CsREV-CsTCP4-CsVND7 module shapes xylem patterns differentially between stem and leaf to enhance tea plant tolerance to drought.

Cultivating drought-tolerant tea varieties enhances both yield and quality of tea plants in northern China. However, the mechanisms underlying their drought tolerance remain largely unknown. Here we identified a key regulator called CsREV, which differentially regulates xylem patterns between leaves and stems, thereby conferring drought tolerance in tea plants. When drought occurs, upregulation of CsREV activates the CsVND7a-dependent xylem vessel differentiation. However, when drought persists, the vessel differentiation is hindered as CsVND7a is downregulated by CsTCP4a. This, combined with the CsREV-promoted secondary-cell-wall thickness of xylem vessel, leads to the enhanced curling of leaves, a characteristic closely associated with plant drought tolerance. Notably, this inhibitory effect of CsTCP4a on CsVND7a expression is absent in stems, allowing stem xylem vessels to continuously differentiate. Overall, the CsREV-CsTCP4-CsVND7 module is differentially utilized to shape the xylem patterns in leaves and stems, potentially balancing water transportation and utilization to improve tea plant drought tolerance.

CsLAZY1 mediates shoot gravitropism and branch angle in tea plants (Camellia sinensis).

BACKGROUND: Branch angle is a pivotal component of tea plant architecture. Tea plant architecture not only affects tea quality and yield but also influences the efficiency of automatic tea plant pruning. However, the molecular mechanism controlling the branch angle, which is an important aspect of plant architecture, is poorly understood in tea plants. RESULTS: In the present study, three CsLAZY genes were identified from tea plant genome data through sequence homology analysis. Phylogenetic tree displayed that the CsLAZY genes had high sequence similarity with LAZY genes from other plant species, especially those in woody plants. The expression patterns of the three CsLAZYs were surveyed in eight tissues. We further verified the expression levels of the key CsLAZY1 transcript in different tissues among eight tea cultivars and found that CsLAZY1 was highly expressed in stem. Subcellular localization analysis showed that the CsLAZY1 protein was localized in the plasma membrane. CsLAZY1 was transferred into Arabidopsis thaliana to investigate its potential role in regulating shoot development. Remarkably, the CsLAZY1 overexpressed plants responded more effectively than the wild-type plants to a gravity inversion treatment under light and dark conditions. The results indicate that CsLAZY1 plays an important role in regulating shoot gravitropism in tea plants. CONCLUSIONS: The results provide important evidence for understanding the functions of CsLAZY1 in regulating shoot gravitropism and influencing the stem branch angle in tea plants. This report identifies CsLAZY1 as a promising gene resource for the improvement of tea plant architecture.

Survival strategies based on the hydraulic vulnerability segmentation hypothesis, for the tea plant [Camellia sinensis(L.) O. Kuntze] in long-term drought stress condition.

Tea plants are important economic perennial crops that can be negatively impacted by drought stress (DS). However, their survival strategies in long-term DS conditions and the accumulation and influence of metabolites and mineral elements (MEs) in their organs, when facing hydraulic vulnerability segmentation, require further investigation. The MEs and metabolites in the leaf, stem, and root after long-term DS (20 d) were examined here, using inductively coupled plasma optical emission spectrometry (ICP-OES) and liquid chromatograph-mass spectrometry (LC-MS). The accumulation patterns of 116 differentially accumulated metabolites (DAMs) and nine MEs were considerably affected in all organs. The concentration of all MEs varied significantly in at least one organ, while the K and Ca levels were markedly altered in all three. Most DAM levels increased in the stem but decreased in the root and leaf, implying that vulnerability segmentation may occur with long-term DS. The typical nitrogen- and carbon-compound levels similarly increased in the stem and decreased in the leaf and root, as the plant might respond to long-term DS by stabilizing respiration, promoting nitrogen recycling, and free radical scavenging. Correlation analysis showed several possible DAM-ME interactions and an association between Mn and flavonoids. Thus, survival strategies under long-term DS included sacrificing distal/vulnerable organs and accumulating function-specialized metabolites and MEs to mitigate drought-induced oxidative damage. This is the first study that reports substance fluctuations after long-term DS in different organs of plants, and highlights the need to use whole plants to fully comprehend stress response strategies.

Blue genome: chromosome-scale genome reveals the evolutionary and molecular basis of indigo biosynthesis in Strobilanthes cusia.

Natural plant dyes have been developed and used across many traditional societies worldwide. The blue pigment indigo has seen widespread usage across South America, Egypt, Europe, India and China for thousands of years, mainly extracted from indigo-rich plants. The utilization and genetic engineering of indigo in industries and ethnobotanical studies on the effects of cultural selection on plant domestication are limited due to lack of relevant genetic and genomic information of dye plants. Strobilanthes cusia (Acanthaceae) is a typical indigo-rich plant important to diverse ethnic cultures in many regions of Asia. Here we present a chromosome-scale genome for S. cusia with a genome size of approximately 865 Mb. About 79% of the sequences were identified as repetitive sequences and 32 148 protein-coding genes were annotated. Metabolic analysis showed that the main indigoid pigments (indican, indigo and indirubin) were mainly synthesized in the leaves and stems of S. cusia. Transcriptomic analysis revealed that the expression level of genes encoding metabolic enzymes such as monooxygenase, uridine diphosphate-glycosyltransferase and ß-glucosidase were significantly changed in leaves and stems compared with root tissues, implying their participation in indigo biosynthesis. We found that several gene families involved in indigo biosynthesis had undergone an expansion in number, with functional differentiation likely facilitating indigo biosynthesis in S. cusia. This study provides insight into the physiological and molecular bases of indigo biosynthesis, as well as providing genomic data that provide the basis for further study of S. cusia cultivation by Asia's traditional textile producers.

Comparative proteomic analysis provides new insights into the specialization of shoots and stolons in bermudagrass (Cynodon dactylon L.).

BACKGROUND: Bermudagrass (Cynodon dactylon L.) is an important turfgrass species with two types of stems, shoots and stolons. Despite their importance in determining the morphological variance and plasticity of bermudagrass, the intrinsic differences between stolons and shoots are poorly understood. RESULTS: In this study, we compared the proteomes of internode sections of shoots and stolons in the bermudagrass cultivar Yangjiang. The results indicated that 376 protein species were differentially accumulated in the two types of stems. Pathway enrichment analysis revealed that five and nine biochemical pathways were significantly enriched in stolons and shoots, respectively. Specifically, enzymes participating in starch synthesis all preferentially accumulated in stolons, whereas proteins involved in glycolysis and diverse transport processes showed relatively higher abundance in shoots. ADP-glucose pyrophosphorylase (AGPase) and pyruvate kinase (PK), which catalyze rate-limiting steps of starch synthesis and glycolysis, showed high expression levels and enzyme activity in stolons and shoots, respectively, in accordance with the different starch and soluble sugar contents of the two types of stems. CONCLUSIONS: Our study revealed the differences between the shoots and stolons of bermudagrass at the proteome level. The results not only expand our understanding of the specialization of stolons and shoots but also provide clues for the breeding of bermudagrass and other turfgrasses with different plant architectures.

Are hydraulic patterns of lianas different from trees? New insights from Hedera helix.

Lianas form long and flexible but disproportionately narrow stems, and thus require particular strategies to maintain the integrity of xylem water transport and ensure supply to large crown areas. The hydraulic architecture of lianas and the respective within-plant coordination of transport efficiency and safety, and the underlying anatomical variations in xylem, are largely unexplored. We analysed Hedera helix, a liana widespread in European temperate forests, with respect to hydraulic and xylem anatomical variations between the main stem and branches, between juvenile and adult life phases, and along the vertical axis. Main stems were significantly less embolism resistant but exhibited a higher hydraulic conductivity than branches. In branches, the cell turgor loss point of leaves decreased, while the embolism resistance and conductivity of xylem, as well as conduit diameters, increased with height. High water-transport capacities allow ivy to compensate for the small cross-section of stems, while the limited resistance to drought-induced xylem dysfunction of the main stem is probably linked to conservative stomatal regulation. Pronounced differences in xylem anatomy, hydraulic efficiency, and safety between the main stem and branches and along the vertical axis are surprisingly similar to those of self-supporting plants, and indicate that the coordination of carbon and water economies requires similar internal adjustments in tall plants.

Do nano-particles cause recalcitrant vulnerability curves in Robinia? Testing with a four-cuvette Cochard rotor and with water extraction curves.

Cavitation resistance is a key trait for characterizing the drought adaption in plants and is usually presented in terms of vulnerability curves. Three principal techniques have been developed to produce vulnerability curves, but curves generated with centrifugation are reported to suffer from artifacts when applied to long-vesseled species. The main cause of this artifact is the issue of open vessels, resulting in a nano-particle effect that may seed premature embolism. We used two methods to test the potential mechanism behind the nano-particle effect in centrifuge-based vulnerability curves. A four-cuvette rotor system based on a traditional Cochard rotor was designed to inhibit effervescence while injecting water, but the recalcitrant vulnerability curves in Robinia could not be eliminated. There may be multiple sources, besides effervescence, of hypothetical nano-particles: they may arise from cut surfaces or they may be always present in the injected water, leading to the premature embolisms. To prevent the entry of the hypothetical nano-particles, water extraction curves in terms of PLV (percentage loss volume of extracted water from stems) vs tensions were constructed. The PLV curves of Robinia showed s-shaped characteristics after subtracting the first Weibull components from water extraction curves, which were not related to the water loss from vessels according to dye staining experiments. The differences between T50 (xylem tension at which 50% of hydraulic conductivity is lost) in mean PLV curve and T50 in percentage loss of conductivity curves determined by the four-cuvette rotor system and by the bench dehydration method were 3.9 MPa and 0.7 MPa, respectively. Hence, PLV curves may be a valid way to measure the cavitation resistance in long-vesseled species with centrifugation. Keeping bark intact in the process of measurement is recommended, otherwise it would increase evaporation from the entire system.

Lilingostrobus chaloneri gen. et sp. nov., a Late Devonian woody lycopsid from Hunan, China.

Lycopsids are a minor component of current terrestrial herbaceous floras. However, lycopsid fossil diversity shows a great diversity and disparity including heterosporous woody plants, e.g. the giant isoetaleans that populated the extensive Pennsylvanian wetlands. The earliest known isoetaleans come from late Devonian localities from China. Here, we describe Lilingostrobus chaloneri gen. et sp. nov., a new isoetalean lycopsid from the Upper Devonian (Famennian) Xikuangshan Formation of China (Hunan Province, South China), which adds to the already impressive diversity of the Devonian lycopsids from China. Lilingostrobus shows an unusual combination of characters. This new plant is pseudoherbaceous, with a possible tufted habit, and consists of narrow axes with rare isotomies. The stem includes small quantities of secondary xylem. Each fertile axis bears one terminal strobilus comprising sporophylls ending in a very long upturned lamina. Microspores and putative megaspores have been found, but whether the plant has mono- or bisporangiate strobili is unknown. Importantly, our cladistic analysis identifies Lilingostrobus as a direct precursor of Isoetales, which provides new insights into the early evolution of lycopsids.

A disturbed auxin signaling affects adventitious root outgrowth in Solanum dulcamara under complete submergence.

Flooding negatively affects the growth and even survival of most terrestrial plants. Upon flooding, the excess water quickly decreases the gas exchange between atmosphere and the submerged plant tissues, which leads to oxygen deficiency resulting in a plant cell energy crisis, and eventually plant death. Solanum dulcamara survives flooding by producing aerenchymatous adventitious roots (ARs) from pre-formed primordia on the stem, which replace the original flood-sensitive root system. However, we found that under complete submergence, AR outgrowth was impaired in S. dulcamara. In the present work, we tried to elucidate the mechanisms behind this phenomenon in particular the involvement of the phytohormones auxin, abscisic acid and jasmonic acid. Abscisic acid (ABA) is a negative regulator of AR outgrowth, but surprisingly the ABA content and signaling were decreased to a similar extent under both partial and complete submergence, suggesting that ABA might not be responsible for the difference in AR outgrowth. Auxin, which is necessary for AR outgrowth, was at similar concentrations in either partially or completely submerged primordia, but complete submergence resulted in a decrease of auxin signaling in the primordia. Application of 1-naphthaleneacetic acid (NAA) to completely submerged plants restored AR outgrowth, implying that auxin response in the rooting tissues of completely submerged plants was reduced. Furthermore, jasmonic acid (JA) concentrations did not differ between partial and complete submergence. To conclude, a disruption in the auxin signaling within S. dulcamara AR primordia may result in the abortion of AR outgrowth under complete submergence.

Cellular injury to 1- to 3+-year-old stems of Camellia sinensis by Tuckerella japonica.

Tuckerella japonica Ehara (Acari: Tuckerellidae) feeds on predigested plant cells beneath exposed periderm tissue of 1- to 3+-year-old stems of Camellia sinensis (L.) O. Kuntze (Theaceae) where longitudinal bark splitting occurs. Control samples from these tissues were compared with areas fed upon by T. japonica adults and immatures to characterize types of cellular injury. Stylet diameters ranged from 1.6 to 2.3 µm and were consistent with observed stylet punctures in the stems. Mite saliva was injected along tracts within the cortical tissue and resulted in cell wall disruption, collapsed cells and, in older tissue, hyperplasia. The range of potential stylet penetration into plant tissues was from 92 to 150 µm. Tuckerella japonica injects saliva in the cortical tissues. The paired stylet lengths would allow for possible injection of saliva into the upper areas of phloem tissue but not in the cambium area of wood exposed by splitting of the outer epidermis.

Etiolated Stem Branching Is a Result of Systemic Signaling Associated with Sucrose Level.

The potato (Solanum tuberosum) tuber is a swollen stem. Sprouts growing from the tuber nodes represent loss of apical dominance and branching. Long cold storage induces loss of tuber apical dominance and results in secondary branching. Here, we show that a similar branching pattern can be induced by short heat treatment of the tubers. Detached sprouts were induced to branch by the heat treatment only when attached to a parenchyma cylinder. Grafting experiments showed that the scion branches only when grafted onto heat- or cold-treated tuber parenchyma, suggesting that the branching signal is transmitted systemically from the bud-base parenchyma to the grafted stem. Exogenous supply of sucrose (Suc), glucose, or fructose solution to detached sprouts induced branching in a dose-responsive manner, and an increase in Suc level was observed in tuber parenchyma upon branching induction, suggesting a role for elevated parenchyma sugars in the regulation of branching. However, sugar analysis of the apex and node after grafting showed no distinct differences in sugar levels between branching and nonbranching stems. Vacuolar invertase is a key enzyme in determining the level of Suc and its cleavage products, glucose and fructose, in potato parenchyma. Silencing of the vacuolar invertase-encoding gene led to increased tuber branching in combination with branching-inducing treatments. These results suggest that Suc in the parenchyma induces branching through signaling and not by excess mobilization from the parenchyma to the stem.

Cactus stem (Opuntia ficus-indica Mill): anatomy, physiology and chemical composition with emphasis on its biofunctional properties.

Cactus stem (Opuntia ficus-indica Mill) is native to Mesoamerica and marketed in different forms such as fresh, frozen or pre-cooked. Worldwide, this vegetable is recognized for its pharmaceutical actions, including its antioxidant, diuretic, anticarcinogenic, anti-inflammatory, anti-diabetic, and anti-hypercholesterolemic properties, as well as their antiviral and antispermatogenic effects. However, not all of these properties have been associated with its chemical composition; therefore, this review aims to present and integrate information available on the physiology and anatomy of cactus stem and its chemical composition, focusing on some of the many factors that determine its biofunctionality. © 2017 Society of Chemical Industry.

Stoichiometric variation of halophytes in response to changes in soil salinity.

Variation in soil salt may change the stoichiometry of a halophyte by altering plant ecophysiology, and exert different influences on various plant organs, which has potentially important consequences for the nutrition of consumers as well as nutrient cycling in a saline ecosystem. Using a greenhouse pot experiment, we investigated the effect of salinity variability on the growth and stoichiometry of different organs of Suaeda glauca and Salicornia europaea - two dominant species of important ecological and economic value in the saline ecosystem. Our results showed that appropriate salt stimulated the growth of both species during the vigorous growth period, while high salt suppressed growth. Na significantly increased with increased salt in the culture, whereas concentrations of other measured elements and K:Na ratio for both species significantly decreased at low salt treatments, and became more gradual under higher salt conditions. Furthermore, with the change of salt in culture, variations in leaf (degenerated leaf for S. europaea, considered as young stem) stoichiometry, except N:P ratio, were large and less in stems (old stems for S. europaea) than in roots, reflecting physiological and biochemical reactions in the leaf in response to salt stress, supported by sharp changes in trends. These results suggest that appropriate saline conditions can enhance biological C fixation of halophytes; however, increasing salt could affect consumer health and decrease cycling of other nutrients in saline ecosystems.

Plant Responses to Brief Touching: A Mechanism for Early Neighbour Detection?

In natural habitats plants can be exposed to brief and light contact with neighbouring plants. This mechanical stimulus may represent a cue that induces responses to nearby plants. However, little is known about the effect of touching on plant growth and interaction with insect herbivores. To simulate contact between plants, a soft brush was used to apply light and brief mechanical stimuli to terminal leaves of potato Solanum tuberosum L. The number of non-glandular trichomes on the leaf surface was counted on images made by light microscope while glandular trichomes and pavement cells were counted on images made under scanning electronic microscope. Volatile compounds were identified and quantified using coupled gas chromatography-mass spectrometry (GC-MS). Treated plants changed their pattern of biomass distribution; they had lower stem mass fraction and higher branch and leaf mass fraction than untouched plants. Size, weight and number of tubers were not significantly affected. Touching did not cause trichome damage nor change their total number on touched terminal leaves. However, on primary leaves the number of glandular trichomes and pavement cells was significantly increased. Touching altered the volatile emission of treated plants; they released higher quantities of the sesquiterpenes (E)-ß-caryophyllene, germacrene D-4-ol and (E)-nerolidol, and lower quantities of the terpenes (E)-ocimene and linalool, indicating a systemic effect of the treatment. The odour of touched plants was significantly less preferred by the aphids Macrosiphum euphorbiae and Myzus persicae compared to odour of untouched plants. The results suggest that light contact may have a potential role in the detection of neighbouring plants and may affect plant-insect interactions.

Alterations of physiology and gene expression due to long-term magnesium-deficiency differ between leaves and roots of Citrus reticulata.

Seedlings of Ponkan (Citrus reticulata) were irrigated with nutrient solution containing 0 (Mg-deficiency) or 1mM MgSO4 (control) every two day for 16 weeks. Thereafter, we examined magnesium (Mg)-deficiency-induced changes in leaf and root gas exchange, total soluble proteins and gene expression. Mg-deficiency lowered leaf CO2 assimilation, and increased leaf dark respiration. However, Mg-deficient roots had lower respiration. Total soluble protein level was not significantly altered by Mg-deficiency in roots, but was lower in Mg-deficient leaves than in controls. Using cDNA-AFLP, we obtained 70 and 71 differentially expressed genes from leaves and roots. These genes mainly functioned in signal transduction, stress response, carbohydrate and energy metabolism, cell transport, cell wall and cytoskeleton metabolism, nucleic acid, and protein metabolisms. Lipid metabolism (Ca(2+) signals)-related Mg-deficiency-responsive genes were isolated only from roots (leaves). Although little difference existed in the number of Mg-deficiency-responsive genes between them both, most of these genes only presented in Mg-deficient leaves or roots, and only four genes were shared by them both. Our data clearly demonstrated that Mg-deficiency-induced alterations of physiology and gene expression greatly differed between leaves and roots. In addition, we focused our discussion on the causes for photosynthetic decline in Mg-deficient leaves and the responses of roots to Mg-deficiency.

Plants adapted to nutrient limitation allocate less biomass into stems in an arid-hot grassland.

Biomass allocation can exert a great influence on plant resource acquisition and nutrient use. However, the role of biomass allocation strategies in shaping plant community composition under nutrient limitations remains poorly addressed. We hypothesized that species-specific allocation strategies can affect plant adaptation to nutrient limitations, resulting in species turnover and changes in community-level biomass allocations across nutrient gradients. In this study, we measured species abundance and the concentrations of nitrogen and phosphorus in leaves and soil nutrients in an arid-hot grassland. We quantified species-specific allocation parameters for stems vs leaves based on allometric scaling relationships. Species-specific stem vs leaf allocation parameters were weighted with species abundances to calculate the community-weighted means driven by species turnover. We found that the community-weighted means of biomass allocation parameters were significantly related to the soil nutrient gradient as well as to leaf stoichiometry, indicating that species-specific allocation strategies can affect plant adaptation to nutrient limitations in the studied grassland. Species that allocate less to stems than leaves tend to dominate nutrient-limited environments. The results support the hypothesis that species-specific allocations affect plant adaptation to nutrient limitations. The allocation trade-off between stems and leaves has the potential to greatly affect plant distribution across nutrient gradients.

Effects of drought stress on growth, solute accumulation and membrane stability of leafy vegetable, huckleberry (Solanum scabrum Mill.).

The present study sought to investigate the factors implicated in growth impairment of huckleberry (a leafy vegetable) under water stress conditions. To achieve this, seedlings of plant were subjected to control, mild stress and severe stress conditions for 30 days. Plant growth, plant water relation, gas exchange, oxidative stress damage, electrolyte leakage rate, mineral content and osmolyte accumulation were measured. Water deficit markedly decreased leaf, stem and root growth. Leaf photosynthetic rate was tremendously reduced by decrease in stomatal conductance under stress conditions. Malondialdehyde (MDA) content markedly increased under mild (82%) and severe (131%) stress conditions, while electrolyte leakage rate (ELR) increased by 59% under mild stress and 3-fold under severe stress. Mineral content in leafwas high in stressed plants, while proline content markedly increased under mild stress (12-fold) and severe stress (15-fold), with corresponding decrease in osmotic potential at full turgor and an increase in osmotic adjustment. These results suggest that maintenance of high mineral content and osmotic adjustment constitute important adaptations in huckleberry under water deficit conditions and that growth depression under drought stress would be mainly caused by increased electrolyte leakage resulting from membrane damage induced by oxidative stress.

Different combinations of morpho-physiological traits are responsible for tolerance to drought in wild tomatoes Solanum chilense and Solanum peruvianum.

Herbaceous species can modify leaf structure during the growing season in response to drought stress and water loss. Evolution can select combinations of traits in plants for efficient water use in restricted environments. We investigated plant traits that mediate adaptation and acclimation to water stress in two herbaceous drought-tolerant species. Anatomical, morphological and physiological traits related to stems and leaves were examined under optimal watering (OW) and a long period of restricted watering (RW) in 11 accessions from three Solanaceae species (Solanum chilense, S. peruvianum and S. lycopersicum). The relationships between these traits were tested using linear regression and PCA. There were significant differences in anatomical traits between the species under both OW and RW, where leaf area correlated with stem diameter. Proline and total carbohydrates accumulated highly in S. chilense and S. peruvianum, respectively, and these osmolytes were strongly correlated with increased osmotic potential. Stomatal density varied between species but not between acclimation treatments, while stomatal rate was significantly higher in wild tomatoes. There was a strong positive relationship between stem growth rate and a group of traits together expressed as total stomatal number. Total stomata is described by integration of leaf area, stomatal density, height and internode length. It is proposed that constitutive adaptations and modifications through acclimation that mediate RW play an important role in tolerance to drought stress in herbaceous plants. The capacity for growth under drought stress was not associated with any single combination of traits in wild tomatoes, since the two species differed in relative levels of expression of various phenotypic traits.

Diurnal patterns in Scots pine stem oleoresin pressure in a boreal forest.

Coniferous tree stems contain large amounts of oleoresin under positive pressure in the resin ducts. Studies in North-American pines indicated that the stem oleoresin exudation pressure (OEP) correlates negatively with transpiration rate and soil water content. However, it is not known how the OEP changes affect the emissions of volatile vapours from the trees. We measured the OEP, xylem diameter changes indicating changes in xylem water potential and monoterpene emissions under field conditions in mature Scots pine (Pinus sylvestris L.) trees in southern Finland. Contrary to earlier reports, the diurnal OEP changes were positively correlated with temperature and transpiration rate. OEP was lowest at the top part of the stem, where water potentials were also more negative, and often closely linked to ambient temperature and stem monoterpene emissions. However, occasionally OEP was affected by sudden changes in vapour pressure deficit (VPD), indicating the importance of xylem water potential on OEP as well. We conclude that the oleoresin storage pools in tree stems are in a dynamic relationship with ambient temperature and xylem water potential, and that the canopy monoterpene emission rates may therefore be also regulated by whole tree processes and not only by the conditions prevailing in the upper canopy.

Shade affects responses to drought and flooding - acclimation to multiple stresses in bittersweet (Solanum dulcamara L.).

Plants exposed to environmental stress often respond by a change in their phenotypic traits. These changes in trait expression may alleviate the negative effect of such stress factors. However, if multiple stresses are present, responses are likely to be less predictable and hence do not necessarily correlate to plant performance. This study tested if this expectation was true, by subjecting Solanum dulcamara plants to various simultaneous stress factors. Plants were grown in well-watered conditions, drought or flooding, and exposed to either full light or shade for 4 weeks. Shoot and root biomass, stem morphological parameters, such as height, number of nodes and length of stem internodes, and leaf traits like length, specific leaf area, chlorophyll content and stomatal conductance were determined. Both variation in light and in water availability typically caused slower growth, and resulted in distinct phenotypic changes in stem, leaf and root traits. However, effects of stresses on the expression of traits were not always additive. Instead, some combined stress responses (e.g. leaf size) appeared to be limited by physical or physiological constraints, whereas other responses were opposite to each other (e.g. root:shoot ratio), resulting in an intermediate phenotype in the combined stress treatment. These data suggest that in natural conditions, where combined stress factors are likely to be present, the optimal phenotype may not necessarily be expressed. Responses of plants to multiple stress factors may therefore not be associated with immediate advantages in terms of increased performance.