Presence of arbuscular mycorrhizas in typically ectomycorrhizal host species from Cameroon and New Zealand

 Ectomycorrhizas (EcM) and arbuscular mycorrhizas (AM) were screened for in saplings of 14 EcM tree species from the N'Dupé and Korup National Park rainforests, SW Cameroon, belonging to Caesalpiniaceae and Uapacaceae. The pattern of EcM and AM colonisation of a dual mycorrhizal species from this rainforest (Uapaca staudtii, Uapacaceae) was compared with dual EcM/AM colonisation of Leptospermum scoparium (Myrtaceae) from New Zealand. Both species were collected in a range of habitats. EcM and AM colonisation differed among species in the Korup National Park rainforest: 12 species belonging to the Caesalpiniaceae (Amherstieae) were consistently EcM, and AM structures occurred occasionally in six of them; two other species belonging to Caesalpiniaceae (Afzelia bipindensis) and Uapacaceae (U. staudtii) were dual mycorrhizal with variable levels of colonisation by both EcM and AM fungi. EcM and AM dual colonisation varied with both habitat and identity of the partners. The presence of EcM fungi in most of the root samples of U. staudtii and a negative relationship between AM and EcM colonisation within the same root system suggested a greater EcM affinity of this species. In contrast, most root samples of L. scoparium were colonised by AM, but only a few by EcM. Genuine dual EcM/AM associations in root samples of U. staudtii where the two mycorrhizal types co-occurred could be attributed to an AM-EcM succession. However, differences between predicted and observed frequencies of genuine dual EcM/AM associations in several samples of both U. staudtii and L. scoparium indicated that other factors influenced dual EcM/AM associations. The results of this study showed the importance of the identity of the host species in determining the pattern of dual EcM and AM colonisation. Accepted: 18 September 1998     

Coinoculation efficacy of ectomycorrhizal fungi on Pinus patula seedlings in a nursery

 The coinoculation efficacy of the ectomycorrhizal fungi Laccaria laccata and Thelephora terrestris on the growth and mycorrhizal development of Pinus patula seedlings was studied and compared to individual inoculation of these fungi in a nursery. The total number of mycorrhizas was higher in seedlings inoculated with the combined inoculum than with the individual inocula. The colonization by T. terrestris was higher than L. laccata when the seedlings were inoculated with the two fungi simultaneously. Coinoculation significantly increased the height and dry weight of the seedlings compared with individual inoculation, both in steam-sterilized and unsterilized soil. Accepted: 12 May 1997     

Fungal ectomycorrhizal community and drought affect root hydraulic properties and soil adherence to roots ofPinus pinaster seedlings

Pinus pinaster seedlings were grown in a sandy dune soil either inoculated withHebeloma cylindrosporum or let to natural colonisation. Six months later, half of the seedlings of both treatments were subjected to a 3-week moderate drought. Root colonisation analysis showed that root tips were colonised to almost 100% independent of the inoculation. DNA determination of the ectomycorrhizal morphotypes showed that inoculated seedlings were extensively mycorrhized byH. cylindrosporum (more than 75%) whereas non-inoculated seedlings were mycorrhized by the exotic speciesThelephora terrestris (50%) andLaccaria bicolor (30%) and to a lesser extent byH. cylindrosporum (20%). Drought did not affect these frequencies. Total plant biomass was not affected by the mycorrhizal status or by drought but the root/shoot biomass ratio as well as the root/leaf surface area ratio were much lower in seedlings extensively colonised byH. cylindrosporum. Root hydraulic conductivity was higher in plants mainly mycorrhized byH. cylindrosporum, showing that this fungus improved the water uptake capacity of the root system as compared toT. terrestris and/orL. bicolor. This positive effect was also found under drought but to a lesser extent.H. cylindrosporum also increased the amount of root-adhering soil as compared to the other fungal symbionts, illustrating the performance of this association in aggregating sandy soil particles and developing the rhizosheath. The origin of the reduced root hydraulic resistance byH. cylindrosporum mycorrhization is discussed for the whole path including soil, soil-root interface and root cortex.     

Mycorrhizal inoculum potentials of pure reclamation materials and revegetated tailing sands from the Canadian oil sand industry

Recent improvements in the management of oil sand tailings used by the Canadian oil sand industry have resulted in the production of composite tailing sands (CT): a new challenging material for reclamation work. Jack pine (Pinus banksiana Lamb.), hybrid poplar (Populus deltoides Bartr. ex Marsh. ×Populus nigra L.) and red clover (Trifolium pratense L.) plants were used in an 8-week greenhouse bioassay to evaluate the mycorrhizal inoculum potential of CT. This inoculum potential was compared with that of three other reclamation materials [common tailing sands (TS), deep overburden (OB) and muskeg peat (MK)], and with three sites reclaimed in 1982 (R82), 1988 (R88) and 1999 (R99). CT was devoid of active mycorrhizal propagules while all other materials showed some level of inoculum potential. Arbuscular mycorrhizal fungi were observed on roots of clover or poplar grown in TS, OB, and all substrates containing peat (MK, R82, R88 and R99). Pine roots were also colonized by vesicle-forming hyphae of an unidentified fine endophyte and by dark septate fungi. Ectomycorrhizas (ECM) were observed on pine and poplar grown in OB, MK, and in soils from the two older reclaimed sites (R82 and R88). Using morpho- and molecular typing, six ECM fungi were identified to the genus or species level: Laccaria sp., Thelephora americana, Wilcoxina sp. (E-strain), Tuber sp. (I-type), a Sebacinoid, and a Pezizales species. Laccaria sp. and Wilcoxina sp. were the most frequently observed ECM species.     

Nitrate reductase activity of nonmycorrhizal Douglas-fir rootlets and of some associated mycorrhizal fungi

Douglas-fir root tips reduced nitrate at much higher rates than seven mycorrhizal fungi, which also differed between species in rate of nitrate reduction. Results are related to nitrogen nutrition of Douglas-fir.Excised, nonmycorrhizal root tips of Douglas-fir reduced nitrate at much higher rates than seven mycorrhizal fungi commonly associated with Douglas-fir. The fungi differed significantly in degree of activity:Cenococcum geophilum, Piloderma bicolor, andRhizopogon vinicolor were highest;Amanita muscaria was intermediate; andHebeloma crustuliniforme, Thelephora terrestris, andLaccaria laccata were lowest.     

Effect of mycorrhizal inocula on the growth of Sitka spruce seedlings in different soils

Summary Different mycorrhizal fungi were tested for their effectiveness in promoting growth of Sitka spruce seedlings, in two contrasting soils, in a glasshouse pot experiment. In nursery soil,Laccaria amethystina significantly improved growth of seedlings in comparison toL. laccata. Seedlings inoculated with a forest isolate ofThelephora terrestris were significantly larger than those inoculated with a nursery isolate when grown in forest soil. The effectiveness ofComplexipes moniliformis in forest soil was poor in comparison to other mycorrhizal fungi. Strains aswell as species of mycorrhizal fungi affect seedling growth differently. These effects are further influenced by soil type.     

Effect of a synthetic pyrethroid on the growth of ectomycorrhizal fungi and mycorrhiza formation in Pinus patula

Th effect of fenvalerate 20% EC (a synthetic pyrethroid) on the growth of two ectomycorrhizal fungi, Thelephora terrestris and Laccaria laccata, was studied in vitro. In the case of T. terrestris, mycelial growth was promoted at lower concentrations of the insecticide but inhibited at higher concentrations; mycelial growth of L. laccata decreased with increase in concentration. The effect of the insecticide at three concentrations (0.25, 0.5, 1.5 ppm) was also studied on ectomycorrhizal formation in 4-month-old seedlings of Pinus patula in nursery trials. At lower concentrations the insecticide had no significant effect on mycorrhizal formation.     

Uptake of arsenic by mushrooms from soil

ABSTRACT

Graphite furnace atomic absorption spectrometry (standard addition method) was used to determine the total arsenic in wild-growing mushrooms after digestion with nitric acid, then with perchloric acid and in associated soils after digestion with mixtures of nitric and hydrofluoric acids in a microwave system. Among 83 species of mushrooms the highest concentrations of arsenic on a dry mass basis were found in Laccaria amethystea (26–125 mg kg^?1), Laccaria laccata (11–33 mg kg^?1), Thelephora terrestris (38 mg kg^?1), Boletus cavipes (11.6 mg kg^?1) and Ramaria botrytis (10 mg kg^?1). Mushroom caps of L. laccata, L. amethystea, and B. cavipes had approximately double the arsenic concentrations found in stems. The arsenic concentrations in caps of L. amethystea and L. laccata were directly proportional to the concentrations in the soils. The concentrations of arsenic in the soils were in the range 6.5–65 mg kg^?1. Among the 19 mushroom caps with arsenic concentrations above the method detection limit of 0.2 mg As/kg dry mass, only L. amethystea and L. laccata had arsenic concentration ratios ‘cap/soil’ higher than 1 (between 1.1 and 1.9). Thelephora terrestris had a ratio of 2.37.     

Mycorrhizal synthesis of four ectomycorrhizal fungi in potted <Emphasis Type="Italic">Populus maximowiczii</Emphasis> seedlings

Ectomycorrhizal (ECM) syntheses between four ECM fungi, Laccaria amethystina, Hebeloma mesophaeum, Thelephora terrestris, and Tomentella sp., and Populus maximowiczii seedlings that are known to form ECM at a denuded area of Mt. Usu were performed in volcanic debris in a controlled growth chamber. The percentage of ECM colonization and seedling growth were determined 3 months after inoculation. Seedlings were successfully colonized by the inoculated ECM fungi with low contamination ratios. Seedling height and biomass were larger in the inoculated seedlings than in the control, although the effects of inoculation on seedling growth varied with the ECM fungus.     

ALLELOPATHIC POTENTIAL OF RASPBERRY FOLIAR LEACHATES ON GROWTH OF ECTOMYCORRHIZAL FUNGI ASSOCIATED WITH BLACK SPRUCE

In vitro bioassays on seven species of ectomycorrhizal fungi associated with black spruce (Picea mariana (Mill.) BSP) were tested under allelopathic action of raspberry (Rubus idaeus L.). Radial growth inhibitions were observed when fungi were confronted with increasing concentrations (0%–0.1%–0.5%–2.5%) of foliar leachates made from May, July, and September material (1986). For the 2.5% treatment, species Paxillus involutus, Laccaria proximo, Laccaria bicolor, Thelephora terrestris, and Cortinarius pseudonapus grew only 6, 8, 32, 42, and 46% of their control, whereas Hebeloma cylindrosporum and Cenococcum geophilum were stimulated. Tree seedlings from raspberry invaded plantation showed mycorrhizal infection rate over 75%, with fine roots mainly colonized by C. geophilum. In combination with efforts of tree improvement and based on the results of this study, it seems important to select a well-adapted my-corrhizae, when interference by raspberry is involved.     

Fungal communities in mycorrhizal roots of conifer seedlings in forest nurseries under different cultivation systems, assessed by morphotyping, direct sequencing and mycelial isolation

Fungi colonising root tips of Pinus sylvestris and Picea abies grown under four different seedling cultivation systems were assessed by morphotyping, direct sequencing and isolation methods. Roots were morphotyped using two approaches: (1) 10% of the whole root system from 30 seedlings of each species and (2) 20 randomly selected tips per plant from 300 seedlings of each species. The first approach yielded 15 morphotypes, the second yielded 27, including 18 new morphotypes. The overall community consisted of 33 morphotypes. The level of mycorrhizal colonisation of roots determined by each approach was about 50%. The cultivation system had a marked effect on the level of mycorrhizal colonisation. In pine, the highest level of colonisation (48%) was observed in bare-root systems, while in spruce, colonisation was highest in polyethylene rolls (71%). Direct internal transcribed spacer ribosomal DNA sequencing and isolation detected a total of 93 fungal taxa, including 27 mycorrhizal. A total of 71 (76.3%) fungi were identified at least to a genus level. The overlap between the two methods was low. Only 13 (13.9%) of taxa were both sequenced and isolated, 47 (50.5%) were detected exclusively by sequencing and 33 (35.5%) exclusively by isolation. All isolated mycorrhizal fungi were also detected by direct sequencing. Characteristic mycorrhizas were Phialophora finlandia, Amphinema byssoides, Rhizopogon rubescens, Suillus luteus and Thelephora terrestris. There was a moderate similarity in mycorrhizal communities between pine and spruce and among different cultivation systems.Electronic supplementary material Supplementary material is available for this article at and accessible for authorised users     

Positive association between mycorrhiza and foliar endophytes in Poa bonariensis, a native grass

The interaction between mycorrhiza and leaf endophytes (Neotyphodium sp.) was studied in three Poa bonariensis populations, a native grass, differing significantly in endophyte infection. The association between endophytes and mycorrhizal fungi colonisation was assessed by analysing plant roots collected from the field. We found that roots from endophyte-infected populations showed a significantly higher frequency of colonisation by mycorrhizal fungi and that soil parameters were not related to endophyte infection or mycorrhiza colonization. In addition, we did not observe significant differences in the number of AM propagules in soils of the three populations sites. We also report the simultaneous development of Paris-type and Arum-type mycorrhiza morphology within the same root systems of P. bonariensis. The co-occurrence of both colonisation types in one and the same root system found in the three populations, which differed in Neotyphodium infection, suggests that foliar endophytes do not determine AM morphology. The percentage of root length colonised by different types of fungal structures (coils, arbuscules, longitudinal hyphae and vesicles) showed significant and positive differences in arbuscular frequency associated with endophyte infection, whereas the much smaller amounts of vesicles and hyphal coils did not differ significantly.     

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.     

Interaction of soil microbes with mycorrhizal fungi in tomato

In this study, the interaction of soil microbes with mycorrhizal fungi (MF) was performed to understand the effect on tomato. A pot and a field experiment were employed to investigate the impact of soil microbes i.e. Fusarium oxysporum f. sp. lycopersici, Trichoderma harzianum, Aspergillus niger and Rhizobium leguminosarum, on AM fungi in pots and field studies. The soils without microbes which treated controls with or without mycorrhizal inoculation were also included. Plant growth and root colonisation were measured 36, 75 and 120?days post inoculation (dpi) in the both pot experiment and field study. Soil microbes’ effects on the growth behaviour of the tomato plant were determined via the shoot and root weight. R. leguminosarum and A. niger did not affect the colonisation ability much, but F. oxysporum f. sp. lycopersici and T. harzianum resulted in the inhibition of AM fungal colonisation in both pot and field studies. Our study provides evidence for the effects of soil microbes on the diversity of AM fungi and their effect on the tomato plants. The higher concentrations of phosphorus and of proteins in the root tissues, previously colonised with AM fungi, point out their effect as biofertilizers, according to the concept of sustainable agriculture.     

Effect of root exudates of mycorrhizal tomato plants on microconidia germination of Fusarium oxysporum f. sp. lycopersici

The effect of root exudates from mycorrhizal and non-mycorrhizal tomato plants on microconidia germination of the tomato pathogen Fusarium oxysporum f. sp. lycopersici was tested. Microconidia germination was enhanced in the presence of root exudates from mycorrhizal tomato plants. Tomato plants were colonised by the arbuscular mycorrhizal fungus Glomus fasciculatum, indicating that alterations of the exudation pattern depended on the degree of root AM colonisation. Testing the exudates from plants with a high and a low P level revealed that the alterations of the root exudates from mycorrhizal plants, resulting in a changed effect on microconidia germination, are not due to an improved P status of mycorrhizal plants.     

The effects of flooding on the formation of ectomycorrhizae in Pinus sylvestris seedlings

Pinus sylvestris seedlings, grown in a vertical petri dish system, were inoculated with five different mycorrhizal fungi. Half of the root system in the petri dish was subjected to periodic flooding, and mycorrhizal colonization was studied. Thelephora terrestris, Laccaria laccata, and Hebeloma crustuliniforme were not sensitive to flooding, whereas Suillus flavidus and S. bovinus were highly sensitive. The latter failed to colonize the root even when flooded for only 2 min per day four times a week.     

Vesicular-arbuscular mycorrhizal inoculum potential affects the growth of Stryphnodendron microstachyum seedlings in a Costa Rican human tropical lowland

This study used a plant bioassay to investigate the vesicular-arbuscular mycorrhizal (VAM) inoculum potential of soil from three vegetation types (fern, secondary forest, and grass) in an abandoned pasture in the tropical humid lowlands at La Selva, in northeastern Costa Rica. Growth, measured as seedling height, number of leaves, and total (above- and belowground) biomass, of Stryphnodendron microstachyum Poepp. et Endl. (Synon. S. excelsum Harms) seedlings was significantly lower when grown in soil inoculum from the fern areas than in soil inoculum from the forest and grass areas. However, S. microstachyum seedlings grown in the fern inoculum had significantly greater VAM colonization than seedlings grown in the forest and grass inoculum. In addition, roots collected from a dominant plant species from each of the three vegetation types showed that the fern (Nephrolepsis biserrata) had significantly greater mycorrhizal colonization than the tree (Pentaclethra macroloba (Willd.) Kuntze or the grass (Brachiaria spp.). The results of this study suggest that differences in mycorrhizal inoculum potential among vegetation types and its effects on seedling growth may have important implications for the restoration and management of degraded lands.     

Differential benefits of arbuscular mycorrhizal and ectomycorrhizal infection of Salix repens

 The functional significance of arbuscular mycorrhiza (AM) and ectomycorrhiza (EcM) for Salix repens, a dual mycorrhizal plant, was investigated over three harvest periods (12, 20 and 30 weeks). Cuttings of S. repens were collected in December (low shoot P) and March (high shoot P). Glomus mosseae (an arbuscular mycorrhizal fungus, AMF) resulted in low AM colonization (<5%), but showed large short-term (<12 weeks) effects on shoot growth and root length. Hebeloma leucosarx (an ectomycorrhizal fungus, EcMF) resulted in high EcM colonization (70%), but benefits occurred over a longer term (>12 weeks). Furthermore, G. mosseae colonization resulted in higher shoot P uptake, shoot growth, root growth and response duration for S. repens collected in December than for those collected in March, whereas with H. leucosarx and the non-mycorrhizal treatment there were no differences between cuttings collected on different dates. Low AMF colonization was effective in the short term for cuttings at both collecting dates. Low AMF colonization of S. repens occurred irrespective of the amount of AMF inoculum used. The intensities and relative amounts of AMF structures in S. repens and Trifolium repens were compared over three harvest periods (12, 20 and 30 weeks) to assess plant species effects on AM colonization patterns. Accepted: 13 October 2000     

Influence of inoculation with a Trichoderma bio‐inoculant on ectomycorrhizal colonisation of Pinus radiata seedlings

Trichoderma species, through mechanisms such as mycoparasitism, antibiosis, induced resistance and competition, are able to suppress the growth and activity of soil microbes and therefore have the potential to influence ectomycorrhizal (ECM) fungal colonisation. ECM colonisation potential of Pinus radiata seedlings inoculated with the commercially available Trichoderma species bio‐inoculant, ArborGuard?, was investigated in a commercial containerised nursery setting and in a separate glasshouse experiment. Application of ArborGuard? to P. radiata seedlings in both the containerised nursery and glasshouse experiment had no detrimental effect on the ability of the naturally occurring ECM fungi to colonise the seedlings. Thelephora terrestris was the only ECM species colonising the P. radiata root tips in the containerised system while Tomentella ellisii was the dominant species found colonising the P. radiata root tips in the glasshouse experiment. No seedling growth promotion was observed with the addition of ArborGuard?. The results show that the Trichoderma bio‐inoculant ArborGuard? does not affect ECM colonisation of P. radiata seedlings by Th. terrestris and T. ellisii in a containerised nursery system.     

Effects of red pine (Pinus resinosa Ait.) mycorrhizoplane-associated actinomycetes onin vitro growth of ectomycorrhizal fungi

Mycorrhizoplane-associated actinomycetes were isolated using an enrichment technique from red pine (Pinus resinosa Ait.) roots of seedlings recently outplanted onto cleared northern hardwood sites in the Upper Peninsula of Michigan, USA. Interactions were assessedin vitro between actinomycete isolates and three commonly occurring ectomycorrhizal fungi (Laccaria bicolor (Maire) Orton,L. laccata (Scop.: Fr.) Berk. and Br., andThelephora terrestris Fr.). Most actinomycete isolates exerted a range of effects on the growth of the three fungus isolates during the four week test period, inhibiting some while stimulating others; several inhibited growth of all three fungus isolates. Mycorrhizoplane-associated actinomycetes show potential for use as coinoculants with selected ectomycorrhizal fungi to optimize the soil microflora for developing seedlings.