Hydathode trichomes actively secreting water from leaves play a key role in the physiology and evolution of root-parasitic rhinanthoid Orobanchaceae

Background and Aims Root hemiparasites from the rhinanthoid clade of Orobanchaceae possess metabolically active glandular trichomes that have been suggested to function as hydathode trichomes actively secreting water, a process that may facilitate resource acquisition from the host plant’s root xylem. However, no direct evidence relating the trichomes to water secretion exists, and carbon budgets associated with this energy-demanding process have not been determined.Methods Macro- and microscopic observations of the leaves of hemiparasitic Rhinanthus alectorolophus were conducted and night-time gas exchange was measured. Correlations were examined among the intensity of guttation, respiration and transpiration, and analysis of these correlations allowed the carbon budget of the trichome activity to be quantified. We examined the intensity of guttation, respiration and transpiration, correlations among which indicate active water secretion.Key Results Guttation was observed on the leaves of 50 % of the young, non-flowering plants that were examined, and microscopic observations revealed water secretion from the glandular trichomes present on the abaxial leaf side. Night-time rates of respiration and transpiration and the presence of guttation drops were positively correlated, which is a clear indicator of hydathode trichome activity. Subsequent physiological measurements on older, flowering plants indicated neither intense guttation nor the presence of correlations, which suggests that the peak activity of hydathodes is in the juvenile stage.Conclusions This study provides the first unequivocal evidence for the physiological role of the hydathode trichomes in active water secretion in the rhinanthoid Orobanchaceae. Depending on the concentration of organic elements calculated to be in the host xylem sap, the direct effect of water secretion on carbon balance ranges from close to neutral to positive. However, it is likely to be positive in the xylem-only feeding holoparasites of the genus Lathraea, which is closely related to Rhinanthus. Thus, water secretion by the hydathodes might be viewed as a physiological pre-adaptation in the evolution of holoparasitism in the rhinanthoid lineage of Orobanchaceae.     

Long-Distance Water Transport in Aquatic Plants

Acropetal mass flow of water is demonstrated in two submerged angiosperms, Lobelia dortmanna L. and Sparganium emersum Rehman by means of guttation measurements. Transpiration is absent in truly submerged plants, but the presence of guttation verifies that long-distance water transport takes place. Use of tritiated water showed that the water current arises from the roots, and the main flow of water is channeled to the youngest leaves. This was confirmed by measurement of guttation, which showed the highest rates in young leaves. Guttation rates were 10-fold larger in the youngest leaf of S. emersum (2.1 [mu]L leaf-1 h-1) compared with the youngest leaf of L. dortmanna (0.2 [mu]L leaf-1 h-1). This is probably due to profound species differences in the hydraulic conductance (2.7 x 10-17 m4 Pa-1 s-1 for S. emersum and 1.4 x 10-19 m4 Pa-1 s-1 for L. dortmanna). Estimates derived from the modified Hagen-Poiseuille equation showed that the maximum flow velocity in xylem vessels was 23 to 84 cm h-1, and the required root pressure to drive the flow was small compared to that commonly found in terrestrial plants. In S. emersum long-distance transport of water was shown to be dependent on energy conversion in the roots. The leaves ceased to guttate when the roots were cooled to 4[deg]C from the acclimatization level at 15[deg]C, whereas the guttation was stimulated when the temperature was increased to 25[deg]C. Also, the guttation rate decreased significantly when vanadate was added to the root medium. The observed water transport is probably a general phenomenon in submerged plants, where it can act as a translocation system for nutrients taken up from the rich root medium and thereby assure maximum growth.     

Evaluation of water stress control with polyethylene glycols by analysis of guttation

The water relations of pepper plants (Capsicum frutescens L.) under conditions conducive to guttation were studied to evaluate the control of plant water stress with polyethylene glycols. The addition of polyethylene glycol 6000 to the nutrient solution resulted in water relations similar to those expected in soil at the same water potentials. Specifically, xylem pressure potential in the root and leaf became more negative during a 24-hour treatment period, while osmotic potential of the root xylem sap remained constant. The decrease in pressure potential was closely correlated with the decrease in osmotic potential of the nutrient solution. In contrast, the addition of polyethylene glycol 400 to the nutrient medium resulted in a reduction of osmotic potential in the root xylem sap; this osmotic adjustment in the xylem was large enough to establish an osmotic gradient for entry of water and cause guttation at a nutrient solution osmotic potential of −4.8 bars. Pressure potential in the root and leaf xylem became negative only at nutrient solution osmotic potentials lower than −4.8 bars. About half of the xylem osmotic adjustment in the presence of polyethylene glycol 400 was caused by increased accumulation of K⁺, Na⁺, Ca²⁺, and Mg²⁺ in the root xylem. These studies indicate that larger polyethylene glycol molecules such as polyethylene glycol 6000 are more useful for simulating soil water stress than smaller molecules such as polyethylene glycol 400.     

Production of recombinant proteins in tobacco guttation fluid

Guttation, the loss of water and dissolved materials from uninjured plant organs, is a common phenomenon in higher plants. By using endoplasmic reticulum signal peptides fused to the recombinant protein sequences, we have generated transgenic tobacco (Nicotiana tabacum L. cv Wisconsin) plants that secrete three heterologous proteins of different genetic backgrounds (bacterial xylanase, green fluorescent protein of jellyfish [Aequorea victoria], and human placental alkaline phosphatase) through the leaf intercellular space into tobacco guttation fluid. Production rates of 1.1 microg/g of leaf dry weight per day were achieved for alkaline phosphatase with this protein comprising almost 3% of total soluble protein in the guttation fluid. Guttation fluid can be collected throughout a plant's life, thus providing a continuous and nondestructive system for recombinant protein production. Guttation fluid has the potential of increasing the efficiency of recombinant protein production technology by increasing yield, abolishing extraction, and simplifying its downstream processing.     

Sap Exudation via the Epidermis of Leaves

Reports of stomatal guttation in Gladiolus and of active waterexudation in Vidafaba could not be confirmed by an experimentaland anatomical investigation. Exudation of water from detachedpine needles is considered to be due to processes distinct fromthose causing guttation. Water soaking of leaves, or the flooding of leaf air spaces,and the wettability of cell walls under root pressure as distinctfrom their wettability by water externally applied are discussedin relation to the experimental results.     

Salt stress aggravates boron toxicity symptoms in banana leaves by impairing guttation

Boron (B) is known to accumulate in the leaf margins of different plant species, arguably a passive consequence of enhanced transpiration at the ends of the vascular system. However, transpiration rate is not the only factor affecting ion distribution. We examine an alternative hypothesis, suggesting the participation of the leaf bundle sheath in controlling radial water and solute transport from the xylem to the mesophyll in analogy to the root endodermis. In banana, excess B that remains confined to the vascular system is effectively disposed of via dissolution in the guttation fluid; therefore, impairing guttation should aggravate B damage to the leaf margins. Banana plants were subjected to increasing B concentrations. Guttation rates were manipulated by imposing a moderate osmotic stress. Guttation fluid was collected and analysed continuously. The distribution of ions across the lamina was determined. Impairing guttation indeed led to increased B damage to the leaf margins. The kinetics of ion concentration in guttation samples revealed major differences between ion species, corresponding to their distribution in the lamina dry matter. We provide evidence that the distribution pattern of B and other ions across banana leaves depends on active filtration of the transpiration stream and on guttation.     

Evidence that root pressure flow is required for calcium transport to head leaves of cabbage

Young cabbage plants (Brassica oleracea L. var. capitata) that were exposed to an atmosphere at 50% relative humidity transpired freely and accumulated significant quantities of ⁴⁵Ca in the leaves. Plants that were enclosed by plastic bags to stop transpiration from all leaves exhibited guttation with the development of root pressure and also accumulated significant quantities of ⁴⁵Ca in the leaves. ⁴⁵Ca accumulation increased in the leaves and tended to decrease in roots and stems with increasing quantities of water transpired or guttated by the plant. When plants were only partially enclosed so that some leaves were covered and the remainder exposed, only the exposed leaves that were transpiring accumulated significant quantities of ⁴⁵Ca. The covered leaves of partially enclosed plants exhibited no guttation and accumulated little ⁴⁵Ca with no measurable ⁴⁵Ca at the margins of the leaves. The results demonstrate that root pressure flow is required to transport adequate amounts of Ca to those tissues in plants that are not undergoing transpirational water loss.     

The uptake of mannitol by higher plants

Experiments with youngHordeum sativum andHelianthus annus plants showed that in the excretion of mannitol in the guttation liquid observed byGroenewegen andMills (1960) after uptake by the root system of plants, the osmotic concentration of mannitol in the nutrient medium and the temperature are significant. The beginning of mannitol excretion during guttation is accelerated considerably by the increase of the osmotic concentration of mannitol in the nutrient medium and the rising temperature. The osmotic concentration of mannitol is also important for the duration of mannitol excretion in the guttation liquid after transfer of the plants into a nutrient medium without mannitol. In the presence of mannitol in the nutrient medium water uptake by the root system and growth are inhibited and the tissues of the organs above ground and of the root system are dehydrated. The inhibitory effect of mannitol on the water uptake by the root system is immediate.     

Identification of guttation fluid proteins: the presence of pathogenesis-related proteins in non-infected barley plants

The main driving force behind water transport in plants is the air's low water potential. In the presence of high humidity, the transpiration process is halted and water transport is mainly sustained by the root pressure. The surplus of water following the removal of essential components (e.g. salts) is excreted by the plant via guttation through the hydathodes. When guttation occurs, the plant surface is wetted. These are the conditions that will allow epiphytic living, motile bacteria to move and to eventually enter the plant's interior via the hydathodes. The question arose as to whether the plant has developed a protection mechanism against motile bacteria in the vicinity of the hydathodes. Such a protection mechanism could use the well known pathogenesis-related (PR) proteins. Indeed, an analysis of the guttation fluid using one- and two-dimensional electrophoresis showed a clustering of approximately 200 proteins, primarily with isoelectric points in the acidic pH. Proteins identified using electrospray ionization mass spectroscopic analysis and western blot analysis belong mostly to the family of PR-proteins suggesting a role in plant protection against invaders. The protein profile of the guttation fluid was remarkably modified by treating plants with methyl jasmonic acid suggesting that the protein composition of the guttation fluid is controlled by internal and/or external stimuli.     

Hydathodal leaf teeth of Chloranthus japonicus (Chloranthaceae) prevent guttation-induced flooding of the mesophyll

Why the leaves of cold temperate deciduous and moisture-loving angiosperms are so often toothed has long puzzled biologists because the functional consequences of teeth remain poorly understood. Here we provide functional and structural evidence that marginal leaf teeth of Chloranthus japonicus, an understory herb, enable the release of guttation sap during root pressure. When guttation from teeth hydathodes was experimentally blocked, we found that the leaf intercellular airspaces became flooded. Measurements of chlorophyll a fluorescence revealed that internal flooding resulted in an inhibition of photosynthesis, most likely through the formation of a film of water within the leaf that reduced CO₂ diffusion. Comparing a developmental series of leaves with and without teeth experimentally covered with wax, we found that teeth did not affect overall leaf stomatal conductance and CO₂ uptake. However, maximal and effective light-saturation PSII quantum yields of teeth were found to be lower or equal to the surrounding lamina throughout leaf ontogeny. Collectively, our results suggest hydathodes and their development on teeth apices enable the avoidance of mesophyll flooding by root pressure. We discuss how these new findings bear on the potential physiological interpretations of models that apply leaf marginal traits to infer ancient climates.     

Photosynthetic adjustment after rehydration in <Emphasis Type="Italic">Annona emarginata</Emphasis>

Reports indicate that Annona emarginata is tolerant to drought and is also used as an alternative rootstock for atemoya under drought conditions. The photosynthetic process can be adjusted after rehydration, resulting on total or partial recovery. The aim of this study was to determine if A. emarginata shows adjusts in gas exchange and the chlorophyll a fluorescence pattern after rehydration. During water deficits, the gas exchange and water content in the leaf decreased. However, after 5 days of rehydration, the water content in the leaf recovered and rehydrated plants presented the water use efficiency better than irrigated plants. Further remaining gas exchange parameters were lower in relation to irrigated plants. In chlorophyll a fluorescence, the rehydrated plants showed higher dissipation of light energy as heat, maintaining high activity of photoprotection. After rehydration, A. emarginata shows a positive correlation between transpiration and CO₂ assimilation rate, which optimize the water use efficiency. Thus, A. emarginata presents adjustments in gas exchange and photochemical process, resulting on a possible long-term photosynthetic acclimation to water deficiency.     

Variations in leaf characteristics of six species of Sagittaria (alismataceae) caused by various water levels

Plants grown from seeds of Sagittaria falcata, S. lancifolia, S. platyphylla, S. rigida, S. isoetiformis and S. papillosa were grown in water-saturated soil or soil submerged to 4.5, 12, 19.5 or 27 cm. Length and width of leaves and petiole lengths were measured at anthesis of the first flower on the first inflorescence produced by each plant. In general, leaf width and length were decreased by submergence, and petiole length increased. The species × water depth interaction was significant for emersed leaf width, leaf length and petiole length except for S. lancifolia leaf length and S. papillosa leaf length and width. These trends indicate genetic differences among, and variability within, taxa. Leaf width, leaf length and petiole length of plants growing in seed source populations were measured. The means from these measurements, when compared to those from experimental plants, indicate that both groups of plants respond similarly to variations in water depth.     

Suppression of bacterial blight by a bacterial community isolated from the guttation fluids of anthuriums

Growth and survival of Xanthomonas campestris pv. dieffenbachiae in guttation fluids (xylem sap exuded from leaf margins) of anthuriums were suppressed by several bacterial strains indigenous to leaves of various anthurium cultivars. Inhibition of growth was not observed in filter-sterilized guttation fluids and was restored to original levels only by reintroducing specific mixtures of bacteria into filter-sterilized guttation fluids. The inhibitory effect was related to the species in the bacterial community rather than to the total numbers of bacteria in the guttation fluids. One very effective bacterial community consisted of five species isolated from inhibitory guttation fluids of two susceptible anthurium cultivars. The individual strains in this community had no effect on the pathogen, but the mixture was inhibitory to X. campestris pv. dieffenbachiae in guttation fluids. The populations of the individual strains remained near the initial inoculum levels for at least 14 days. The effect of the five inhibitory strains on reducing disease in susceptible anthurium plants was tested by using a bioluminescent strain of X. campestris pv. dieffenbachiae to monitor the progression of disease in leaves nondestructively. Invasion of the pathogen through hydathodes at leaf margins was reduced by applying the strain mixture to the leaves. When the strain mixture was applied directly to wounds created on the leaf margins, the pathogen failed to invade through the wounds. This bacterial community has potential for biological control of anthurium blight.     

Stachybotrys spp. and the guttation phenomenon

The formation of guttation droplets is a long-known property of various fungi. However, their composition, biological function and metabolism in fungi have hardly attracted deeper research interest. The highly toxic mould Stachybotrys (S.) chartarum chemotype S is supposed to play—amongst other factors such as endotoxins and microbial volatile organic compounds (MVOCs)—an important role in indoor air toxicity, mainly after water damage. The way of toxins becoming airborne and leading to exposure via inhalation, however, is still under discussion. We hypothesised that guttation may be a factor for exudation of toxins into the environment. Therefore, selected isolates (n?=?15) of our own culture collection of Stachybotrys spp. (S. chartarum chemotype S, S. chartarum chemotype A, S. chlorohalonta) originating from various habitats were cultivated on malt extract agar for 3 weeks. All strains but one produced different amounts of guttation droplets, which were collected quantitatively and subjected to various independent analytical techniques like ELISA, effect-based bioassay (MTT cell culture test) and tandem mass spectrometry (LC-MS/MS). Actually, the toxigenic isolates (n?=?5) produced highly toxic guttation droplets, which was confirmed by all methods. The concentration of macrocyclic trichothecenes, such as satratoxin G and H, ranged between the LOD and 7,160 ng/ml exudate and 280 and 4,610 ng/ml as determined by LC-MS/MS, respectively. According to our knowledge, the ability of S. chartarum to produce toxic exudates is reported for the first time, which possibly plays an important role regarding its toxic potential in indoor environments.     

Ecophysiological responses of Caragana korshinskii Kom. under extreme drought stress: Leaf abscission and stem survives

Caragana korshinskii Kom. is a perennial xerophytic shrub, well known for its ability to resist drought. In order to study ecophysiological responses of C. korshinskii under extreme drought stress and subsequent rehydration, diurnal patterns of gas exchange and chlorophyll (Chl) fluorescence parameters of photosystem II as well as Chl content were analyzed. Plant responses to extreme drought included (1) leaf abscission and using stem for photosynthesis, (2) improved instantaneous water-use efficiency, (3) decreased photosynthetic rate and partly closed stomata owing to leaf abscission and low water status, (4) decreased maximum photochemical efficiency of photosystem II (PSII) (variable to maximum fluorescence ratio, F_v/F_m), quantum efficiency of noncyclic electron transport of PSII, and Chl a and Chl b. Four days after rehydration, new leaves budded from stems. In the rewatered plants, the chloroplast function was restored, the gas exchange and Chl fluorescence returned to a similar level as control plant. The above result indicated that maintaining an active stem system after leaf abscission during extreme drought stress may be the foundation which engenders these mechanisms rapid regrowth for C. korshinskii in arid environment.     

Detrimental effect of rust infection on the water relations of bean

Bean plants (Phaseolus vulgaris L.) infected with the rust Uromyces phaseoli became unusually susceptible to drought as sporulation occurred. Under the conditions used (1,300 ft-c, 27 C, and 55% relative humidity) such plants wilted at soil water potentials greater than −1 bar, whereas healthy plants did not wilt until the soil water potential fell below −3.4 bars. Determinations of leaf water and osmotic potentials showed that an alteration in leaf osmotic potential was not responsible for the wilting of diseased plants. When diffusive resistance was measured as a function of decreasing leaf water content, the resistance of healthy leaves increased to 50 sec cm⁻¹ by the time relative water content decreased to 70%, whereas the resistance of diseased leaves remained less than 8 sec cm⁻¹ down to 50% relative water content. Apparently, water vapor loss through cuticle damaged by the sporulation process, together with the reduction in root to shoot ratio which occurs in diseased plants, upset the water economy of the diseased plant under mild drought conditions.     

Improving growth, flower yield, and water relations of snapdragon (Antirhinum majus L.) plants grown under well-watered and water-stress conditions using arbuscular mycorrhizal fungi

The influence of arbuscular mycorrhizal (AM) fungus Glomus deserticola (Trappe and John) on plant growth, nutrition, flower yield, water relations, chlorophyll (Chl) contents and water-use efficiency (WUE) of snapdragon (Antirhinum majus cv. butterfly) plants were studied in potted culture under well-watered (WW) and water-stress (WS) conditions. The imposed water stress condition significantly reduced all growth parameters, nutrient contents, flower yield, water relations, and Chl pigment content and increased the electrolyte leakage of the plants comparing to those of nonstressed plants. Regardless of the WS level, the mycorrhizal snapdragon plants had significantly higher shoot and root dry mass (DM), WUE, flower yield, nutrient (P, N, K, Mg, and Ca) and Chl contents than those nonmycorrhizal plants grown both under WW or WS conditions. Under WS conditions, the AM colonization had greatly improved the leaf water potential (??_w), leaf relative water content (RWC) and reduced the leaf electrolyte leakage (EL) of the plants. Although the WS conditions had markedly increased the proline content of the leaves, this increase was significantly higher in nonmycorrhizal than in mycorrhizal plants. This suggests that AM colonization enhances the host plant WS tolerance. Values of benefit and potential dry matter for AM-root associations were highest when plants were stressed and reduced under WW conditions. As a result, the snapdragon plants showed a high degree of dependency on AM fungi which improve plant growth, flower yield, water relations particularly under WS conditions, and these improvements were increased as WS level had increased. This study confirms that AM colonization can mitigate the deleterious effect of water stress on growth and flower yield of the snapdragon ornamental plant.     

模拟酸雨对西洋杜鹃生理生态特性的影响

采用盆栽方法,研究了不同pH值条件下的模拟酸雨对西洋杜鹃生长及其叶片主要生理生化特征的影响。结果表明:在酸雨胁迫下,随着酸雨胁迫的增强,西洋杜鹃叶片受害程度逐渐加重;叶片丙二醛含量逐渐升高;可溶性蛋白含量先升后降;过氧化氢酶(CAT)、过氧化物酶(POD)和超氧化物歧化酶(SOD)活性均呈先升后降的单峰曲线变化,其中pH值为4.3的处理下西洋杜鹃叶片CAT和SOD活性最高,pH值为3.0的处理下其POD活性最高;其叶绿素含量逐渐下降;叶片净光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)及水分利用率(WUE)均呈先升后降的趋势,而胞间CO2浓度(Ci)则持续下降,pH值为4.3的处理下其Pn、Tr、Gs和WUE均达到最高。研究表明,大致可以认为pH值≤3.0是酸雨对西洋杜鹃造成隐形伤害的阀值,而酸雨灾害严重地区的降水pH值为2.0—4.0左右,说明西洋杜鹃可以在酸雨灾害较重的地区生长,可作为酸雨灾害严重地区园林绿化及植被构建的物种之一。     

Overexpression of Tamarix albiflonum TaMnSOD increases drought tolerance in transgenic cotton

Drought is a major environmental stress that limits cotton (Gossypium hirsutum L.) production worldwide. TaMnSOD plays a crucial role as a peroxidation scavenger. In this study, TaMnSOD cDNA of Tamarix albiflonum was overexpressed in the cotton cultivar fy11 by Agrobacterium tumefaciens-mediated transformation. The transformed plants were assessed by gDNA PCR, RT-PCR and DNA gel blot analysis. The physiological and biochemical characters of two independent transgenic lines and control plants were tested and compared, and the morphological traits (biomass, root and lateral root length, leaf number) were also detected after recovery from water-withholding stress. When water was withheld from pot-grown 6-week-old seedlings for 18 days (watering to 8 % of field capacity), transgenic cotton plants accumulated more proline and soluble sugar than wild-type plants (WT). The activity of antioxidant enzymes such as superoxide dismutase and peroxidase was enhanced in transgenic plants under drought stress. Cell membrane integrity was also considerably improved under water stress, as indicated by reduced malondialdehyde content relative to control plants. Furthermore, net photosynthesis, stomatal conductance and transpiration rate were increased in transgenic plants compared with wild type. Transgenic cotton showed increases in biomass as well as root and leaf systems compared with WT after 2 weeks recovery from stress. These results suggest that TaMnSOD transgenic cotton plants acquired improved drought tolerance through enhanced development of the root and leaf system and the regulation of superoxide scavenging.     

Distribution of hydroxamic acids in zea mays tissues

The hydroxamic acid content of leaves of cereals correlates well with resistance to aphids. In maize these compounds were absent from xylem exudates and guttation drops. Lateral veins of leaves of 7-day-old maize plants contained 8 mmol/kg fr. wt. while the entire leaf contained only 4.2 mmol/kg fr. wt. In leaves of 20-day-old plants, these amounts decreased by ca one-third. In mesocotyls, the cortex and central vascular cylinder contained 1.3 and 2.2 mmol/kg fr. wt, respectively. In 12-day-old wheat plants, the complete leaves and their veins contained 2.4 and 6.4 mmol/kg fr. wt respectively. Thus, the concentration of hydroxamic acid was always higher in the vascular bundles.