The allelopathic potential of alfalfa root medicagenic acid glycosides and their fate in soil environments

Summary The allelopathic effect of alfalfa (Medicago media Pers.) and red clover (Trifolium pratense L.) root saponins on winter wheat seedling growth and the fate of these chemicals in soil environments were studied. Seed germination, seedling and test fungus growth were suppressed by water and by alcohol extracts of alfalfa roots, and by crude saponins of alfalfa roots, indicating that medicagenic acid glycosides are the inhibitor. Powdered alfalfa roots inhibited wheat seedling growth when added to sand. At concentrations as low as 0.25% (w/w) the root system was completely destroyed whereas seedling shoots suffered little damage. Red clover roots caused some wheat growth inhibition when incorporated to sand, but their effect was much lower than in the alfalfa root treatment. Soil textures had a significant influence on the inhibitory effect of alfalfa roots. The inhibition of seedling growth was more pronounced on light than on heavy soils. This was attribted to the higher sorption of inhibitors by heavy soils. Incubation of alfalfa roots mixed into loose sand, coarse sand, loamy sand and clay loam for a period of 0–8 days resulted in decreased toxicity to bothT. viride and wheat seedlings. This decrease occurred more quickly in heavier soils than in loose sand, due to the hydrolysis of glycosides by soil microorganisms. Soil microbes were capable of detoxifying medicagenic acid glycosides by partial hydrolysis of sugar chain to aglycone. These findings illustrate the importance of medicagenic acid glycosides as an inhibitor of wheat seedling growth, and of their fate in different soil environments.     

Survival of the external mycelium of a VAM fungus in frozen soil over winter

The present investigation examines (1) whether the external VAM mycelium survives winter freezing to act as a source of inoculum in the spring, and (2) whether soil disturbance reduces the infectivity of the external VAM mycelium following freezing of the soil. Sealed pouches of fine nylon mesh were placed in pots containing soil inoculated with a Glomus species. The mesh was impervious to roots but not to hyphae. Following two 3-week growth cycles of maize in the pots, the pouches were transplanted to the field. Pouches were removed from the field once during the 4 months when the soil was frozen, and once after spring thaw. Measurements were made of VAM spore density, hyphal length and viability in the pouches. Bioassays for infectivity were conducted on all pouches. Some VAM hyphae survived freezing and remained infective following winter freezing, in the absence of plant roots. Soil disturbance did not reduce the infectivity of hyphae following exposure to freezing temperatures. We observed a change in the distribution of viable cytoplasm within hyphae over winter, which we hypothesize represents an adaptation allowing hyphae to survive freezing temperatures. We suggest that the effect of disturbance on hyphal infectivity may be related to this seasonal change in the distribution of hyphal viability.     

Soil disturbance and infection of Trifolium repens roots by vesicular-arbuscular mycorrhizal fungi

Removal and storage of the surface layers of soil is known to decrease the infectivity of vesicular-arbuscular mycorrhizal (VAM) fungi. Previous studies have mostly examined the effects of profound soil disturbance on the infectivity of VAM fungi. This study examined the effects of increasing degrees of topsoil disturbance on the infectivity of VAM fungi in two sites on sandstone soils in southeastern Australia. Intact soil blocks (20×20×15 cm) were taken from each of the two sites. Increasing degrees of topsoil disturbance were achieved by cutting the blocks longitudinally into four (dist. 1), nine (dist. 2), and 25 (dist. 3) equal portions. Seeds of Trifolium repens L. were sown into the blocks and harvested 14, 21, 28, 35 and 42 days after sowing. At each sampling date, total root length, root length colonised by VAM fungi and shoot dry mass were measured. VAM colonisation had commenced by 14 days in the roots of seedlings grown in intact, dist. 1, and dist. 2 soil blocks. The initiation of VAM colonisation was delayed by up to 6 weeks for seedlings grown in the dist. 3 soil blocks. The low (i.e. dist. 1) and intermediate (i.e. dist. 2) degrees of soil disturbance did not cause a delay in the initiation of VAM, bud did significantly reduce the proportion of root length colonised by VAM fungi after 21 days. After 21 days, shoot dry mass was significantly less in the seedlings grown in the dist. 3 soil blocks though not in the low and intermediate disturbance treatments. It is concluded that the most severe experimental disturbance probably disturbed the external hyphal network and root fragments (containing hyphae and vesicles), which in turn temporarily reduced the infective potential of the fungus to zero. The observed delay in the initiation of VAM in the most disturbed blocks can, therefore, be explained by the time required for hyphae to grow from other propagules in the soil which survived the disturbance event.     

The epidemiology of tomato brown root rot

In the absence of nematodes, three different symptoms of disease, parts of the brown root rot complex (BRR), occurred on tomato roots surviving in soils infested with GSF (= grey sterile fungus) and Colletotrichum atramentarium (Berk. & Br.) Taubenh. In heavily infested soils brown lesions occurred throughout cropping, appearing within a week of planting. Corkiness and black dot, caused by GSF and C. atramentarium respectively, rarely occurred until the third month after planting but towards the end of the season the incidence of black dot sometimes suddenly increased greatly. Observations of crops growing in plots treated with different soil partial sterilants suggested that GSF was more damaging than C. atramentarium. Yield was not related to the incidence of black dot but was inversely proportional to the occurrences of brown lesions and corkiness. The relation with brown lesions was significant within 8 weeks of planting, when most brown lesions gave cultures of GSF, but later more of these lesions gave cultures of C. atramentarium than of GSF. Pathogenicity tests with pure cultures of GSF and C. atramentarium were done on agar media and by artificially infesting partially sterilized soils. Roots of undamaged seedlings on agar media developed 10 mm. brown lesions within 2 weeks of inoculating 10-day-old tomatoes with most GSF cultures isolated from: (1) rotted roots of Lycopersicon esculentum, Solanum capsicastrum, Capsicum annuum var. longum and C. frutescens; (2) browned zones of Lycopersicon hirsutum roots; and (3) apparently healthy roots of Cucumis sativus. After inoculation with C. atramentarium, small (c. 2 mm.) pink lesions developed, whereas none formed using Pyrenochaeta spp. In soil tests the greater root damage done by GSF, including root loss, was reflected in decreased aerial growth and smaller fruit yields; C. atramentarium affected neither. In the second year of soil infestation GSF decreased yields during 6 weeks of picking from 1.96 kg. in the uninoculated controls to 1.02 kg./plant. The pattern of damage done by GSF changed as plants aged. In soil, brown lesions occurred within a few days of planting but corkiness did not appear for 2–3 months, when stem lesions and leaf yellowing often developed simultaneously. A 50% root loss after 21 weeks did not affect fruit yields whereas a 40% loss within 11 weeks of planting was reflected by a 45% yield decrease.     

Influence of Arbuscular Mycorrhizal (AM) Symbiosis on Phosphorus Leaching through Soil Cores

Two experiments with soil cores were carried out to investigate the effects of arbuscular mycorrhizal (AM) fungal colonization on mobility of phosphorus (P) during leaching of repacked columns of a soil with a loamy sand texture. Trifolium subterraneum plants inoculated with an AM fungus or not inoculated were grown in cores with low or high P concentrations for 8 or 10 weeks in the glasshouse. Cores were then irrigated with 2500 mL water and the leachate collected. Plant growth and the amounts of P removed by plants, remaining in soil as available P and removed dissolved in leachate were measured. Mycorrhizal fungal colonization and development of external hyphae were also determined. Inoculation and/or P application significantly increased plant growth and plant P removal and decreased P leaching. In low P soils AM fungal colonization significantly increased plant P uptake and decreased soil available P and total dissolved P in leachates. Lower P leaching from cores with AM plants under low P conditions was related to enhancement of plant growth and to scavenging and removal of P from the soil by roots and/or external hyphae. When P was applied AM effects were not observed and available P remaining in the soil after leaching was much higher, regardless of AM fungal colonization.     

Studies of iron transport by arbuscular mycorrhizal hyphae from soil to peanut and sorghum plants

 The influence of an arbuscular mycorrhizal (AM) fungus on phosphorus (P) and iron (Fe) uptake of peanut (Arachis hypogea L.) and sorghum (Sorghum bicolor L.) plants was studied in a pot experiment under controlled environmental conditions. The plants were grown for 10 weeks in pots containing sterilised calcareous soil with two levels of Fe supply. The soil was inoculated with rhizosphere microorganisms only or with rhizosphere microorganisms together with an AM fungus (Glomus mosseae [Nicol. & Gerd.] Gerdemann & Trappe). An additional small soil compartment accessible to hyphae but not roots was added to each pot after 6 weeks of plant growth. Radiolabelled P and Fe were supplied to the hyphae compartment 2 weeks after addition of this compartment. After a further 2 weeks, plants were harvested and shoots were analysed for radiolabelled elements. In both plant species, P uptake from the labelled soil increased significantly more in shoots of mycorrhizal plants than non-mycorrhizal plants, thus confirming the well-known activity of the fungus in P uptake. Mycorrhizal inoculation had no significant influence on the concentration of labelled Fe in shoots of peanut plants. In contrast, ⁵⁹Fe increased in shoots of mycorrhizal sorghum plants. The uptake of Fe from labelled soil by sorghum was particularly high under conditions producing a low Fe nutritional status of the plants. These results are preliminary evidence that hyphae of an arbuscular mycorrhizal fungus can mobilise and/or take up Fe from soil and translocate it to the plant. Accepted: 6 March 1998     

The Influence of Glomus mosseae on Tylenchulus semipenetrans-Infected and Uninfected Citrus limon Seedlings

Greenhouse studies have shown that when rough lemon (Citrus limon) seedlings infected with TyIenchulus semipenetrans were transplanted into soil infested with Glomus mosseae, the mycorrhizal fungus infection increased seedling growth compared to nonntycorrhizal seedlings. Tylenchulus semipenetrans significantly suppressed seedling growth below that of mycorrhizal seedlings. Histological observations of nematode-free mycorrhizal roots showed that hyphae penetrated the epidermis and invaded the cortex, giving rise to arbuscules and vesicles. Nematode infection sites in T. semipenetrans-infected roots grown in soil infested with G. mosseae did not show evidence of vesicle development in the cortex but did show arbuscule development.     

Infektionsstrukturen von Microdochium bolleyi an Wurzeln und Koleoptilen von Gerste

Infection structures of Microdochium bolleyi on barley roots and coleoptiles Microdochium bolleyi grows within roots and coleoptiles of barley either leading to a damage of the tissue or to a symptomless infection. The present work investigated whether these different types of infection are correlated with different infection structures of the fungus. Two types of hyphae were formed infectmg either the rhizodermis or the epidermis. The fungus spread within the cortex only, not affecting the endodermal cells or the vascular tissue. The extend to which the roots were damaged depended on the infection density. Differences in the structure of hyphae formed in symptomless or necrotic parts of the roots were not observed. In both, roots and coleoptiles, hyphae were detected in dead cells only.     

Influence of arbuscular mycorrhizal fungi on soil structure and aggregate stability of a vertisol

The influence of arbuscular mycorrhizal (AM) fungi on aggregate stability of a semi-arid Indian vertisol was studied in a pot experiment in which Sorghum bicolor (L.) was grown as test plant for 10 weeks. Pasteurized soil inoculated with AM fungi was studied with pasteurized and unpasteurized soils as references. A part of the soil in each pot was placed in nylon mesh bags to separate effects of roots and hyphae. The sorghum plants were planted outside the mesh bags which permitted AM hyphae to enter while excluding roots. Aggregate stability of the soil was determined by wet-sieving and turbidimetric measurements. Development of the AM fungi was quantified as colonized root length and external hyphal length. Soil exposed to growth of roots and hyphae (outside mesh bags) showed aggregates with larger geometric mean diameter (GMD) in pasteurized soil inoculated with AM fungi than in pasteurized uninoculated soil. There was no significant difference in GMD of the inoculated, pasteurized soil and the unpasteurized soil. No significant effects of inoculation or plant growth were found in pasteurized soil exposed to hyphal growth only (inside the mesh bags). However, the unpasteurized soil had significantly higher GMD than the pasteurized soil, irrespective of plants and inoculum. Turbidimetric measurements of soil exposed to roots and hyphae (outside mesh bags) showed the highest aggregate stability for the inoculated pasteurized soil. These results demonstrate that AM fungi contribute to the stabilization of soil aggregates in a vertisol, and that the effect is significant after only one growing season. The effect was associated with both AM hyphae and the stimulation of root growth by AM fungi. The contribution from plant roots and AM hyphae to aggregate stability of different size fractions is discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Soil aggregation status and rhizobacteria in the mycorrhizosphere

Soil aggregation is a dynamic process in which plants and the soil microbiota play a major role. This experiment was conducted to determine whether the effects of mycorrhizae on the stability of water-stable soil aggregates (WSA) and on selected groups of soil microorganisms are interrelated. Soil containers consisting of four compartments were utilized. Two compartments on each side of a solid barrier were separated by a 43 m screen that permitted the passage of hyphae, but not of roots. The roots of Sorghum bicolor plants were split over the center barrier, and the roots on one side were inoculated with an arbuscular-mycorrhizal (AM) fungus. This design produced mycorrhizosphere soils (M) by AM roots or hyphosphere (H) soils by AM hyphae in the two compartments on the one side of the barrier, and rhizosphere soils (R) by nonAM roots or root- and hypha-free bulk soil (S) in the two compartments on the other side. At harvest (10 wk), there were significant differences in WSA between soils in the order: M>R>H>S, and WSA stability was significantly correlated with root or hyphal length. Numbers of colony-forming units of the microflora (total bacteria, actinomycetes, anaerobes, P solubilizers, and nonAM fungi) were in general not correlated with root or hyphal length, but in some cases were significantly correlated with WSA. Bacteria isolated from the water-stable soil-aggregate fraction tended to be more numerous than from the unstable fraction. The difference was significant in the M soil for total bacteria and P solubilizing bacteria. NonAM fungi were more numerous in the unstable fraction of the M soil. The data show that the root and fungal components of mycorrhizae enhance WSA stability individually and additively in concert, and suggest that they affect microorganism numbers indirectly by providing a favorable and protective habitat through the creation of habitable pore space in the WSA.     

板栗褐缘叶枯病协同致病菌Ophiognomonia castaneae的生活史

采用室外定点观察,子实体诱导及rDNA ITS、MS204、tef1-α 3种分子标记进行系统发育分析等方法,对板栗褐缘叶枯病Phomopsis castaneae-mollissimae的协同致病菌板栗蛇孢日规壳Ophiognomonia castaneae的生活史进行了研究。结果表明,每年7月下旬至8月初叶片发病初期很少分离到O. castaneae,随着病斑扩大该菌的分离频率逐渐增大,至发病后期其分离频率可高达78.5%,甚至可超过致病菌P. castaneae-mollissimae,10月下旬板栗落叶背面的病斑上开始形成O. castaneae的分生孢子盘,11月下旬开始形成O. castaneae的子囊壳原基,次年5、6月越冬落叶背面的病斑上长出子囊壳;带病斑的叶片经室内外诱导,0-25℃范围均可产生成熟子囊壳;湿度是决定子囊壳能否形成的关键因素,强光照不利于子囊壳的产生;分离物的菌丝体在PDA培养基上培养,易产生分生孢子;将分离物分为两种交配型,相互交配后6个月所有处理均未长出该菌的有性型子实体。室外定点观察及rDNA ITS、MS204、tef1-α 3种分子标记表明分离物和病斑上的子囊孢子及其萌发菌丝为O. castaneae的不同生长发育阶段。     

Effects of wheat volunteers and blackgrass in set-aside following a winter wheat crop on soil infectivity and soil conduciveness to take-all

Two experiments were carried out in France in which disease indices were used to evaluate the effects of wheat volunteers and blackgrass (Alopecurus myosuroides) on soil infectivity and soil conduciveness to take-all caused by Gaeumannomyces graminis var. tritici. Soil infectivity was evaluated by measuring the disease index on susceptible wheat plants grown on soil samples collected from the field. Soil conduciveness to the disease was obtained by measuring disease indices on plants grown on soil samples to which different amounts of take-all fungus inoculum were added. One experiment (Expt. 1) was carried out using soils from farmers' fields (two fields in 1994 and two in 1995); soil infectivity and soil conduciveness were evaluated for three experimental situations: bare soil, soil with wheat volunteers and soil with blackgrass plants. In 1994 the soil infectivity was zero in bare soil, high with the wheat cover, and intermediate with the blackgrass cover. In 1995 the soil infectivity was uniformly low for all three conditions. Soils bearing wheat were less conducive than bare soil, soils bearing blackgrass and bare soils were similarly conducive. A second experiment (Expt. 2) carried out in 1995 compared the soil infectivity and soil conduciveness to take-all of soils planted with wheat or blackgrass in set-aside land after periods of wheat monoculture of 0–6 yr. The soil infectivity was low for all treatments. The soil was more conducive after blackgrass than after wheat. In both cases, the soil conduciveness was less when the monoculture had continued for more than 4 yr. The decline was less after blackgrass than after wheat. Thus, whenever set-aside is set up during the increase phase of the disease in fields with cereal successions, abundant wheat volunteers might hinder the expected positive effect of a break in cereal successions on take-all development. The presence of blackgrass in a set-aside field, with significant soil infectivity and high soil conduciveness, might increase the risks of take-all development in a wheat crop following set-aside.     

Long-term orchard groundcover management systems affect soil microbial communities and apple replant disease severity

Apple replant disease (ARD) is a soil-disease syndrome of complex etiology that affects apple tree roots in replanted orchards, resulting in stunted tree growth and reduced yields. To investigate whether different groundcover management systems (GMSs) influence subsequent ARD severity, we grew apple seedlings in an outdoor nursery in pots containing orchard soil from field plots where four GMSs had been maintained for 14 years in an orchard near Ithaca, NY, USA. The GMS treatments were: (1) pre-emergence herbicide (Pre-H), bare soil strips maintained by applying tank-mixed glyphosate, norflurazon and diuron herbicides annually; (2) post-emergence herbicide (Post-H), sparse weed cover maintained by applying glyphosate in May and July each year; (3) mowed sod grass (Mowed Sod); and (4) bark mulch (Mulch). Soils were also sampled from the grass drive lane maintained between the trees in the orchard (Grass Lane). Sampled soils (Orchard soil) were either pasteurized or left untreated, placed into 4-L pots, and planted with one apple seedling per pot. After 3 months of growth, soil (Bioassay soil) and apple tree roots (Bioassay roots) were sampled from each pot and microbial populations colonizing samples were characterized. Seedling growth was reduced in soils sampled from all four GMS treatments compared to the Grass Lane soils. Among the GMS treatments, seedling biomass was greater in Pre-H than in the Post-H soil. Soil microbial communities and nutrient availability differed among all four GMS treatments and the Grass Lane. Root-lesion (Pratylenchus sp.) nematode populations were higher in the Mowed Sod than in the other GMS treatments. Soil bacterial and fungal community composition was assessed in Orchard and Bioassay soils and Bioassay roots with a DNA fingerprinting method (T-RFLP). Redundancy analysis indicated that soils sampled from the different GMS treatments differentially influenced seedling biomass. A clone library of 267 soil bacteria was developed from sampled Orchard soils and Bioassay roots. These communities were dominated by Acidobacteria (25% of sequences), Actinobacteria (19%), δ-Proteobacteria (12%), β-Proteobacteria (10%), and these ratios differed among the GMS soils. Members of the family Comamonadaceae were detected only in tree-row soil, not in the Grass Lanes. The dominant sequences among 145 cloned fungi associated with apple seedling roots were Fusarium oxysporum (16% of sequences), an uncultured soil fungus submitted under DQ420986 (12%), and Rhodotorula mucilaginosa (9%). In a redundancy analysis, factors including fungal and oomycete community compositions, soil respiration rates, population sizes of culturable bacteria and fungi, soil organic matter content, and nutrient availability, were not significant predictors of apple seedling biomass in these soils. Different GMS treatments used by apple growers may influence subsequent ARD severity in replanted trees, but edaphic factors commonly associated with soil fertility may not reliably predict tree-root health and successful establishment of replanted orchards.     

Perithecial Formation in Gelasinospora reticulispora: IV. Action Spectra for the Photoinduction

Action spectra for photoinduction of perithecia after different lengths of dark period were determined with apically growing mycelium of a sordariaceous fungus Gelasinospora reticulispora. When hyphae were exposed to monochromatic light in near ultraviolet and visible regions, reciprocity between intensity and exposure time was observed within the range of incident energy used. The resulting action spectrum determined after a dark period of 48 hours showed a peak at 460 nm with shoulders at 420 and 480 nm and another peak at 370 nm, indicating minima at 410, 430, and 470 nm. After 72 hours darkness the spectrum was very similar to the above, except that the major peak shifted to 450 nm and the near ultraviolet region was somewhat less effective. In both cases, wavelengths longer than 520 nm showed no effect.     

Vesicular-arbuscular mycorrhizal fungi,particularly Glomus tenue,in Venezuelan bromeliad epiphytes

The mycorrhizal status of water-impounding tank bromeliad epiphytes from three locales differing in altitude and moisture regime within Venezuelan cloud forest was examined. Species of vesicular-arbuscular mycorrhizal (VAM) fungi found in arboreal soils were compared to VAM fungi found in terrestrial soils. Sixteen of the 19 epiphytes examined for the presence of VAM fungi had roots with infection stages; 14 of these specimens showed growth of the fine endophyte Glomus tenue. Fine endophyte was the only VAM fungus found associated with epiphytes in the driest locale studied, while coarse VAM fungi (Gigaspora and Scutellospora spp.) were found at sampling locales receiving more moisture. Root infection was usually composed of intercellular hyphae and peletons; few arbuscules were observed. However, abundant extracellular hyphae were often observed tangled about roots in arboreal soil. It is concluded that epiphytic bromeliads probably benefit, at least periodically, from VAM fungi scavenging for sporadically available nutrients in arboreal soils. Glomus tenue may be particularly important as a colonizing VAM fungus in drier sites of Venezuelan cloud forest. The species composition of VAM fungi in arboreal soils was different to that of terrestrial soils sampled directly under epiphytic bromeliad perches, suggesting that VAM fungi species associated with bromeliads are dispersed to their hosts by vagile animal vectors.     

Growth and survival of Verticillium chlamydosporium goddard,a parasite of nematodes,in soil

Selective media for the isolation of the nematophagous fungus Verticillium chlamydosporium, are described. These enabled densities > 500 colony forming units (CFU) g^‐1 soil to be reliably estimated. However, there was little relationship between estimates of the Verticillium biomass in a sterilized soil and the numbers of CFU which developed on the selective media. The growth and survival of the fungus infield soils were studied and estimates of the numbers of CFU in soils in which cyst‐nematode multiplication was suppressed were greater than in those in which the nematode multiplied. Isolates of the fungus differed in their ability to proliferate in soil, but some increased rapidly from applications of chlamydospores or a mixture of hyphae and conidia in alginate granules containing wheat bran. The energy source (wheat bran) was essential for the establishment of the fungus from granular applications. Numbers of CFU greatly exceeded those of chlamydospores, but there was considerable variation in the relationship in different soils. Some isolates of V. chlamydosporium proliferated in soil and survived in considerable numbers for at least 3 months. Hence, pre‐cropping applications of the fungus should survive long enough to kill nematode eggs and females that develop on roots of spring‐sown crops.     

Use of a root assessment tray for the detection of take-all on lateral roots of wheat seedlings

A root assessment tray was designed for the meticulous assessment of take-all on wheat seedling roots from soil bioassays. Subsequently, the detection of lateral root infections (in addition to the more obvious infections on main axes of seminal roots) resulted in increased estimates of propagule numbers of the take-all fungus (Gaeumannomyces graminis var.tritici) for 196 of the 368 soil samples bioassayed in a field study conducted in Western Australia between 1984 and 1986.     

Impact of the exodermis on infection of roots byFusarium culmorum

Patterns of infection withFusarium culmorum (W G Smith) Saccardo were observed in seedling roots of barley (Hordeum vulgare L.), wheat (Triticum aestivum L.), maize (Zea mays L.) and asparagus (Asparagus officinalis L). Apical regions of the main roots were not infected. Since penetration into the root occurred several days after inoculation and the roots were growing during the experiment, these regions had apparently not been in existence long enough to be infected. In older regions of barley, wheat and asparagus, hyphae entered through the tips of lateral roots. In barley and wheat, which had not developed any suberin lamellae in their subepidermal layer, infection occurred randomly over the remainder of the root. In maize, the fungus penetrated the epidermis at many sites but did not breach the exodermis in which all cells possessed both Casparian bands and suberin lamellae. Maize roots, therefore, sustained only minimal infections. In asparagus, the fungus grew through the short (passage) cells but never the long cells of the exodermis. In doing so, it penetrated cells possessing Casparian bands but lacking suberin lamellae. The results support the hypothesis that suberin lamellae provide effective barriers to the growth ofF. culmorum hyphae.     

Identification of host plants and description of sclerotia of the truffle <Emphasis Type="Italic">Mattirolomyces terfezioides</Emphasis>

The truffle, Mattirolomyces terfezioides, is a hypogeous ascomycete with uncertain host relationships. The fungus has been regularly collected on sandy soils in the Carpathian Basin. During the study of the natural host plants of the fungus, strange, amorphous, belowground hyphal aggregates incorporating soil and sand particles have been found attached to the surface of the roots. The fruitbodies of M. terfezioides develop from these hyphal aggregates. This structure, similar to that formed by morels, could be interpreted as a sclerotium. Sclerotia were found both on roots of woody and herbaceous plants. To detect the roots colonized by M. terfezioides, a species-specific polymerase chain reaction was developed. Seven natural hosts of the fungus were identified by this method. No specificity regarding taxa or life form of the plants was found. The colonization of the roots by the septate hyphae of M. terfezioides was weak, particularly compared to the colonization by arbuscular–mycorrhizal fungi. This suggests that this fungus is not the dominant fungal partner of these plants. Therefore, using M. terfezioides as the only inoculum may be inappropriate in truffle cultivation experiments. Nevertheless, further in vitro experiments are needed to develop reliable knowledge on the still ambiguous symbiotic strategy of this fungus.     

Characterisation of arbuscular mycorrhizal fungi colonisation in cluster roots of shape Hakea verrucosa F. Muell (Proteaceae), and its effect on growth and nutrient acquisition in ultramafic soil

Ultramafic soils at Bandalup Hill (Western Australia) are characterised by high concentrations of Ni and low levels of P. Amongst the plant species that can sustain such hostile conditions, Hakea verrucosa F. Muell from a non-mycorrhizal family (Proteaceae) would be expected to rely on cluster roots to access P. However, the acidification of ultramafic soils by cluster roots might increase the dissolution of soil Ni, and therefore its availability to plants. Symbiosis with mycorrhizal fungi, on the other hand, might help to reduce the uptake of Ni by H. verrucosa. Therefore, the aim of this study was to investigate the mycorrhizal status of H. verrucosa, and assess any contribution from mycorrhizal fungi to its growth and nutrient status. Seedlings of H. verrucosa were first grown in undisturbed ultramafic soil cores from Bandalup Hill for 8 weeks to assess the presence of mycorrhizal fungi in their roots. In a second experiment, H. verrucosa seedlings were grown in the same ultramafic soil that was either steamed or left untreated. Seedlings were inoculated with an arbuscular mycorrhizal (AM) fungal consortium from Bandalup Hill. Fungal hyphae, vesicles, as well as intracellular arbuscules and hyphal coils were observed in the cluster roots of H. verrucosa in both experiments. In the first experiment, 57% of the root length was colonized by AM fungi. Seedlings had high (between 1.4 and 1.9) shoot to root ratios and their roots had very few root hairs, despite growing in P-deficient soil. Steaming of the ultramafic soil increased the growth of seedlings and their nutrient uptake. Inoculation with AM fungi reduced the seedling growth in steamed ultramafic soil; however, it increased their shoot P and K concentration and also the shoot K content. The shoot Ni concentration of seedlings was not affected by the presence of AM fungi.