Occurrence of the wattle wilt pathogen,Ceratocystis albifundus on native South African trees

Ceratocystis albifundus causes the disease known as wattle wilt of non‐native Acacia mearnsii trees in South Africa, Uganda and Kenya. Infection results in rapid wilt and death of susceptible trees and stem cankers on more tolerant trees. It has been suggested that C. albifundus is indigenous to southern Africa, possibly having spread from native Protea spp. to non‐native A. mearnsii and A. decurrens trees. Although C. albifundus has been collected from Protea spp., these reports are based on limited records for which only aged herbarium specimens exist. During surveys of wound‐infecting fungi on native tree species in South Africa, a fungus resembling C. albifundus was collected from Protea gaguedi, Acacia caffra, Burkea africana, Combretum molle, C. zeyheri, Faurea saligna, Ochna pulchra, Ozoroa paniculosa and Terminalia sericea. The identity of the fungus was confirmed as C. albifundus, using comparisons of DNA sequence data for the ITS and 5.8S gene of the rRNA operon. In pathogenicity trials, lesions were produced on C. molle and A. caffra, with some trees beginning to die at the termination of the experiment. This study represents the first report of C. albifundus from native tree species in South Africa and provides unequivocal evidence that the fungus occurs naturally on native Protea spp. The wide host range of C. albifundus, as well as its abundance on these indigenous hosts lends further support to the view that it is a native African pathogen.     

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.     

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.     

Effect of season and different fungi on phenolics in response to xylem wounding and inoculation in Eucalyptus nitens

Pot‐grown and plantation‐grown Eucalyptus nitens trees (approximately 2 and 3 years old, respect‐ively) were experimentally wounded and inoculated with different fungi and in different seasons. Decay lesion development and defence zones were assessed. Two zones were described, a narrow brown decay interface (interface reaction zone, IRZ) and a diffuse zone beyond this being either pale brown or purple (reaction zone, RZ). The total phenol levels in the reaction zone were determined. Selected phenolics (pedunculagin, tellimagrandin 1, tetragalloylglucose, pentagalloylglucose and catechin) were quantified by liquid chromatography–mass spectrometry (LC–MS). A range of fungi (mainly decay‐causing) were used to inoculate wounds and the results indicated that more extensive decay lesions were generally associated with greater production of soluble phenols in response. Sterile inoculations and weakly aggressive fungi were associated with no or little xylem discoloration, whereas aggressive fungi elicited more discoloration and phenolic accumulation in advance of infection. This indicates that phenol accumulation is not a generalized response to wounding, but a variable response due to the interaction between microorganisms and sapwood. In plantation‐grown trees examined 6 months after wounding, purple reaction zones were commonly associated with large decay lesions. Seasonal differences in decay column area caused by Ganoderma applanatum were not significant 1 month after wounding and inoculation.     

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.     

Impact of bark wounds on sapwood in Norway spruce and silver fir

Bark wounds by damage during harvesting are a serious problem in forestry due to fungi infection and wood deterioration. This paper presents results of an investigation about the influence of the wounds on the internal structure of such injured stems. In an experiment, bark wounds were artificially created at the stem base of Norway spruce [Picea abies (L.) Karst.] and silver fir (Abies alba Mill.). 2 years later, the injured stems along with undamaged controls were cut and the trunk portion below breast height subjected to computer tomographic (CT) analysis. Analysis of the CT-images revealed a substantial impact of wounding on sapwood properties in spruce: directly adjacent to the wound surface in all examined trees, a large disturbance zone was detected affecting on average 17% of the potential sapwood area. With increasing distance from the wound, the size of this disturbance zone diminished, but was still detectable in all trees at breast height ca. 1 m above the bark wound.     

Bark phenolics of American beech (Fagus grandifolia) in relation to the beech bark disease1

The amount of total extractable phenols was determined for bark sections obtained from behind cankers naturally induced by Nectria coccinea var. faginata and from behind mechanically inflicted wounds on stems of American beech. Healthy bark from susceptible trees was found to contain a similar level of phenolics as bark from trees determined to be resistant to the beech bark disease. Six months after wounding, wound-altered bark from susceptible trees was found to be lower in phenols than woundaltered bark from resistant trees. Inoculation of wounds with N. coccinea var. faginata resulted in decreased phenolic levels in bark sections nearest the wound surface, and increased phenolic levels in sections nearest the vascular cambium, several mm distant. Phenolic levels in injured or infected bark appear to follow similar patterns as those resulting from injury or infection of xylem tissues.     

Fungi associated with Ips typographus on Picea abies in southern Poland and their succession into the phloem and sapwood of beetle‐infested trees and logs

This study dealt with the species distribution and frequency of fungi associated with the bark beetle Ips typographus (Scolytidae) on spruce trees of various states of health in southern Poland. The spruce trees were assessed by their degree of defoliation and damage of their crowns and trunks after attack by I. typographus. The state of health of trees from which samples were obtained was related to varying stages of brood development of I. typographus. Fungi were isolated from phloem taken from and around insect galleries and the sapwood underneath brood systems. Samples were taken from ‘healthy‐looking’, weakened, wind‐fallen and wind‐broken trees as well as from trap trees. The mycobiota associated with I. typographus was quite diverse in respect of the number of detected species, 65 fungal taxa were obtained from the phloem of trees infested by I. typographus, and 36 taxa occurred in the sapwood underneath insect galleries. The spectrum of fungi mainly consisted of ascomycetes and anamorphic fungi. The ophiostomatoid fungi were represented by 14 species and were the most numerously represented group in all niches examined. The most frequent ophiostomatoid species were Ceratocystis polonica, Ophiostoma ainoae, O. bicolor, O. penicillatum, O. piceae and O. piceaperdum. The frequency of occurrence of ophiostomatoid fungi differed significantly between the phloem and sapwood as well as in relation to the varying states of health of the spruce trees. These quantitative differences in the mycobiota of I. typographus between spruce trees belonging to different health categories can be explained by successional patterns of fungal colonization of host tissues following attack by I. typographus. The pathogenic species C. polonica was the primary invader, occurring most frequently in the sapwood of ‘healthy‐looking’ trees. Ophiostoma bicolor, O. penicillatum and O. piceaperdum also occurred during the early stages of brood development of I. typographus on ‘healthy‐looking’ trees, but they mainly colonized the phloem. In contrast, O. ainoae, O. minuta and O. piceae likely follow the aforementioned species as secondary and tertiary invaders into the phloem and the sapwood of spruce trees.     

Pruning quality affects infection of Acacia mangium and A. crassicarpa by Ceratocystis acaciivora and Lasiodiplodia theobromae

Pruning (singling) is a common silvicultural practice in commercial Acacia plantations because these trees tend to have multiple stems. The wounds resulting from pruning are susceptible to infection by pathogens. Ceratocystis acaciivora and Lasiodiplodia theobromae have been shown recently to be important pathogens of A. mangium in Indonesia, where they are commonly associated with wounds on trees. The aim of this study was to determine the impact of different wound types on infection of A. mangium and A. crassicarpa by these two pathogens. Isolates of C. acaciivora and L. theobromae, found to be the most pathogenic in a prior study, were used to inoculate pruning wounds. Results showed that pruning conducted in a manner to reduce stem damage, resulted in lower levels of fungal infection. Where pruning resulted in tearing of the bark, there were higher levels of infection and disease occurred even without artificial inoculation. Inoculation of pruning wounds with C. acaciivora and L. theobromae showed that both fungi have the potential to cause disease. However, C. acaciivora was most virulent. Results of this study showed conclusively that careful pruning will result in lower levels of disease in young A. mangium and A. crassicarpa plantations.     

国外黑荆树资源利用状况及我国发展的前景

黑荆树是一种周期短、用途广、效益高的好树种.它原产于澳大利亚,现已在南美和亚洲广泛种植和利用.世界现有黑荆树人工林40多万公顷.黑荆树栲胶年产量11万多吨,占总产量的20%.本文着重介绍了南非和巴西的经验且结合国情认为我们应借鉴国外经验改变传统经营方式走发展工业人工林的道路.     

Wood‐rotting basidiomycetes are a minor component of fungal communities associated with Acacia hybrid trees grown for sawlogs in South Vietnam

Acacia hybrid (Acacia mangium × Acacia auriculiformis) clones are widely planted in Vietnam with a total of approximately 400,000 ha to meet the demand for pulpwood, sawn timber and wood chip exports. Silvicultural techniques such as pruning and thinning have been applied to improve productivity and sawlog quality of Acacia hybrid plantations. However, those techniques may also create opportunities for wood decay fungi to enter the Acacia hybrid stems through wounds and cause stem defects that reduce sawlog quality and the value of the plantation. The presence of fungal decay agents in Acacia hybrid trees was examined in two Vietnamese plantations. In July 2011, just prior to a second thinning, discoloured wood samples were taken from a three‐year‐old Acacia hybrid plantation at Phan Truong Hai for the isolation of fungi. In July 2012, approximately 18 months after pruning and thinning treatments, discoloured wood samples were taken from a three‐year‐old Acacia hybrid plantation at Nghia Trung for the isolation of fungi. DNA sequencing of the rDNA ITS identified the isolates. In May 2015, approximately 4 years after thinning and fertilizer treatments, discoloured and decayed wood samples were taken from the above (7‐year‐old) Acacia hybrid plantation at Phan Truong Hai for fungal identification. DNA was extracted directly from discoloured and decayed wood samples and fungal rDNA ITS amplicons sequenced on a Roche 454 sequencer. The results showed that silvicultural treatments did not affect the fungal communities associated with discoloured and decayed wood of Acacia hybrid plantation at Phan Truong Hai. A total of 135 fungal species or OTUs (operational taxonomic units) were identified, including 82 members of Ascomycota and 52 Basidiomycota.     

Response of maple sapwood to injury and infection

In sapwood challenge experiments in Acer rubrum, columns of discolouration initiated by wounding and inoculation with pioneer fungi (Cephalosporium sp., Phialophora sp.) were similar in size to untreated wounds. Inoculation with decay fungi (Pleurotus ostreatus, Trametes versicolor) produced larger columns of wound-initiated discolouration. The removal of bark around a bore wound caused a significantly larger column to form compared to the sum of the columns inititiated by separate wounds. Stage-I discoloured wood, not associated with obviously rotted wood, had concentrations of mobile cations and soluble phenols similar to sapwood. Stage-II discoloured wood, spatially associated with rotted wood, was frequently bounded by a chemically distinct boundary layer and the discoloured wood contained significantly greater concentrations of mobile cations and soluble phenols than stage-I discoloured wood.     

Fungi in bark peeling wounds of Picea abies in central Sweden

A total of 210 Norway spruce trees with stem wounds resulting from bark peeling by moose (Alces alces) were examined in three 45–50-year-old stands that contained 20-30% of damaged trees. Injured stems were between 8 and 40 cm diameter at breast height (d.b.h.) and showed 1 to 23-year-old wounds. Wounds varied in size from 2 to 4815 cm². The size of injury correlated negatively with the age of the injury (r=-0.24; p=0.004), and positive correlation was established between the age of the injury and d.b.h. of the wounded tree (r=0.50; p=0.0001). Each wound was sampled once using an increment borer and fungi were isolated. Among basidiomycetes, Stereum sanguinolentum was the most common (isolated from 26.7% of the damaged stems). Other common species were Cylindrobasidium evolvens (23.8%), Amylostereum areolatum (5.2%), A. chailletii (0.5%), Heterobasidion annosum (5.2%), Peni-ophorapithya (1.4%), Sistotrema brinkmannii (1.0%). The ascomycete Nectria fuckeliana was the most common among all fungi (present in 35.7% of bark peeling wounds). The frequency of S. sanguinolentum infection correlated positively with the age of the injury (r=0.27; p=0.001) and the opposite relationship was revealed for C. evolvens (r=?0.30; p=0.0001). Furthermore, C. evolvens infection correlated positively with the wound size (r=0.30; p=0.0001) and negatively with the tree d.b.h. (r=?0.20; p=0.004). A positive correlation was found between tree d. b. h. and the occurrence in stems of H. annosum (r=0.23; p=0.001 and N. fuckeliana (r=0.23; p=0.0006). The spruce bark beetle Dendroctonus micans attacked 14.8% of wounded trees. Presence in stems of N. fuckeliana was associated significantly with the D. micans attack (r=0.190; p =0.006; χ² test: p=0.01). Except for a negative correlation between infections of S. sanguinolentum and H. annosum (r=-0.140; p=0.04), no significant relationship between fungal species was found.     

Penetration of Picea sitchensis root bark by Armillaria mellea,Armillaria ostoyae and Heterobasidion annosum

Penetration of root bark tissues of Picea sitchensis by Armillaria ostoyae, Armillaria mellea and Heterobasidion annosum was examined in the absence of wounds, in superficial wounds (rhytidome tissues removed to expose the secondary phloem) and in wounds to the depth of the vascular cambium (deep wounding). Both species of Armillaria penetrated bark without prior wounding, but neither species formed rhizomorphs in this treatment. Armillaria ostoyae penetrated to 39 cell layers in depth by 48 days after inoculation of unwounded bark, whereas A. mellea penetrated 25 cell layers in the same time. Armillaria mellea penetrated superficial wounds significantly more rapidly than did A. ostoyae. Both species produced rhizomorphs within wounded host tissues. Inoculation of deep wounds with Armillaria resulted in a greater depth of bark necrosis with A. mellea than with A. ostoyae. In the absence of wounding, H. annosum failed to penetrate root bark tissues, but in both superficial and deep wounds hyphae penetrated beyond the ligno–suberized boundary zone (LSZ) by 12 days after inoculation. Where no inoculations were made, superficial or deep wounding led within 25 days to the restoration of a structurally continuous LSZ, and by day 48 the wound periderm (WP) was fully differentiated. In inoculated wounds, however, formation of the LSZ and WP was delayed or inhibited in most trees, particularly following inoculation with A. ostoyae or A. mellea. Suberization in the LSZ and WP remained diffuse and discontinuous 48 days after inoculation. Moreover, the presence of WP did not prevent further penetration of the tissues by the pathogens. Variations between trees in the depth of pathogen penetration were noted, possibly indicating differing susceptibilities of individual host genotypes. The possible host factors involved in resistance to penetration of root bark tissues by Armillaria and Heterobasidion are discussed.     

Pathogenicity of ophiostomatoid fungi on Picea abies in Slovenia

Pathogenic fungi can survive and develop in living plants, often causing diseases in the host. Some theories speculate that pathogenic ophiostomatoid fungi provide benefits to its vectors – bark beetles – by overcoming the tree's defence mechanisms. This study reports the results of an experiment in south‐eastern Europe in which mature and seedling Norway spruce trees were artificially inoculated with various ophiostomatoid fungi. The aim of the experiment was to determine the relative virulence of ophiostomatoid fungi by assessing the ability of the fungi to stimulate host tree defence mechanisms through inoculation experiments. Experiments were performed by inoculation of Picea abies in seedling and mature trees. The following fungi were used in low‐density and seedling inoculations: Ophiostoma ainoae, O. brunneo‐ciliatum, Grosmannia cucullata and an unidentified Leptographium sp., O. bicolor, O. fuscum, O. piceae, G. penicillata and G. piceiperda. Endoconidiophora polonica was used in mass and seedling inoculations. Various characteristics such as host vitality, blue stain, lesion and resin outflow were measured before and after the trees were felled. E. polonica caused blue stain, induced large lesions and killed some of the mature trees and seedlings, confirming earlier reports that it is a strong wound pathogen. Only E. polonica, Leptographium sp. and O. ainoae caused blue stains in the sapwood of inoculated seedlings. In low‐density inoculations, G. piceiperda induced intense necrosis and had higher values for all the characteristics monitored. Some of the other ophiostomatoid fungi showed a moderate level of pathogenicity. Fungi with the capacity to stimulate a host defence mechanism could play a role in the establishment of bark beetle populations.     

Chemical alterations induced by <Emphasis Type="Italic">Pycnoporus sanguineus</Emphasis>/<Emphasis Type="Italic">Aspergillus flavipes</Emphasis> co-cultures in wood from different tree species

Chemical alterations following inoculation of Acacia mearnsii, Eucalyptus dunnii, E. grandis, and E. macarthurii with a Pycnoporus sanguineus/Aspergillus flavipes co-culture were investigated. Several wood chemical parameters were measured using standard methods from the pulp and paper industry. The data were described and analyzed using univariate as well as multivariate statistical techniques. Boxplots and in particular biplots show clearly how the chemical composition of each tree species was differently affected by the co-culture. Lignin content was significantly decreased in A. mearnsii, while E. dunnii showed a decrease in cellulose content. The results, therefore, indicate that the manner in which wood is degraded by a specific fungal co-culture depends on the tree species involved. This phenomenon should be considered when selecting fungi for bio-pulping.     

Variation of heartrot,sapwood infection and polyphenol extractives with provenance of Acacia mangium

Infection of heartwood by decay fungi (heartrot) is a concern for growers of Acacia mangium for solid‐wood products as the incidence can be high in some regions of Indonesia. Variation of heartrot incidence for different provenances of A. mangium was determined using two field trials in Sumatra, Indonesia. In a Riau Province trial of 21 provenances, the effect of provenance was statistically significant for natural heartrot incidence, which ranged from 1.6% to 27.2%. In a smaller trial using artificial inoculation in South Sumatra, heartwood infection incidence ranged from 39.4% to 70.8% across six provenances and both wound type and provenance were statistically significant factors. There was also significant variation in sapwood infection length related to provenance. Wood extractives (yield, total phenols, protein‐precipitable tannin and 2,3‐trans‐3,4′,7,8‐tetrahydroxyflavanone) were quantified from a subsample of trees for each trial. However, no significant differences in extractive concentration were detectable according to provenance and evidence for a relationship between heartwood extractives and heartrot incidence was generally poor. While further studies need to be completed to establish the basis for heartrot incidence, results from these trials allow for recommendations on provenance selection to reduce heartrot incidence and provide information for further genetic selection programmes.     

Effects of mass inoculation on induced oleoresin response in intensively managed loblolly pine

Oleoresin flow is an important factor in the resistance of pines to attack by southern pine beetle, Dendroctonus frontalis Zimm., and its associated fungi. Abiotic factors, such as nutrient supply and water relations, have the potential to modify this plant-insect-fungus interaction; however, little is known of the effects of inoculation with beetle-associated fungi on oleoresin flow. We observed that constitutive and induced resin yield in loblolly pine, Pinus taeda L., were affected by either fungal inoculation (with the southern pine beetle-associated fungus Ophiostoma minus (Hedgcock) H. & P. Sydow) or silvicultural treatment. The effects of mass wounding (400 wounds m(-2)) and mass wounding and inoculation with O. minus were assessed by comparison with untreated (control) trees. The treatments were applied to trees in a 2 x 2 factorial combination of fertilizer and irrigation treatments. Fertilization did not significantly affect constitutive resin yield. Even as long as 105 days post-treatment, however, mass-inoculated trees produced higher induced resin yields than control or wounded-only trees, indicating a localized induced response to fungal inoculation. We noted no systemic induction of host defenses against fungal colonization. Although beetles attacking previously attacked trees face a greater resinous response from their host than beetles attacking trees that had not been previously attacked, the effect of an earlier attack may not last more than one flight season. Despite mass inoculations, O. minus did not kill the host trees, suggesting that this fungus is not a virulent plant pathogen.     

Microorganisms which invade Picea abies in seasonal stem wounds

Wounds were experimentally created on the stems of 55–60 years old Picea abies. Infection, spread, and interrelationship of microorganisms, mainly fungi, were studied after 1, 2, and 4 years. The effect on the invading Hymenomycetes of wounding season, number of years since wounding, tree diameter class, height above ground, size and depth of the wound is discussed.     

Susceptibility of Eucalyptus grandis and Acacia mearnsii seedlings to five Phytophthora species common in South African plantations

Eucalyptus grandis and its hybrids, as well as Acacia mearnsii, are important non‐native trees commonly propagated for forestry purposes in South Africa. In this study, we conducted pathogenicity trials to assess the relative importance of five commonly isolated Phytophthora spp. (Phytophthora alticola, P. cinnamomi, P. frigida, P. multivora and P. nicotianae) from the plantation environment on E. grandis and A. mearnsii seedlings. Overall E. grandis was more susceptible to the tested Phytophthora spp. than A. mearnsii. Phytophthora cinnamomi was the only pathogen that had a significant negative effect on both the host tree species, leading to a reduction in root and shoot weight as well as to death in the case of E. grandis. Phytophthora alticola and P. nicotianae exclusively affected E. grandis and A. mearnsii, respectively. This study updated the current knowledge on the pathogenicity of Phytophthora spp. on two important non‐native commercially propagated tree species from South Africa.