Ophiostomatoid fungi associated with three spruce-infesting bark beetles in Slovenia

Context

Ophiostomatoid fungi can severely affect the health and economic value of Norway spruce trees (Picea abies). Although the diversity of ophiostomatoid species and their associations with insects have been well-investigated in central and northern Europe, little is known about the conditions in south-eastern Europe.

Aim

This study aims to study the assemblages of ophiostomatoid fungi associated with three bark beetle species (Ips typographus, Ips amitinus, and Pityogenes chalcographus) that infect Norway spruce in Slovenia.

Methods

Bark beetles were sampled in four phytogeographic regions in Slovenia. The fungi found on the bark beetles were identified based on morphology, DNA sequence comparisons of ITS regions and phylogenetic analysis. The species compositions of the fungal associates of the three insect species were compared and the pairwise associations of the occurrence of the fungal species were analysed.

Results

Thirteen different species were found. The most commonly encountered fungal associates of the beetles were Ophiostoma bicolor, Ophiostoma brunneo-ciliatum, Grosmannia piceiperda, Ophiostoma ainoae, Ceratocystiopsis minuta, and Grosmannia penicillata. The composition of the fungal associates differed among the bark beetle species, but not among the phytogeographic regions.

Conclusions

This study confirms that ophiostomatoid species are common associates of the investigated bark beetle species. Many ophiostomatoid species have strong host associations. I. typographus and P. chalcographus can act as effective vectors for O. bicolor, O. ainoae, G. piceiperda and O. brunneo-ciliatum, whereas I. amitinus often carries G. piceiperda and C. minuta in Slovenian forests.     

Occurrence of pathogens in associated living bark beetles (Col., Scolytidae) from different spruce stands in Austria

Pathogen occurrence was studied in 16?099 adult specimens of 10 different bark beetle species, which live associated on Norway spruce (Picea abies (L.) Karst.). Beetles (mainly Ips typographus L. and Pityogenes chalcographus L.) were collected from 6 different localities in Austria (4 secondary spruce stands and 2 natural forest type stands, 9 sampling plots in total) in elevations between 400?m and 1600?m. Various viral, protozoan, and fungal pathogens could be diagnosed with a light microscope in the examined beetles. Numerous pathogen species were known from former studies, some pathogens were totally new or could be found in a new host species beside their type host. The most dominant pathogen species were Protozoa, Gregarina cf. typographi, Malamoeba cf. scolyti, and Chytridiopsis cf. typographi. Over the whole investigation period, the highest pathogen diversity with eight pathogen species was found in I. typographus. Differences were observed in the pathogen complex of each beetle species from the different collection sites and in different years of investigation. Several species showed an overlapping in their host range and infected various bark beetle species. Furthermore, pathogen occurrence and prevalence differed in bark beetles from 4 different sampling plots in an area (one locality) within a distance of a few kilometres.     

Lipophilic extractives of the inner and outer barks from six different <Emphasis Type="Italic">Pinus</Emphasis> species grown in Indonesia

The chemical compositions of the dichloromethane extracts of inner and outer barks from six Pinus species (P. elliotii, P. oocarpa, P. caribeae, P. merkusii, P. montezumae, and P. insularis) grown in Indonesia were investigated by GC and GC–MS. Generally, the amounts of extractive contents were higher in the inner bark than in the outer bark except for P. merksuii. Fatty acids, monoterpenes, sesquiterpenes, resin acids, triterpenoids, and steroids were detected and quantified. Inner and outer barks differed not only in content of these compounds but also in their composition. Fatty acids and alcohols were the major classes of lipophilic compounds in the outer bark of P. caribeae, P. insularis, and P. montezumae. Steroids and triterpenoids were the dominant compounds identified in the inner bark of P. elliotii, P. insularis, and P. merkusii. Resin acids were the most abundant group in the inner bark of P. oocarpa whereas monoterpenes and sesquiterpenes were recorded in minor quantities in both bark layers of all species.     

Symptoms and fungi associated with elm trees decline in Iran

Elm (Ulmus, Ulmaceae) is one of the most popular and important trees in urban and landscape areas in Iran. A severe decline of elm trees has recently been observed in some areas of Kerman and Shiraz cities. However, the identity of the causal agents has not yet been investigated. The purpose of this study, carried out in the years 2012–2014, was to isolate and identify fungal trunk pathogens associated with decline symptoms observed in elm trees. Samples were collected from trunks and branches of trees showing various disease symptoms and internal wood lesions. Fungal isolations were made from discolored or decayed wood tissue. In Kerman some elm trees were attacked by the beetle Aeolesthes sarta; therefore, samples were also collected from larvae and adults of beetles associated with trees showing wood damage. Fungal isolates were identified by morphological, cultural and molecular characteristics. Thirteen fungal species, Phaeoacremonium (P.) minimum, P. parasiticum, P. sicilianum, P. alvesii, P. fraxinopennsylvanicum, Spencermartinsia viticola, Dothiorella (Do.) sarmentorum, Neoscytalidium hyalinum, Diatrype (Di.) whitmanensis, Cosmospora viridescens, Phoma (Pho.) herbarum, Phellinus (Phe.) tuberculosus and Inonotus levis were identified. Phaeoacremonium parasiticum was the only species isolated from both larvae and adult beetles. Pathogenicity trials were performed on detached shoots of elm under greenhouse conditions. Dothiorella sarmentorum was the most virulent species based on the length of wood necrosis. This study is the first report of P. sicilianum, C. viridescens and Phe. tuberculosus in Iran. In addition, most of the isolated species are reported for the first time on elm trees in the world.     

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.     

Regeneration patterns of the late-successional <Emphasis Type="Italic">Abies alba</Emphasis> Mill.: inhibition in monospecific stands and colonization in mixed stands

Key message

In Abies alba Mill. stands and mixed stands of A. alba and Picea abies L. (H. Karst), microsites neighbouring the trunks of adult trees were more conducive to A. alba regeneration. Although at the stand level, the effect of Fagus sylvatica L. was positive; the local effect of the adult F. sylvatica neighbourhood was insignificant. Hence, forming mixed stands with a fine-grained mosaic of admixed species might better facilitate natural regeneration of A. alba than monospecific stands.

Context

The establishment of natural regeneration in Abies alba Mill. stands is a slow, spatially heterogeneous and stochastic process. Recent studies based on inventory data indicate that A. alba more readily regenerates in mixed stands than in monospecific stands.

Aims

The objective was to examine how this positive association evidenced at the stand level operates on the scale of microsites with contrasting local species composition and stand density.

Methods

In 8 monospecific and 22 mixed stands with Fagus sylvatica L. or Picea abies L. (H. Karst), microsites with a contrasting density of A. alba seedlings were selected and compared in terms of local species composition, stand density, canopy characteristics and topsoil properties.

Results

In A. alba stands, seedling density was positively associated with the proximity of adult trees. In mixed stands of A. alba and P. abies, adult trees of both species exerted a positive effect on A. alba regeneration, but the P. abies neighbourhood influenced regeneration occurrence more strongly than the A. abies neighbourhood. In mixtures with F. sylvatica, however, the effect of local stand density and local species composition was not evidenced at all.

Conclusion

Although at the stand level, P. abies and F. sylvatica exert a positive effect on A. alba regeneration, on the microsite scale, their influences differ. In stands with a dominance of A. alba, the hampered seedling establishment in gaps may be considered an inhibitive effect that facilitates the emergence of other species.

     

Stimulation of tree defenses by Ophiostomatoid fungi can explain attack success of bark beetles on conifers

? Our aim is to present why the hypothesis, that Ophiostomatoid fungi play an important role in the establishment of most bark beetle species on living conifers, is valuable.

? After summarizing knowledge about the relationships of bark beetles with conifers and fungi, we conclude that controversy results from misinterpretations when using fungal pathogenicity to demonstrate the role of Ophiostomatoid fungi in beetle establishment on host trees.

? We demonstrate that fungal pathogenicity is not the right parameter to appreciate the role of fungus in beetle establishment on host trees. We argue that artificial low density inoculations that allow the appreciation of fungus ability to stimulate tree defenses and thus to help beetles in overcoming tree resistance must be used in complement to mass inoculations. In both cases, results must be expressed in terms of tree defense stimulation rather than in terms of tree killing.

1. Fungal species stimulating tree defenses are generally not those that grow the best in the sapwood.
2. We argue that beetle development in the phloem, fungal invasion of the sapwood and phloem, and tree death, occur after tree defenses are exhausted, and that any fungus present in the beetle gallery could thus potentially invade the sapwood after defense exhaustion.

? We conclude that stimulation of the tree defense reactions in both the phloem and the superficial sapwood is a real benefit brought by fungi to the beetles during the first phase of establishment (overcoming tree resistance).

? Considering the origin of the bark beetle fungus associations attacking living trees and their general functioning based on stimulation of tree defenses, we develop three hypotheses:

1. any beetle species would be helped in its establishment in a given tree species by developing an association, even loosely, with a fungus species belonging to the Ophiostomatoid flora of that tree species;
2. the necessity of a considerably low level of tree resistance for fungus extension into the tree is the selection pressure that has led fungi to develop their intrinsic ability to stimulate tree defenses, through their ability to grow into the phloem. This association can be completed by antagonistic fungal species controlling extension of the previous fungal species in the tree tissues;
3. Beetle species using the strategy of overcoming tree resistance are associated with a fungal complex, of which species could assume three roles regarding relationships between beetles and trees: 1- to stimulate tree defenses in the phloem and superficial sapwood, 2- to grow into the sapwood after tree resistance is overcome, and 3- to control phloem extension of the first other two categories. Bringing nutrients to the beetle progeny can be a fourth role.

? We propose that bark beetle — Ophiostomatoid associations can be categorized, based on associations’ frequency and complexity while taking into account beetle aggressiveness. We show that a close correspondence exists between beetles’ aggressiveness and the ability of their main associated fungal species to stimulate the defenses of their host tree.

? We conclude with suggesting that most sapwood invading fungi might be “cheaters” which have taken advantage of the efficiency of the relationship between beetles and fungi that stimulate tree defenses.

     

Modelling the predisposition of Norway spruce to <Emphasis Type="Italic">Ips typographus</Emphasis> L. infestation by means of environmental factors in southern Finland

New measures for effective monitoring and controlling of bark beetle infestations are needed as a response to intensified outbreaks caused by the climate change. Various environmental factors affect tree health and susceptibility, as well as stand predisposition to bark beetles. European spruce bark beetle Ips typographus L. abundance and outbreak frequency in Finland has significantly increased during the last decade. The ability to identify sites under a high risk of infestation would facilitate adaptation to this new situation and help target limited forest health management resources. Accordingly, our goal was to investigate the importance of various stand, soil and topographic characteristics in the assessing predisposition of Norway spruce dominated urban forest in southern Finland to I. typographus infestations. Information on the environmental factors was assessed in the field in 2014 and derived from a digital elevation model. Ips typographus infestation intensity was classified into three infestation index classes based on tree-wise symptoms of resin flow, discoloration and defoliation. Cumulative logit link models were utilized for investigating stand-level infestation probability. The best explanatory factors were aspect, slope, site type and soil texture. Models with the highest cumulative probabilities for severe infestation were linked with eastern aspect, moderate steep slope and rich site type fertility (0.72) and eastern aspect, shallow soil and rich site type fertility (0.71). Higher soil C/N ratios with east aspect and rich site type fertility was associated with an increased risk of severe infestation in a third model. The lowest risk was associated with southern and southwestern aspects, fine soil texture, moderate site fertility and gentle slopes.     

Patterns of within-stem variations in wood specific gravity and water content for five temperate tree species

? Key message

Intensive measurements of basic specific gravity and relative water content of lumens show that within-stem variations strongly depend on species and cannot be summarised through the typical patterns reported in the literature; breast height measurements are not always representative of the whole stem.

? Context

Knowledge of the distribution of wood properties within the tree is essential for understanding tree physiology as well as for biomass estimations and for assessing the quality of wood products.

? Aims

The radial and vertical variations of basic specific gravity (BSG) and relative water content of lumens (RWC _L ) were studied for five species: Quercus petraea/robur, Fagus sylvatica, Acer pseudoplatanus, Abies alba and Pseudotsuga menziesii. The observations were compared with typical patterns of variations reported in the literature.

? Methods

Wood discs were sampled regularly along tree stems and X-rayed in their fresh and oven-dry states.

? Results

At breast height, BSG was found to clearly increase radially (pith to bark) for two species and to decrease for one species. For F. sylvatica and A. alba, the radial variations of BSG were rather U-shaped, with in particular inner wood areas showing respectively lower and higher BSG than the corresponding mature wood. RWC _L increased generally from inner to outer area but wet sapwood was clearly distinguishable only for the coniferous species. Vertical variations of BSG and RWC _L were strongly dependant on the species with usually non-linear patterns.

? Conclusion

The observed variations of BSG were only partially in agreement with the reported typical radial patterns. Despite the vertical variations, the mean BSG of a cross-section at breast height appeared to be a good estimator of the mean BSG of the whole stem (although the difference was statistically significant for coniferous species), whereas breast height measurement of RWC _L was not representative of the whole stem.

     

Factors affecting field performance of pheromone traps for tobacco beetle, <Emphasis Type="Italic">Lasioderma serricorne</Emphasis>, and tobacco moth, <Emphasis Type="Italic">Ephestia elutella</Emphasis>

Tobacco beetle, Lasioderma serricorne (F.) (Coleoptera: Anobiidae), is one of the most serious insect pests of stored tobacco, and traps baited with the female-produced sex pheromone, serricornin, are used for monitoring the pest. In two trapping experiments carried out in tobacco warehouses in Greece, two commercially available trap and lure systems for L. serricorne were found to be equally effective in terms of the numbers of beetles trapped. In contrast to previous reports, anhydroserricornin was unattractive and lures containing serricornin and anhydroserricornin were less attractive than lures containing serricornin only. The sex pheromone of the other main insect pest of tobacco, Ephestia elutella (Hübner) (Lepidoptera: Pyralidae), could be added to the lures without affecting the attractiveness of either pheromone to their respective species. Lures remained attractive for at least 4 weeks under field conditions, and, in laboratory tests, release of pheromone could still be detected after 30 days at 27 °C. The stereoisomeric composition of the serricornin in the two commercial lures was similar with high proportions of the attractive (4S,6S,7S)-isomer. The proportion of the (4S,6S,7R)-isomer was low, and this is known to reduce the attractiveness.     

Diversity of ophiostomatoid fungi associated with the large pine weevil, Hylobius abietis, and infested Scots pine seedlings in Poland

Context

Bark beetles are known to be associated with fungi, especially the ophiostomatoid fungi. However, very little is known about role of pine weevils, e.g., Hylobius abietis, as a vector of these fungi in Europe.

Aims

The aims of our study were to demonstrate the effectiveness of H. abietis as a vector of ophiostomatoid fungi in Poland and to identify these fungi in Scots pine seedlings damaged by weevil maturation feeding.

Methods

Insects and damaged Scots pine seedlings were collected from 21 reforestation sites in Poland. The fungi were identified based on morphology, DNA sequence comparisons for two gene regions (ITS, β-tubulin) and phylogenetic analyses.

Results

Sixteen of the ophiostomatoid species were isolated and identified. In all insect populations, Leptographium procerum was the most commonly isolated fungus (84 %). Ophiostoma quercus was also found at a relatively high frequency (16 %). Other ophiostomatoid fungi were found only rarely. Among these rarer fungi, four species, Leptographium lundbergii, Ophiostoma floccosum, Ophiostoma piliferum and Sporothrix inflata, were isolated above 3 %. L. procerum was isolated most frequently and was found in 88 % of the damaged seedlings. S. inflata was isolated from 26 %, while O. quercus occurred in 10 % of the seedlings.

Conclusion

This study confirmed that L. procerum and O. quercus were common associates of H. abietis, while others species were found inconsistently and in low numbers, indicating causal associations. H. abietis also acted as an effective vector transmitting ophiostomatoid species, especially L. procerum and S. inflata, to Scots pine seedlings.     

Ophiostomatoid fungi isolated from three different pine species in Argentinian Patagonia

Pine plantations in Argentinian Patagonia cover ca. 95,000 ha in Chubut, Río Negro and Neuquén provinces. Exotic bark beetles (Orthotomicus laricis, Hylastes ater and Hylurgus ligniperda) commonly occur in freshly cut logs, stumps and slash. These beetles are vectors of “ophiostomatoid” fungi which include primary tree pathogens as well as important agents of blue stain. The aim of this study was to identify these beetle‐associated fungi. Sawing mills and pine plantations were surveyed three consecutive years. Fungal isolates from stained logs, processed wood and insect galleries were identified based on morphological and DNA sequence comparisons of ITS and β‐tubulin gene regions. Two Grosmannia, one Graphilbum and three Ophiostoma species were identified. Ophiostoma piliferum and O. peregrinum sp. nov. were the most frequently isolated taxa. O. peregrinum occurred in all provinces, colonizing different conifer species and, interestingly, also the native broadleaved species Nothofagus dombeyi. Pine plantation forestry in southern South America includes Argentina, Brazil, Chile, Paraguay and Uruguay. Emerging data from Argentina, Chile and Uruguay revealed some coincidences between these countries, but also several differences, probably, as a result of multiple introduction events.     

Ecological implications of mammal feces buried in snow through dung beetle activities

In early spring, snow-buried mammal feces simultaneously emerge on the ground with the melting of the snow across regions with heavy snowfall. Here, we evaluated the ecological implications of this phenomenon for the cool-temperate forest ecosystem in terms of the resource use of dung beetles (Coleoptera: Scarabaeidae), which play key roles in secondary seed dispersal and nutrient cycling. During May 2012 and 2013, we conducted cafeteria experiments in heavy snowfall regions with different types of forest cover in northern Japan by using pitfall traps baited with the snow-buried feces of Japanese macaques (Macaca fuscata), Japanese serows (Capricornis crispus), and Japanese hares (Lepus brachyurus angustidens). From the experiments, we identified 12 dung beetle species, indicating that the snow-buried feces could act as a valuable resource for vernal beetles. Most of the beetles had obvious fecal preferences. The snow-buried feces of serows were widely available resources for most vernal species, including dwellers and tunnelers, contributing to the biomass of those species. Although dung beetles using snow-buried feces in young broadleaf forests with rich feces supplies did not always exhibit the highest species richness, tunnelers frequently emerged in those forests, and Phelotrupes preferred macaque feces. This finding could have important implications for plant regeneration, as it relates to time-lagged mammal–beetle interactions. Endozoochorous seeds dispersed by macaques in the autumn have insufficient opportunities for secondary seed dispersal by beetles as the beetles are inactive in autumn, but those seeds are protected against predation by snow during winter and are safely preserved under soil by vernal tunnelers.     

Effects of <Emphasis Type="Italic">Verticillium nonalfalfae</Emphasis> on <Emphasis Type="Italic">Ailanthus altissima</Emphasis> and associated indigenous and invasive tree species in eastern Austria

The naturally occurring Verticillium nonalfalfae shows promise for biocontrol of the highly invasive Tree of Heaven (Ailanthus altissima), but might also bear a risk for non-target tree species. In this study, we conducted inoculations on potted seedlings of A. altissima as well as on eight indigenous and two invasive tree species associated with Tree of Heaven in Austria. Although vascular discolourations developed in all inoculated tree species, V. nonalfalfae was reisolated from Ailanthus and eight of the ten non-target-species, whereas typical disease symptoms and mortality only occurred on A. altissima. Results confirmed high susceptibility (S) of A. altissima to V. nonalfalfae but indicated tolerance (T) of Acer campestre, Acer pseudoplatanus and Quercus robur, possible resistance (PR) of Fraxinus excelsior, Populus nigra, Tilia cordata, Ulmus laevis and Ulmus minor and resistance (R) of Fraxinus pennsylvanica and Robinia pseudoacacia to this potential biocontrol agent. Results from seedling inoculations were confirmed by cursory field observations in Ailanthus-inoculated forest stands, where admixed A. campestre, A. pseudoplatanus, F. excelsior, Populus alba, R. pseudoacacia and U. laevis canopy trees remained asymptomatic, while mortality was induced in Ailanthus.     

Hymenopterous parasitoids of the cherry bark tortrix, <Emphasis Type="Italic">Enarmonia formosana</Emphasis> (Scopoli) in central-east Europe (Hymenoptera,Ichneumonoidea; Lepidoptera,Tortricidae)

Strategies targeting the biological control of the cherry bark tortrix (=CBT), Enarmonia formosana (Scopoli) in North America have also included the search for potentially useful native hymenopterous parasitoids of the pest in Europe. The search for such agents undertaken in the central-east Europe (Czech Republic, Slovak Republic, Hungary) resulted in determination of several ichneumonid species (Campoplex cf. dubitator Horst., Liotryphon sp., Lissonotus sp., Pimpla spuria (Grav.), Pimpla turionellae (L.), and some others, and a braconid (Apanteles britannicus Wilk.) in 2001–2002.     

<Emphasis Type="Italic">Ophiostoma</Emphasis> species associated with bark beetles infesting three <Emphasis Type="Italic">Abies</Emphasis> species in Nikko,Japan

Ophiostoma species were isolated from bark beetles and Abies mariesii, A. veitchii and A. homolepis attacked by the beetles in Nikko, Tochigi, central Honshu, Japan. One to two Ophiostoma species were frequently isolated from each species of bark beetle. Ophiostoma subalpinum was the most common associate of Cryphalus montanus. Ophiostoma sp. B as well as O. subalpinum was a common fungus associated with Polygraphus proximus. Ophiostoma europhioides was isolated from Dryocoetes hectographus and D. autographus as one of the common associates. Ophiostoma sp. J and Ophiostoma sp. S were frequently isolated from D. autographus and D. striatus, respectively. These fungi seem to have specific relationships with particular bark beetles. Ophiostoma sp. B, Ophiostoma sp. J and Ophiostoma sp. S have unique morphological characteristics and appear to be new species. Five trees of A. veitchii, approximately 43 years old, were inoculated with five Ophiostoma species to assess the relative virulence of the fungi. Ophiostoma subalpinum, Ophiostoma sp. B, and O. europhioides had relatively higher virulence than the other species studied.     

Diversity and distribution of soil micro-fungi along an elevation gradient on the north slope of Changbai Mountain

Elevational changes in patterns of diversity are important to understanding of the influence of global changes, yet few studies have addressed the distribution of microorganisms, e.g. soil micro-fungi. We studied the diversity of the forest soil micro-fungi in four vegetation belts along an elevation gradient on the north slope of Changbai Mountain in Changbai National Nature Reserve.The four belts were characterized as coniferous–deciduous mixed forest, coniferous forest, Erman's birch forest, and alpine tundra. We estimated the quantity and distribution of the fungal species in each belt and calculated three indices,viz. Shannon–Wiener diversity(H'), Pielou's evenness(J'),and Margalef's abundance(E), to depict fungal species diversity. A total of 932 strains were recorded and identified, representing 53 genera, and 108 species. Among these, Penicillium, Aspergillus, Trichoderma, Mucor, Rhizopus and Fusarium were the dominant genera. With increasing elevation, the quantity of fungi and values of H',E, and J' gradually declined.     

Prior contact of <Emphasis Type="Italic">Pinus tabulaeformis</Emphasis> with ectomycorrhizal fungi increases plant growth and survival from damping-off

Pinus tabulaeformis (Chinese pine) is a widely planted conifer species in northern China and is used for soil and water conservation on the Loess Plateau. Due to its strong reliance on ectomycorrhizae and low survival rate from damping-off during seedling cultivation and in nurseries, we explored the early influence of three ectomycorrhizal fungi (Handkea utriformis, Suillus lactifluus, and Suillus tomentosus) on the growth, root morphological characters, root vitality, and survival of P. tabulaeformis seedlings from subsequent damping-off in a pot experiment. In addition, the in vitro suppression of three ectomycorrhizal fungi on the damping-off pathogen (Fusarium solani) was evaluated and observed using a scanning electron microscope. We found that all three ectomycorrhizal fungi could colonize more than 40% of the roots of P. tabulaeformis, promote plant shoot and root growth, increase the proportion of large-diameter roots, improve root vitality, and increase survival rates 3 months after inoculation. All three ectomycorrhizal fungi suppressed the growth of F. solani to a different extent in vitro. Ectomycorrhizal fungal hyphae proliferated and wrapped around the hyphae of F. solani when the hyphae encountered each other. The colonization rate of P. tabulaeformis roots was highly correlated with root vitality and root growth parameters, while the survival rate of P. tabulaeformis seedlings was highly correlated with the colonization rate of P. tabulaeformis roots, root vitality and growth. Our results suggest that early contact between P. tabulaeformis and ectomycorrhizal fungi could improve plant growth and resistance against damping-off.     

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.     

Selection of arbuscular mycorrhizal fungal strains to improve <Emphasis Type="Italic">Casuarina equisetifolia</Emphasis> L. and <Emphasis Type="Italic">Casuarina glauca</Emphasis> Sieb. tolerance to salinity

Key message

Selection of the best salt-tolerant combination of Casuarina sp. and arbuscular mycorrhizal fungi (AMF) is one of the key criteria for successful setup of saline land rehabilitation program.

Context

Land salinization is a serious problem worldwide that mainly leads to soil degradation and reduces crop productivity. These degraded areas could be rehabilitated by planting salt-tolerant species like Casuarina glauca Sieb. and Casuarina equisetifolia L. These are pioneer plants, able to form symbiotic associations with arbuscular mycorrhizal fungi (AMF), ectomycorrhizal fungi (EMF), and nitrogen-fixing bacteria.

Aims

The aim of this study was to select the highest salt-tolerant combination of Casuarina/AMF that can be used for the rehabilitation of lands degraded by salinity.

Methods

C. equisetifolia and C. glauca were grown in sandy sterile soil in the greenhouse and inoculated separately with Rhizophagus fasciculatus (Thaxt.) C. Walker & A. Schüßler, Rhizophagus aggregatus (N.C. Schenck & G.S. Sm.) C. Walker, and Rhizophagus intraradices (N.C. Schenck & G.S. Sm.) C. Walker & A. Schüßler. After confirming the establishment of a symbiosis, the plants were watered with gradually increasing concentrations of saline solution. After harvest, size and biomass of the seedlings, root colonization by AMF, and AMF metabolic activities were evaluated.

Results

A larger growth was obtained in the two species when the individuals were inoculated with R. fasciculatus. Root colonization rates did not differ among fungal species, but fungal metabolic activities were higher in mycorrhizal roots of C. glauca plants inoculated with R. fasciculatus.

Conclusion

Among the three mycorrhizal fungi, R. fasciculatus was more efficient in association with Casuarinaceae species under salt stress. Our results suggest that selection of appropriate fungal strains is crucial to improve plant performance in saline soils.