyll cycle de-epoxidation is strongly inversely related to photosynthetic rates, and (iii) vitamin E accumulation is sharply induced upon severe chlorophyll degradation.

Biostimulants may be particularly interesting for application in agricultural and horticultural crops since they can exert a growth-promoting effect on roots. This may be important for promoting longitudinal and lateral root growth and therefore increasing belowground vegetative growth, which may in turn lead to improved aboveground vegetative growth and increased yields. Here, we examined the effects and mechanism of action of an enzymatically hydrolyzed animal protein-based biostimulant (Pepton) on the root growth of tomato plants, with an emphasis on its possible role on chorismate-derived hormones (auxin, salicylic acid, and melatonin). Tomato plants growing in hydroponic systems were exposed to either nutrient stress conditions (experiment 1) or suboptimal temperatures (experiment 2) in a greenhouse, and the concentration of auxin, salicylic acid, and melatonin in roots were measured just prior and after the application of the biostimulant. Results showed that the application of Pepton exerted a growth-promoting effect on roots in plants growing under suboptimal conditions, which might be associated with enhanced salicylic acid levels in roots. The extent of effects of this enzymatically hydrolyzed animal protein-based biostimulant might strongly depend on the growth conditions and stage of root system development. It is concluded that an enzymatically hydrolyzed animal protein-based biostimulant (Pepton) may exert a positive effect enhancing primary and lateral root growth of tomato plants growing under suboptimal conditions, by stimulating the biosynthesis of specific hormonal pathways, such as salicylic acid under stress.

Biostimulants may promote growth or alleviate the negative effects of abiotic stress on plant growth eventually resulting in enhanced yields. We examined the mechanism of action of an enzymatically hydrolyzed animal protein-based biostimulant (Pepton), which has previously been shown to benefit growth and yield in several horticultural crops, particularly under stressful conditions. Tomato plants were exposed to well-watered and water-stressed conditions in a greenhouse and the hormonal profiling of leaves was measured during and after the application of Pepton. Results showed that the Pepton application benefited antioxidant protection and exerted a major hormonal effect in leaves of water-stressed tomatoes by increasing the endogenous content of indole-3-acetic acid (auxin), trans-zeatin (cytokinin), and jasmonic acid. The enhanced jasmonic acid content may have contributed to an increased production of tocochromanols because plastochromanol-8 concentration per unit of chlorophyll was higher in Pepton-treated plants compared to controls. In conclusion, the tested Pepton application may exert a positive effect on hormonal balance and the antioxidant system of plants under water stress in an economically important crop, such as tomato plants.

Mediterranean ecosystems harbor a very important part of Earth’s biodiversity, and they are a conservation priority due to the effects of global change. Here, we examined the performance of the semi-deciduous shrub Cistus albidus under Mediterranean conditions during winter, including changes in leaf angle governed by diurnal, seasonal, and positional effects and their relationship with winter photoinhibition and photoprotection. We found marked diurnal variations in leaf angle during the day in autumn, which disappeared as the photoperiod shortened during winter due to a progressive decrease in the predawn leaf angle from November to January. During this period, auxin contents decreased, while those of melatonin increased, and the Fv/Fm ratio, chlorophyll, and tocopherol contents kept unaltered, thus indicating the absence of photoinhibitory damage. A marked decrease in the leaf angle toward the shoot apex occurred during winter, which was associated with slightly higher Fv/Fm ratios. An analysis of the inter-individual variability and sun orientation effects on leaf movements in a population growing in the Montserrat mountains revealed a very marked variability of 46.8% in the leaf angle, while Fv/Fm ratio showed a variation of 7.5% only. West orientation, which was associated with reduced leaf temperatures, but with no differences in the photosynthetic photon flux density, led to enhanced tocopherol contents, while leaf angle, Fv/Fm ratio, chlorophyll, auxin, and melatonin contents kept unaltered. It is concluded that (1) C. albidus has very effective and fine-regulated photoprotection mechanisms, including an adequate orientation of decussate leaves as part of the developmental program, (2) leaf angle is modulated on a diurnal and seasonal basis, thus contributing to prevent photoinhibition as a first line of defense, and (3) enhanced tocopherol contents help withstand combined high light with low temperature stress in C. albidus growing at high elevation.

Climacteric and non-climacteric fruits are differentiated by the ripening process, in particular by the involvement of ethylene, high respiration rates and the nature of the process, being autocatalytic or not, respectively. Here, we focus on the biosynthesis, metabolism and function of three compounds (auxin, salicylic acid and melatonin) sharing not only a common precursor (chorismate), but also regulatory functions in plants, and therefore in fruits. Aside from describing their biosynthesis in plants, with a particular emphasis on common precursors and points of metabolic diversion, we will discuss recent advances on their role in fruit ripening and the regulation of bioactive compounds accumulation, both in climacteric and non-climacteric fruits.

Vitamin E inhibits the propagation of lipid peroxidation and helps protecting photosystem II from photoinhibition, but little is known about its possible role in plant response to Pi availability. Here, we aimed at examining the effect of vitamin E deficiency in Arabidopsis thaliana vte mutants on phytohormone contents and the expression of transcription factors in plants exposed to contrasting Pi availability. Plants were subjected to two doses of Pi, either unprimed (controls) or previously exposed to low Pi (primed). In the wild type, α-tocopherol contents increased significantly in response to repeated periods of low Pi, which was paralleled by increased growth, indicative of a priming effect. This growth-stimulating effect was, however, abolished in vte mutants. Hormonal profiling revealed significant effects of Pi availability, priming and genotype on the contents of jasmonates and salicylates; remarkably, vte mutants showed enhanced accumulation of both hormones under low Pi. Furthermore, expression profiling of 1,880 transcription factors by qRT-PCR revealed a pronounced effect of priming on the transcript levels of 45 transcription factors mainly associated with growth and stress in wild-type plants in response to low Pi availability; while distinct differences in the transcriptional response were detected in vte mutants. We conclude that α-tocopherol plays a major role in the response of plants to Pi availability not only by protecting plants from photo-oxidative stress, but also by exerting a control over growth- and defense-related transcriptional reprogramming and hormonal modulation.

In Mediterranean-type ecosystems plants are exposed to several adverse environmental conditions throughout the year, ranging from drought stress during the warm and dry summers to chilling stress due to the typical drop in temperatures during winters. Here we evaluated the ecophysiological response, in terms of photoinhibition and photoprotection, of the dioecious Mediterranean palm, Chamaerops humilis to seasonal variations in environmental conditions. Furthermore, we considered as well the influence of plant size, maturity and sexual dimorphism. Results showed evidence of winter photoinhibition, with a marked decrease of the Fv/Fm ratio below 0.7 between January and March, which was coincident with the lowest temperatures. During this period, the de-epoxidation state of the xanthophyll cycle and zeaxanthin levels increased, which might serve as a photoprotection mechanism, owing the full recovery from winter photoinhibition during spring. Furthermore, mature plants showed lower chlorophyll levels and higher β-carotene levels per unit of chlorophyll than juvenile plants, and females displayed lower leaf water contents and higher photoinhibition than males during summer, probably due to increased reproductive effort of females. However, neither low temperatures during winter nor reproductive events in females during the summer led to irreversible damage to the photosynthetic apparatus. We conclude that (i) the Mediterranean dwarf palm, C. humilis, suffers from photoinhibition during winter, but this is transient and does not lead to irreversible damage, and (ii) females from this plant species are more sensitive than males to photoinhibition during reproductive events.