Localization of ferruginol,a diterpene phenol,in <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> heartwood by time-of-flight secondary ion mass spectrometry

Time-of-flight secondary ion mass spectrometry (TOF-SIMS) was applied to the investigation of heartwood extractives in Sugi (Cryptomeria japonica). Sugi heartwood tissue generated secondary ions that were not produced from sapwood tissue by TOF-SIMS. Among the peculiar ions generated from heartwood, two positive ions of m/z 285 and 301 were remarkable due to their appearance in a larger mass range and with a high intensity. These two ions were not generated from heartwood tissue preextracted with n-hexane, and the n-hexane extract of Sugi heartwood produced both ions. Gas chromatography-mass spectrometry of the n-hexane extract demonstrated that ferruginol, a diterpene phenol, the molecular weight of which is 286, constituted one of the predominant constituents of the extract. Authentic ferruginol also generated both ions by TOF-SIMS. The molecular formula of the m/z 285 ion generated from Sugi heartwood tissue was estimated to be C₂₀H₂₉O, which corresponds well with that of ferruginol, i.e. C₂₀H₃₀O, by peak identification. All these results strongly suggest that the m/z 285 ion generated from Sugi heartwood tissue originated significantly from ferruginol in Sugi heartwood. By TOF-SIMS imaging, the m/z 285 ion was detected uniformly in the tracheid cell walls, in the cell walls of the axial parenchyma cells and ray parenchyma cells, and also inside these parenchyma cells. These results indicate that ferruginol was distributed almost evenly in Sugi heartwood tissue.     

Preparation of antibody against agatharesinol, a norlignan, using a hapten-carrier conjugate.

In order to immunolabel heartwood extractives in Japanese cedar (Sugi, Cryptomeria japonica), we attempted to prepare antibodies against agatharesinol, a major norlignan of these heartwood extractives. Agatharesinol by itself is not antigenic due to its low-M(r), and thus was covalently bound to bovine serum albumin in order to synthesize an antigenic hapten-carrier conjugate (artificial antigen). The number of agatharesinol molecules per artificial antigen molecule was estimated as 27-28 by quantifying Lys in an acid hydrolysate of the artificial antigen by HPLC. Reaction between the artificial antigen and serum obtained from a rabbit immunized with the artificial antigen was competitively inhibited by agatharesinol, indicating the successful production of anti-agatharesinol antibodies. Inhibition by sequirin C, another major norlignan in Sugi, was weaker than that by agatharesinol. Furthermore, an EtOAc soluble fraction, which contains mainly norlignans, inhibited the reaction more strongly than any of the other fractions of Sugi heartwood extractives. Thus, the antiserum we have produced reacts most strongly with agatharesinol and recognizes norlignans almost selectively among Sugi heartwood extractives.     

The function of intercellular spaces along the ray parenchyma in sapwood, intermediate wood, and heartwood of Cryptomeria japonica (Cupressaceae)

? Premise of the study: Intercellular spaces along ray parenchyma (ISRP) are common in many conifer xylems, but their function is uncertain because the in-situ structural network among ISRP, ray parenchyma, and tracheids has not been evaluated. Analysis of water distribution in ISRP from sapwood to heartwood is needed to elucidate the function of ISRP in sapwood, intermediate wood, and heartwood. ? Methods: We used cryo-scanning electron microscopy, x-ray photography, and water content measurement in xylem to analyze the presence of liquids in ISRP, ray parenchyma, and tracheids from sapwood to heartwood in Cryptomeria japonica (Cupressaceae). ? Key results: In sapwood, almost all ISRP were empty. "Cingulate-cavitated regions", which lose water along the tangential direction within one annual ring, formed in the earlywood tracheids, and their frequency increased toward the inner annual rings, whereas ray parenchyma cells were alive and not involved in the partial cavitation. In intermediate wood, almost all ISRP and earlywood tracheids and many of the ray cells were empty, and only some latewood tracheids retained liquid in their lumina. The ISRP were connected with tracheids via gas-filled ray parenchyma cells. ? Conclusions: The ISRP work as a pathway of gas for aspiration of ray parenchyma cells in sapwood. On the other hand, the occurrence of a gas network between ISRP, ray parenchyma, and tracheids facilitates cavitation of tracheids, resulting in the generation of low-moisture, intermediate wood.     

What is disjunctive xylem parenchyma? A case study of the African tropical hardwood Okoubaka aubrevillei (Santalaceae)

The morphological variation and structure-function relationships of xylem parenchyma still remain open to discussion. We analyzed the three-dimensional structure of a poorly known type of xylem parenchyma with disjunctive walls in the tropical hardwood Okoubaka aubrevillei (Santalaceae). Disjunctive cells occurred among the apotracheal parenchyma cells and at connections between axial and ray parenchyma cells. The disjunctive cells were partly detached one from another, but their tubular structures connected them into a continuous network of axial and ray parenchyma. The connecting tubules had thick secondary walls and simple pits with plasmodesmata at the points where one cell contacted a tubule of another cell. The imperforate tracheary elements of the ground tissue were seven times longer than the axial parenchyma strands, a fact that supports a hypothesis that parenchyma cells develop disjunctive walls because they are pulled apart and partly separated during the intrusive growth of fibers. We discuss unresolved details of the formation of disjunctive cell walls and the possible biomechanical advantage of the wood with disjunctive parenchyma: the proportion of tissue that improves mechanical strength is increased by the intrusive elongation of fibers (thick-walled tracheids), whereas the symplastic continuum of the parenchyma is maintained through formation of disjunctive cells.     

Living cells in wood. 1. Absence,scarcity and histology of axial parenchyma as keys to function

The diversity of expression in axial parenchyma (or lack of it) in woods is reviewed and synthesized with recent work in wood physiology, and questions and hypotheses relative to axial parenchyma anatomy are offered. Cell shape, location, abundance, size, wall characteristics and contents are all characteristics for the assessment of the physiological functions of axial parenchyma, a tissue that has been neglected in the consideration of how wood histology has evolved. Axial parenchyma occurrence should be considered with respect to mechanisms for the prevention and reversal of embolisms in tracheary elements. This mechanism complements cohesion–tension‐based water movement and root pressure as a way of maintaining flow in xylem. Septate fibres can substitute for axial parenchyma (‘axial parenchyma absent’) and account for water movement in xylem and for the supply of carbohydrate abundance underlying massive and sudden events of foliation, flowering and fruiting, as can fibre dimorphism and the co‐occurrence of septate fibres and axial parenchyma. Rayless woods may or may not contain axial parenchyma and are informative when analysing parenchyma function. Interconnections between ray and axial parenchyma are common, and so axial and radial parenchyma must be considered as complementary parts of a network, with distinctive but interactive functions. Upright ray cells and more numerous rays per millimetre enhance interconnection and are more often found in woods that contain tracheids. Vesselless woods in both gymnosperms and angiosperms have axial parenchyma, the distribution of which suggests a function in osmotic water shifting. Water and photosynthate storage in axial parenchyma may be associated with seasonal changes and with succulent or subsucculent modes of construction. Apotracheal axial parenchyma distribution often demonstrates storage functions that can be read independently of osmotic water shifting capabilities. Axial parenchyma may serve to both enhance mechanical strength or, when parenchyma is thin‐walled, as a tissue that adapts to volume change with a change in water content. Other functions of axial parenchyma (contributing resistance to pathogens; a site for the recovery of physical damage) are considered. The diagnostic features of axial parenchyma and septate fibres are reviewed in order to clarify distinctions and to aid in cell type identification. Systematic listings are given for particular axial parenchyma conditions (e.g. axial parenchyma ‘absent’ with septate fibres substituting). A knowledge of the axial parenchyma information presented here is desirable for a full understanding of xylem function. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 177 , 291–321.     

Agatharesinol biosynthesis-related changes of ray parenchyma in sapwood sticks of Cryptomeria japonica during cell death

Planta - The work demonstrates a relationship between the biosynthesis of the secondary metabolite, agatharesinol, and cytological changes that occur in ray parenchyma during cell death in sapwood...     

Vessel diameter is related to amount and spatial arrangement of axial parenchyma in woody angiosperms

Parenchyma represents a critically important living tissue in the sapwood of the secondary xylem of woody angiosperms. Considering various interactions between parenchyma and water transporting vessels, we hypothesize a structure–function relationship between both cell types. Through a generalized additive mixed model approach based on 2,332 woody angiosperm species derived from the literature, we explored the relationship between the proportion and spatial distribution of ray and axial parenchyma and vessel size, while controlling for maximum plant height and a range of climatic factors. When factoring in maximum plant height, we found that with increasing mean annual temperatures, mean vessel diameter showed a positive correlation with axial parenchyma proportion and arrangement, but not for ray parenchyma. Species with a high axial parenchyma tissue fraction tend to have wide vessels, with most of the parenchyma packed around vessels, whereas species with small diameter vessels show a reduced amount of axial parenchyma that is not directly connected to vessels. This finding provides evidence for independent functions of axial parenchyma and ray parenchyma in large vesselled species and further supports a strong role for axial parenchyma in long‐distance xylem water transport.     

Differences in patterns of cell death between ray parenchyma cells and ray tracheids in the conifers <Emphasis Type="Italic">Pinus densiflora</Emphasis> and <Emphasis Type="Italic">Pinus rigida</Emphasis>

Differences in patterns of cell death between ray parenchyma cells and ray tracheids in the conifers Pinus densiflora and Pinus rigida were clarified. Differentiation and cell death of ray tracheids occurred successively and both were related to the distance from the cambium. In this respect, they resembled those of longitudinal tracheids. Thus, the cell death of short-lived ray tracheids could be characterized as time-dependent programmed cell death. In contrast, ray parenchyma cells survived for several years or more, and no successive cell death occurred, even within a single radial line of cells in a ray. Thus, the features of death of the ray parenchyma cells were different from those of ray tracheids. Cell death occurred early in ray parenchyma cells that were in contact with ray tracheids. The initiation of secondary wall thickening occurred earlier in ray parenchyma cells that were in contact with ray tracheids in Pinus densiflora than in others. In addition, localized thickening of secondary walls occurred only in ray parenchyma cells that were in contact with ray tracheids in Pinus rigida. Moreover, no polyphenols were evident in such cells in either species. Therefore, ray parenchyma cells that were in contact with ray tracheids appeared not to play a role in the formation of heartwood extractives. Our observations indicate that short-lived ray tracheids might affect the pattern of differentiation and, thus, the functions of neighboring long-lived ray parenchyma cells in conifers.     

Bordered pits in ray cells and axial parenchyma: the histology of conduction, storage, and strength in living wood cells

Bordered pits occur in walls of living ray cells of numerous species of woody dicotyledons. The occurrence of this feature has been minimally reported because the pits are relatively small and not easily observed in face view. Bordered pits are illustrated in sectional view with light microscopy and with scanning electron microscopy in face view for dicotyledonous and gnetalean woods. Bordered pits are more numerous and often have prominent borders on tangential walls of procumbent ray cells, but also occur on radial walls; they are approximately equally abundant on tangential and horizontal walls of upright cells, suggesting parallels to cell shape in flow pathway design. Axial parenchyma typically has secondary walls thinner than those of ray cells, but bordered pits or large simple pit areas occur on some cross walls of parenchyma strands. There is no apparent correlation between the phylogenetic position of species and the presence of borders in ray cells or axial parenchyma. Bordered pits represent a compromise between maximal mechanical strength and maximal conductive capability. High rates of flow of sugar solutions may occur if starch in ray cells or axial parenchyma is mobilized for sudden osmotic enhancement of the conductive stream or for rapid development of foliage, flowers, or fruits. Measurement of the secondary wall thickness of ray cells may offer simple inferential information about the role that rays play in the mechanical strength of woods. © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 157–168.     

The accumulation pattern of ferruginol in the heartwood-forming Cryptomeria japonica xylem as determined by time-of-flight secondary ion mass spectrometry and quantity analysis

Background and Aims

Heartwood formation is a unique phenomenon of tree species. Although the accumulation of heartwood substances is a well-known feature of the process, the accumulation mechanism remains unclear. The aim of this study was to determine the accumulation process of ferruginol, a predominant heartwood substance of Cryptomeria japonica, in heartwood-forming xylem.

Methods

The radial accumulation pattern of ferruginol was examined from sapwood and through the intermediate wood to the heartwood by direct mapping using time-of-flight secondary ion mass spectrometry (TOF-SIMS). The data were compared with quantitative results obtained from a novel method of gas chromatography analysis using laser microdissection sampling and with water distribution obtained from cryo-scanning electron microscopy.

Key Results

Ferruginol initially accumulated in the middle of the intermediate wood, in the earlywood near the annual ring boundary. It accumulated throughout the entire earlywood in the inner intermediate wood, and in both the earlywood and the latewood in the heartwood. The process of ferruginol accumulation continued for more than eight annual rings. Ferruginol concentration peaked at the border between the intermediate wood and heartwood, while the concentration was less in the latewood compared wiht the earlywood in each annual ring. Ferruginol tended to accumulate around the ray parenchyma cells. In addition, at the border between the intermediate wood and heartwood, the accumulation was higher in areas without water than in areas with water.

Conclusions

TOF-SIMS clearly revealed ferruginol distribution at the cellular level. Ferruginol accumulation begins in the middle of intermediate wood, initially in the earlywood near the annual ring boundary, then throughout the entire earlywood, and finally across to the whole annual ring in the heartwood. The heterogeneous timing of ferruginol accumulation could be related to the distribution of ray parenchyma cells and/or water in the heartwood-forming xylem.     

In vitro hydroxylation of a norlignan: From agatharesinol to sequirin C and metasequirin C with a microsomal preparation from Cryptomeria japonica

In vitro hydroxylation of the norlignan agatharesinol to sequirin C and metasequirin C was demonstrated for the first time. After incubating agatharesinol with a microsomal preparation from the heartwood side of the intermediate wood of Cryptomeria japonica in the presence of cofactors, the aromatic ring-monohydroxylated derivatives of agatharesinol, sequirin C and metasequirin C, were formed. Although hydroxylation hardly occurred in the absence of cofactors, it could be initiated by adding NADPH or NADH, and was enhanced by further adding FAD or FMN. When microsomal preparations from the sapwood or from the sapwood side of the intermediate wood were used, hydroxylation did not occur. This in vitro conversion of the norlignans indicates that the hydroxylation of agatharesinol to sequirin C and metasequirin C is part of the in vivo biosynthetic pathway of norlignans. Another C. japonica norlignan, sugiresinol, which is a side chain-cyclized isomer of agatharesinol, does not seem to be accepted as a substrate, because hardly any hydroxysugireinol was formed after similar incubation with the enzyme.     

The cytoskeleton facilitates a three-dimensional symplasmic continuum in the long-lived ray and axial parenchyma cells of angiosperm trees

The microtubule (MT), microfilament (MF) and myosin components of the cytoskeleton were studied in the long-lived ray and axial parenchyma cells of the secondary xylem (wood) and secondary phloem of two angiosperm trees, Aesculus hippocastanum L. (horse-chestnut) and Populus tremula L. x P. tremuloides Michx. (hybrid aspen), using indirect immunofluorescence localisation and transmission electron microscopy. MTs and MFs were bundled and oriented axially (parallel to the cell's long axis) within all parenchyma cell types after they had fully differentiated. Additionally, actin and myosin were immunolocalised at the thin-walled membranes of the pits, which linked cells in neighbouring files of both ray and axial parenchyma, and at the pits between axial and ray parenchyma cells themselves. Anti-callose antibody immunolocated the plasmodesmata at the pit membranes, and in the same pattern as that of anti-myosin. Ray cells are important symplasmic pathways between the xylem and the phloem throughout the life of trees. We hypothesise that the MT and MF components of the cytoskeleton in the ray and axial parenchyma cells are involved in the transport of materials within those cells, and, in association with the acto-myosin of plasmodesmata at pit fields, are also important in intercellular transport. Thus, the symplasmic coupling between ray cells, between axial parenchyma cells, and between axial parenchyma and ray cells represents an extensive three-dimensional communication pathway permeating the tree from the phloem through the cambium into the wood. We suggest that this cytoskeletal pathway has an important role in delivery of photosynthate, and mobilised reserves, to the actively dividing cambium, and in the movement of materials to sites of reserve deposition, principally within the wood. This pathway could also have an important role in co-ordinating developmental processes throughout the tree.     

Wood anatomy and heartwood formation in Neem (Axadirachta indica A. Juss.)

The wood of Azadirachta indica is diffuse porous. Axial parenchyma is paratracheal banded or vasicentric. Rays are uniseriate to multiseriate and heterocellular with procumbent and upright cells. There is a strong negative correlation between vessel member length and diameter. The vulnerability and mesomorphic values are different in two trees of almost the same age growing in the same locality. The vessel member wall has spiral thickenings on its inner surface. Axial and ray parenchyma cells and sometimes vessels and fibres of the heartwood show the presence of extractives. The necrobiosis of the parenchyma cells occurs at the heartwood boundary. Senescence and death of parenchyma cells are associated with depletion of starch grains and accumulation of extractives. There is a climacteric rise in succinate dehydrogenase and acid phosphatase activities at the sapwood-heartwood interface which may be associated with heartwood formation. Peroxidase activity is greater near the cambial zone, indicating its probable role in lignification.     

Living Cells in Wood 3. Overview; Functional Anatomy of the Parenchyma Network

The very different evolutionary pathways of conifers and angiosperms are very informative precisely because their wood anatomy is so different. New information from anatomy, comparative wood physiology, and comparative ultrastructure can be combined to provide evidence for the role of axial and ray parenchyma in the two groups. Gnetales, which are essentially conifers with vessels, have evolved parallel to angiosperms and show us the value of multiseriate rays and axial parenchyma in a vessel-bearing wood. Gnetales also force us to re-examine optimum anatomical solutions to conduction in vesselless gymnosperms. Axial parenchyma in vessel-bearing woods has diversified to take prominent roles in storage of water and carbohydrates as well as maintenance of conduction in vessels. Axial parenchyma, along with other modifications, has superseded scalariform perforation plates as a safety mechanism and permitted angiosperms to succeed in more seasonal habitats. This diversification has required connection to rays, which have concomitantly become larger and more diverse, acting as pathways for photosynthate passage and storage. Modes of growth such as rapid flushing, vernal leafing-out, drought deciduousness and support of large leaf surfaces become possible, advantaging angiosperms over conifers in various ways. Prominent tracheid-ray pitting (conifers) and axial parenchyma/ray pitting to vessels (angiosperms) are evidence of release of photosynthates into conductive cells; in angiosperms, this system has permitted vessels to survive hydrologic stresses and function in more seasonal habitats. Flow in ray and axial parenchyma cells, suggested by greater length/width ratios of component cells, is confirmed by pitting on end walls of elongate cells: pits are greater in area, more densely placed, and are often bordered. Bordered pit areas and densities on living cells, like those on tracheids and vessels, represent maximal contact areas between cells while minimizing loss of wall strength. Storage cells in rays can be distinguished from flow cells by size and shape, by fewer and smaller pits and by contents. By lacking secondary walls, the entire surfaces of phloem ray and axial phloem parenchyma become conducting areas across which sugars can be translocated. The intercontinuous network of axial parenchyma and ray parenchyma in woods is confirmed; there are no “isolated” living cells in wood when three-dimensional studies are made. Water storage in living cells is reported anatomically and also in the form of percentile quantitative data which reveal degrees and kinds of succulence in angiosperm woods, and norms for “typically woody” species. The diversity in angiosperm axial and ray parenchyma is presented as a series of probable optimal solutions to diverse types of ecology, growth form, and physiology. The numerous homoplasies in these anatomical modes are seen as the informative results of natural experiments and should be considered as evidence along with experimental evidence. Elliptical shape of rays seems governed by mechanical considerations; unusually long (vertically) rays represent a tradeoff in favor of flexibility versus strength. Protracted juvenilism (paedomorphosis) features redirection of flow from horizontal to vertical by means of rays composed predominantly or wholly of upright cells, and the reasons for this anatomical strategy are sought. Protracted juvenilism, still little appreciated, occurs in a sizeable proportion of the world’s plants and is a major source of angiosperm diversification.     

Ultrastructural Evidence That Intracellular Ice Formation and Possibly Cavitation Are the Sources of Freezing Injury in Supercooling Wood Tissue of Cornus florida L

Although cellular injury in some woody plants has been correlated with freezing of supercooled water, there is no direct evidence that intracellular ice formation is responsible for the injury. In this study we tested the hypothesis that injury to xylem ray parenchyma cells in supercooling tissues is caused by intracellular ice formation. The ultrastructure of freezing-stress response in xylem ray parenchyma cells of flowering dogwood (Cornus florida L.) was determined in tissue prepared by freeze substitution. Wood tissue was collected in the winter, spring, and summer of 1992. Specimens were cooled from 0 to -60[deg]C at a rate of 5[deg]C h-1. Freezing stress did not affect the structural organization of wood tissue, but xylem ray parenchyma cells suffered severe injury in the form of intracellular ice crystals. The temperatures at which the ice crystals were first observed depended on the season in which the tissue was collected. Intracellular ice formation was observed at -20, -10, and -5[deg]C in winter, spring, and summer, respectively. Another type of freezing injury was manifested by fragmented protoplasm with indistinguishable plasma membranes and damaged cell ultrastructure but no evidence of intracellular ice. Intracellular cavitation may be a source of freezing injury in xylem ray parenchyma cells of flowering dogwood.     

Cell wall structure in the xylem parenchyma ofCryptomeria

Summary Cell wall structure in ray and axial parenchyma cells in the wood ofCryptomeria was shown to be typically crossed polylamellate and dissimilar to the characteristically layered wall of fibers and tracheids. Ray cells differed from axial cells in terms of form and also in the relative inclination of crossed microfibrillar helices in the cell wall. This feature was reflected by positive birefringence in ray cells and negative birefringence in axial cells. Localized wall thickenings,viz. transverse bars in ray cells and longitudinal ribs in axial cells, also displayed crossed polylamellate structure. This observation contrasts with the exclusively longitudinal microfibrillar orientation previously reported for longitudinal ribs in elongated parenchyma cells of primary tissue. On the basis of similar microfibrillar orientations between outer and inner wall lamellae, the cell walls ofCryptomeria parenchyma were judged to be predominantly secondary.Lignin was heterogeneously distributed in lamellate fashion and a high concentration characterized the thin middle lamella. Both types of parenchyma suggested a higher lignin content than adjacent longitudinal tracheids.     

Detection and features of wetwood in Quercus mongolica var. grosseserrata

To elucidate the location of water in wetwood, variations in moisture content within a tree of Quercus mongolica var. grosseserrata were observed macroscopically by determination of moisture content and by soft x-ray photography. Then the typical wetwood, which had been precisely located by such macroscopic analysis, was examined further by cryo-scanning electron microscopy. The moisture content varied considerably within the tree trunk, and typical wetwood was detected in limited regions of the heartwood. The distribution of the wetwood did not always correspond to the discoloration of the wood. In the wetwood, almost all the lumina of vessels and fibers were filled with water, while the lumina of ray and axial parenchyma cells were often cavitated. Intercellular spaces produced by deformation of cell walls and radial shakes were filled with water. In addition, tyloses were collapsed and torn in the wetwood, and they did not block the vessel lumina. These results suggest the possibility that vessels and fibers might serve as effective pathways for the accumulation of water in the wetwood.     

Death and decay in the trees of Mango (Mangifera indica L.)

Death and decay of trees of Mango (Mangifera indica L.) var. Kesar due to fungal infection was studied histologically. Fungal infection in the trees was observed due to various reasons like mechanical injuries in the stem, pruning of the branches, through the inflorescences, attack of Ambrosia beetle and termites. In the initial stage, fungal spores get settled on the flowers due to presence of nectar, followed by their germination and entry of the hypha into peduncle, which gradually spreads into younger branches. The inflorescences were first attacked by Fusarium moniliformis followed by other fungi like Alternaria, Chetomium sp., Aspregillus ellipticus, Aspregillus niger, etc. Fungal mycelia gradually invade the xylem tissues from the top of the branches and spread basipetally ultimately causing death of the infected branches. During monsoon, the crevices on the surface of bark of the healthy plants supported the growth of fungi like Pleurotus, Auricularia, Xyleria, Daldinia sp., and Polyporous sp. The removal of bark from such infected trees revealed minute holes on the surface of the woody cylinder made by Ambrosia beetles. During wet season fungal mycelia makes an easy entry into the xylem through the wounded portion of the stem or pruned branches. Initial entry of the hyphae into xylem was seen through the ray cells. Then the hyphae enter into the lumen of axial elements lining the ray cells through pits and intracellular spaces. The vessel elements located in the xylem (transition zone) between healthy and infected portion were filled with tyloses while axial and ray parenchyma showed heavy accumulation of tannin contents. On the other hand, the infected xylem was also found devoid of reserve metabolites while in normal trees, axial and ray parenchyma showed heavy accumulation of starch grains.