Immunohistochemical localization of agatharesinol,a heartwood norlignan,in Cryptomeria japonica

Localization of a heartwood norlignan, agatharesinol, in Sugi (Japanese cedar, Cryptomeria japonica D. Don, Taxodiaceae) was investigated by immunohistochemistry. Immuno light microscopy showed that the contents of ray parenchyma cells were immunostained in heartwood but not in sapwood. The staining of the heartwood tissue was competitively inhibited by agatharesinol but not by other Sugi heartwood extractives, and was, furthermore, markedly reduced by pre-extraction of the tissue with MeOH. These results indicated that the staining can be ascribed to the immunolabeling of agatharesinol in situ. The accumulations over the inner surface of some tracheid cell walls adjacent to the ray parenchyma cells were also immunolabeled, while the contents in axial parenchyma cells were not. In conclusion, agatharesinol was localized in the ray parenchyma cells in Sugi heartwood, and differences between the chemical structure of the contents of ray and axial parenchyma cells were also suggested.     

Natural durability of selected larch and Scots pine heartwoods in laboratory and field tests

The aim of this study was to compare natural durability of Siberian larch heartwood grown in Siberia and Sweden as well as European larch and Scots pine heartwood grown in Sweden. The study was based on standard in- and above ground tests lasting 12 years but laboratory decay tests with white and brown rot fungi was also included. Field test results showed that Siberian larch heartwood from Siberia was the most durable among the studied heartwoods with a decay index of 60 after 12 years in Simlångsdalen (Sweden), while European larch heartwood grown in Sweden, was decayed to failure before the end of the test. Scots pine heartwood was found to perform similarly to Siberian larch from Siberia. No relationship could be established between natural durability of examined heartwoods and their water absorption behavior; however, strong correlation to the total amount of extractives was observed. Scots pine and Siberian larch heartwood from Siberia had 12.7 and 19.6% total extractives content respectively but the extractives composition differs. The study revealed also that lignin and monosaccharide content could not explain the variations in decay resistance of the studied heartwoods. No similarities in the natural durability revealed by laboratory and field tests were observed.     

Parenchyma cell respiration and survival in secondary xylem: does metabolic activity decline with cell age?

Sapwood respiration often declines towards the sapwood/heartwood boundary, but it is not known if parenchyma metabolic activity declines with cell age. We measured sapwood respiration in five temperate species (sapwood age range of 5-64 years) and expressed respiration on a live cell basis by quantifying living parenchyma. We found no effect of parenchyma age on respiration in two conifers (Pinus strobus, Tsuga canadensis), both of which had significant amounts of dead parenchyma in the sapwood. In angiosperms (Acer rubrum, Fraxinus americana, Quercus rubra), both bulk tissue and live cell respiration were reduced by about one-half in the oldest relative to the youngest sapwood, and all sapwood parenchyma remained alive. Conifers and angiosperms had similar bulk tissue respiration despite a smaller proportion of parenchyma in conifers (5% versus 15-25% in angiosperms), such that conifer parenchyma respired at rates about three times those of angiosperms. The fact that 5-year-old parenchyma cells respired at the same rate as 25-year-old cells in conifers suggests that there is no inherent or intrinsic decline in respiration as a result of cellular ageing. In contrast, it is not known whether differences observed in cellular respiration rates of angiosperms are a function of age per se, or whether active regulation of metabolic rate or positional effects (e.g. proximity to resources and/or hormones) could be the cause of reduced respiration in older sapwood.     

Novel gene expression profiles define the metabolic and physiological processes characteristic of wood and its extractive formation in a hardwood tree species,Robinia pseudoacacia

Wood is of critical importance to humans as a primary feedstock for biofuel, fiber, solid wood products, and various natural compounds including pharmaceuticals. The trunk wood of most tree species has two distinctly different regions: sapwood and heartwood. In addition to the major constituents, wood contains extraneous chemicals that can be removed by extraction with various solvents. The composition and the content of the extractives vary depending on such factors as, species, growth conditions, and time of year when the tree is cut. Despite the great commercial and keen scientific interest, little is known about the tree-specific biology of the formation of heartwood and its extractives. In order to gain insight on the molecular regulations of heartwood and its extractive formation, we carried out global examination of gene expression profiles across the trunk wood of black locust (Robinia pseudoacacia L.) trees. Of the 2,915 expressed sequenced tags (ESTs) that were generated and analyzed in the current study, 55.3% showed no match to known sequences. Cluster analysis of the ESTs identified a total of 2278 unigene sets, which were used to construct cDNA microarrays. Microarray hybridization analyses were then performed to survey the changes in gene expression profiles of trunk wood. The gene expression profiles of wood formation differ according to the region of trunk wood sampled, with highly expressed genes defining the metabolic and physiological processes characteristic of each region. For example, the gene encoding sugar transport had the highest expression in the sapwood, while the structural genes for flavonoid biosynthesis were up-regulated in the sapwood-heartwood transition zone. This analysis also established the expression patterns of 341 previously unknown genes.     

Xanthones and triterpenes of Calophyllum tomentosum

The bark, branch timber, sapwood and heartwood extractives of Calophyllum tomentosum contain friedelin, friedelan-3β-ol, betulinic acid, taraxero     

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.     

长白落叶松人工林心材变化规律

依据黑龙江省孟家岗林场49株人工落叶松1179个圆盘和轮盘数据,分析了心材半径的纵向变化规律.结果表明: 心材半径随树高增高而逐渐减小,与树干外形基本一致,其中去皮半径(XR)、胸径(DBH)及形成层年龄(CA)与心材半径之间关系较显著,利用逐步回归分析建立落叶松心材半径(HR)和面积(HA)模型:HR=b₁+b₂XR²+b₃CA+b₄XR, HA=b₁+b₂DBH·XR+b₃CA+b₄DBH·XR².应用AIC、BIC、对数似然值以及似然比检验等模型评价指标,对利用样地、样木效应拟合的心材半径和面积模型进行比较.当考虑样木效应拟合心材半径和面积模型时,将b₁、b₂、b₃作为混合参数得出的模型最好.混合模型的预测精度高于基本模型.在应用上,总体心材半径和面积可以通过混合模型来预测.采用Beta回归模型模拟了心材比例,模型中各参数均显著,决定系数较高,模型模拟效果较好.     

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.     

Volatile constituents of Cupressus stephensonii heartwood

Nonpolar volatile extractives of Cupressus stephensonii heartwood amounting to 1·3% (drywood weight basis) were analyzed for their constituents and the main component was found to be carvacrol (78%). Tropolones (17%) were composed largely of β-thujaplicin and nootkatin with γ-thujaplicin in secondary quantities. Acids were low (1·7%). Neutral constituents (3·4%) contained α-pinene (8%), 4-terpinenol (27%), and methyl 4-trans-dehydrogeranate (45%).     

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.     

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.     

Formation and properties of some wood extractives

Chemical and other data concerning the location and site of formation of heartwood extractives are discussed. The biological conditions in the transition zone adjacent to the heartwood boundary are briefly described and some properties of extractives are given. The classes of polyphenolic compounds found in the wood of Eucalyptus species are listed together with the details of ellagic acid derivatives which are the most common class in this genus.     

Chemical factors affecting the brown-rot decay resistance of Scots pine heartwood

The cell wall chemistry (amount of hemicellulose, f-cellulose, and total lignin) and the concentration of extractives (total acetone-soluble extractives, resin acids, pinosylvins and the total phenolics quantified as tannin acid equivalents) were studied in brown-rot resistant and susceptible juvenile heartwood of Scots pine (Pinus sylvestris L.). The study material consisted of a total of 18 trees from two 34-year-old progeny trials at Korpilahti and Kerimäki. The trees were selected from among 783 trees whose decay rate had previously been screened in a laboratory test using a brown-rot fungus, Coniophora puteana. Samples from neither location showed any significant difference in the concentration (mg/cm³) of hemicellulose, f-cellulose and total lignin between the decay resistant and susceptible trees. At both locations only the concentration of total phenolics was higher in the decay-resistant heartwood than in the decay-susceptible heartwood. At Korpilahti, the amount of acetone-soluble extractives and the concentration of pinosylvin and its derivatives were higher in the resistant than in the susceptible trees.     

人工促成檀香结香的研究

在自然情况下生长的檀香（ＳａｎｔａｌｕｍａｌｂｕｍＬ．）植株,约１０龄左右开始形成具芳香的心材（通称结香）,约３０—４０年方可砍伐利用．作者采用两年生的幼树,施用植物生长抑制剂ＰＧＩ１进行促成结香试验,结果证明采用１％３ｍｌ生长激素处理的植株,其檀香油和檀香醇的含量一般较对照和用水处理的植株高１—２倍．     

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.     

Contributions to the Wood Anatomy of the Rubioideae (Rubiaceae)

Damnacanthus , Lasianthus, Saldinia, and Trichostachys are also included. Wood anatomical characters are compared with recent phylogenetic insights into the study group on the basis of molecular data. The observations demonstrate that the delimitation and separation of several taxa from the former Coussareeae/Morindeae/Prismatomerideae/Psychotrieae aggregate is supported by wood anatomical data. The Coussareeae can be distinguished from the other Rubioideae by their scanty parenchyma, septate libriform fibres, and the combination of uniseriate and very high multiseriate rays with sheath cells. Axial parenchyma bands and fibre-tracheids characterise Gynochtodes and some species of Morinda (Morindeae s.str.), but the latter genus is variable with respect to several features (e.g. vessel groupings and axial parenchyma distribution). Wood data support separation of Rennellia and Prismatomeris from Morindeae s.str.; vessels in both genera are exclusively solitary and axial parenchyma is always diffuse to diffuse-in-aggregates. Damnacanthus differs from the Morindeae alliance by the occurrence of septate fibres, absence of axial parenchyma, and the occasional presence of fibre wall thickenings. There are interesting similarities between members of the Lasianthus clade and the Pauridiantheae/Urophyleae group such as the sporadic occurrence of spiral thickenings in axial parenchyma cells. Received 26 January 2001/ Accepted in revised form 6 June 2001     

Polyphenols of Eucalyptus sideroxylon wood

Twenty-three major components were detected in the methanol extractives of the heartwood of Eucalyptus sideroxylon. The components identified include resveratrol, resveratrol-β-glucoside, 3,3′-di- and 3,3′,4-tri-o-methylellagic acids and their glucosides. The 3,3′-di-o-methylellagic acid 4′-glucoside isolated had properties significantly different from those previously reported for this compound. Also present were gailic acid, catechin, ellagic acid, an unidentified stilbene, the ellagitannins D-6 and D-13, polymerized leucocyanidin and an oily material. The sapwood contained gailic acid, small amounts of ellagitannins and ellagic acids and traces of other components. The heartwood extractives of related eucalypt species were also examined.