Parasitic plant-host interaction between the holoparasite Cytinus hypocistis and the shrub Cistus albidus in their natural Mediterranean habitat: local and systemic hormonal effects.

Mediterranean-type ecosystems provide a unique opportunity to study parasitic plant-host interactions, such as the relationship between the dominant shrub Cistus albidus L. and the root holoparasitic plant Cytinus hypocistis L. We examined this interaction (i) locally, by measuring the hormonal profiling of the interaction zone between the holoparasitic plant and the host, and (ii) systemically, by examining the hormonal profiling and physiological status of leaves from infested and uninfested plants. Furthermore, we explored how temporal variation (seasonal effects) and geographical location influenced the systemic hormonal and physiological response of leaves. Results shed light on tissue-related variations in hormones, suggesting the parasite exerted a sink effect, mainly influenced by cytokinins. Jasmonates triggered a defense response in leaves, far from the infestation point, and both jasmonates and abscisic acid (ABA) appeared to be involved in the tolerance to holoparasitism when plants were simultaneously challenged with summer drought. Parasitism did not have any major negative impact on the host, as indicated by physiological stress markers in leaves, thus indicating a high tolerance of the shrub C. albidus to the root holoparasitic plant C. hypocistis. Rather, parasitism seemed to exert a priming-like effect and some compensatory effects were observed (increased chlorophyll contents) in the host under mild climatic conditions. We conclude that (i) cytokinins, jasmonates and ABA play a role at the local and systemic levels in the response of C. albidus to the biotic stress caused by C. hypocistis, and that (ii) seasonal changes in environmental conditions and geographical location may impact holoparasitic plant-host interactions in the field, modulating the physiological response.

Application of a Biostimulant (Pepton) Based in Enzymatic Hydrolyzed Animal Protein Combined With Low Nitrogen Priming Boosts Fruit Production Without Negatively Affecting Quality in Greenhouse-Grown Tomatoes.

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 < 0.001) fruit production (due to an increase in the number of fruits), tended to increase (p = 0.057) by 20% the amount of TSS and increased (p < 0.05) the contents of lycopene (by 90%) and vitamin E (by 40%). Pepton displayed a tendency, almost significant, to improve (p = 0.054) total fruit production both in control and NEC plants, thus showing an additive effect to low N priming in boosting fruit production. Pepton maintained fruit quality in terms of sugar accumulation, total acidity and the contents of carotenoids, vitamins C and E. Pepton-related improvement in fruit production seemed to be related, at least partially, to an increased accumulation of cytokinins and photosynthetic pigments in leaves, which might favor vegetative vigor and ultimately fruit yield. In conclusion, Pepton application was effective in improving the yield of greenhouse tomatoes showing additive effect with low N priming, without negatively affecting fruit quality.

Vitamin E protects from lipid peroxidation during winter stress in the seagrass Cymodocea nodosa.

MAIN CONCLUSION: Adjustments in the antenna size and α-tocopherol contents provide protection from sustained damage in leaves of a seagrass, while low vitamin E contents appear to be enough to protect rhizomes (which appear to be more cold tolerant than leaves). Despite low temperatures can adversely affect the proper growth and development of marine angiosperms, by, among other processes, increasing reactive oxygen species production and causing oxidative damage to lipid membranes, the role of vitamin E in seagrasses, such as Cymodocea nodosa has not been explored thus far. Here, we aimed to better understand the possible role of this chain-breaking (peroxyl radical-trapping) antioxidant in response to low temperatures, and most particularly in relation to the occurrence of photo-inhibition and lipid peroxidation. Low temperatures caused an important desiccation of leaves, but not of rhizomes, which were much more tolerant to cold stress than leaves. Cold stress during winter was associated with chlorophyll loss and transient photo-inhibition, as indicated by reversible reductions in the Fv/Fm ratio. Adjustments in pigment antenna size and vitamin E contents per unit of chlorophyll during winter may help protect the photosynthetic apparatus from sustained photo-inhibitory damage and lipid peroxidation events in leaves. Rhizomes also accumulated significant amounts of vitamin E, although to a much lesser extent than leaves, and kept protected from lipid peroxidation during winter, as indicated by malondialdehyde contents, a product from secondary lipid peroxidation. It is concluded that vitamin E can help protect both leaves and rhizomes from lipid peroxidation, although cold stress during winter can cause transient photo-inhibition of the photosynthetic apparatus, in C. nodosa.

A Dual Role for Abscisic Acid Integrating the Cold Stress Response at the Whole-Plant Level in Iris pseudacorus L. Growing in a Natural Wetland.

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.

Leaf size modulation by cytokinins in sesame plants.

Phytohormones play important roles in controlling leaf size and in the modulation of various stress responses, including drought. In this study, hormone profiling analyses by ultra high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (UHPLC-MS/MS) was performed in leaves collected at three stages of active leaf growth to unravel which phytohormones modulate leaf size in sesame (Sesamum indicum L.) plants, an important oil-rich crop. Furthermore, endogenous contents of phytohormones were measured in parallel to various stress markers in sesame plants exposed to mild water deficit conditions by withholding water in potted plants for one week. Results revealed a major role of cytokinins and auxin in the modulation of leaf growth in sesame plants (which increased by 21.5 and 2.1-fold, respectively, with leaf growth), as well as a putative antagonistic response between jasmonic acid and salicylic acid during leaf development. Furthermore, growth arrest during water deficit stress appeared to be modulated by cytokinins, the endogenous contents of which decreased (by 48%) in parallel with ABA increases (by 59%). Reductions in the contents of the active cytokinin trans-zeatin occurred in parallel with increases in isopentenyladenine contents under drought, which suggests a partial metabolic limitation in cytokinin biosynthesis in leaves upon water deficit stress. These results provide useful information for the hormonal modulation of leaf size and the improvement of leaf growth and production in sesame plants through manipulation of the levels of key regulatory phytohormones.

Linking jasmonates with vitamin E accumulation in plants: a case study in the Mediterranean shrub Cistus albidus L.

MAIN CONCLUSION: Jasmonic acid positively modulates vitamin E accumulation, but the latter can also partly influence the capacity to accumulate the jasmonic acid precursor, 12-oxo-phytodienoic acid, in white-leaved rockrose (Cistus albidus L.) plants growing in their natural habitat. This study suggests a bidirectional link between chloroplastic antioxidants and lipid peroxidation-derived hormones in plants. While vitamin E is well known for its antioxidant properties being involved in plant responses to abiotic stress, jasmonates are generally related to biotic stress responses in plants. Studying them in non-model plants under natural conditions is crucial for the knowledge on their relationship, which will help us to better understand mechanisms and limits of stress tolerance to implement better conservation strategies in vulnerable ecosystems. We studied a typical Mediterranean shrub, white-leaved rockrose (Cistus albidus) under natural conditions during three winters and we analyzed both α and γ-tocopherol, and the three main jasmonates forms 12-oxo-phytodienoic acid (OPDA), jasmonic acid (JA), and jasmonoyl-isoleucine (JA-Ile). We found that JA contents positively correlated with vitamin E accumulation, most particularly with γ-tocopherol, the precursor of α-tocopherol (the most active vitamin E form). This finding was confirmed by exogenous application of methyl jasmonate (MeJA) in leaf discs under controlled conditions, which increased γ-tocopherol when applied at 0.1 mM MeJA and α-tocopherol at 1 mM MeJA. Furthermore, a complementary meta-analysis study with previously published reports revealed a positive correlation between JA and vitamin E, although this relationship turned to be strongly species specific. A strong negative correlation was observed, however, between total tocopherols and OPDA (a JA precursor located in chloroplasts). This antagonistic effect was observed between α-tocopherol and OPDA, but not between γ-tocopherol and OPDA. It is concluded that (i) variations in jasmonates and vitamin E due to yearly, inter-individual and sun orientation-driven variability are compatible with a partial regulation of vitamin E accumulation by jasmonates, (ii) vitamin E may also exert a role in the modulation of the biosynthesis of OPDA, with a much smaller effect, if any, on other jasmonates, and (iii) a trade-off in the accumulation of vitamin E and jasmonates might occur in the regulation of biotic and abiotic stress responses in plants.

Holoparasitic plant-host interactions and their impact on Mediterranean ecosystems.

Although photosynthesis is essential to sustain life on Earth, not all plants use sunlight to synthesize nutrients from carbon dioxide and water. Holoparasitic plants, which are important in agricultural and natural ecosystems, are dependent on other plants for nutrients. Phytohormones are crucial in holoparasitic plant-host interactions, from seed germination to senescence, not only because they act as growth and developmental regulators, but also because of their central role in the regulation of host photosynthesis and source-sink relations between the host and the holoparasitic plant. Here, we compile and discuss current knowledge on the impact and ecophysiology of holoparasitic plants (such as the broomrapes Orobanche sp. and Phelipanche sp.) that infest economically important dicotyledonous crops in Mediterranean agroecosystems (legumes [Fabaceae], sunflowers [Helianthus sp.], or tomato [Solanum lycopersicum] plants). We also highlight the role of holoparasitic plant-host interactions (such as those between Cytinus hypocistis and various shrubs of the genus Cistus) in shaping natural Mediterranean ecosystems. The roles of phytohormones in controlling plant-host interactions, abiotic factors in parasitism, and the biological significance of natural seed banks and how dormancy and germination are regulated, will all be discussed. Holoparasitic plants are unique organisms; improving our understanding of their interaction with hosts as study models will help us to better manage parasitic plants, both in agricultural and natural ecosystems.

Differential accumulation of tocochromanols in photosynthetic and non-photosynthetic tissues of strawberry plants subjected to reiterated water deficit.

Tocochromanols are a group of lipid-soluble antioxidants that include tocopherols, tocotrienols and plastochromanol-8. Here, we examined a putative differential accumulation of tocochromanols in photosynthetic and non-photosynthetic tissues (including leaves and whole fruits) of strawberry (Fragaria x ananassa cv. Albion) plants and evaluated their endogenous variations in response to a reiterated water deficit during a vegetative (non-productive) and a fruiting (productive) period. In addition, we evaluated the concentration of tocochromanols in achenes (true fruits) and flesh of strawberries (whole fruits) at the white and full-red stages both under optimal and stress conditions. Results showed that leaves mainly accumulated α-tocopherol, with plastochromanol-8 and γ-tocopherol being present at low amounts. In contrast, whole fruits did not accumulate plastochromanol-8, γ-tocopherol being the major tocochromanol in the achenes (true fruit) and α-tocopherol in the flesh. While α-tocopherol content in leaves increased up to seven-fold after 12 weeks of stress during the fruiting period, it kept unaltered during the vegetative period. Neither plastochromanol-8 nor γ-tocopherol contents increased in leaves of stressed plants. During the fruiting period, γ-tocopherol content increased in whole fruits of stressed plants (most of it being accumulated in the achenes). Among the compounds examined, the flesh of strawberries accumulated α-tocopherol only, both at the white and full-red stages. It is concluded that (i) α-tocopherol is the major tocochromanol in leaves, while γ-tocopherol is the major tocochromanol in achenes (ii) reiterated water deficit promotes the accumulation of α-tocopherol in leaves and γ-tocopherol in fruits, (iii) α-tocopherol not only accumulates in photosynthetic tissues (leaves and whole fruits at green stages), but also in non-photosynthetic tissues (flesh of whole fruits at the white and full-red stages), and (iv) achenes (true fruits) of strawberry plants are an extraordinary rich source of tocopherols.

An Enzymatically Hydrolyzed Animal Protein-Based Biostimulant (Pepton) Increases Salicylic Acid and Promotes Growth of Tomato Roots Under Temperature and Nutrient Stress.

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.

An overview of plant-based natural biostimulants for sustainable horticulture with a particular focus on moringa leaf extracts.

The horticulture sector is facing various challenges in the near future. Aside from maintaining or even improving yields, sustainable horticulture production is crucial to achieve food security. Reducing the reliance on agro-chemicals and/or increasing the efficiency of use under a changing climate is crucial. Natural biostimulants can play an important role in this regard, increasing production at a relatively low cost sustainably. Natural biostimulant feedstocks include leaf, root or seed extracts, either individually or in combination with others. Their positive effect on horticultural production is mostly due to plant growth-enhancing bioactive compounds such as phytohormones, amino acids, and nutrients. Here we review recent progress made in research and applications on plant-derived extracts with an emphasis on the use of these renewable biochemicals as biostimulants in sustainable horticulture. Moringa leaf extracts in particular have been shown to improve seed germination, plant growth and yield, nutrient use efficiency, crop and product quality traits (pre- and post-harvest), as well as tolerance to abiotic stresses. Although horticulture production relies on synthetic fertilisers to maintain and improve production, the use of plant-derived biostimulants such as moringa leaf extracts may be an option to reduce quantities needed and thus contribute in achieving global food security sustainably.

Contrasting patterns of hormonal and photoprotective isoprenoids in response to stress in Cistus albidus during a Mediterranean winter.

MAIN CONCLUSION: Seasonal accumulation of hormonal and photoprotective isoprenoids, particularly α-tocopherol, carotenoids and abscisic acid, indicate their important role in protecting Cistus albidus plants from environmental stress during a Mediterranean winter. The high diurnal amounts of α-tocopherol and xanthophylls 3 h before maximum light intensity suggest a photoprotective response against the prevailing diurnal changes. The timing to modulate acclimatory/defense responses under changing environmental conditions is one of the most critical points for plant fitness and stress tolerance. Here, we report seasonal and diurnal changes in the contents of isoprenoids originated from the methylerythritol phosphate pathway, including chlorophylls, carotenoids, tocochromanols, and phytohormones (abscisic acid, cytokinins, and gibberellins) in C. albidus during a Mediterranean winter. Plants were subjected not only to typically low winter temperatures but also to drought, as shown by a mean plant water status of 54% during the experimental period. The maximum PSII efficiency, however, remained consistently high (Fv/Fm > 0.8), proving that C. albidus had efficient mechanisms to tolerate combined stress conditions during winter. While seasonal α-tocopherol contents remained high (200-300 µg/g DW) during the experimental period, carotenoid contents increased during winter attaining maximum levels in February (minimum air temperature ≤ 5 °C for 13 days). Following the initial transient increases of bioactive trans-zeatin (about fivefold) during winter, the increased abscisic acid contents proved its important role during abiotic stress tolerance. Diurnal amounts of α-tocopherol and xanthophylls, particularly lutein, zeaxanthin and neoxanthin including the de-epoxidation state, reached maximum levels as early as 2 h after dawn, when solar intensity was 68% lower than the maximum solar radiation at noon. It is concluded that (1) given their proven antioxidant properties, both α-tocopherol and carotenoids seem to play a crucial role protecting the photosynthetic apparatus under severe stress conditions; (2) high seasonal amounts of abscisic acid indicate its important role in abiotic stress tolerance within plant hormones, although under specific environmental conditions, accumulation of bioactive cytokinins appears to be involved to enhance stress tolerance; (3) the concerted diurnal adjustment of α-tocopherol and xanthophylls as early as 3 h before maximum light intensity suggests that plants anticipated the predictable diurnal changes in the environment to protect the photosynthetic apparatus.

Leaf Orientation as Part of the Leaf Developmental Program in the Semi-Deciduous Shrub, Cistus albidus L.: Diurnal, Positional, and Photoprotective Effects During Winter.

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 F v/F m 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 F v/F m 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 F v/F m 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, F v/F m 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.

Hormonal Effects of an Enzymatically Hydrolyzed Animal Protein-Based Biostimulant (Pepton) in Water-Stressed Tomato Plants.

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.