The influence of temperature on the susceptibility of vine weevil, Otiorhynchus sulcatus (Fabricius) (Coleoptera: Curculionidae), larvae to Metarhizium anisopliae (Deuteromycotina: Hyphomycetes)

Mortality of Otiorhynchus sulcatus (Fabricius) larvae at 10^oC, 15^oC, 20^oC and 25^oC following treatment with 10⁷conidia ml“¹suspensions of six Metarhizium anisopliae (Metschnikoff) Sorokin isolates was temperature dependent. In all cases, the LT₅₀s were inversely related to temperature, but the nature of this response varied between isolates. Strain 101-82 was the most virulent isolate at 25^oC with an LT₅₀ of 3.7 days, but it was the least virulent isolate at 15^oC and it failed to kill any O. sulcatus larvae at 10^oC. In contrast, strain 159-83 had the lowest virulence at both 20^oC and 25^oC, whereas it was the most virulent isolate at 10^oC with an LT₅₀ of 20.0 days. The mortality rates followed a similar pattern and were positively related to temperature in all cases with the exception of strain 159-83 at 25^oC. Mycosis development was examined on larvae treated with strain 275-86 and significant differences were obtained between all four temperatures. Sporulation commenced after approximately 2.75 days at 25^oC, but took nearly 11 days at 10^oC. The infection rates also varied between temperatures; sporulation occurred on 98% of the treated larvae at 25^oC, but only on 93%, 87% and 49% of the larvae at 20^oC, 15^oC and 10^oC, respectively. The results of these bioassays demonstrate that temperature has a significant effect on the virulence of M. anisopliae. The differences between fungal strains also emphasises the importance of selecting isolates for specific situations on the basis of their temperature profiles.     

The effects of cadmium and the cadmium-zinc interaction on the axenic growth of ectomycorrhizal fungi

Eleven strains of ectomycorrhizal fungi belonging to seven species have been cultured on a cadmium-contaminated growth medium in order to determine their in vitro cadmium tolerance. Four strains were collected from a zinc and cadmium-polluted soil. Radial growth rate was a sensitive parameter to detect Cd toxicity. A wide differential response to Cd was obtained between the individual species. A clear relation between Cd tolerance and site origin of the isolates did not exist, although such a relationship was found when strains are compared within one species. Cd-sensitive and Cd-tolerant strains of Suillus bovinus were studied in more detail. Two isolates were grown on media with combinations of two non-toxic zinc concentrations and three cadmium levels. Adding a higher Zn concentration to the medium resulted in a reduction of the toxic effect of Cd. This antagonistic effect also resulted in a lowered Cd concentration in the mycelium.     

The entomopathogenic nematode Steinernema kraussei and Metarhizium anisopliae work synergistically in controlling overwintering larvae of the black vine weevil,Otiorhynchus sulcatus,in strawberry growbags

Previously, the combination of reduced rate of entomopathogenic nematodes (EPN) and fungus caused additive or synergistic mortality to third-instar black vine weevil (BVW), Otiorhynchus sulcatus. In this study, we examined this interaction in unheated glasshouses during winter and compared a combination of commercial formulation of a cold-tolerant EPN, S. kraussei (Nemasys L?) and fungus Metarhizium anisopliae strain V275 against overwintering third-instar BVW. The combination of M. anisopliae with S. kraussei at a rate of 1×10¹⁰ conidia+250,000 nematodes/growbag resulted in additive or synergistic effects, providing 100% control of overwintering larvae.     

Effect of ectomycorrhizal fungi on nitrogen mineralisation and the growth of Scots pine seedlings in natural peat

The aim of this study was to investigate the potential of different isolates of ectomycorrhizal (EM) fungi to enhance the growth of Pinus sylvestris seedlings in five natural peat substrates with different nitrogen concentrations, and the effect of the Scots pine seedlings and fungal inoculum on the formation of dissolved inorganic and organic nitrogen in peat. Utilization of different organic nitrogen compounds by microbial community in the peat was also investigated using Biolog MT MicroPlates. Inoculation of the seedlings with EM fungi enhanced seedling growth. Piloderma croceum increased root growth especially, whereas Lactarius rufus increased needle growth and Suillus variegatus I, II and III improved both root and needle growth. All the EM fungi also significantly affected stem growth. Nitrogen concentration of the peat did not affect seedling growth as much as the EM fungi. At the lowest peat N concentration (1.17%) NH ₄ ⁺ mineralisation was lower and DON (dissolved organic nitrogen) accumulation higher than at higher peat N concentrations. The EM fungal isolates had different effects on NH ₄ ⁺ and DON accumulation/degradation in peat. The EM fungal isolates significantly increased NH ₄ ⁺ formation in the peat, whereas L. rufus and P. croceum had an opposite effect on DON accumulation. S. variegatus I significantly decreased the DON concentrations during peat incubation. The N concentration of the peat slightly affected the utilization of amino acids and polyamines by the microbial community, whereas inoculation with S. variegatus I, II or III had no effect.     

Distribution of insect pathogenic soil fungi in Switzerland with special reference to Beauveria brongniartii and Metharhizium anisopliae

Between 1998 and 2000 soil samples from 82fields in north, east, central and south westof Switzerland were analysed for presence ofinsect pathogenic soil fungi using a selectivemedium and the Galleria bait method. Thedetection rates with either method were verysimilar. 96% of the fields distributed overall examined regions contained Metarhiziumanisopliae. The presence of Beauveriabrongniartii was limited to sites colonised byits host, Melolontha melolontha. It wasalso present at a site where M.melolontha disappeared about 40 years ago. Onthe other hand, B. brongniartii was notfound in four fields containing M.melolontha populations. Beauveriabassiana, Paecilomyces fumosoroseus and Conidiobolus sp. were other entomopathogenicfungi isolated during this survey. Differencesin the presence of M. anisopliae betweenarable fields and adjacent meadows as well asbetween orchards and meadows are discussed.     

Effect of brassicaceous plants on the survival and infectivity of insect pathogenic fungi

Studies were conducted to investigate potentialinteractions between brassicaceous plants, theisothiocyanates they produce and insectpathogenic fungi. Studies in vitro showedthat 100 ppm of 2-phenylethyl isothiocyanatecompletely inhibited growth of Metarhiziumanisopliae and Tolypocladiumcylindrosporum. T. cylindrosporum wassignificantly inhibited by 10 ppm 2-phenylethylisothiocyanate, but not by 1 ppm. M.anisopliae was not inhibited by 10 or 1 ppm2-phenylethyl isothiocyanate. The effect offreshly grated rutabaga on virulence of M.anisopliae to Galleria mellonella larvaewas also tested, resulting in reduced infectionby M. anisopliae. To study the effect ofplants in the Brassicaceae under morefield-like conditions fungi were added to soilwithout plants or at the bases of two differentplant species, Barbarea vulgaris and Eruca vesicaria sativa. Soil was sampled at 5,10 and 15 days to determine the concentrationof colony forming units (CFUs). Treatments withplants did not have fewer CFUs than thecontrol. Instead significantly fewer CFUsresulted when M. anisopliae was added topots with soil only than to pots with plants.There were no significant differences betweenthe two plant species in the number of CFUs atday 5. For days 10 and 15, however,significantly more M. anisopliae CFUswere found in pots containing E. vesicariasativa than B. vulgaris. T.cylindrosporum was less affected by thedifferent treatments than M. anisopliae.Our studies demonstrated that whileisothiocyanates can inhibit insect pathogenicfungi in Petri dishes, when using a morerealistic fungus/plant/soil microcosm no fungalinhibition was found.     

外生菌根菌对油松幼苗抗氧化酶活性及根系构型的影响

研究盆栽条件下,接种褐环乳牛肝菌(Suillus luteus)和红汁乳菇(Lactarius hatsudake)对油松(Pinus tabulaeformis)苗木生长,抗氧化酶活性和根系构型的影响。结果表明:(1)褐环乳牛肝菌和红汁乳菇均可与油松合成外生菌根,并显著提高苗木的苗高、地径、鲜重和干重。(2)接种褐环乳牛肝菌苗木针叶抗氧化酶(SOD、CAT、POD)活性分别比接种红汁乳菇苗木高14.77%、20.77%、34.68%;接种褐环乳牛肝菌苗木根系抗氧化酶(SOD、CAT、POD)活性分别比接种红汁乳菇苗木高8.54%,4.34%,33.31%;接种处理苗木抗氧化酶活性均显著高于不接种处理。接种处理苗木根长、表面积、体积、平均直径、根尖数、分叉数显著高于不接种处理;接种处理间苗木只有根尖数存在显著差异;接种褐环乳牛肝菌的油松根系分支成80°—90°的一级侧根数占23.81%,显著多于接种红汁乳菇和对照苗木。研究表明,接种褐环乳牛肝菌和红汁乳菇均能促进油松苗木生长,提高油松体内抗氧化酶活性,扩大苗木根系的吸收范围,其中褐环乳牛肝菌各方面的促进效果要优于红汁乳菇。     

Laboratory investigations on the potential of entomopathogenic fungi for biocontrol of Helicoverpa armigera (Lepidoptera: Noctuidae) larvae and pupae

The susceptibility of third instar Helicoverpa armigera to seven strains of three entomopathogenic fungal species, i.e. Metarhizium anisopliae, Beauveria bassiana and Paecilomyces fumosoroseus, was tested under laboratory conditions using the larval immersion method. High efficacies ranging from 68 to 100% corrected mortality were recorded with more profound effects in treatments with B. bassiana and P. fumosoroseus strains. The median lethal concentration (LC₅₀) for L3 was 6.0×10⁵ in M. anisopliae 79, 1.5×10⁵ in B. bassiana 124 and 4.2×10⁴ in P. fumosoroseus 14. These three strains were further used to characterize the age-dependent mortality of different larval stages (L2-L5) and the effect against pupae of H. armigera. Larval stages did not differ in their mortality but differed i in median lethal time, with shorter values recorded in the second instar. Tested fungi also caused a high reduction between 74.4 and 100% in the emergence of pupae using the soil inoculation method and the pupal immersion technique. All three fungal species, especially P. fumosoroseus, have a high potential for biocontrol of H. armigera larvae and also as a soil treatment targeting the pupae.     

Survival of inoculated Laccaria bicolor in competition with native ectomycorrhizal fungi and effects on the growth of outplanted Douglasfir seedlings

The survival, development and mycorrhizal efficiency of a selected strain of Laccaria bicolor along with naturally occurring ectomycorrhizal fungi in a young plantation of Douglas fir was examined. Symbionts were identified and their respective colonization abilities were determined. Eight species of symbiotic fungi, which may have originated in adjacent coniferous forests, were observed on the root systems. Mycorrhizal diversity differed between inoculated (5 taxa) and control (8 taxa) seedlings. Ectomycorrhizal fungi which occurred naturally in the nursery on control seedlings (Thelephora terrestris and Suillus sp.) did not survive after outplanting. Both inoculated and naturally occurring Laccaria species, as well as Cenococcum geophilum, survived on the old roots and colonized the newly formed roots, limiting the colonization by other naturally occurring fungi. Other fungi, such as Paxillus involutus, Scleroderma citrinum and Hebeloma sp. preferentially colonized the old roots near the seedling's collar. Russulaceae were found mainly in the middle section of the root system. Mycorrhizal colonization by Laccaria species on inoculated seedlings (54%) was significantly greater than on controls (13%) which were consequently dominated by the native fungi. Significant differences (up to 239%) were found in the growth of inoculated seedlings, especially in root and shoot weight, which developed mainly during the second year after outplanting. Seedling growth varied with the species of mycorrhizae and with the degree of root colonization. Competitiveness and effectiveness of the introduced strain on improving growth performances of seedlings are discussed.     

Synergistic effect of soil pathogenic fungi and nematodes reducing bioprotection of Arbuscular mycorrhizal fungi on the grass Leymus arenarius

We examined the role ofarbuscular mycorrhizal fungi (AMF) in thebioprotection of the sand dune grass Leymus arenarius against soil fungi andnematodes. Six soil fungi (Fusariumnivale, Fusarium sp., Cladosporiumherbarum, Cladosporium sp., Phomasp., Sporothrix sp.) and four species ofnematodes (Pratylenchoidesmagnicauda, Paratylenchusmicrodorus, Rotylenchus goodeyi, Merlinius joctus) were isolated from a coastalsand dune in Iceland where a population of L. arenarius was declining in vigour. Acommercial AMF inoculum (Microbio Ltd. England)containing Glomus caledonium, G.fasciculatum, and G. mossae was used.Seedlings of L. arenarius were grownunder controlled conditions in sterile sand andsubjected to the following treatments: (1)control, (2) + AMF, (3) + AMF + soil fungi, (4)+ AMF + nematodes, (5) + AMF + nematodes + soilfungi, (6) + soil fungi, (7) + soil fungi +nematodes, (8) + nematodes. Mycorrhizal plantshad significantly the highest root dry weightof all treatments. Mycorrhizal plants hadsignificantly higher leaf dry weight thanplants in other treatments, with the exceptionof AMF inoculated plants exposed to nematodes. Mycorrhizal plants exposed to soil fungi andnematodes had significantly higher growthparameters except total number of leaves thanAMF inoculated plants exposed to both soilfungi and nematodes. Mycorrhizal plantssubjected to a dual application of soil fungiand nematodes did not vary significantly in anygrowth parameters from plants without AMF thatwere exposed to a dual application of soilfungi and nematodes. This suggests asynergistic effect of soil fungi and nematodesthat break down the protection of AMF againstpathogens. The results are discussed inrelation to plant dynamics of sand duneecosystems.     

Spatial structure and the effects of host and soil environments on communities of ectomycorrhizal fungi in wooded savannas and rain forests of Continental Africa and Madagascar

Mycorrhizal fungi play a key role in mineral nutrition of terrestrial plants, but the factors affecting natural distribution, diversity and community composition of particularly tropical fungi remain poorly understood. This study addresses shifts in community structure and species frequency of ectomycorrhizal (EcM) fungi in relation to host taxa, soil depth and spatial structure in four contrasting African ecosystems. We used the rDNA and plastid trnL intron sequence analysis for identification of fungi and host plants, respectively. By partitioning out spatial autocorrelation in plant and fungal distribution, we suggest that African EcM fungal communities are little structured by soil horizon and host at the plant species and family levels. These findings contrast with patterns of vegetation in these forests and EcM fungal communities in other tropical and temperate ecosystems. The low level of host preference indirectly supports an earlier hypothesis that pioneer Phyllanthaceae may facilitate the establishment of late successional Fabaceae and potentially other EcM host trees by providing compatible fungal inoculum in deforested and naturally disturbed ecosystems of tropical Africa.     

The influence of vegetation type,soil properties and precipitation on the composition of soil mite and microbial communities at the landscape scale

Aim We used a landscape‐scale study of birch invasion onto heather moorland to determine the consistency of changes in vegetation type and soil properties and in the community composition of five soil organism groups. Our aim was to determine whether the degree to which soil organisms respond to natural changes and/or induced changes (e.g. changes in land‐use type and climate) in habitat is consistent across trophic and taxonomic groups in the context of conservation policies for birch woodland and heather moorland. Location Mainland Scotland. Methods We sampled mesostigmatid mites, oribatid mites, fungi, bacteria and archaea in adjacent patches of birch woodland (dominated by Betula pubescens) and heather moorland (dominated by Calluna vulgaris) at 12 sites for which annual rainfall ranged between 713 and 2251 mm. Differences in community composition were visualized using non‐metric multidimensional scaling based on Bray–Curtis dissimilarities. The factors contributing to differences between habitats within sites were explored using general linear models and those among sites using redundancy analysis. Results The communities of all groups differed between habitats within sites, but only the oribatid mites and fungi differed consistently between habitats across sites. Within sites, dissimilarity in fungal communities was positively related to the difference in C. vulgaris cover between habitats, whereas dissimilarities in bacteria and archaea were positively related to differences in soil pH and C:N ratio between habitats, respectively. Main conclusions The influence of vegetation type and soil properties differed between groups of soil organisms, albeit in a predictable manner, across the 12 sites. Organisms directly associated with plants (fungi), and organisms with microhabitat and resource preferences (Oribatida) were strongly responsive to changes in habitat type. The response of organisms not directly associated with plants (bacteria, archaea) depended on differences in soil properties, while organisms with less clear microhabitat and resource preferences (Mesostigmata) were not strongly responsive to either vegetation type or soil properties. These results show that it is possible to predict the impact of habitat change on specific soil organisms depending on their ecology. Moreover, the community composition of all groups was related to variation in precipitation within the study area, which shows that external factors, such as those caused by climate change, can have a direct effect on belowground communities.     

The effects of toxic metals,content of nutrients and inoculation with mycorrhizal fungi on the level of phenolics in roots and growth of Scots pine seedlings

The effects of two substrates and several strains of mycorrhizal fungi on the content of soluble phenolics in roots and growth of Scots pine seedlings was investigated. The first substrate was fertile and contaminated with copper, zinc and lead, whereas the second one displayed nutrient deficiency. The dry weights of needles, trunks, roots and the total biomass were higher in groups of seedlings inoculated with mycorrhizal fungi on fertile and polluted substrate. Inoculation of pine seedlings on this substrate resulted in a decrease in concentration of phenolics in roots and except for seedlings inoculated with Laccaria laccata negatively influenced the above-ground part: root ratio.     

Effects of isolates of ectomycorrhizal fungi and endophytic Mycelium radicis atrovirens that were dominant in soil from disturbed sites on growth of Betula platyphylla var. japonica seedlings

Effects of ectomycorrhizal fungi and endophytic Mycelium radicis atrovirens Melin (MRA) on growth of Betula platyphylla var. japonica seedlings were investigated under aseptic culture conditions. Three isolates of ectomycorrhizal fungi and two isolates of MRA were used. One MRA isolate was Phialocephala fortinii. Previous field work revealed that these isolates were dominant on the roots of B. platyphylla var. japonica seedlings grown in a mineral subsoil that had been exposed by the removal of surface soil. After a 100-day incubation, the growth of the seedlings was significantly enhanced by the colonization of these ectomycorrhizal fungal isolates as compared with uninoculated seedlings. In contrast, the growth of seedlings was retarded by the colonization of the MRA isolates. The growth of seedlings that were co-inoculated with ectomycorrhizal fungi and MRA was similar to that of uninoculated seedlings in most cases. These results suggest that ectomycorrhizal fungi have a beneficial effect on the growth of B. platyphylla var. japonica seedlings and that they suppress the deleterious effect of MRA. Thus, these ectomycorrhizal fungi probably have an important role in establishing B. platyphylla var. japonica seedlings during the initial stage of re-vegetation following site disturbance by the removal of surface soil.     

The effects of elevated CO2 and temperature on the resistance of winter-dormant birch seedlings (Betula pendula) to hares and voles

The effects of elevated CO₂ and temperature on the resource allocation pattern and resistance against mammalian herbivores of silver birch (Betula pendula Roth) were studied. Birch seedlings were grown through two growing seasons in closed‐top chambers exposed to four different treatments: ambient CO₂ and temperature, elevated atmospheric CO₂ (700 ppm) and ambient temperature, elevated temperature (+3°C above ambient) and ambient CO₂, and a combination of elevated CO₂ and temperature. After winter hardening of the seedlings, the growth of the seedlings was measured and the concentration of secondary compounds such as phenolics and papyriferic acid determined. The top parts of the stem were fed to hares, and the basal parts of the same stems were offered to voles. Elevated CO₂ increased the height and basal diameter of the shoots, shoot biomass and total biomass of the seedlings but did not have any effect on secondary chemistry. Elevated temperature increased the height and shoot biomass, but did not have a significant effect on the total biomass of the seedlings. Elevated temperature decreased the concentration of condensed tannins and their precursor, (+)‐catechin, in the top part of the stems, but only the concentration of (+)‐catechin in the basal part of the stems. There were no significant interactive effects between CO₂ and temperature on phenolics in the stems, while the concentration of papyriferic acid showed significant interaction in the top part of the stems. This indicates high accumulation of papyriferic acid in ambient CO₂ under increased temperature. Consequently, elevated temperature increased the resistance of birch against hares, but did not affect the resistance of the basal parts of the same birches to voles. Our results indicate that the predicted climatic change will not necessarily lead to increased browsing damage by the mountain hare and the field vole to silver birch.     

The influence of soil moisture and flooding on formation of VA-endo- and ectomycorrhizae in Populus and Salix

Native mixtures of extomycorrhizal fungi were found to infect Populus and Salix roots primarily in very moist but well drained soils in both the field and in controlled experiments (0 to –0.2 MPa), whereas native mixtures of VA-endomycorrhizal fungi infected roots over a much wider range of soil moisture (flooded to –3.4 MPa). Although a moisture gradient experiment showed endomycorrhizal formation was greater in moist soil than in very dry or flooded soils, this pattern was reversed in field transects along drainage gradients. Infection by VA-endomycorrhizal fungi in the field was the lowest where infection by ectomycorrhizal fungi was high, which suggests possible antagonism among the fungal symbionts. The narrow moisture range for ectomycorrhizal formation, and antagonism among endo- and ectomycorrhizal fungi, apparently combine to produce the mycorrhizal distributions found in nature.