Blue stain fungi associated withTomicus piniperda (Coleoptera: Scolytidae) on Japanese red pine

A number of various species of blue-stain fungi were isolated fromTomicus piniperda adults at various stages of development, as well as from the galleries, pupal chambers and sapwood underneath galleries on Japanese red pine. This study was an attempt to identify the species, composition of blue-stain fungi associated withT. piniperda, the frequency of occurrence of the fungi, and their role in the sapwood-staining of Japanese red pine in Tsukuba City, central Japan. Among the seven species of blue-stain fungi isolated, an undescribed species ofOphiostoma together withO. minus were the dominant species and closely associated withT. piniperda. These two species occurred on newly emerging adults more frequently than the overwintered adults.Hormonema dematioides was also associated with the beetle, however, its frequency of occurrence from the emerged new adults was very low. Although the two other species,O. ips andGraphium sp. were also isolated from emerged beetles, the frequency of these fungi from gallery systems suggested that they were accidentally carried byT. piniperda. Leptographium wingfieldii, known to be associated with the beetle in Europe, was also isolated at a very low frequency and the fungus seemed not to be closely associated with the beetle.Ophiostoma sp. andO. minus appear to be the most important causes of blue-stain of Japanese red pine sapwood after infestation byT. piniperda.     

杨树溃疡病潜伏侵染的研究

在陕西省杨树溃疡病(Dothiorella gregaria Sacc.)是一种严重的病害,分布普遍,危害130多个杨树品种、杂交种和无性系,其中有些树种如箭杆杨、北京杨、陕林2号杨、箭×钻杨等受害更为严重,常导致整株死亡,造成重大损失。据研究杨树溃疡病菌有潜伏侵染的特性,但在陕西省杨树苗木上未见其发病,有无潜伏侵染现象存在,更不清楚,为了弄清这一问题,我们于1987—1988年,对来自三个苗圃的24种杨树苗木     

Nutritional and pathogenic fungi associated with the pine engraver beetle trigger comparable defenses in Scots pine

Conifer bark beetles are often associated with fungal complexes whose components have different ecological roles. Some associated species are nutritionally obligate fungi, serving as nourishment to the larvae, whereas others are pathogenic blue-stain fungi known to be involved in the interaction with host defenses. In this study we characterized the local and systemic defense responses of Scots pine (Pinus sylvestris L.) against Ophiostoma brunneo-ciliatum Math. (a blue-stain pathogen) and Hyalorhinocladiella macrospora (Franke-Grosm.) Harr. (a nutritional fungus). These fungi are the principal associates of the pine engraver beetle, Ips acuminatus (Gyll.). Host responses were studied following inoculation with the fungi, singly and as a fungal complex, and by identifying and quantifying terpenoids, phenolic compounds and lignin. Although the length of the necrotic lesions differed between control (wound) and fungal treatments, only two compounds (pinosylvin monomethyl ether and (+)-α-pinene) were significantly affected by the presence of the fungi, indicating that Scots pine has a generic, rather than specific, induced response. The fact that both nutritional and blue-stain fungi triggered comparable induced defense responses suggests that even a non-pathogenic fungus may participate in exhausting host plant defenses, indirectly assisting in the beetle establishment process. Our findings contribute to the further development of current theory on the role of associated fungal complexes in bark beetle ecology.     

我国杨树溃疡病研究进展

杨树溃疡病主要由真菌侵染所致,该真菌是一类世界性分布的真菌,寄主范围十分广泛,可危害多种林木和果树的树干及果实,引起枯枝、溃疡、流胶和果腐等。另外,还可以引起根腐,导致整株树的枯死。发生病害的林木,用材林材性降低,经济林产量降低,造成严重危害和经济损失。文中对其病害分布、致病机理、抗病机制以及抗病育种等进行了综述。     

Use of apigenin from Cordia dichotoma in the treatment of colitis

Cordia dichotoma f. (Boraginaceae) is a small deciduous tree from India. The bark of was used in the treatment of ulcerative colitis (UC) and colic pain traditionally hence present work was undertaken to identify the phytoconstituent responsible for this activity. Apigenin is isolated by column chromatography from methanol fraction of crude methanol extract of C. dichotoma bark. Structure of apigenin is established by various spectroscopic studies. Apigenin (5 mg/kg, p.o.) showed significant healing and reduction in inflammatory enzymes when screened for UC. It can be concluded that apigenin from C. dichotoma bark may be responsible for the treatment of UC.     

Lesion development in stems of rough‐ and smooth‐barked Eucalyptus nitens following artificial inoculations with canker fungi

A study of lesion development in stems of Eucalyptus nitens following artificial inoculations with canker fungi was carried out on 16‐year‐old plantation trees. In a first trial cambium bark wounds on smooth‐ and rough‐barked trees were inoculated with the mycelium of nine species of canker fungi, including Endothia gyrosa. In a second trial spores or mycelium of E. gyrosa were applied directly onto undamaged or superficially wounded bark surfaces. Infection subsequent to artificial inoculation via wounding (whatever the wounding technique or type of inoculum) resulted in significantly larger external lesions (mean lesion area up to 35.6 cm² 20 months after inoculation) on smooth bark compared with those on rough bark (up to 19.0 cm²). Microscopic studies of infected rough and smooth bark suggest that, once smooth bark is compromised by wounding and artificial inoculation, the particular anatomical structure of smooth bark may offer less mechanical resistance to post‐penetration hyphal spread in comparison with rough bark. It is suggested that at a pre‐penetration stage under natural conditions spores of E. gyrosa more easily infect rough bark via cracks associated with this type of bark but not present in smooth bark.     

Antioxidant activity of extracts from <Emphasis Type="Italic">Calocedrus formosana</Emphasis> leaf,bark, and heartwood

Calocedrus formosana Florin (Cupressaceae) is an endemic tree to Taiwan. To evaluate the antioxidant activity of the plant extracts from heartwood, bark, and leaf of C. formosana, assays for 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and superoxide anion scavenging activities, as well as prevention of DNA strand cleavage were performed in this study. Similar IC₅₀ values against the DPPH radical were found for the heartwood and bark extracts at approximately 23µg/ml. Moreover, the heartwood extract exhibited the highest inhibitory activity against superoxide radicals among the test samples; a 2.3-fold lower value of IC₅₀ for superoxide radical inhibition was found in the heartwood extract relative to that of (+)-catechin. Much less effect on inhibition of DPPH and superoxide radicals was found from the leaf extract of C. formosana. More than 70% of superoxide radicals were inhibited in the presence of 10µg/ml heartwood extract, whereas only 15% inhibition was obtained from the leaf extract. The heartwood extract, at a dose of approximately 0.5mg/ml, apparently completely prevented the X174 supercoiled DNA cleavage induced by ultraviolet photolysis of H₂O₂, as judged by agarose gel electrophoresis. This report also suggests that the antioxidant activities of the plant extracts of C. formosana are in good correlation with their phenolic contents.     

Antimutagenic, antioxidant and antimicrobial properties of Maytenus krukovii bark

The hydroalcoholic extract of Maytenus krukovii bark was investigated for its in vitro mutageno-protective activities by means of the Ames Salmonella/microsome assay. The extract showed an inhibitory effect in both T98 and T100 strains against the mutagenic activity of promutagen 2-aminoanthracene but was not protective against directly acting mutagens sodium azide and 2-nitrofluorene. When tested as a radical scavenger and antioxidant it produced a dose-dependent inhibition. The extract did not show significant antibacterial properties, and was weakly active against dermatophyte and phytopathogenic fungi, but inhibited the growth of phytopathogen Pithyum ultimum.     

Induced resistance of Norway spruce,variation of phenolic compounds and their effects on fungal pathogens

Three clones of Norway spruce (Picea abies) were studied for their response to mass‐inoculation with the blue‐stain fungus Ceratocystis polonica. The effect of different pretreatments (fungal inoculation and wounding) before mass‐inoculation was investigated for their possible role in an acquired resistance reaction. Pretreated trees showed enhanced resistance to the subsequent mass‐inoculation relative to control trees that received no pretreatment. Furthermore, the fungal colonization of inoculated trees was less than that of wounded trees. The phenolic content of the bark, analysed by RP‐HPLC, was compared in trees receiving different treatments. Trees inoculated with C. polonica had higher average concentration of (+)‐catechin, taxifolin and trans‐resveratrol than wounded trees. Both inoculated and wounded trees had higher average concentrations of these compounds than control trees. The effect of the phenolic extract of Norway spruce bark on the growth of the root rot fungus Heterobasidion annosum and the blue‐stain fungi C. polonica and Ophiostoma penicillatum were investigated in vitro. Heterobasidion annosum was not negatively affected, and the extracts had fungistatic effects on the blue‐stain fungi. The growth of O. penicillatum was more inhibited than the growth of the more aggressive C. polonica.     

Endothia gyrosa associated with severe stem cankers on plantation grown Eucalyptus nitens in Tasmania,Australia

Severe stem cankers in Eucalyptus nitens, from a 14-year-old mixed provenance plantation, were associated with infection by Endothia gyrosa, present in its teleomorph state. Surveys of incidence among canker severity classes were carried out in a thinned and pruned stand and an adjacent unthinned and unpruned stand within the affected plantation. No differences in incidence among the canker severity classes were found between the thinned/pruned and unthinned/unpruned stands or between different crown dominance classes within the unthinned/unpruned stand. However, the incidence among canker severity classes was strongly associated with bark roughness with 97% of rough-barked trees developing either annual cankers or cankers causing cambial damage. Stem cankers were found on only 11% of trees with smooth bark. Bark roughness in E. nitens was shown to differ significantly between provenances. Deployment of provenances prone to rough bark in routine plantation establishment may pose a risk of damaging stem canker outbreaks.     

Screening for condensed tannin-degrading fungi with a synthetic <Superscript>14</Superscript>C-labeled compound

To find fungi that are potent for degradation of condensed tannin, a two-step screening was used. This involved measurement of fungal growth rate on Japanese cedar (Cryptomeria japonica) bark, followed by determination of [¹⁴C]-labeled CO₂ generated from fungal degradation of synthetic [¹⁴C]-labeled condensed tannin model. In the first screening, 75 strains of wood rot fungi were tested, and 19 strains effectively decreased bark weight and/or the weight of the methanol-soluble fraction. For the second screening, [¹⁴C]-labeled condensed tannin model compound was synthesized in 11.8% yield based on radioactivity measurements. Over the incubation period, Coriolus hirsutus K-2671, Lentinus edodes Is, and Lampteromyces japonicus Nn showed higher cumulative [¹⁴C]-labeled CO₂ emissions than the other strains and mineralized the [¹⁴C]-labeled condensed tannin model compound by 3.7%, 3.0%, and 3.0%, respectively. Fractionation of the methanol extracts from the medium by gel permeation chromatography after fungal treatment suggested that fungi that can induce the emission of significant levels of [¹⁴C]-labeled CO₂ can extensively depolymerize condensed tannins.     

Ursachen und Ausmaß von Stammfäulen in Fichtenbeständen auf verschiedenen Standorten

Summary In 9 Norway spruce stands from different growth regions in Southern Germany, which were provided for clear cutting and where considerable damage by red rot was supposed, on experimental plots all trees before felling were surveyed respecting species, diameter, tree-class and soundness. After felling, the stems were tested for fungus attack, the types of rot (root rot or wound rot) were registered, the degree of the damage was determined and the responsible wood-destroying fungi were identified. In all localities almost the same species of fungi were found, but their quote differed considerably according to the predominating type of rot. At some of the investigated stands spruce was not very resistant to root rot, at others—in spite of good nitrogen nutrition—the diseases only resulted from wounds; rots starting from the roots were seldom found. These differences observed in several stands are discussed. Statistical calculations have shown, that the correlation between the degree of decay on the stump and the height of the rot depends on the site. Therefore an estimate of the amount of rotten wood on account of the extent of decay on the stump, certainly is possible for individual stands, but not in universal view.      

Growth inhibition activities of Sugi bark components against Heterosigma akashiwo

In our ongoing efforts to develop new uses for wood-based waste streams, the growth inhibition activities of extracts obtained from Sugi (Cryptomeria japonica) bark were examined against Heterosigma akashiwo, otherwise known as red tide plankton. The Sugi bark was separated into its outer and inner barks and then extracted sequentially with hexane, ethyl acetate, and methanol. Strong inhibitory activities against H. akashiwo were observed in the tests involving the hexane extract from the inner and outer barks, as well as the ethyl acetate extract from the inner bark. Gas chromatography mass spectroscopy (GC–MS) analysis revealed that cubebol, phyllocladanol, 6,7-dehydroferruginol, ferruginol, and sugiol were the main components in the active extracts. These components themselves were then tested for their growth inhibition activities against H. akashiwo. Cubebol and ferruginol showed potent inhibitory activities, whereas phyllocladanol, 6,7-dehydroferruginol, and sugiol were only weakly active. Taken together, these results suggested that the Sugi bark extracts could be used as inhibition reagents against red tide plankton.     

The threshold of successful attack by Ips typographus on Picea abies: A field experiment

The spruce bark beetle, Ips typographus, was induced to attack Norway spruce by means of pheromone dispensers. The degree of attack on each tree was recorded and the trees were later categorized as surviving or dying, according to the degree of sapwood blue-staining caused by the attacks. A threshold of successful attack was observed; i.e. above a certain number of attacks the trees were successfully invaded by the beetles and their mutualistic blue-stain fungi. The height of this threshold increased with increasing tree vigour, measured as the relative increment of the sapwood cross-sectional area.     

Antibacterial activity of Alstonia scholaris and Leea tetramera

The crude methanolic extracts of the leaves, stem and root barks of Alstonia scholaris and Leea tetramera on partitioning (petrol, dichloromethane, ethyl acetate, butanol) gave fractions exhibiting improved and broader spectrum of antibacterial activity. Especially the butanol fractions of A. scholaris and the root bark of L. tetramera. None of the fractions were active against the fungi tested.     

Bark consumption by voles in relation to geographical origin of tree species

Voles have caused more damage by bark consumption to Picea abies and Pinus contorta seedlings from southern than those from northern origins when planted in northern Scandinavia. In field and laboratory experiments, indigenous Scandinavian tree species from different origins and of various phenological stages were tested for susceptability to vole attacks. Seedlings from northern sites and late phenological stages were most severely attacked by both bank voles Clethrionomys glareolus and field voles Microtus agrestis, while a few aspen clones differed independently of origin. Bark consumption by the two vole species was strongly and negatively related to the concentration of crude fibre in the examined twigs. Bank voles also reacted positively to the content of nitrogen‐free extracts. Differences in preferences for seedlings from different geographical regions appear to result from differences in the bark at the end of growth and in the frost‐hardening process in autumn. Plants transferred northwards probably will not harden in a climatically appropriate way, will contain low concentration of fibre in winter, and will be a preferred food for voles.     

Host responses to natural infection by Cytonaema sp. in the aerial bark of Eucalyptus globulus

The chemical and anatomical host responses to natural fungal infection by Cytonaema sp. in the aerial bark of 3‐year‐old Eucalyptus globulus plantation trees were examined. The lesion margin (LM) of the canker‐infected bark was characterized by the formation of a layer of dark extractives visible to the naked eye. Chemical analysis of the LM by gas chromatography‐mass spectrometry (GC‐MS), high‐performance liquid chromatography (HPLC) with ultraviolet detection and HPLC‐MS using negative ion electrospray ionisation indicated the presence of a range of compounds including hydrolysable tannins, polymeric proanthocyanidins, flavonoid glycosides, formlyated phloroglucinol compounds and volatile terpenes. These compounds were either undetectable in healthy tissue or present at significantly lower concentrations than in the LM. The LM of the canker‐infected bark was morphologically distinct from healthy phloem, its characteristics varying depending on severity of canker infection. In superficial infections in which only the phloem was affected, the following LMs were observed: (i) a continuous wound periderm of multiple layers, or (ii) an incompletely differentiated and discontinuous wound periderm. In cases of severe canker infections in which the vascular cambium had been killed, the new phloem formed subsequently contained traumatic oil glands in addition to the responses observed for superficial canker infections. All LMs were characterized by the formation of new parenchyma cells that stained positive for the presence of polyphenols. The significance of the chemical and structural responses as defence mechanisms against fungi causing stem canker is discussed.     

Effects of oleoresins and monoterpenes on in vitro growth of fungi associated with pine decline in the Southern United States

As a means of exploring pine resistance to root disease and declines, the effects of host plant secondary metabolites on the growth of root colonizing fungi associated with three diseases/declines of southern pines – loblolly pine decline, littleleaf disease and annosum root rot were tested. The associated fungi –Leptographium huntii, L. serpens, L. terebrantis, L. procerum, Heterobasidion annosum and Phytophthora cinnamomi– were grown in saturated atmospheres or in direct contact with, pure monoterpenes and crude oleoresin collected from the four southern pines (Pinus taeda, P. eschinata, P. palustris and P. elliotti) for 7 day. Fungal growth was measured at 3, 5 and 7 day. Root‐infecting fungi differed significantly in sensitivity to crude oleoresin and pure monoterpenes. All fungi tested were inhibited, to some extent, by the resins tested. H. annosum and P. cinnamomi were strongly inhibited by all the monoterpenes tested. The ophiostomatoid fungi were significantly less affected by the compounds tested. L. huntii and L. serpens were less inhibited by monoterpenes than either L. terebrantis or L. procerum. These fungal growth studies show that the kind and amount of secondary metabolite produced by the host plant have a profound effect on tree pathogens. Alterations of tree physiology may have implications for defenses against tree pathogens as well as to the ecology and management of forest ecosystems. Difference in incidence of root disease observed in the field may be explained by the ability of the fungus to tolerate these host defense mechanisms.     

Comparison of terpenes in extracts from the resin and the bark of the resinous stem canker of Chamaecyparis obtusa and Thujopsis dolabrata var. hondae

A monoterpene and 15 diterpenes were isolated from the ethyl acetate extracts of the bark-glued resin from the resinous stem canker ofThujopsis dolabrata var.hondae Makino. A monoterpene (nezukone20) and 4 diterpenes (acetyl torulosol5, acetyl isocupressic acid8, acetyl abietinol11, and 7-methoxytotarol18) were characteristic constituents of the ethyl acetate extracts but were absent in then-hexane extracts from the resinous stem canker ofT. dolabrata var.hondae. These terpenes were first isolated fromT. dolabrata var.hondae andT. dolabrata. The available literature suggests that diterpene18 is a new compound. The resinous stem canker ofChamaecyparis obtusa Endlicher contained larger amounts of manool1,trans-communic acid6, and ferruginol12 and smaller amounts of isocupressic acid7 and abietinol10 than the resinous stem canker ofT. dolabrata var.hondae. The concentration of18 was less than 2% in the extracts, and the resinous stem canker ofC. obtusa lacked this compound. The resinous stem cankers ofC. obtusa andT. dolabrata var.hondae provided extracts 15.6 and 4.96 times, respectively, heavier than the healthy ones. Large differences in the ratios and compositions of terpenes were also observed between the resinous stem canker and the healthy trees. Terpenes isolated from the extracts contained many kinds of diterpene, especially the labdane-type diterpenes, in these diseased trees. These results suggest that the presence of labdane-type diterpenes is closely associated with the resinous stem canker or the causal fungi of this disease.Part of this report was presented at the 50th annual meeting of the Japan Wood Research Society, Kyoto, April 2000     

Ophiostomatoid fungi associated with bark beetles onAbies veitchii in wave-regenerated forests

Relationships between tree mortality and bark beetle infestation onAbies veitchii at the wave-regenerated forest in Mt. Asahi, Okuchichibu area were investigated. Most of the firs with green needles and newly developed current year’s shoots in the dieback zone were heavily infested by bark beetles before the death of the trees. After heavy infestation of beetles, about half of the infested firs died within the year, and the other half died in the next year. When the species composition of bark beetles and associated ophiostomatoid fungi were investigated in Mt. Asahi and also at a typical wave-regenerated forest in Mt. Shimagare, Yatsugatake area,Cryphalus montanus andC. piceae were dominant beetle species for Mt. Asahi and Mt. Shimagare, respectively.Ophiostoma subalpinum andO. europhioides were dominant fungal species at both wave-generated forests. BecauseO. subalpinum was detected more frequently from deeper areas of sapwood thanO. europhioides, it was suggested that the fungal species may accelerate the death of stressed firs in wave-regenerated forests. Contribution No. 169, Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba.