Changes In Xylem Sap Composition Research Articles

Multi-omics analysis of xylem sap uncovers dynamic modulation of poplar defenses by ammonium and nitrate.

Xylem sap is the major transport route for nutrients from roots to shoots. In the present study, we investigated how variations in nitrogen (N) nutrition affected the metabolome and proteome of xylem sap and the growth of the xylem endophyte Brennaria salicis, and we also report transcriptional re-wiring of leaf defenses in poplar (Populus × canescens). We supplied poplars with high, intermediate or low concentrations of ammonium or nitrate. We identified 288 unique proteins in xylem sap. Approximately 85% of the xylem sap proteins were shared among ammonium- and nitrate-supplied plants. The number of proteins increased with increasing N supply but the major functional categories (catabolic processes, cell wall-related enzymes, defense) were unaffected. Ammonium nutrition caused higher abundances of amino acids and carbohydrates, whereas nitrate caused higher malate levels in xylem sap. Pipecolic acid and N-hydroxy-pipecolic acid increased, whereas salicylic acid and jasmonoyl-isoleucine decreased, with increasing N nutrition. Untargeted metabolome analyses revealed 2179 features in xylem sap, of which 863 were differentially affected by N treatments. We identified 124 metabolites, mainly from specialized metabolism of the groups of salicinoids, phenylpropanoids, phenolics, flavonoids, and benzoates. Their abundances increased with decreasing N, except coumarins. Brennaria salicis growth was reduced in nutrient-supplemented xylem sap of low- and high- NO3- -fed plants compared to that of NH4+ -fed plants. The drastic changes in xylem sap composition caused massive changes in the transcriptional landscape of leaves and recruited defenses related to systemic acquired and induced systemic resistance. Our study uncovers unexpected complexity and variability of xylem composition with consequences for plant defenses.

Regulation of leaf hydraulics: from molecular to whole plant levels.

The water status of plant leaves is dependent on both stomatal regulation and water supply from the vasculature to inner tissues. The present review addresses the multiple physiological and mechanistic facets of the latter process. Inner leaf tissues contribute to at least a third of the whole resistance to water flow within the plant. Physiological studies indicated that leaf hydraulic conductance (Kleaf) is highly dependent on the anatomy, development and age of the leaf and can vary rapidly in response to physiological or environmental factors such as leaf hydration, light, temperature, or nutrient supply. Differences in venation pattern provide a basis for variations in Kleaf during development and between species. On a short time (hour) scale, the hydraulic resistance of the vessels can be influenced by transpiration-induced cavitations, wall collapses, and changes in xylem sap composition. The extravascular compartment includes all living tissues (xylem parenchyma, bundle sheath, and mesophyll) that transport water from xylem vessels to substomatal chambers. Pharmacological inhibition and reverse genetics studies have shown that this compartment involves water channel proteins called aquaporins (AQPs) that facilitate water transport across cell membranes. In many plant species, AQPs are present in all leaf tissues with a preferential expression in the vascular bundles. The various mechanisms that allow adjustment of Kleaf to specific environmental conditions include transcriptional regulation of AQPs and changes in their abundance, trafficking, and intrinsic activity. Finally, the hydraulics of inner leaf tissues can have a strong impact on the dynamic responses of leaf water potential and stomata, and as a consequence on plant carbon economy and leaf expansion growth. The manipulation of these functions could help optimize the entire plant performance and its adaptation to extreme conditions over short and long time scales.

Comparison of xylem sap mineral concentrations between kiwifruit shoot types using spittlebugs for non-destructive sampling of sap.

Excreta of the meadow spittlebug [Philaenus spumarius L. (Homoptera: Cercopidae)] feeding on leaves and pedicels of kiwifruit [Actinidia deliciosa (A. Chev.) C.F. Liang et A.R. Ferguson var. deliciosa 'Hayward'] were collected from insects from two different positions in the vine: from long, non-terminating axillary shoots producing fruit that are high in Ca2+ and low in K+ and from short-terminating axillary shoots producing fruit that are low in Ca2+ and high in K+. The Ca2+, Mg2+, K+ and P concentrations in the excreta were determined, and found to be similar to those in the xylem sap. Daily and seasonal changes in xylem sap composition were compared in excreta collected from the two different shoot types. On average, Ca2+ and Mg2+ concentrations were higher and K+ and P concentrations were lower in xylem sap collected from pedicels on long, non-terminating axillary shoots than in sap collected from pedicels on short-terminating shoots. Differences in the mineral concentration between these two shoot types may therefore be due to differences in the xylem sap mineral concentration reaching the fruit. There was no measurable gradient in xylem sap composition within the parent shoots that could explain the differences between sap composition of the two axillary shoot types. Long, non-terminating shoots had higher leaf area, were more exposed, had higher stomatal conductance and rates of transpiration, and more negative leaf water potentials than short-terminating shoots. The higher xylem sap Ca2+ and Mg2+ concentrations of long shoots were therefore associated with higher rates of water transport to the long shoots. Xylem sap concentration differences between these two shoots types may have been because of differential loading or unloading of minerals between shoot types, associated with differences in transpiration rate or shoot growth rates. The higher transpiration rate of long shoots may cause phloem immobile minerals such as Ca2+ to accumulate to higher levels at cation exchange sites in the shoot apoplast, resulting in increased xylem sap concentrations arriving at the fruit.

Do increases in xylem sap pH and/or ABA concentration mediate stomatal closure following nitrate deprivation?

Stomatal conductance (g(s)) of pepper (Capsicum annuum L.) plants decreased during the second photoperiod (day 2) after withholding nitrate (N). Stomatal closure of N-deprived plants was not associated with a decreased shoot water potential (Psi(shoot)); conversely Psi(shoot) was lower in N-supplied plants. N deprivation transiently (days 2 and 3) alkalized (0.2-0.3 pH units) xylem sap exuded from de-topped root systems under root pressure, and xylem sap expressed from excised shoots by pressurization. The ABA concentration of expressed sap increased 3-4-fold when measured on days 2 and 4. On day 2, leaves detached from N-deprived and N-supplied plants showed decreased transpiration rates when fed an alkaline (pH 7) artificial xylem (AX) solution, independent of the ABA concentration (10-100 nM) supplied. Thus changes in xylem sap composition following N deprivation can potentially close stomata. However, the lower transpiration rate of detached N-deprived leaves relative to N-supplied leaves shows that factors residing within N-deprived leaves also mediate stomatal closure, and that these factors assume greater importance as the duration of N deprivation increases.

Hormonal Interactions and Stomatal Responses

Both environmental and hormonal factors and their interactions affect stomatal behavior. Methodologies for identifying hormonal interactions affecting stomatal function are reviewed. Although there is abundant evidence that abscisic acid (ABA) closes stomata, evidence that the other classical plant hormones (auxins, cytokinins, ethylene, gibberellins) in isolation alter stomatal response often comes from exogenous applications to detached epidermes and leaves, rather than correlation of endogenous concentrations with stomatal conductance (gs). Evidence for hormonal interactions comes from isolated tissues with exogenous hormones supplied at nonphysiological concentrations, or from variation in stomatal response to xylem ABA concentration in planta. The roles of hormonal changes in causing stomatal closure following changes in soil environment are considered. Although soil drying induces multiple changes in xylem sap composition, analysis of stomatal responses suggests a dominant role for increased endogenous ABA concentrations and relatively little evidence of roles for other hormones. A similar picture emerges from studies of soil compaction. Although soil flooding decreases ABA export from the root system, there is some evidence that apoplastic ABA accumulation elicits stomatal closure. Stomatal closure following nitrogen deprivation does not appear to involve ABA and may provide a suitable experimental system to investigate roles for other hormones. The availability of mutant or transgenic lines with altered hormone homeostasis or sensitivity provides opportunities to screen for altered stomatal behavior in response to different environments, and may provide new evidence that hormonal interactions are important in the control of stomatal behavior.

Influence of Soil Compaction on Xylem Sap Composition in Barley ( Hordeum vulgare L.)

Summary Barley ( Hordeum vulgare L. cv. Steptoe) was grown at various soil bulk densities under non-limiting water and nutrient conditions to assess the role of changes in xylem sap pH and the concentrations of nitrate, calcium, potassium, magnesium and sodium as potential root-to-shoot signal(s) mediating piant responses to soil compaction. Significant reductions in leaf conductance, growth and changes in xylem sap composition were observed at a bulk density of 1.7 g cm −3 (severe compaction) when compared with the uncompacted 1.1 g cm −3 control treatment (low compaction). Xylem sap calcium concentration was increased, while stomatal conductance was reduced in severely compacted soil at 6, 12 and 18 days after emergence (DAE). The concentration of magnesium in the xylem sap was also increased in the 1.7 g cm −3 treatment at 6 and 12 DAE, while that of potassium was up to 3-fold lower than in the 1.1 g cm −3 treatment. The most notable effect of severe compaction on the ionic composition of the xylem sap was a substantial decrease in nitrate concentration relative to uncompacted control plants at 6 and 12 DAE, which was associated with the observed reductions in leaf area. pH and sodium levels were both unaffected by the compaction treatments examined. The role of changes in the ionic concentration of xylem sap as a root-to-shoot signal during compaction stress is evaluated.

Nitrogen Nutrition and Xylem Sap Composition of Peanut (Arachis hypogaea L. cv Virginia Bunch)

The principal forms of amino nitrogen transported in xylem were studied in nodulated and non-nodulated peanut (Arachis hypogaea L.). In symbiotic plants, asparagine and the nonprotein amino acid, 4-methyleneglutamine, were identified as the major components of xylem exudate collected from root systems decapitated below the lowest nodule or above the nodulated zone. Sap bleeding from detached nodules carried 80% of its nitrogen as asparagine and less than 1% as 4-methyleneglutamine. Pulse-feeding nodulated roots with (15)N(2) gas showed asparagine to be the principal nitrogen product exported from N(2)-fixing nodules. Maintaining root systems in an N(2)-deficient (argon:oxygen, 80:20, v/v) atmosphere for 3 days greatly depleted asparagine levels in nodules. 4-Methyleneglutamine represented 73% of the total amino nitrogen in the xylem sap of non-nodulated plants grown on nitrogen-free nutrients, but relative levels of this compound decreased and asparagine increased when nitrate was supplied. The presence of 4-methyleneglutamine in xylem exudate did not appear to be associated with either N(2) fixation or nitrate assimilation, and an origin from cotyledon nitrogen was suggested from study of changes in amount of the compound in tissue amino acid pools and in root bleeding xylem sap following germination. Changes in xylem sap composition were studied in nodulated plants receiving a range of levels of (15)N-nitrate, and a (15)N dilution technique was used to determine the proportions of accumulated plant nitrogen derived from N(2) or fed nitrate. The abundance of asparagine in xylem sap and the ratio of asparagine:nitrate fell, while the ratio of nitrate:total amino acid rose as plants derived less of their organic nitrogen from N(2). Assays based on xylem sap composition are suggested as a means of determining the relative extents to which N(2) and nitrate are being used in peanuts.