Contribution of active defense responses in the limitation of fungal spread in the sapwood of living sugi (Cryptomeria japonica) tree

The participation of active defense responses in the limitation of fungal spread in the sapwood of living sugi tree was determined. Isolates ofGuignardia cryptomeriae were inoculated to living trees, fresh logs, frozen and thawed logs and autoclaved logs, and the spread of wood discoloration and fungal hyphae was assessed. In autoclaved logs and freeze-treated logs, wood discoloration and reaction zone formation were not observed, and fungal hyphae of both of virulent and avirulent isolates grew rapidly in the wood. In fresh logs, wood discoloration and the formation of a pale reaction zone and transition zone were observed. Virulent isolates spread rapidly, but the spread of avirulent isolates was inhibited. In living trees, wood discoloration was observed and reaction zones were formed at lesion margins. These results indicated that constitutive defense mechanisms had no effect on the spread of both of virulent and avirulent isolates, and that active defense responses were important in the limitation of spread of fungal hyphae, hence in the limitation of wood discoloration. Further, it appeared that avirulent, but not virulent, races ofG. cryptomeriae were easily inhibited even by weak host responses. This study was presented at 105th and 106th Annual Meeting of the Japanese Forestry Society.     

Discoloured and non‐conductive sapwood among six Fagaceae species inoculated with Raffaelea quercivora

Mass mortality of oak trees has been occurring in Japan since the late 1980s. The fungus Raffaelea quercivora has been frequently isolated from discoloured sapwood in dead or wilting trees and inoculation experiments have shown it to be capable of causing wilting and xylem discoloration in several oak species, notably Quercus crispula and Q. serrata. In this study, we inoculated seedlings of six Fagaceae species with R. quercivora and, after 56 days, measured the vertical length of the discoloration and the areas of discoloured and non‐conducting sapwood on stem cross‐sections. The sapwood discoloration and the water non‐conduction areas were larger in Q. crispula and Q. serrata than in the other species.     

Xylem dysfunction in Yezo spruce (Picea jezoensis) after inoculation with the blue‐stain fungus Ceratocystis polonica

The blue‐stain fungus Ceratocystis polonica is pathogenic to Norway spruce (Picea abies) in Europe, as well as to Yezo spruce (Picea jezoensis) and Sachalin spruce (Picea glehnii) in Japan. The wilting mechanism in P. jezoensis saplings after inoculation with C. polonica was examined based on anatomical studies of the phloem and xylem of periodically harvested trees. In addition, the course of sap ascent in the trunks was traced by injection of acid fuchsin solution at harvest. As an initial external symptom, needle discolouration was observed. In dye conduction tests, xylem dysfunction in the xylem of inoculated trees became obvious. The dehydrated xylem area (dry zone) had extended more than 20 cm above the inoculation wounds, within 1 month after inoculation. When the sap flow to the branches had nearly stopped, the leaves began to discolour. Hyphae of C. polonica colonized the ray tissue around the inoculation wounds, but were absent at the front of the dry zones. Defence reactions occurred in ray parenchyma cells adjacent to the penetrating hyphae. It is suggested that secondary metabolites, which are formed by the ray cells and epithelial cells of resin canals, are involved in the obstruction of sap flow. Limited necrotic lesions of the phloem and cambium were not associated with foliar symptoms. It is proposed that the dry zone formation caused by C. polonica is the main mechanism leading to tree death.     

Disease biology and histopathology of bronze leaf disease: a threat to cultivated aspens,white poplars and their hybrids

Bronze leaf disease (BLD) of species and hybrids of aspens, grey and white poplars (Genus Populus, section Populus) is a systemic disease incited by the fungus Apioplagiostoma populi (E.K. Cash & A.M. Waterman) M.E. Barr that results in a characteristic bronze to dark brown pigmentation of infected leaves in late summer and early fall. Branches on affected trees die over a period of several years, disfiguring trees and eventually leading to mortality of susceptible trees planted in landscapes. In this study, we describe the histopathology of the disease and fungus development relative to external symptoms on affected trees. A. populi hyphae were observed in leaf vessels, lateral veins and xylem vessels of stems and roots of affected trees. Hyphae were also observed in veins and vessels of asymptomatic leaves. Xylem vessels of growth rings on dead branches were completely occluded with A. populi near the vascular cambium and together with the ray parenchyma were stained a bronze colour. Evidence of putative toxin damage to mesophyll cells, not colonized by hyphae, was observed and may contribute to the characteristic tissue staining observed. The development of the fungus was consistent with previous reports, but in this study, we report additional details including perithecial development and ascospore release from predominantly abaxial leaf surfaces. We also report for the first time the isolation and growth of the pathogen in pure culture and systemic infection of roots.     

Environmental effects on the color of sugi (Cryptomeria japonica D. Don) heartwood

Sugi (Cryptomeria japonica D. Don) is one of the most important planted trees in Taiwan. The normal heartwood of sugi has a red to pinkish rose color. Unfortunately, this pleasant appearance is susceptible to discoloration after environmental exposures. This discoloration is a serious defect that decreases the value of sugi products. The objective of this study was to investigate the effects of environmental factors such as moisture, oxygen, and lights of different wavelengths on the discoloration of sugi. The results show that under the combined effects of light and oxygen the color of heartwood changes from red (or rosepink) to bluish green. The color further darkens and eventually turns to black with the presence of moisture. As for the effect of light wavelengths on the discoloration of sugi heartwood, it was found that the red color was enhanced after being irradiated with light of wavelengths above 600 nm. Botha ^* andb ^* values increased significantly as a result of such exposure.     

广西融水特色红心杉木优树材质性状变异规律研究

[目的]通过测定和分析广西融水种源红心杉木优树材质性状指标,以了解该地区红心杉木材性状变异规律。[方法]以广西融水种源60株20年生的红心杉木优树为研究材料,测定单株材积、红心率、基本密度、组织比量、管胞性状、微纤丝角等10个材质性状指标,分析各材性性状指标分布和变异规律及性状间的相关关系。[结果]表明:红心率、基本密度、木射线比量、管胞比量、管胞长、管胞宽、管胞长宽比等性状数据分布服从正态分布。融水两个地区红心杉优树轴向薄壁细胞比量的变异系数分别为35.08%和44.97%,变异较大。管胞比量变异系数分别为3.28%和3.56%,变异较小。红心率、木射线比量、管胞长、管胞宽、微纤丝角等性状差异极显著(P0.01),轴向薄壁细胞比量差异显著(P0.05)。10个材质性状间存在12对表型显著相关。[结论]测定的10个性状均是连续性数量性状。早、晚材的管胞长度、管胞宽度和管胞长宽比等性状数据呈从心材至边材逐渐增加的规律。作为重要经济性状的红心率与木射线比量呈极显著正相关(P0.01),单株材积与轴向薄壁细胞比量呈显著正相关(P0.05),而单株材积与基本密度的相关性不显著,这使红心杉木生长量与材性相结合的遗传改良成为可能。     

Wood properties of juvenile and mature heartwood in Robinia pseudoacacia L.

The aim of this study is to characterise the properties of juvenile and mature heartwood of black locust (Robinia pseudoacacia L.). Content, composition and the subcellular distribution of heartwood extractives were studied in 14 old-growth trees from forest sites in Germany and Hungary as well as in 16 younger trees of four clone types. Heartwood extractives (methanol and acetone extraction) were analysed by HPLC-chromatography. UV microspectrophotometry was used to topochemically localise the extractives in the cell walls. The natural durability of the juvenile and mature heartwood was analysed according to the European standard EN 350-1. Growth as well as chemical analyses showed that, based on extractives content, the formation of juvenile wood in black locust is restricted to the first 10–20 years of cambial growth. In mature heartwood, high contents of phenolic compounds and flavonoids were present, localised in high concentrations in the cell walls and cell lumen of axial parenchyma and vessels. In juvenile wood, the content of these extractives is significantly lower. Juvenile wood had a correspondingly lower resistance to decay by Coniophora puteana (brown rot fungus) and Coriolus versicolor (white rot fungus) than mature heartwood.     

Fomannoxin,a phytotoxic metabolite of Fomes annosus: in vitro production,host toxicity and isolation from naturally infected Sitka spruce heartwood

The effect of different culture conditions on growth and production of fomannoxin and the sesquiterpene phytotoxin fomannosin by a single isolate of Fomes annosus was studied. Whereas fomannoxin was produced by actively growing hyphae, fomannosin production was associated with the declinning growth phase of the fungus. The presence of fomannoxin in chloroform extracts from the stained stein heartwood zone of Fomes annosus diseased Sitka spruce trees was established. Fomannosin however was not detected. Toxicity of fomannoxin to germinating seeds and young seedlings of Sitka spruce was demonstrated.     

Ceratocystis smalleyi colonization of bitternut hickory and host responses in the xylem

Colonization of Carya cordiformis sapwood by Ceratocystis smalleyi and subsequent host defence responses following artificial inoculation were investigated using anatomical and histological techniques. Hyphae of C. smalleyi were observed in all sapwood xylem features confirming the ability of the pathogen to invade and colonize the xylem tissues of the host species. The fungus was isolated from within and at the margins of discoloured sapwood areas at 2 and 12 months after inoculation. General host defence responses that included vessel occlusion with gels or tyloses, lipid accumulation, and production of phenolic compounds were observed in xylem tissues of inoculated C. cordiformis stems. Pectic substances, lipids, and to a rare extent, phenolic compounds were detected in vascular gels. The lipid‐rich barriers observed likely prevent lateral expansion of the fungus in the sapwood. Furthermore, lack of fungus sporulation within vessels may restrict axial spread of the fungus. C. smalleyi appears to be a limited vascular wilt pathogen of bitternut hickory based on these observations and previously reported sap flow reduction correlated with multiple infections in artificially inoculated trees.     

Extracellular peroxidase reaction at hyphal tips of white-rot fungus <Emphasis Type="Italic">Phanerochaete crassa</Emphasis> WD1694 and in fungal slime

The distribution of an extracellular peroxidase reaction by white-rot fungus Phanerochaete crassa WD1694 was visualized by peroxidase activity staining. The extracellular peroxidase reaction occurred at the hyphal tips and in the fungal slime filling the gaps between the hyphae. We investigated whether the peroxidase reaction occurred from the hyphal tips or in the slime. The hyphal tips were observed by phase-contrast microscopy, which showed that slime did not exist around the hyphal tips. Time-course observation of hyphal tips showed that peroxidase staining became thick and intense at the tips that did not have fungal slime. Daily observation of the peroxidase staining revealed that the staining was first observed at the hyphal tips. Furthermore, strongly stained hyphae were observed in the stained slime. These results suggested that an active species that oxidizes a peroxidase substrate is first produced at the tips of the hyphae, and then occurs in the slime via diffusion when slime exists around the hyphae. Our results show that the extracellular peroxidase reaction that is important to lignin biodegradation by white-rot fungi occurs directly at the tips of the hyphae and in the slime. Part of this report was presented at the 50th Lignin Symposium, October 19–20, 2005, Nagoya, Japan     

Temporally and spatially controlled death of parenchyma cells is involved in heartwood formation in pith regions of branches of <Emphasis Type="Italic">Robinia pseudoacacia</Emphasis> var. <Emphasis Type="Italic">inermis</Emphasis>

Focussing on four types of parenchyma cell around pith regions of branches of Robinia pseudoacacia L. var. inermis, we examined the timing and role of cell death during heartwood formation. Large parenchyma cells that were located in the inner part of the pith died within a year. By contrast, other parenchyma cells died within 4 years, with the timing of cell death depending on the type of cell. Axial parenchyma cells of the xylem close to the pith died first. Then, small parenchyma cells died in the perimedullary zone in the outer part of the pith. Finally, ray parenchyma cells in the xylem close to the pith died. Variations in the autofluorescence of cell walls, which might have been due to deposition of heartwood substances, were observed first in xylem ray parenchyma cells and small parenchyma cells in the perimedullary zone. Our results indicate that the initiation of heartwood formation occurs within 4 years in pith regions of branches in Robinia pseudoacacia L. var. inermis. Moreover, it appears that not only xylem ray parenchyma cells but also small parenchyma cells in the perimedullary zone might be involved in the synthesis of heartwood substances.     

Susceptibility of eliteAcacia mearnsii families to Ceratocystis wilt in south Africa

Ceratocystis albofundus is a recently described pathogen infectingAcacia mearnsii in South Africa, and it causes a disease known as Ceratocystis wilt. Symptoms of the disease include die-back, gummosis and wilting of infected trees. In order to select trees tolerant to this fungus, susceptibility tests were conducted on trees representing fourteen families ofA. mearnsii. A virulent isolate ofC. albofundus was selected and inoculated into the stems of twelve-month-old plants in a plantation. Lesion lengths, in the bark, and disease development were assessed after 6 weeks. All fourteen families ofA. mearnsii were susceptible to infection byC. albofundus. Considerable variation was, however, noticed between individual trees within the same family and the incorporation of disease tolerant trees into breeding programmes is proposed.     

Factors influencing the formation of heartwood discolouration in sycamore (<Emphasis Type="Italic">Acer pseudoplatanus</Emphasis> L.)

Sycamore (Acer pseudoplatanus L.) is a tree species with highly valuable wood. Similar to European beech (Fagus sylvatica L.), the stemwood of sycamore can be devalued due to the discolouration i.e. brown heart (hearwood discolouration, hereafter). This paper aims to establish the influence of tree traits and site characteristics on the formation of heartwood discolouration and the possibility of predicting the occurrence of heartwood discolouration. For this purpose, 351 sycamore trees from 38 sites in Slovenia, most of them on carbonate bedrock and in the mountain vegetation belt, were analysed. Stem analysis was carried out on all the trees and the age, diameter at breast height (dbh), height of crown base, forking and crown size, were established. On the stump and upper fronts of the first and second logs, the maximum heartwood discolouration diameter was measured. Using logistic regression, the positive influence of age, relative crown length and average diameter increment on the formation of heartwood discolouration was ascertained. Conversely, the probability of heartwood discolouration was diminished by diameter increment in the last 20 years. In the less productive sites, the probability of formation of heartwood discolouration was relatively high in small diameter trees, but it increased slowly with diameter. Tree forking also contributed to a large extent of the heartwood discolouration. After the diameter at breast height achieves 45 cm, the formation of heartwood discolouration on the first log is highly probable. As in beech, the heartwood discolouration increases along the stem axis up to a height of 6–8 m, and decreases in the higher parts of the tree.     

Peroxidase activity, isoenzymes and histological localisation in sapwood and heartwood of Scots pine (Pinus sylvestris L.)

Peroxidase activity and isoenzymes of fresh wood samples of the third shoot of 12-year old trees and from the sapwood, transition zone and heartwood of c. 60-year old stems of Scots pine (Pinus sylvestris L.) were investigated. Wood samples were ground at −30°C, extracted, and the extracts concentrated c. 20-fold for peroxidase activity assays (guaiacol method) and for IEF-PAGE. At least 11 major isoenzymes could be found in the gels. Even the heartwood contained some peroxidase isoenzymes. Isoenzyme patterns of the juvenile wood did not change with the season. However, juvenile wood showed the highest peroxidase activity at the end of the growing season. Peroxidase activity decreased from the outer sapwood towards the heartwood. Thin sections of different wood zones stained for peroxidase revealed activity in ray parenchyma and resin canal epithelial cells. Intensive staining was localised in the bordered pits of vertical and ray tracheids, and in the end walls of ray parenchyma cells.     

Wundfäule in Fichtenwaldungen mit alten Schälschäden

Wound decays in spruce stands following bark stripping . 2083 trees in 91 sample plots on four trial areas (24 ha) were examined for bark stripping and decay. Only 25% of trees showed no injury. Of the injured trees, 73% showed some degree of decay, 10% discoloration. and 17% neither decay nor discoloration. Of the wood destroying fungi isolated Fomes annosus was dominant and appeared to be the most important and widespread of the wound inhabiting fungi in the test area. Besides the Basidiomycetes a considerable number of other fungi was isolated. The most frequent was Cylindrocarpon cylindroides var. tenue.     

Discoloured wood of aspen caused by increment boring

Decay resulting from increment borings varies with tree species. Trembling aspen (Populus tremuloidcs Michx.) in Newfoundland is seriously affected by this particular wound. Three years after 20 trees had been bored, none had healed probably because of a pronounced split of the bark. The wood discoloration extended horizontally to about 2 cm on each side and vertically half metre above and below the wound. Peniophora polygonia (Pers. ex Fr.) Bourd. et Galz. was the most common fungus isolated while Polyporus adustus Willd. ex Fr. and several imperfect fungi and bacteria were also present.     

Xyleborus glabratus attacks and systemic colonization by Raffaelea lauricola associated with dieback of Cinnamomum camphora in the southeastern United States

Laurel wilt, caused by Raffaelea lauricola, is responsible for extensive mortality of redbay and other American members of the Lauraceae in the southeastern United States. Raffaelea lauricola is a mycangial symbiont of the redbay ambrosia beetle (Xyleborus glabratus), and the beetle and fungus were accidentally introduced from Asia. Branch dieback of camphortree (Cinnamomum camphora), an Asian member of the Lauraceae, has been occasionally observed in areas where laurel wilt has decimated redbay populations, and R. lauricola was isolated from such camphortrees. However, the role of X. glabratus and R. lauricola in this branch dieback remains unclear. Examination of camphortrees on Jekyll Island, Georgia showed that healthy‐appearing trees and those with branch dieback had been attacked by X. glabratus, but the trees with branch dieback had four times as many beetle attacks. Raffaelea lauricola was routinely isolated from discoloured xylem near beetle tunnels in healthy trees and those with dieback. Single‐point inoculations with R. lauricola on stems of mature, healthy camphortree trees failed to induce wilt‐like symptoms or branch dieback, although areas of discoloration were scattered throughout the xylem, and R. lauricola was reisolated irregularly at various heights in some inoculated trees. In growth chamber experiments, single‐point inoculations with R. lauricola resulted in systemic colonization but no wilt symptoms or branch dieback in camphortree saplings. In contrast, inoculations at multiple points along the stem (simulating multiple attacks by the vector) caused branch dieback and wilt‐like symptoms, including a brownish, diffuse discoloration of the xylem. Camphortree appears to be more resistant than American species of Lauraceae to the vascular wilt caused by R. lauricola. The fungus does colonize camphortrees systemically, however, and can apparently cause branch dieback. This suggests that the fungus may provide brood material for X. glabratus in Asia as it does in the southeastern United States.     

Histology of current shoots of Japanese cedar (Cryptomeria japonica D. don) inoculated withCercospora sequoiae Ellis et Everhart

The histological reaction of current shoots of the Japanese cedar,Cryptomeria japonica, inoculated withCercospora sequoiae was investigated. Wound periderm was formed on current shoots of the cedar on and after the 4th day after inoculation. In the cells of the cork layer, lignin and suberin were deposited, and a phenolic substance was accumulated. In other inoculated samples, plasmolysis, contraction of the cells and necrosis of the tissues were also observed. On the 32nd day, wound perioderm and phellogen were observed in samples of the tissue inside the necrotic tissues in response to hyphal invasion. Occurrence of wound periderm and phellogen in tissues of the needles was confirmed in the inoculated samples although no hyphae were observed in and around the tissues. These results suggest that this fungus causes necrosis of host tissues and induces defense responses. A part of this paper was presented at the 106th Annual Meeting of the Japanese Forestry Society (1995).     

Profile of bordered pit aspiration in Cryptomeria japonica using confocal laser scanning microscopy: pit aspiration and heartwood color

Nine trees of Cryptomeria japonica from six elite tree clones with a broad range of heartwood colors were selected. The profiles of pit aspiration percentage (ASP) of earlywood and latewood from pith to bark for green and air-dry conditions were determined to study the relationship between heartwood color and pit aspiration. Confocal laser scanning microscopy (CLSM) observations showed that the ASP of earlywood was low in sapwood and high in heartwood in the green condition. Pit aspiration increased in intermediate wood when compared with sapwood. On the other hand, latewood pits did not aspirate during heartwood formation. Comparing the air-dry condition with the green condition, sapwood pits aspirated during drying in both earlywood and latewood; however, there was no significant difference in pit aspiration of heartwood. There was no significant difference between samples with red and black heartwoods for ASP. The difference in ASP between individual trees was larger than that by heartwood color. The general advantage of CLSM over light microscopy is that serial optical sections along the Z axis can be obtained for any moisture condition, without the need for thin sectioning or embedding.     

Genetic variation in Picea abies (L.) Karst. and Fomes annosus (Fr.) Cke. shown by growth of the fungus on wood dust

Growth of Fomes annosus is studied by a test based on growth of previously starved hyphae on sawdust of heartwood of Picea abies as the only nutrition. Our observations show that the method can be used to distinguish beween different spruce individuals or clones, between different Fomes annosus strains and between different growing sites of the trees.