First report of Ceratocystis platani in Albania

The fungus Ceratocystis platani was detected in various localities of the Gjirokastër prefecture in southern Albania, where it was causing widespread mortality on Platanus orientalis trees. The identification of the fungus was based on both morphological characteristics in culture and DNA sequencing. The pathogenicity of C. platani was confirmed in inoculations on seedlings of P. orientalis. To our knowledge, this is the first report of C. platani in Albania.     

Genotype distribution of Raffaelea quercivora in the oak galleries and its composition in the mycangia of Platypus quercivorus

Raffaelea quercivora is the pathogenic fungus that causes Japanese oak wilt. The female monogynous ambrosia beetle, Platypus quercivorus, carries this fungus in mycangia on the pronotum. These beetles bore galleries in oak trees with their partners to produce offspring, and they deposit fungus on the gallery walls from their mycangia. The offspring mature in the gallery, before loading the fungal pathogen and flying from the gallery to other healthy trees. To investigate the unloading and loading modes of the fungus within the gallery, we developed four polymorphic microsatellite markers for R. quercivora and identified the fungal genotypes in the galleries and mycangia of the beetles. Small wood chips were sampled at 5–10‐mm intervals from the walls of five galleries in a dead Quercus serrata tree. The pronota were also sampled from five female adult beetles. The genotypes of the R. quercivora isolates from the wood chips and pronota were identified using the microsatellite makers. The genotypic analysis showed that each gallery was inhabited patchily by 5–10 genotypes of R. quercivora, and the mycangia of each beetle contained 3–6 genotypes. These results indicate that diverse R. quercivora genotypes are unloaded repeatedly from the mycangia of female beetles onto the gallery wall, which results in their patchy distribution on the walls. When the offspring leave the host tree, the fungal clones that proliferate in the walls are also loaded repeatedly into the mycangia of the mature beetles.     

Ceratocystis platani is killing plane trees in Istanbul (Turkey)

Ceratocystis platani was detected at several locations in the European side of Istanbul, causing severe dieback and mortality, mainly on Platanus × acerifolia imported from Italy approximately 160 years ago, but also on Platanus orientalis. In the work reported here, the causal agent of the dieback and mortality was identified based on morphological characteristics in culture and internal transcribed spacer (ITS) sequencing, confirming the presence of C. platani in Turkey. All analysed C. platani isolates from Istanbul shared the same ITS sequence with European C. platani strains available in the NCBI, but differed from other species previously ascribed to the Ceratocystis fimbriata species complex. Pathogenicity of C. platani was proven by inoculation onto 25‐cm‐long and 4–9 mm diam. P. orientalis twigs and the causal agent reisolated from symptomatic tissues. In a survey of 976 Platanus in streets and parks of Istanbul, 26.5% of trees showed symptoms of C. platani infections, whilst 5.6% were dead. The proportion of symptomatic trees was higher where management interventions, such as topping or pruning and felling infected trees had been carried out.     

Presence and viability of Ceratocystis lukuohia in ambrosia beetle frass from Rapid ʻŌhiʻa Death‐affected Metrosideros polymorpha trees on Hawaiʻi Island

Rapid ?ōhi?a Death (ROD) is a fungal disease of ?ōhi?a lehua (Myrtaceae: Metrosideros polymorpha) caused by Ceratocystis lukuohia and C. huliohia. ROD is the aetiological agent of widespread mortality of this important tree on Hawai?i Island, but its epidemiology remains unclear. We investigated the prevalence and viability of C. lukuohia in ambrosia beetle frass in ROD‐affected ?ōhi?a trees. A total of 200 frass traps were placed onto C. lukuohia‐infected ?ōhi?a at four locations on the east side of Hawai?i Island. Frass was collected and screened for the presence of C. lukuohia DNA using a diagnostic qPCR assay. In addition, frass samples were screened for viability by carrot baiting. All trapped beetles were of the genus Xyleborus, with the majority being the non‐native X. ferrugineus. Of the frass samples tested, 62% contained C. lukuohia DNA and 17% of carrot baits were positive for the fungus. These results indicate that ambrosia beetle frass releases C. lukuohia into the environment. We discuss the potential role infested frass could play in the ROD pathosystem.     

Ophiostoma tsotsi and Ophiostoma quercus associated with Platypus cylindrus F. (Coleoptera: Curculionidae) in cork oak stands in Tunisia

Ambrosia beetles, including Platypus cylindrus, commonly act as vectors of Ophiostomatales. In an investigation of fungi associated with P. cylindrus, several species of Ophiostomatales were identified, including two species of Ophiostoma s. str. These two species were characterized based on comparisons of DNA sequences of the internal transcribed spacer regions including the 5.8S gene of rDNA and β‐tubulin gene. Two distinct clades were discernable in the phylogenetic trees, supported by high bootstrap values. One clade included the species Ophiostoma tsotsi, reported herein for the first time in Tunisia, and the species Ophiostoma quercus.     

Discrimination among host tree species by the ambrosia beetle Platypus quercivorus

The ambrosia beetle Platypus quercivorus is a vector for the fungus that causes Japanese oak wilt, and susceptibility to infestation by P. quercivorus varies by tree species. We postulated that P. quercivorus discriminates among host tree species differing in susceptibility to attack. To test this postulate, we counted the number of flying male beetles (NFM), the number of holes bored by male beetles (NH), and the number of flying female beetles (NFF) per unit area of bark surface in three fagaceous tree species: Quercus crispula (with high susceptibility to infestation) and Q. serrata and Castanea crenata (both with low susceptibility). NFM and NH were used to calculate the proportion of male beetles that bored holes out of those that flew to the tree (PBM). We used generalized additive models to predict NFM, NFF, and PBM. The locations of trees, expressed as x and y coordinates, numbers of weeks after the first male beetle’s flying (WEEK), diameters of trees 130 cm above ground (DBH), and tree species (SP) were incorporated into the models as candidate explanatory variables. The best-fit models for NFM and NFF included WEEK and DBH and the effect of location; SP was not included in the models. For PBM, the best-fit model included WEEK, DBH, and SP. The results indicate that male P. quercivorus prefer Q. crispula to Q. serrata and C. crenata and that selection is made before boring holes on trees, but that P. quercivorus do not discriminate among host species when they fly to trees.     

Types of frass produced by the ambrosia beetle <Emphasis Type="Italic">Platypus quercivorus</Emphasis> during gallery construction,and host suitability of five tree species for the beetle

We studied the biology of the ambrosia beetle Platypus quercivorus in the logs of five tree species to determine: (1) the relationship between the amount of frass produced by beetles and tunnel length, (2) the relationship between frass shape and the morphological characteristics of P. quercivorus mouthparts, and (3) the suitability of five tree species for P. quercivorus reproduction. Five logs each from healthy Quercus crispula, Q. serrata, Castanea crenata, Sorbus japonica, and Cryptomeria japonica trees were used in this experiment. The results showed that there was a linear relationship between the amount of frass and tunnel length. Whenever powdery frass was produced, larvae were found in the gallery in the log, while fibrous frass was present only in galleries that contained just adults. The mouthparts of adults were completely sclerotized, which likely accounts for the fibrous frass production. Host preference of P. quercivorus was examined at two stages using five tree species. The first stage is digging initiation, which concerns male preference for digging a tunnel. More holes were made by males on S. japonica and Q. serrata logs, while fewer holes were made on C. japonica logs. The second stage is characterized by female orientation, mating, and progeny development. Platypus quercivorus could complete its lifecycle only in the two Quercus spp.     

A low <Emphasis Type="Italic">Platypus quercivorus</Emphasis> hole density does not necessarily indicate a small flying population

The ambrosia beetle Platypus quercivorus causes mass mortality of Fagaceae trees in Japan, and tree species differ in their susceptibility to P. quercivorus. We hypothesized that interspecific differences in susceptibility are caused by differences in beetle infestation patterns, that is, how many beetles fly to a tree and how many of those arriving bore holes. To examine how tree susceptibility is related to these parameters, two tree species with different degrees of susceptibility were studied (highly susceptible Quercus crispula and less susceptible Q. salicina). Specifically, we measured the number of male beetles per unit area that flew to the host trees (NFM) and the density of holes bored by male beetles (DH). From these two values, we calculated the proportion of male beetles flying to a host tree that bored holes (PBM). These parameters were compared for the two Quercus species. Although the two species did not differ in NFM in 2003, PBM was markedly lower for Q. salicina than for Q. crispula and DH was significantly lower for Q. salicina than for Q. crispula. Thus, the lower susceptibility of Q. salicina is partly explained by its low PBM.     

Volatile organic compounds (VOC) as biomarkers for detection of Ceratocystis platani

Ceratocystis platani causes canker stain of plane trees, and it represents a serious disease of Platanus spp. both in the United States and Europe. Current chemical or biological controls do not effectively manage C. platani, so new preventive methods need to be developed in order to limit this pathogen spreading. In this work, we have characterized the main volatile organic compounds (VOC) emitted in vitro from pure cultures of C. platani and other common pathogenic fungal species of hosts plants growing in the same ecosystems as plane trees. We found that C. platani emitted a similar blend of VOC compared with phylogenetically similar species C. populicola. In particular, C. platani was characterized by emission of isoamyl acetate and isobutyl acetate while C. populicola by ethyl acetate and isobutyl acetate, which were not released by any of the other out‐group fungal species grown on the same medium. Moreover, following a targeted approach based on the main VOC found in vitro, we have successfully validated in vivo that VOC uniquely emitted by C. platani (i.e. isobutyl acetate along with isoamyl alcohol) were released from the bark of plane trees following C. platani inoculation. Our results highlight the possibility to exploit VOC emitted specifically by C. platani as biomarkers to recognize Platanus x acerifolia plants infected by this pathogen.     

Isolation and characterization of 10 new microsatellite loci in the ambrosia beetle <Emphasis Type="Italic">Platypus quercivorus</Emphasis>

The ambrosia beetle, Platypus quercivorus, is an insect vector for the causal agent of Japanese oak wilt disease, Raffaelea quercivora. We have developed 10 new polymorphic microsatellite markers from Type 1, one of the genetic types of the beetle, using a biotin-enrichment procedure. Fifty-three adult beetles were used for characterization of the markers. The number of alleles per locus ranged from two to nine (average 4.9). The observed and expected heterozygosity ranged from 0.49 to 0.77 and from 0.44 to 0.80, respectively. None of the loci deviated significantly from Hardy–Weinberg equilibrium. No linkage disequilibrium was detected in any pairwise comparisons of loci. Most of the markers successfully amplified in other genetic types of the beetle. These markers could become useful tools for population genetic studies and for parentage and sibship analysis for this beetle.     

Field trap test for bioassay of synthetic (1<Emphasis Type="Italic">S</Emphasis>,4<Emphasis Type="Italic">R</Emphasis>)-4-isopropyl-1-methyl-2-cyclohexen-1-ol as an aggregation pheromone of <Emphasis Type="Italic">Platypus quercivorus</Emphasis> (Coleoptera: Platipodidae)

An ambrosia beetle, Platypus quercivorus, is a vector of Japanese oak wilt, which causes massive mortality of oak trees in Japan. Previous studies have identified (1S,4R)-4-isopropyl-1-methyl-2-cyclohexen-1-ol ((−)-IMCH) as the major component of the aggregation pheromone of P. quercivorus. We tested the ability of synthetic (−)-IMCH to attract P. quercivorus adults. The synthetic compound attracted 14.4 times as many P. quercivorus, of both sexes, as control traps. Control and (−)-IMCH-baited traps both captured 3.32 times as many males as females, probably because of the nature of Lindgren funnel traps and of male-biased emergence sex ratio early in the season of adult flights. (−)-IMCH was proved to be a major component of the aggregation pheromone of P. quercivorus.     

Horizontal niche differentiation of ambrosia beetles (Coleoptera: Scolytidae and Platypodidae) within the standing trunk of <Emphasis Type="Italic">Quercus serrata</Emphasis>

Niche differentiation, in terms of time and space, has been reported within a community of ambrosia beetles (Coleoptera: Scolytidae and Platypodidae) in a tree trunk. Two platypodid ambrosia beetles, Platypus quercivorus (Murray) and Platypus calamus Blandford, utilize a similar height range with respect to the Quercus serrata Murray (Fagaceae) trunk. The sapwood and heartwood (i.e., wood types) are known to differ in their physical and chemical characteristics. The objective of this study was to determine the differences among ambrosia beetles, in terms of wood type preferences, in a Q. serrata tree trunk. We analyzed the horizontal distribution patterns of ambrosia beetles within two mature Q. serrata tree trunks. Platypus calamus and the other ambrosia beetles [P. quercivorus, Ambrosiodmus lewisi (Blandford), Ambrosiophilus atratus Eichhoff, and Xyleborus sp.] showed different horizontal distribution patterns within the wood. The former tended to utilize the heartwood, whereas the latter utilized the sapwood. These results suggest that the wood type can be regarded as one of the niche dimensions of ambrosia beetles in a Q. serrata trunk.     

Responses ofQuercus sapwood to infection with the pathogenic fungus of a new wilt disease vectored by the ambrosia beetlePlatypus quercivorus

Quercus serrata andQ. crispula wilt during the summer in wide areas along the Sea of Japan. Mass attacks of trees by an ambrosia beetle (Platypus quercivorus) are characteristic before appearance of the wilting symptoms. This study investigated the pathogenic effects of a fungus detected specifically in the wilting trees. This hyphomycete fungus,Raffaelea sp., has a distribution that correlates with the discolored xylem area called wound heartwood in which vessels are dysfunctional. Tylosis formation around the hyphae indicates vessel dysfunction. In areas with discoloration, the fungal hyphae were invading living ray parenchyma cells from the vessel lumen. As a protective reaction the ray cells exuded yellow substances into the vessels, but these substances seemed ineffective against the fungal activity, probably because the fungus disperses along the beetle's gallery before enough substance can accumulate. It should allow wide discoloration in sapwood. Cambium was not necrotic around the fungus. The cytological process in the host was as follows: (1) synthesis of secondary metabolites by the stimuli of oak fungus; (2) exudation of yellow substances into vessels; and (3) dysfunction of vessels and wound heartwood formation. In regard to wilting of trees, the pathogenicity of the fungus should be assessed by its ability to stop sap flow.     

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.     

Ambrosia beetles (Coleoptera: Curculionidae) can directly transmit the fungal pathogens responsible for Rapid ʻŌhiʻa Death

The ecologically and culturally vital tree species, ʻōhiʻa lehua (Metrosideros polymorpha), is threatened by the fungal pathogens Ceratocystis lukuohia and Ceratocystis huliohia, the causal agents of the disease complex called Rapid ʻŌhiʻa Death (ROD). Four invasive ambrosia beetle (Coleoptera: Curculionidae: Scolytinae) species in the Xyleborini tribe colonize ROD Ceratocystis-infested ‘ōhiʻa and produce inoculum through their frass; however, the potential for direct transmission of the ROD fungi by these beetles was unknown. We fulfilled Leach's rules to support insect transmission of ROD by documenting the visitation of these ambrosia beetles to healthy ‘ōhiʻa trees, culturing C. lukuohia and C. huliohia from the ROD-associated beetles using three different collection methods at multiple study sites, and challenging healthy ʻōhiʻa seedlings with beetles that were exposed to both C. lukuohia and C. huliohia cultures. We documented all four invasive ROD-associated ambrosia beetle species including Xyleborinus saxesenii, Xyleborus affinis, Xyleborus ferrugineus, and Xyleborus perforans to regularly visit healthy ʻōhiʻa trees on sticky traps. Viable Ceratocystis propagules were isolated from all species, and C. lukuohia was most commonly isolated of the two ROD-causing fungi. Consistently across all collection techniques, ROD Ceratocystis spp. were detected on just under 3% of all assayed beetles, with the highest detection rate from X. affinis. All four beetle species were capable of directly transmitting both pathogens to healthy ʻōhiʻa seedlings with a high rate of transfer. Ceratocystis spp. are highly virulent pathogens in trees, and a single inoculation can result in tree death, therefore any direct transmission is a cause for concern. After meeting the criteria of Leach's rules, we propose that Xi. saxesenii, X. affinis, X. ferrugineus, and X. perforans are vectors of C. lukuohia and C. huliohia, particularly in areas of high ROD pressure and tree stress.     

Investigation of the mating‐type distribution of Raffaelea quercus‐mongolicae in South Korea

Raffaelea quercus‐mongolicae is a mycangial symbiont of a wood‐boring ambrosia beetle, Platypus koryoensis (Coleoptera: Curculionidae: Platypodidae), which is believed to be associated with oak mortality in South Korea. Since the first discovery of the disease in 2004, oak mortality has continued to spread across the country, and the pathogen has had a significant impact on forest ecosystems in the country; however, little is known regarding the biology of the fungus. The aim of this study was to develop markers to determine the mating types of individual isolates of R. quercus‐mongolicae collected across the country and then determine the mating‐type ratio in natural populations. To achieve this, partial sequences of MAT1‐1–3 and MAT1‐2–1 genes were amplified using degenerate primers and cloned. The new primer sets, which were specific to each mating‐type idiomorph, identified isolates as MAT1‐1 and MAT1‐2 mating types. The roughly equal frequencies of the two mating types suggest that sexual reproduction might have occurred in natural populations of R. quercus‐mongolicae in South Korea.     

Types of bark beetle (Coleoptera: Scolytidae) infestation in spruce forest stands affected by air pollution, bark beetle outbreak and honey fungus (Armillaria mellea)

583 spruce stands in an area affected by air pollution and bark beetle outbreak in Eastern Slovakia were studied in 1996. According to bark beetle infestation of dominant and codominant trees, stands were classified into following types of spruce stand decline:Ips typographus-A,Ips typographus-B,Polygraphus poligraphus, I. typographus/P. poligraphus—A,I. typographus/P. poligraphus—B. The presence of attacked trees in forest edges, bark beetle spots and forest interior was the key important factor for the classification. Data from forest inventory and forest management evidence together with data on types of spruce stands decline were used in further analyses. Results shows that the distribution of forest stands classified into different types or uninfested stands is related mainly to host size and site quality. The percentage of spruce, exposition of stands and stand density showed significant effects. The mechanisms of spreading of studied bark beetle outbreak could be explained by direct effects of stress of trees caused by an abrupt increase of level of solar irradiation and by weakening of trees by the honey fungus.     

Genetic diversity and population structure of Raffaelea quercus‐mongolicae,a fungus associated with oak mortality in South Korea

Raffaelea quercus‐mongolicae is a fungus associated with oak wilt and deemed to cause extensive oak mortality in South Korea. Since the discovery of this fungus on a dead Mongolian oak (Quercus mongolica) in 2004, the mortality continued to spread southwards in South Korea. Despite continued expansion of the disease and associated significant impacts on forest ecosystems, information is lacking about the origin and genetic diversity of R. quercus‐mongolicae. Restriction‐site‐associated DNA (RAD) sequencing was used to assess genetic diversity and population structure among five populations (provinces) of R. quercus‐mongolicae in South Korea. In total, 179 single nucleotide polymorphisms (SNPs) were identified among 2,639 RAD loci across the nuclear genome of the 54 R. quercus‐mongolicae isolates (0.0012 SNPs per bp), which displayed an overall low expected heterozygosity and no apparent population structure. The low genetic diversity and no apparent population structure among South Korean populations of this ambrosia beetle‐vectored fungus support the hypothesis that this fungus was introduced to South Korea.     

Characteristics of leaf areas of plantations in semiarid hills and gully loess regions

The objectives of our study were to explore the relationship of leaf area and stand density and to find a convenient way to measure stand leaf areas. During the 2004 growing season, from May to October, we used direct and indirect methods to measure the seasonal variation of the leaf areas of tree and shrub species. The trees were from Robinia pseudoacacia stands of four densities (3333 plants/hm², 1666 plants/hm², 1111 plants/hm², and 833 plants/hm²) and Platycladus orientalis stands of three densities (3333 plants/hm², 1666 plants/hm², and 1111 plants/hm²). The shrub species were Caragana korshinskii, Hippophae rhamnoides, and Amorpha fruticosa. Based on our survey data, empirical formulas for calculating leaf area were obtained by correlating leaf fresh weight, diameter of base branches, and leaf areas. Our results show the following: 1) in September, the leaf area and leaf area index (LAI) of trees (R. pseudoacacia and P. orientalis) reached their maximum values, with LAI peak values of 10.5 and 3.2, respectively. In August, the leaf area and LAI of shrubs (C. korshinskii, H. rhamnoides, and A. fruticosa) reached their maximum values, with LAI peak values of 1.195, 1.123, and 1.882, respectively. 2) There is a statistically significant power relation between leaf area and leaf fresh weight for R. pseudoacacia. There are significant linear relationships between leaf area and leaf fresh weight for P. orientalis, C. korshinskii, H. rhamnoides, and A. fruticosa. Moreover, there is also a significant power relation between leaf area and diameter of base branches for C. korshinskii. There are significant linear relations between leaf area and diameter of base branches of H. rhamnoides and A. fruticosa. 3) In the hills and gully regions of the Loess Plateau, the LAIs of R. pseudoacacia stand at different densities converged after the planted stands entered their fast growth stage. Their LAI do not seem to be affected by its initial and current density. The same is true for P. orientalis stands. However, the leaf area of individual trees is negatively and linearly related with stand density. We conclude that, in the hills and gully regions of the Loess Plateau, the bearing capacity of R. pseudoacacia and P. orientalis stands we studied have reached their maximum limitation, owing to restricted access to soil water. Therefore, in consideration of improving the quality of single trees, a stand density not exceeding 833 and 1111 plants/hm² is recommended for R. pseudoacacia and P. orientalis, respectively. In consideration of improving the quality of the entire stands, the density can be reduced even a little more. __________ Translated from Journal of Plant Ecology (Chinese Version), 2008, 32 (2): 440–447 [译自: 植物生态学报]     

Extractives of Quercus crispula sapwood infected by the pathogenic fungus Raffaelea quercivora II: isolation and identification of phenolic compounds from infected sapwood

There has been a mass mortality of oak trees in the area along the coast of the Japan Sea. This phenomenon is caused by the ambrosia beetle Platypus quercivorus, which carries the ambrosia fungus Raffaelea quercivora. Extractives of a necrotic brownish coloration formed in the infected sapwood of Quercus crispula were investigated. The methanol extract of the damaged sapwood of Q. crispula was concentrated in vacuo and centrifuged to yield precipitates and the supernatant. The precipitates were subjected to Sephadex LH-20 column chromatography and preparative HPLC to describe a novel ellagic acid derivative (1). The 10 % methanol water-soluble part of the supernatant was subjected to medium-pressure ODS column chromatography and preparative HPLC, respectively, to analyze a known lignan (2). Sulfuric acid hydrolysis of (1) yielded an ellagic acid and a gallic acid. NMR and LC-TOF/MS indicated that an ellagic acid and a gallic acid bonded to a xylose with glycosidic and ester bonds, respectively. Compound (1) was identified as 4,5-dihydroxy-6-(3,7,8–trihydroxy-5,10-dihydro–chromeno[5,4,3-cde]chromen-2-yloxy)-tetrahydro-pyran-3-yl ester, and compound (2) was identified as (–)-lyoniresinol. The presence of (–)-lyoniresinol from damaged sapwood indicated that infection of R. quercivora may cause the formation of a pseudo-heartwood in the sapwood of Q. crispula.