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Sequential leaf senescence and correlatively controlled increases in xylem flow resistance.

In bean, Phaseolus vulgaris L. (Contender), the directly measured hydraulic resistance of the xylem pathway between roots and primary leaf pulvinal junctions increased rapidly and progressively from 21 to 28 days after planting. These increases in xylem resistance (+390%) were specifically located in the pulvinal junction of the primary leaf. Moreover, they occurred just prior to the onset of primary leaf yellowing. Developmental increases in xylem hydraulic flow resistance and stomatal resistance, as well as subsequent primary leaf yellowing, were completely prevented by detopping the shoots above the primary leaves at 21 days. Thus, the onset of these senescence-associated symptoms was correlatively controlled. In short-term investigations of the mechanisms involved, flow between petiole and cut tip of excised leaves was rapidly reduced by infiltration of 20 picomoles of soluble dextran into the xylem. Moreover, imbibition of approximately 120 picomoles of dextran by excised leaves increased stomatal resistances. A programmed secretion of hormonal concentrations of similar polysaccharides into specific xylem sections in vivo might provide a mechanism for regulating the partitioning of essential xylem supplies between leaves, thus inducing sequential leaf senescence.

Journal Article↗

Nitric oxide production by the differentiating xylem of Zinnia elegans.

Nitric oxide (NO) is currently regarded as a signal molecule involved in plant cell differentiation and programmed cell death. Here, we investigated NO production in the differentiating xylem of Zinnia elegans by confocal laser scanning microscopy to answer the question of whether NO is produced during xylem differentiation. Results showed that NO production was mainly located in both phloem and xylem regardless of the cell differentiation status. However, there was evidence for a spatial NO gradient inversely related to the degree of xylem differentiation and a protoplastic NO burst was associated with the single cell layer of pro-differentiating thin-walled xylem cells. Confirmation of these results was obtained using trans-differentiating Z. elegans mesophyll cells. In this system, the scavenging of NO by means of 2-phenyl-4,4,5,5-tetramethyl imidazoline-1-oxyl-3-oxide (PTIO) inhibits tracheary element differentiation but increases cell viability. These results suggest that plant cells, which are just predetermined to irreversibly trans-differentiate in xylem elements, show a burst in NO production, this burst being sustained as long as secondary cell wall synthesis and cell autolysis are in progress.

Apoptosis↗

Fastidious xylem-limited bacterial plant pathogens.

Numerous bacteria have been isolated from within plants, and many reported from xylem, but only three species of xylem-limited bacteria (XLB) that are fastidious in cultural requirements, are plant pathogens, and exclusively occupy xylem, have been well characterized. Two XLB, Xylella fastidiosa and Pseudomonas syzygii, are transmitted by sucking insects that feed on xylem sap but are not transmitted mechanically from plant to plant. In contrast, Clavibacter xyli is mechanically transmitted to plants by cutting tools. All of these XLB occupy a highly specialized yet diverse ecological niche: the water-conducting systems of an extremely wide range of plant hosts. A variety of detection methods are available as diagnostic aids; each method has advantages and disadvantages; no single method is best for all uses. Molecular and genetic comparisons of strains of XLB lag behind progress being made for many other plant-pathogenic bacteria, but such studies are needed to answer important questions: (a) How do XLB move from cell to cell within plants? (b) What are the physiological and genetic bases of plant host specificity for XLB? (c) Why are only xylem-feeding specialists vectors of X. fastidiosa (and probably P. syzygii), when many leafhoppers feed regularly (but not continuously) on xylem?

Journal Article↗

Analysis of xylem sap proteins from Brassica napus.

BACKGROUND: Substance transport in higher land plants is mediated by vascular bundles, consisting of phloem and xylem strands that interconnect all plant organs. While the phloem mainly allocates photoassimilates, the role of the xylem is the transport of water and inorganic nutrients from roots to all aerial plant parts. Only recently it was noticed that in addition to mineral salts, xylem sap contains organic nutrients and even proteins. Although these proteins might have important impact on the performance of above-ground organs, only a few of them have been identified so far and their physiological functions are still unclear. RESULTS: We used root-pressure xylem exudate, collected from cut Brassica napus stems, to extract total proteins. These protein preparations were then separated by high-resolution two-dimensional gel electrophoresis (2-DE). After individual tryptic digests of the most abundant coomassie-stained protein spots, partial peptide sequence information was deduced from tandem mass spectrometric (MS/MS) fragmentation spectra and subsequently used for protein identifications by database searches. This approach resulted in the identification of 69 proteins. These identifications include different proteins potentially involved in defence-related reactions and cell wall metabolism. CONCLUSION: This study provides a comprehensive overview of the most abundant proteins present in xylem sap of Brassica napus. A number of 69 proteins could be identified from which many previously were not known to be localized to this compartment in any other plant species. Since Brassica napus, a close relative of the fully sequenced model plant Arabidopsis thaliana, was used as the experimental system, our results provide a large number of candidate proteins for directed molecular and biochemical analyses of the physiological functions of the xylem under different environmental and developmental conditions. This approach will allow exploiting many of the already established functional genomic resources, like i.e. the large mutant collections, that are available for Arabidopsis.

Brassica napus↗

Global comparative transcriptome analysis identifies gene network regulating secondary xylem development in Arabidopsis thaliana.

Our knowledge of the genetic control of wood formation (i.e., secondary growth) is limited. Here, we present a novel approach to unraveling the gene network regulating secondary xylem development in Arabidopsis, which incorporates complementary platforms of comparative-transcriptome analyses such as "digital northern" and "digital in situ" analysis. This approach effectively eliminated any genes that are expressed in either non-stem tissues/organs ("digital northern") or phloem and non-vascular regions ("digital in situ"), thereby identifying 52 genes that are upregulated only in the xylem cells of secondary growth tissues as "core xylem gene set". The proteins encoded by this gene set participate in signal transduction, transcriptional regulation, cell wall metabolism, and unknown functions. Five of the seven signal transduction-related genes represented in the core xylem gene set encode the essential components of ROP (Rho-related GTPase from plants) signaling cascade. Furthermore, the analysis of promoter sequences of the core xylem gene set identified a novel cis-regulatory element, ACAAAGAA. The functional significances of this gene set were verified by several independent experimental and bioinformatics methods.

Arabidopsis↗

Structure of lignins in developing xylem of Norway spruce.

The developing xylem in a Norway spruce (Picea abies) clone was investigated during a growth season and compared to lignin from sapwood of the same tree clone. Klason and acid-soluble lignin contents were determined as well as the carbohydrate monomer distribution and protein content. By analyzing lignin thioacidolysis products, it was shown that only guaiacyl units could be detected in the materials, and the relative amount of beta-O-4' bonds was assessed. Monomeric and selected dimeric lignin products were identified by mass spectrometry. The specimens were embedded and thin sections examined by microscopy to determine the state of cell differentiation in the samples. In the spring and early summer, growth was very rapid and the intention was to collect tissue in which exclusively the middle lamella/primary cell wall had begun to lignify. Combining data regarding Klason lignin, protein content and carbohydrate monomer distribution with microscopy, it was found that the developing xylem sample from mid-June contained lignin from exclusively middle lamella/primary wall. The Klason lignin content in the developing xylem during the growth season was 20%, 5% and 10% in April, June and August, respectively. Thioacidolysis showed that the lignin had more condensed structures than lignin from the reference Norway spruce clone wood. Mass spectrometry showed that the developing xylem specimens from June and August contained more lignin structures with end-groups than the reference sample. These results suggest that lignification in the cambial layer and early developing xylem may take place more in a bulk fashion during the summer.

Lignin↗

Effect of partial rootzone drying on the concentration of zeatin-type cytokinins in tomato (Solanum lycopersicum L.) xylem sap and leaves.

Decreased cytokinin (CK) export from roots in drying soil might provide a root-to-shoot signal impacting on shoot physiology. Although several studies show that soil drying decreases the CK concentration of xylem sap collected from the roots, it is not known whether this alters xylem CK concentration ([CK(xyl)]) in the leaves and bulk leaf CK concentration. Tomato (Solanum lycopersicum L.) plants were grown with roots split between two soil columns. During experiments, water was applied to both columns (well-watered; WW) or one (partial rootzone drying; PRD) column. Irrigation of WW plants aimed to replace transpirational losses every day, while PRD plants received half this amount. Xylem sap was collected by pressurizing detached leaves using a Scholander pressure chamber, and zeatin-type CKs were immunoassayed using specific antibodies raised against zeatin riboside after separating their different forms (free zeatin, its riboside, and nucleotide) by thin-layer chromatography. PRD decreased the whole plant transpiration rate by 22% and leaf water potential by 0.08 MPa, and increased xylem abscisic acid (ABA) concentration 2.5-fold. Although PRD caused no detectable change in [CK(xyl)], it decreased the CK concentration of fully expanded leaves by 46%. That [CK(xyl)] was maintained and not increased while transpiration decreased suggests that loading of CK into the xylem was also decreased as the soil dried. That leaf CK concentration did not decline proportionally with CK delivery suggests that other mechanisms such as CK metabolism influence leaf CK status of PRD plants. The causes and consequences of decreased shoot CK status are discussed.

Solanum lycopersicum↗

Interspecific variation in xylem vulnerability to cavitation among tropical tree and shrub species.

In tropical moist forests, seasonal drought limits plant survival, productivity and diversity. Drought-tolerance mechanisms of tropical species should reflect the maximum seasonal water deficits experienced in a particular habitat. We investigated stem xylem vulnerability to cavitation in nine tropical species with different life histories and habitat associations. Stem xylem vulnerability was scored as the xylem water potential causing 50 and 75% loss of hydraulic conductivity (P50 and P75, respectively). Four shade-tolerant shrubs ranged from moderately resistant (P50=-1.9 MPa for Ouratea lucens Kunth. Engl.) to highly resistant to cavitation (P50=-4.1 MPa for Psychotria horizontalis Sw.), with shallow-rooted species being the most resistant. Among the tree species, those characteristic of waterlogged soils, Carapa guianensis Aubl., Prioria copaifera Griseb. and Ficus citrifolia Mill., were the most vulnerable to cavitation (P50=-0.8 to -1.6 MPa). The wet-season, deciduous tree, Cordia alliodora (Ruiz and Pav.) Oken., had resistant xylem (P50=-3.2 MPa), whereas the dry-season, deciduous tree, Bursera simaruba (L.) Sarg. was among the most vulnerable to cavitation (P50=-0.8 MPa) of the species studied. For eight out of the nine study species, previously reported minimum seasonal leaf water potentials measured in the field during periods of drought correlated with our P50 and P75 values. Rooting depth, deciduousness, soil type and growth habit might also contribute to desiccation tolerance. Our results support the functional dependence of drought tolerance on xylem resistance to cavitation.

Seasons↗

Translocation of nitrogen in the xylem of field-grown cherry and poplar trees during remobilization.

Studies of small trees growing in pots have established that individual amino acids or amides are translocated in the xylem sap of a range of tree species following bud burst, as a consequence of nitrogen (N) remobilization from storage. This paper reports the first study of N translocation in the xylem of large, deciduous, field-grown trees during N remobilization in the spring. We applied 15N fertilizer to the soil around 10-year-old Prunus avium L. and Populus trichocharpa Torr. & Gray ex Hook var. Hastata (Dode) A. Henry x Populus balsamifera L. var. Michauxii (Dode) Farwell trees before bud burst to label N taken up by the roots. Recovery of unlabeled N in xylem sap and leaves was used to demonstrate that P. avium remobilizes N in both glutamine (Gln) and asparagine (Asn). Sap concentrations of both amides rose sharply after bud burst, peaking 14 days after bud burst for Gln, and remaining high some 45 days for Asn. There was no 15N enrichment of either amide until 21 days after bud burst. In the Populus trees, nearly all the N was translocated in the sap as Gln, the concentration of which peaked and then declined before the amide was enriched with 15N, 40 days after bud burst. Xylem sap of clonal P. avium trees was sampled at different positions in the crown to assess if the amino acid and amide composition of the sap varied within the crown. Sap was sampled during remobilization (when the concentration of Gln was maximal), at the end of remobilization and at the end of the experiment (68 days after bud burst). Although the date of sampling had a highly significant effect on sap composition, the effect of position of sampling was marginal. The results are discussed in relation to N translocation in adult trees and the possibility of measuring N remobilization by calculating the flux of N translocation in the xylem.

Amides↗

Monolignol oxidation by xylem peroxidase isoforms of Norway spruce (Picea abies) and silver birch (Betula pendula).

We partially purified peroxidase isoform fractions from xylem extracts of a gymnosperm, Norway spruce (Picea abies (L.) Karst.), and an angiosperm, silver birch (Betula pendula Roth.), to determine the participation of xylem-localized peroxidases in polymerization of different types of lignin in vivo. Several peroxidase fractions varying in isoelectric point values from acidic to basic were tested for their ability to catalyze the oxidation of the monolignols coniferyl alcohol, sinapyl alcohol and p-coumaryl alcohol in vitro. All of the xylem peroxidases extracted from Norway spruce and most of those from silver birch showed the highest rate of oxidation with coniferyl alcohol in the presence of hydrogen peroxide. The exception was an acidic peroxidase fraction (pI 3.60-3.65) from silver birch that exhibited higher oxidation activity for sinapyl alcohol than for coniferyl alcohol. For the xylem enzyme fractions extracted from silver birch, the ability to oxidize the artificial phenolic substrate syringaldazine coincided with high specific activity for sinapyl alcohol. Therefore, we conclude that the acidic, neutral and basic xylem peroxidases of Norway spruce all function in the synthesis of guaiacyl-type lignin, whereas in silver birch the acidic peroxidases preferentially oxidize sinapyl subunits. The latter provides a mechanism for synthesis of guaiacyl-syringyl lignin typical of tracheid cell walls in angiosperm trees.

Betula↗

Xylella fastidiosa infection and ethylene exposure result in xylem and water movement disruption in grapevine shoots.

It is conventionally thought that multiplication of the xylem-limited bacterium Xylella fastidiosa (Xf) within xylem vessels is the sole factor responsible for the blockage of water movement in grapevines (Vitis vinifera) affected by Pierce's disease. However, results from our studies have provided substantial support for the idea that vessel obstructions, and likely other aspects of the Pierce's disease syndrome, result from the grapevine's active responses to the presence of Xf, rather than to the direct action of the bacterium. The use of magnetic resonance imaging (MRI) to observe the distribution of water within the xylem has allowed us to follow nondestructively the development of vascular system obstructions subsequent to inoculation of grapevines with Xf. Because we have hypothesized a role for ethylene produced in vines following infection, the impact of vine ethylene exposure on obstruction development was also followed using MRI. In both infected and ethylene-exposed plants, MRI shows that an important proportion of the xylem vessels become progressively air embolized after the treatments. The loss of xylem water-transporting function, assessed by MRI, has been also correlated with a decrease in stem-specific hydraulic conductivity (K(S)) and the presence of tyloses in the lumens of obstructed water conduits. We have observed that the ethylene production of leaves from infected grapevines is greater than that from healthy vines and, therefore, propose that ethylene may be involved in a series of cellular events that coordinates the vine's response to the pathogen.

Ethylenes↗

Functional coordination between leaf gas exchange and vulnerability to xylem cavitation in temperate forest trees.

We examined functional coordination among stem and root vulnerability to xylem cavitation, plant water transport characteristics and leaf traits in 14 co-occurring temperate tree species. Relationships were evaluated using both traditional cross-species correlations and phylogenetically independent contrast (PIC) correlations. For stems, the xylem tension at which 50% of hydraulic conductivity was lost (psi50) was positively associated (P < 0.001) with specific conductivity (K(S)) and with mean hydraulically weighted xylem conduit diameter (D(h-w)), but was only marginally (P = 0.06) associated with leaf specific conductivity (K(L)). The PIC correlation for each of these relationships, however, was not statistically significant. There was also no relationship between root psi50 and root K(S) in either cross-species or PIC analysis. Photosynthetic rate (A) and stomatal conductance (g(s)) were strongly and positively correlated with root psi50 in the cross-species analysis (P < 0.001), a relationship that was robust to phylogenetic correction (P < 0.01). A and g(s) were also positively correlated with stem psi50 in the cross-species analysis (P = 0.02 and 0.10, respectively). However, only A was associated with stem psi50 in the PIC analysis (P = 0.04). Although the relationship between vulnerability to cavitation and xylem conductivity traits within specific organs (i.e. stems and roots) was weak, the strong correlation between g(s) and root psi50 across species suggests that there is a trade-off between vulnerability to cavitation and water transport capacity at the whole-plant level. Our results were therefore consistent with the expectation of coordination between vulnerability to xylem cavitation and the regulation of stomatal conductance, and highlight the potential physiological and evolutionary significance of root hydraulic properties in controlling interspecific variation in leaf function.

Biological Transport↗

[Anatomical and water physiological plasticity of Grewia biloba var. parviflora leaf and secondary xylem].

Based on the anatomical observations of leaf and secondary xylem as well as the measurements of leaf water physiological parameters, this paper studied the anatomical and water physiological plasticity of Grewia biloba var. parviflora growing in different succession stage communities. The results showed that G. biloba var. parviflora leaf was characterized by thin bifacial with thin cuticle and few stoma, indicating that it was mesophyte anatomically, while the secondary xylem had typical xeromorphic traits, such as short and thin vessel, high vessel frequency, low percentage single pores, and short fibers and rays. G. biloba var. parviflora had high plasticity in the anatomical structure and water physiological features of leaf and secondary xylem, and the plasticity index was in the order of secondary xylem anatomical structure (0. 24) > water physiological traits (0. 19) > leaf anatomical structure (0. 18). Compared with those growing in mixed forest and Platycladus orientalis forest, the individuals of G. biloba var. parviflora in shrub communities had the xeromorphic traits in the aspects of (1) their secondary xylem had shorter vessel elements, higher vessel frequency, less single porous percentage, lower rays, higher relative conductivity and lower vulnerability index, and (2) their leaf had lower water potential, lower water content and free water content, higher bound water content, high ratio of bound to free water content, and less specific leaf area. The phenotypic plasticity, both anatomical and physiological, made G. biloba var. parviflora tolerate to the drought at earlier succession stages and better adapt to the mesophytic condition at later stages, and consequently, become a widely distributed and dominant species in mixed forests.

Adaptation, Physiological↗

Properties of the K+ inward rectifier in the plasma membrane of xylem parenchyma cells from barley roots: effects of TEA+, Ca2+, Ba2+ and La3+.

Xylem parenchyma cells are situated around the (apoplastic) xylem vessels and are involved in the control of the composition of the xylem sap by exporting and resorbing solutes. We investigated properties of the K+ inward rectifier in the plasma membrane of these cells by performing patch clamp experiments on protoplasts in the whole-cell configuration. Inward currents were sensitive to the K+ channel blocker TEA+ at a high concentration (20 mM). Barium, another "classical" K+ channel blocker, inhibited K+ currents with a Ki of about 1.3 mM. In contrast to guard cells, the cytosolic Ca2+ level proved to be ineffective in regulating the K+ conductance at hyperpolarization. External Ca2+ blocked currents weakly in a voltage-dependent manner. From instantaneous current-voltage curves, we identified a binding site in the channel pore with an electrical distance of about 0.2 to 0.5. Lanthanum ions reduced the inward current in a voltage-dependent manner and simultaneously displaced the voltage at which half of the channels are in the open state to more positive values. This finding was interpreted as resulting from a sum of two molecular effects, an interaction with the mouth of the channel that causes a reduction of current, and a binding to the voltage sensor, leading to a shielding of surface charges and, subsequently, a modulation of channel gating. A comparison between the K+ inward rectifier in xylem parenchyma cells, guard cells and KAT1 from Arabidopsis leads to the conclusion that these rectifiers form subtypes within one class of ion channels. The ineffectiveness of Ca2+ to control K+ influx in xylem parenchyma cells is interpreted in physiological terms.

Barium↗

Cellular machinery of wood production: differentiation of secondary xylem in Pinus contorta var. latifolia.

The objectives of this study were to define cell structure during pine secondary xylem development and to integrate this information with current knowledge of the biochemistry and physiology of secondary cell wall biosynthesis in gymnosperms. Lodgepole pine (Pinus contorta var. latifolia Englem.) cambium and secondary xylem were cryofixed using high pressure freezing and freeze-substitution which allowed excellent preservation of the cell structure of developing secondary xylem and enabled high-resolution transmission electron microscopic viewing of these cells for the first time. In contrast to their precursors in the adjacent cambial zone, developing tracheids were active in secondary wall deposition, with abundant cortical microtubules and developing bordered pits. These cells were also characterized by unusual Golgi structures: the trans-Golgi network was highly developed and the associated vesicles were large and darkly stained. These unusual Golgi structures persisted throughout the period of xylem maturation until programmed cell death occurred. Immuno-cytochemistry and enzyme-gold probes were used to investigate the distribution of key secretory products (mannans) and a lignification-associated enzyme (coniferin beta-glucosidase) during xylogenesis. Mannans were localized to the secondary cell wall, the trans-Golgi cisternae and trans-Golgi network vesicles of developing xylem. Coniferin beta-glucosidase was found only in the secondary cell wall. The cell wall localization of coniferin beta-glucosidase, the enzyme responsible for cleaving glucose from coniferin to generate free coniferyl alcohol, provides a mechanism to de-glucosylate monolignols in muro. A two-step model of lignification of conifer tracheids is proposed. First, Golgi-mediated secretion deposits monolignols into the cell wall, where they polymerize in cell corners and middle lamella. Secondly, cell lysis releases stored, vacuolar monolignol glucosides into the wall where they are deglucosylated and their polymerization is influenced by the wall environment including the lignin deposited earlier.

Cell Differentiation↗

Phenylcoumaran benzylic ether reductase, a prominent poplar xylem protein, is strongly associated with phenylpropanoid biosynthesis in lignifying cells.

It has previously been shown (D.R. Gang et al., 1999, J Biol Chem 274: 7516-7527) that the most abundant protein in the secondary xylem of poplar (Populus trichocarpa cv. 'Trichobel') is a phenylcoumaran benzylic ether reductase (PCBER), an enzyme involved in lignan synthesis. Here, the distribution and abundance of PCBER in poplar was studied at both the RNA and protein level. The cellular expression pattern was determined by immunolocalization of greenhouse-grown plants as well as of a field-grown poplar. Compared to other poplar tissues, PCBER is preferentially produced in the secondary xylem of stems and roots and is associated with the active growth period. The protein is present in all cells of the young differentiating xylem, corresponding to the zone of active phenylpropanoid metabolism and lignification. In addition, PCBER is located in young differentiating phloem fibers, in xylem ray parenchyma, and in xylem parenchyma cells at the growth-ring border. Essentially the same expression pattern was observed in poplars grown in greenhouses and in the field. The synthesis of PCBER in phenylpropanoid-synthesizing tissues was confirmed in a bending experiment. Induction of PCBER was observed in the pith of mechanically bent poplar stems, where phenylpropanoid metabolism is induced. These results indicate that the products of PCBER activity are synthesized mainly in lignifying tissues, suggesting a role in wood development.

Antibody Formation↗

Relationship between xylem ion concentration and bean growth responses to short-term salinisation in spring and summer.

Bean plants, Phaseolus vulgaris L. cv. Contender, were grown in the spring and summer seasons to study the relationship between xylem Na+/Cl-, transpiration rate, and salt tolerance. Eight-day-old seedlings were transplanted to 50% modified Hoagland solution with 1 mM NaCl. Four days after transfer, one of two treatments was applied: a control of 1 mM NaCl or a treatment of 25 mM NaCl every two days to reach a final treatment concentration of 75 mM NaCl. Plants were sampled on the fourth day after the final salt concentration was reached, eight days after the salinisation treatment began. Relative growth rate was 2.6-fold greater in summer than in spring. However, while no differences were found between treatments in spring, summer salt-treated plants had growth rates that were 31% lower than those of controls. In summer, CO2 assimilation, stomatal conductance, and transpiration rate of salinised plants declined with respect to controls. Leaf Na+ and trifoliolate leaf Cl- were higher in salt-treated plants in summer, although root Na+ was significantly higher in spring. Moreover, in summer salinity inhibited Ca2+ and K+ uptake and changed its distribution. Summer salt-treated plants had an average of 17-fold higher xylem Na+ during the daily cycle, while xylem Cl-, only in the afternoon, showed higher values (1.5-fold) compared to spring-grown plants. Our results suggest that the faster growth response to salt in summer-grown bean was at least partly due to an increase in xylem Na+ independent of the transpiration rate and possibly related to an increase in xylem Na+ influx or/and Na+ recirculation.

Ions↗

Chemical composition and enzymatic degradability of xylem and nonxylem walls isolated from alfalfa internodes.

During plant maturation, degradability of alfalfa (Medicago sativa L.) stems declines due to accumulation of highly lignified xylary tissue. Xylem and nonxylem tissues dissected from lower alfalfa internodes were analyzed for cell wall constituents and degradability. Cell walls comprised 740 mg g(-1) of xylem and 533 mg g(-1) of nonxylem tissues. Xylem tissues contributed about 60% of the cell wall mass in internodes. Xylem walls contained 28% lignin, 4% pectin, 29% hemicellulose, and 39% cellulose as compared to 15% lignin, 25% pectin, 30% hemicellulose, and 30% cellulose in nonxylem walls. Fungal enzymes hydrolyzed 22 and 73% of the structural carbohydrates in xylem and nonxylem walls, respectively. In both cell wall fractions, the release of xylose was 56-90% lower than that of other sugars, indicating that lignin preferentially restricted xylan degradation in secondary walls and xyloglucan degradation in primary walls. Elucidation of lignin-xylose interactions may reveal strategies for improving fiber degradability of alfalfa.

Cell Wall↗