Pathogenicity of Colletotrichum dematium isolated from current-year beech seedlings exhibiting damping-off

The pathogenicity of Colletotrichum dematium was evaluated as the agent causing post-emergence damping-off of current-year beech (Fagus crenata) seedlings. Current-year beech seedlings inoculated with C. dematium died within 20 days after both wound and non-wound inoculation, indicating that this fungus is pathogenic to current-year seedlings. C. dematium was frequently isolated from hypocotyls exhibiting damping-off at five different sites. The incidence of isolation was high and little variation occurred between sites and years. C. dematium was also detected in all litter samples examined although the incidence of detection differed depending on sites and/or detection methods. These results strongly indicate that C. dematium plays an important role in rapid seedling death during the early stages of beech regeneration.     

Characterization of Cylindrocarpon‐like species associated with litter in the old‐growth beech forests of Central Europe

Cylindrocarpon‐like fungi are globally distributed plant pathogens and have a wide range of host species. However, very little is known about the species that live in the topsoil of forests and their potential role in reducing the natural regeneration of tree species, particularly of forest trees that produce abundant fruit only once every few years. To enhance our understanding of the species that inhabit the topsoil, we studied the diversity and pathogenicity of Cylindrocarpon‐like fungi inhabiting the litter in old‐growth mixed‐beech forests in the Carpathians (Poland) and in the Alps (Austria), and in a managed beech stand in the Krakowsko‐Cz?stochowska Highland (Poland). The fungi inhabiting the beech litter were investigated using beechnuts and pine seedlings as bait. Isolates were identified based on morphology and DNA sequencing. The pathogenicity of the most common species was investigated by inoculating beech germinants. A wide range of Cylindrocarpon‐like fungi were associated with the beech litter: 718 cultures representing 12 species were isolated. Five taxa were identified down to species level: namely Ilyonectria crassa, I. pseudodestructans, I. rufa, Neonectria candida and N. obtusispora, and seven species were identified to genus level (Neonectria or Ilyonectria species). Ilyonectria destructans, which is considered to be the sexual morph of ‘Cylindrocarpon destructans’, was not found. There were qualitative and quantitative differences between the different forest sites in terms of Neonectria and Ilyonectria species composition in beech litter. The isolation frequency and species richness of Cylindrocarpon‐like fungi were greatest in beech litter taken from old‐growth mixed‐beech forests. Neonectria and Ilyonectria species were capable of killing beech germinants, suggesting that they may play a negative role in natural beech regeneration.     

Photosynthesis and biomass allocation of beech (Fagus crenata) and dwarf-bamboo (Sasa kurilensis) in response to contrasting light regimes in a Japan Sea-Type beech forest

Regeneration of beech (Fagus crenata) forests depends on the formation of canopy gaps. However, in Japan Sea-type beech forests, a dwarf bamboo (Sasa kurilensis) conspicuously occupies sunny gaps. Therefore,F. crenata seedlings must escape the severe interference ofS. kurilensis in the gaps and persist beneath a closed canopy of the beech forest. We hypothesized that the growth ofF. crenata seedlings in the understory would be favored by their being more plastic thanS. kurilensis in photosynthetic and morphological traits, which would support the matter production ofF. crenata seedlings in a wide range of light availabilities. To examine this hypothesis, the photosynthetic-light response of individual leaves and the biomass allocation in aboveground parts (i.e., the culm/foliage ratio) were surveyed at sites with contrasting light availabilities in a Japan Sea-type beech forest in central Japan. InF. crenata, photosynthetic light utilization efficiency at relatively low light was greater, and the dark respiration rate was smaller in the leaves of seedlings (10 cm in height) beneath the closed canopy than in the leaves of saplings at the sunny forest edge. The culm/foliage (C/F) ratio of theF. crenata seedlings at the shady site was small, suggesting effective matter-production beneath the beech canopy. On the other hand,S. kurilensis both in the gap and beneath the beech canopy showed low plasticity in photosynthesis and the culm/foliage ratio. Because the shoot density ofS. kurilensis was smaller beneath the beech canopy than in the gap, the light availability at the bottom of theS. kurilensis layer was greater beneath the beech canopy. These results suggest that the photosynthetic productivity of theF. crenata seedlings would be enough for the seedlings to survive in the understory with a low density ofS. kurilensis shoots beneath the closed beech canopy.     

Virulence of Fusarium oxysporum and F. commune to Douglas‐fir (Pseudotsuga menziesii) seedlings

Fusarium species can cause damping‐off and root rot of young conifer seedlings, resulting in severe crop and economic losses in forest nurseries. Disease control within tree nurseries is difficult because of the inability to characterize and quantify Fusarium spp. populations with regard to disease potential because of high variability in isolate virulence. Fusarium isolates were collected from healthy and diseased seedlings of Douglas‐fir (Pseudotsuga menziesii) and western white pine (Pinus monticola) from a nursery in Idaho, USA. Molecular markers such as DNA sequences (mitochondrial small subunit and nuclear translation elongation factor 1‐alpha) and amplified fragment length polymorphism were used to identify isolates as either F. oxysporum or F. commune. In addition, diagnostic primers were developed to detect and distinguish F. commune from F. oxysporum. In vitro and greenhouse virulence tests were completed on Douglas‐fir germinants and seedlings. For Douglas‐fir germinants and seedlings, F. oxysporum isolates generally caused less severe symptoms, whereas most F. commune isolates caused mortality through damping‐off. This is the first report of direct evidence that F. commune can cause damping‐off disease on Douglas‐fir seedlings under greenhouse conditions.     

The effect of nitrogen fertilization on fungistatic phenolic compounds in roots of beech (Fagus sylvatica) and Norway spruce (Picea abies)

The effect of nitrogen fertilization on fungistatic phenolic compounds in fine roots of beech and Norway spruce growing in afforestation plots was analysed. The plots were situated at two sites in Switzerland on acidic soil with low base saturation. For 9 years, the trees have been treated with dry ammonium nitrate to give 0, 10, 20, 40, 80, 160 kg N ha^?1 year^?1, respectively. The phenolic compounds responded differently to fertilization. Fine roots of beech showed a significant decrease of (?)‐epicatechin and piceatannol with increasing nitrogen fertilization. The concentration of protocatechuic acid was increased with fertilization. Roots of fertilized Norway spruce showed significantly decreased concentrations of 4‐hydroxyacetophenone and piceatannol. The mycelial growth of three isolates each of Heterobasidion annosum s.l. and Cylindrocarpon destructans was tested on agar media containing various phenolic compounds in concentrations found in fine roots of Norway spruce (Picea abies) and beech (Fagus sylvatica). All three H. annosum isolates were inhibited by p‐coumaric acid and (?)‐epicatechin. Two isolates were inhibited by another four phenolic compounds (p‐hydroxybenzoic acid, 4‐hydroxyacetophenone, piceatannol and protocatechuic acid), one by (+)‐catechin. Two of three C. destructans isolates were inhibited by all phenolic compounds except for (+)‐catechin which affected only one isolate, one isolate did not respond at all.     

Pathogenicity of fungi associated with ash dieback symptoms of one‐year‐old Fraxinus excelsior in Montenegro

In addition to Hymenoscyphus fraxineus, two fungi identified as Diaporthe eres aff. and Fusarium sambucinum aff. were also isolated from necrotic bark lesions on declining one‐year‐old Fraxinus excelsior in a forest stand in Montenegro. To examine their involvement in ash decline, a pathogenicity test was performed using under bark inoculations on one‐year‐old Fraxinus excelsior. Hymenoscyphus fraxineus was included as comparison. All three fungal species proved highly pathogenic towards one‐year‐old seedlings although lesion sizes differed significantly between the different species. Hymenoscyphus fraxineus was most aggressive, followed by F. sambucinum aff., while D. eres aff. caused the smallest lesions. This study demonstrates for the first time the ability of isolates in the D. eres and F. sambucinum species complexes to cause decline on one‐year‐old common ash seedlings.     

Microbial communities in roots of Pinus sylvestris seedlings with damping‐off symptoms in two forest nurseries as determined by ITS1/2 rDNA sequencing

A methodological molecular procedure, which included extraction and cloning of the ITS1/2 rDNA of root‐associated organisms with subsequent transformation and sequencing of representative clones, was effective for detection, discrimination and determination of the frequency of the main damping‐off pathogens in roots of Pinus sylvestris seedlings growing in different forest‐tree nursery soils and exhibiting different rates of disease progress. Roots exhibiting slower damping‐off progression were colonized by Fusarium oxysporum, Neonectria radicicola (Ascomycota) and Pythium spp. (Oomycota), which comprised 50% of the microbial community. Roots exhibiting faster damping‐off progression were dominated by Thanatephorus cucumeris (Basidiomycota), which comprised 80% of the microbial community. The microbial community was more diverse in roots with slower damping‐off progression (14 species) than in roots with faster disease progression (seven species).     

First report of Calonectria montana causing damping‐off disease on pine and spruce seedlings in Europe

In 2015–2016, damping‐off symptoms were observed on Pinus sylvestris and Picea abies seedlings in four forest nurseries in Poland. Some Calonectria isolates were obtained from symptomatic seedlings by culturing pieces of necrotic tissue on malt extract agar.Morphological and molecular analyses revealed that isolates represent Calonectria montana, a recently described species from natural forests in China. The isolates represent the first detection of this species outside of Asia. Pathogenicity tests demonstrated that C. montana is a severe pathogen of P. sylvestris seedlings.     

Association of seed‐borne Fusarium species on Pinus ponderosa with germination and seedling viability in Argentina

Pinus ponderosa seedlots from eight seed orchards in Western Patagonia, Argentina, were evaluated for the presence of Fusarium species, and isolates obtained were subjected to pathogenicity tests. The following species were isolated: Fusariumoxysporum, Fusarium equiseti, Fusarium verticillioides, Fusarium incarnatum, Fusarium acuminatum, Fusarium graminearum and Fusarium proliferatum. With the exception of F. proliferatum and F. graminearum all Fusarium species were recovered from P. ponderosa seedlings with symptoms of damping‐off. Five Fusarium species were re‐isolated from necrotic roots of seedlings showing chlorosis (F. oxysporum, F. equiseti, F. incarnatum, F. acuminatum and F. verticillioides). High damping‐off losses occurred in Fusarium‐contaminated seed of P. ponderosa and low disease incidence occurred on less contaminated seed. The role of seed‐borne inoculum in disease spread within forest nurseries is discussed.     

Root diseases in bareroot and container nurseries of the Pacific Northwest: epidemiology, management, and effects on outplanting performance

In forest and conservation nurseries in the Pacific Northwest USA, seedling production can be limited by root diseases caused by fungi in the genera Fusarium Link:Fr., Cylindrocarpon Wollenw., Phytophthora de Barry, and Pythium Pringsh. Fusarium, Cylindrocarpon, and Pythium are the most ubiquitous, whereas incidence of Phytophthora is mostly associated with coastal bareroot nurseries. All of these root pathogens are encouraged by water saturated soils or media. Seedlings infected with Fusarium, Phytophthora, or Pythium often appear chlorotic or necrotic with extensive root decay. Cylindrocarpon often causes serious root decay without shoot symptoms. The best approach to mitigate losses from these diseases is to use a holistic integrated pest management program. This program should combine chemical controls with cultural practices, particularly those that increase soil permeability and drainage and reduce potential sources of inoculum, especially by disinfesting seeds and containers reused for crops. In general, we found that seedlings meeting nursery specifications for outplanting on forest soil (proper height, root collar diameter, healthy shoot color, lack of disease symptoms) but having these disease organisms on their root systems perform as well as non-infected seedlings.     

Effects of ectomycorrhizal fungi on damping‐off and induction of pathogenesis‐related proteins in Pinus tabulaeformis seedlings inoculated with Amanita vaginata

The formation of mycorrhiza on Pinus tabulaeformis (Pt) seedlings by ectomycorrhizal fungi (ECMF), the protective effect of the ECMF on damping‐off caused by Rhizoctonia solani and the expression of pathogenesis‐related proteins (PR‐proteins) induced by Amanita vaginata and/or R. solani in Pt seedlings were investigated. The results indicated that: (i) among the 10 ECMF tested, five species (Gomphidius viscidus, Suillus laricinus, Suillus tomentosus, A. vaginata and Basidiomycetes no. 058) formed mycorrhiza with Pt seedlings. There was a positive correlation between mycorrhizal colonization levels and protective effects (r = 0.9207). Among the five species, A. vaginata had the highest colonization level and the best protective effect against damping‐off; however, G. viscidus had no protective effect even though it had a high mycorrhization level. (ii) Chitinases, β‐1,3‐glucanases and PR‐proteins in the seedlings were not induced by A. vaginata colonization, but were induced when A. vaginata‐mycorrhizal seedlings were attacked by R. solani. It is possible that chitinases, β‐1,3‐glucanases and 108, 97, 78, 44, 37 and 14‐kDa PR‐proteins induced both by A. vaginata and R. solani protect Pt seedlings from R. solani infection.     

Seed and microsite limitation for seedling recruitment of <Emphasis Type="Italic">Quercus wutaishanica</Emphasis> on Mt. Ziwuling,Loess Plateau,China

The relative influence of seed and microsite availability in the seedling emergence and recruitment of Liaodong oak (Quercus wutaishanica), which dominates the native hardwood forest in Loess Plateau, was examined by seed sowing experiments. Experiments were performed in 216 plots (30 × 30 cm), located in three typical stands (Pinus tabulaeformis plantation, Liaodong oak forest and grassland, 72 plots per stand). A seed augmentation experiment was performed in an oak forest and a conifer plantation, and a seed introduction experiment was carried out in a grassland. Three main factors were considered: seed addition, disturbance and shrub cover. The seedling number and growth in every plot were recorded continuously over 3 years. GLM multivariate was used to analyze the relationships between seedling emergence/recruitment and the explanatory variables in every stand. The results showed that seed addition in undisturbed or disturbed plots always caused significantly increased seedling emergence and recruitment in oak and conifer forests. It demonstrated that the recruitment of Liaodong oak was limited by seed and microsite availability under closed forest, and litter is one of the factors leading to microsite limitation. However, in the grassland, no treatments improved recruitment, indicating that the grassland was not suitable for seedling recruitment because of intense light and soil drought. In the conifer forest, more recruited seedlings, and lower herbivory on seedling leaves, as well as thicker stem basal diameters, indicated that the P. tabulaeformis plantation was safer for Liaodong oak seedling establishment. Unexpectedly, shrub cover did not affect the recruitment of Liaodong oak in the three stands.     

红花尔基沙地樟子松天然林枯立木特征分析

[目的]了解沙地樟子松天然纯林中枯立木的数量及空间结构特征,探究枯立木形成的原因,为樟子松林的保护和经营提供依据。[方法]在沙地樟子松天然纯林中设置2块1 hm~2的大样地,用全站仪对样地中所有胸径大于5 cm的立木进行定位并进行全面调查;对调查样地的基本特征,枯立木的数量特征及径级分布进行了分析,提出了用于表达林分中枯立木微环境的活立木比的概念,并采用林分空间结构参数一元分布和二元分布分析方法,对枯立木与其最近4株相邻木的关系进行分析。[结果]2块不同密度的樟子松天然纯林下更新幼苗和枯立木数量相差较大,密度较小(样地1)的样地更新幼苗和枯立木较少,而密度较大的样地(样地2)中枯立木达到200棵,林下更新幼苗数量达到15 280株·hm~(-2);樟子松天然纯林样地内枯立木主要以小径级木为主,胸径集中在11 cm以下;样地1枯立木径级连续分布,幅度较窄;样地2中的枯立木径级幅度较宽,但在20 22 cm缺刻,有2株大于23 cm的枯立木;2块样地中枯立木的分布格局均为随机分布,样地1中枯立木周围的4株相邻立木大多为活立木,且胸径较枯立木大;样地2中,只有一半的枯立木周围的最近4株立木为活立木,且有三分之一以上的枯立木胸径不是最小的,枯立木有连续分布的现象。2块样地中枯立木的角尺度-大小比数二元分布特征的差异不明显,而角尺度-活立木比二元分布特征和大小比数-活立木比二元分布特征差异明显,样地1中枯立木的最近4株随机分布于其周围的相邻木为活立木且胸径大于枯立木的比例明显高于样地2,而枯立木最近4株随机分布于其周围的相邻木有枯立木的比例明显小于样地2。[结论]樟子松天然纯林枯立木以小径级林木为主,枯立木的数量与林分密度相关,林木竞争是林木死亡的主要原因,密度过大也会产生病虫害,因此,对天然樟子松纯林要进行适度经营,保持合理密度。     

Extent and causes of mortality in Pinus sylvestris advance growth in northern Sweden following overstorey removal

Mortality of Scots pine (Pinus sylvestris L.) advance growth seedlings following overstorey removal was studied in nine stands in north Sweden. In each stand, one 40 × 40 m plot was established for each of four different intensities of overstorey removal: 0% (control), 60%, 85% and 100%. Seedlings were monitored on one 18 × 18 m sub‐plot at the centre of each plot for two or three growing seasons following overstorey removal. For seedlings taller than 100 mm, mortality and number of injured seedlings after two growing seasons increased significantly with increasing basal area removal. Seedlings ≤ 100 mm showed the same trend, though not statistically significant. For the height interval > 100 ≤ 500 mm, mortality was not significantly influenced by seedling height. Most of the mortality and the injuries to seedlings were caused by Hylobius abietis (L.) and Pissodes spp. It is concluded that insect damage to released Scots pine advance growth in northern Sweden is a common problem.     

Temporal and spatial dynamics of an old-growth beech forest in western Japan

Dendrochronological approaches enable us to understand forest stand dynamics by estimation of disturbance history and age structure. The present study was conducted in an old-growth beech forest in a forest reserve in western Japan. Increment cores were taken for tree ring analysis from all canopy trees in a 50 m × 130 m study plot. Radial growth release criteria were developed to identify significant growth releases in each tree ring series and to characterize the disturbance history of the study site. The age structure of the forest was indicative of continuous establishment by Fagus crenata and simultaneous establishment by Magnolia obovata. A variety of low-intensity disturbances were identified in each decade, especially after the 1900s, but the occurrence of high-intensity catastrophic disturbance was rare, and likely played an important role in maintaining species diversity in the existing forest canopy. The results also suggest that F. crenata regenerates gradually before and after both large- and small-scale disturbances, whereas M. obovata and Betula grossa regenerate only after large-scale catastrophic disturbances.     

Conifer seedling root fungi and root dieback in Finnish nurseries

Fungi were isolated from the roots and growth substrate of bare‐rooted and containerized Pinus sylvestris and Picea abies nursery seedlings displaying a root dieback. Isolations were also made from visually healthy seedlings. The potential pathogenicity of all isolated species was determined in laboratory trials.

Cylindrocarpon spp., Fusarium spp. and Trichoderma viride were frequently isolated. The isolation frequency of a uninucleate Rhizoctonia‐like fungus, Pythium spp. and Phytophthora imdulatum from diseased containerized seedlings and their pathogenicity in tests suggest that these fungi are likely involved in the root dieback disease in containers. The pathogenic Rhizoctonia‐like fungus in addition to Pythium spp. was also isolated from bare‐rooted seedlings. In greenhouse tests Pythium spp. were more pathogenic to 4‐week‐old Scots pine seedlings grown before transplantation in unsterile substrate than to those seedlings grown axenically in agar. External factors are considered to have some role in the expression of disease.     

Establishment and growth of coniferous seedlings in an altered forest floor after long-term exclusion of deer

We evaluated differences between the forest floors and the establishment and growth of coniferous seedlings in fenced (13 years) and unfenced plots on Mt Ohdaigahara where the sika deer (Cervus nippon) population density is high. Large coniferous seedlings (height > 0.05 m) were less abundant in the unfenced plot, as a result of deer browsing. Small coniferous seedlings (height < 0.05 m), however, were more abundant in the unfenced plot, where most seedlings of Abies homolepis were found on bare ground and those of Picea jezoensis var. hondoensis were found on buttresses and fallen logs. The large area of bare ground in the unfenced plot was caused by deer browsing. Deer therefore have an indirect effect on the emergence and growth of small coniferous seedlings by modifying the forest floor.     

Armillaria ostoyae associated with dying 60‐year‐old Scots pines in northern Turkey

We investigated an Armillaria root disease centre in a 60‐year‐old, naturally regenerated Pinus sylvestris stand located at 1250 m altitude in Sinop province in northern Turkey. Based on sequence analysis of the internal transcribed spacer region of the rDNA, the mycelial fans sampled from the infected trees were Armillaria ostoyae. Multilocus genotyping indicated that the fans belonged to a single genet, at least 0.2 ha in size. Drought as a predisposing factor is discussed. The study area received 50–70 and 25–50% of the normal precipitation in May, June and August in 2009 and in August and September in 2010, respectively. Moreover, earlier excavation work to prepare a forest road had probably disrupted the water table in the affected stand.     

European pear rust on Juniperus excelsa L. in south‐western Turkey

In this study, the frequency of European pear rust (Gymnosporangium fuscum) lesions on Juniperus excelsa was investigated both on seedlings in a nursery and on forest trees in six juniper stands located in south‐western Turkey. In the nursery, lesions occurred on 0.7% of 5510 1‐year‐old seedlings and 29% of 200 2‐year‐old potted seedlings. In each of the juniper forests, 50 trees were chosen within a square of approximately 400 m² in size. In general, rust lesions were common on the trees. On average, 49.0% of the trees had lesions; on 17.3% of the trees, the lesions were located on the trunk, on 43.3% on the branches and on 11.7% on both. In lesion‐bearing trees, the average number of lesions was 1.47–3.59 and the maximum number of lesions per tree were six to 11. Most of the lesions (85.1%) were on branches. There were significant differences between the stands, which can be attributed to different age of the trees, site factors and the presence of alternate hosts. The rust identified from stem lesions in forest trees and from nursery seedlings was European pear rust (G. fuscum), but possible co‐occurrence of other Gymnosporangium spp. cannot be excluded.     

Effects of altitudinal gradient on species composition of naturally regenerated trees in <Emphasis Type="Italic">Larix kaempferi</Emphasis> plantations in central Japan

Species composition and community structure of naturally regenerated trees in Larix kaempferi plantations and natural forests were compared in relation to altitudinal gradient. Fifty-nine L. kaempferi plantations and 26 natural forest stands including old-growth and secondary forests were selected from 1300 to 2000 m a.s.l. in Mt Kushigata of central Japan. Vegetation plots (10 × 10 m) were established in each stand (85 plots in total). Species composition differed significantly between forest types, particularly on high altitude plots. Some species were significantly biased to natural forests, while no species were biased to plantations. Although some species that responded significantly to altitude were common to both forest types (e.g., Acer ukurunduese, Castanea crenata, and Fraxinus lanuginose f. serrata), some differed between forest types (e.g., Fagus japonica, Prunus maximowiczii, and P. nipponica). Thus, plantation management altered species responses to altitude. Altitude should be considered when planning and carrying out forest ecological restoration in plantations, because species composition of naturally regenerated trees has already been changed by past plantation management. In particular, attention should be paid to ecological restoration at high altitude.