Mycorrhizal mediation of plant N acquisition and residue decomposition: Impact of mineral N inputs

Mycorrhizas are ubiquitous plant–fungus mutualists in terrestrial ecosystems and play important roles in plant resource capture and nutrient cycling. Sporadic evidence suggests that anthropogenic nitrogen (N) input may impact the development and the functioning of arbuscular mycorrhizal (AM) fungi, potentially altering host plant growth and soil carbon (C) dynamics. In this study, we examined how mineral N inputs affected mycorrhizal mediation of plant N acquisition and residue decomposition in a microcosm system. Each microcosm unit was separated into HOST and TEST compartments by a replaceable mesh screen that either prevented or allowed AM fungal hyphae but not plant roots to grow into the TEST compartments. Wild oat (Avena fatua L.) was planted in the HOST compartments that had been inoculated with either a single species of AM fungus, Glomus etunicatum, or a mixture of AM fungi including G. etunicatum. Mycorrhizal contributions to plant N acquisition and residue decomposition were directly assessed by introducing a mineral ¹⁵N tracer and ¹³C‐rich residues of a C₄ plant to the TEST compartments. Results from ¹⁵N tracer measurements showed that AM fungal hyphae directly transported N from the TEST soil to the host plant. Compared with the control with no penetration of AM fungal hyphae, AM hyphal penetration led to a 125% increase in biomass ¹⁵N of host plants and a 20% reduction in extractable inorganic N in the TEST soil. Mineral N inputs to the HOST compartments (equivalent to 5.0 g N m^?2 yr^?1) increased oat biomass and total root length colonized by mycorrhizal fungi by 189% and 285%, respectively, as compared with the no‐N control. Mineral N inputs to the HOST plants also reduced extractable inorganic N and particulate residue C proportion by 58% and 12%, respectively, in the corresponding TEST soils as compared to the no‐N control, by stimulating AM fungal growth and activities. The species mixture of mycorrhizal fungi was more effective in facilitating N transport and residue decomposition than the single AM species. These findings indicate that low‐level mineral N inputs may significantly enhance nutrient cycling and plant resource capture in terrestrial ecosystems via stimulation of root growth, mycorrhizal functioning, and residue decomposition. The long‐term effects of these observed alterations on soil C dynamics remain to be investigated.     

Arbuscular mycorrhizal fungi can transfer substantial amounts of nitrogen to their host plant from organic material

Nitrogen (N) capture by arbuscular mycorrhizal (AM) fungi from organic material is a recently discovered phenomenon. This study investigated the ability of two Glomus species to transfer N from organic material to host plants and examined whether the ability to capture N is related to fungal hyphal growth. Experimental microcosms had two compartments; these contained either a single plant of Plantago lanceolata inoculated with Glomus hoi or Glomus intraradices, or a patch of dried shoot material labelled with (15)N and (13)carbon (C). In one treatment, hyphae, but not roots, were allowed access to the patch; in the other treatment, access by both hyphae and roots was prevented. When allowed, fungi proliferated in the patch and captured N but not C, although G. intraradices transferred more N than G. hoi to the plant. Plants colonized with G. intraradices had a higher concentration of N than controls. Up to one-third of the patch N was captured by the AM fungi and transferred to the plant, while c. 20% of plant N may have been patch derived. These findings indicate that uptake from organic N could be important in AM symbiosis for both plant and fungal partners and that some AM fungi may acquire inorganic N from organic sources.     

Tracking carbon from the atmosphere to the rhizosphere

Turnover rates of arbuscular mycorrhizal (AM) fungi may influence storage of soil organic carbon (SOC). We examined the longevity of AM hyphae in monoxenic cultures; and we also used ¹³C incorporation into signature fatty acids to study C dynamics in a mycorrhizal symbiosis involving Glomus intraradices and Plantago lanceolata. ¹³C enrichment of signature fatty acids showed rapid transfer of plant assimilates to AM fungi and a gradual release of C from roots to rhizosphere bacteria, but at a much slower rate. Furthermore, most C assimilated by AM fungi remained 32 days after labelling. These findings indicate that ¹³C labelled fatty acids can be used to track C flux from the atmosphere to the rhizosphere and that retention of C in AM fungal mycelium may contribute significantly to SOC.     

Cooccurring plants forming distinct arbuscular mycorrhizal morphologies harbor similar AM fungal species

Arbuscular mycorrhizal (AM) fungal spores were isolated from field transplants and rhizosphere soil of Hedera rhombea (Miq) Bean and Rubus parvifolius L., which form Paris-type and Arum-type AM, respectively. DNA from the spore isolates was used to generate molecular markers based on partial large subunit (LSU) ribosomal RNA (rDNA) sequences to determine AM fungi colonizing field-collected roots of the two plant species. Species that were isolated as spores and identified morphologically and molecularly were Gigaspora rosea and Scutellospora erythropa from H. rhombea, Acaulospora longula and Glomus etunicatum from R. parvifolius, and Glomus claroideum from both plants. The composition of the AM fungal communities with respect to plant trap cultures was highly divergent between plant species. Analysis of partial LSU rDNA sequences amplified from field-collected roots of the two plant species with PCR primers designed for the AM fungi indicated that both plants were colonized by G. claroideum, G. etunicatum, A. longula, and S. erythropa. G. rosea was not detected in the field-collected roots of either plant species. Four other AM fungal genotypes, which were not isolated as spores in trap cultures from the two plant species, were also found in the roots of both plant species; two were closely related to Glomus intraradices and Glomus clarum. One genotype, which was most closely related to Glomus microaggregatum, was confined to R. parvifolius, whereas an uncultured Glomeromycotan fungus occurred only in roots of H. rhombea. S. erythropa was the most dominant fungus found in the roots of H. rhombea. The detection of the same AM fungal species in field-collected roots of H. rhombea and R. parvifolius, which form Paris- and Arum-type AM, respectively, shows that AM morphology in these plants is strongly influenced by the host plant genotypes as appears to be the case in many plant species in natural ecosystems, although there are preferential associations between the hosts and colonizing AM fungi in this study.     

Variation in aluminum resistance among arbuscular mycorrhizal fungi

Arbuscular mycorrhizal (AM) fungi mediate interactions between plants and soils, and are important where nutrient or metal concentrations limit plant growth. Variation in fungal response to edaphic conditions may influence the effectiveness of the plant-mycorrhizal association in some soil environments. Andropogon virginicus (broomsedge) colonizes disturbed sites in the eastern United States, including acidic mine soils where aluminum (Al) is phytotoxic, and Al resistance in broomsedge has been associated with colonization by the AM fungus Glomus clarum. In the present study, inter- and intra-specific variation to confer Al resistance to broomsedge was assessed among selected species of AM fungi. Broomsedge seeds were grown in sand culture inoculated with one of five isolates of three species of fungi (G. clarum, Acaulospora morrowiae, and Scutellospora heterogama). Plants were exposed to 0 or 400 µM Al in nutrient solution and harvested after 4 or 9 weeks of growth. Mean infection percentage, plant biomass, and plant tissue Al and phosphorus (P) concentrations were measured. G. clarum conferred the greatest Al resistance to broomsedge, with the lowest variability among isolates for colonization and growth inhibition by Al [tolerance indices (TI) between 22.4 and 92.7%]. Broomsedge plants colonized by A. morrowiae were consistently the most sensitive to Al, with little variation among isolates (TI between 1.6 and 12.1%). Al resistance by S. heterogama isolates was intermediate and wide-ranging (TI between 3.9 and 40.0%). Across all AM fungal isolates, resistance was associated with high rates of colonization and low tissue Al concentrations of broomsedge plants. The functional diversity in Al resistance displayed by these AM fungi reflect variation in acclimation mechanisms operating in the mycorrhizal symbiosis under environmental stress.     

Phytate utilization of maize mediated by different nitrogen forms in a plant–arbuscular mycorrhizal fungus–phosphate-solubilizing bacterium system

A pot experiment was conducted to investigate the organic phosphorus (P) (phytate) utilization of Zea mays L. with different nitrogen (N) forms (NH₄⁺ and NO₃^?) when both arbuscular mycorrhizal (AM) fungus (Funelliformis mosseae) and phosphate-solubilizing bacterium (PSB, Pseudomonas alcaligenes) are present. The soil was supplied with either KNO₃ or (NH₄)₂SO₄ (200 mg kg^?1 N) with or without phytin (75 mg P kg^?1). Results showed that the application of NH₄⁺ to the soil in a plant–AM fungus–PSB system decreased rhizosphere pH and increased phosphatase activity. It also enhanced the mineralization rate of phytin, which resulted in the release of more inorganic P. The application of NO₃^? promoted mycorrhizal colonization and hyphal length density in the soil. The inorganic P in the hyphosphere decreased, but more P was transferred to the plant through the mycorrhizal hyphae. Hence, in addition, the application of the two different N forms did not significantly alter the content of plant P. The plant supplied with different N fertilizers acquired P through different mechanisms associated with other microbes. NH₄⁺ application promoted phytin mineralization by decreasing soil pH, whereas NO₃^? application increased inorganic P uptake by strengthening the mycorrhizal pathway.     

Interactions between Lotus japonicus genotypes and arbuscular mycorrhizal fungi

Abstract

Interactions between three genotypes (Ljsym 71-1, Ljsym 71-2 and Ljsym 72) of Lotus japoicus and one isolate from each of four species of arbuscular mycorrhizal fungi (Glomus sp. R-10, Glomus intraradices, Glomus etunicatum, and Gigaspora margarita) were investigated and compared with the wild-type ‘Gifu’ B-129. All the three genotypes showed no or defective internal colonization after inoculation with these AM fungi. In Ljsym72 mutant, the AM fungi produced deformed appressoria on the root surface, but failed to form any internal structures (internal hyphae, arbuscules and vesicles) except only in Glomus intraradices. The Ljsym71-1 and Ljsym71-2 mutants had more deformed appressoria and occasionally formed internal hyphae, arbuscules and vesicles, depending on AM fungi used. Wild-type ‘Gifu’ (nod⁺myc⁺) plants had typical colonization. The colonization of mutants by several fungi varied and provides a basis for studying recognition and compatibility between plants and mycorrhizal fungal species. These mutants also will be useful in studies of the genetics of the symbiosis between plant species and AM fungi.     

Soil�Cstrain compatibility: the key to effective use of arbuscular mycorrhizal inoculants?

Consistency of response to arbuscular mycorrhizal (AM) inoculation is required for efficient use of AM fungi in plant production. Here, we found that the response triggered in plants by an AM strain depends on the properties of the soil where it is introduced. Two data sets from 130 different experiments assessing the outcome of a total of 548 replicated single inoculation trials conducted either in soils with a history of (1) high input agriculture (HIA; 343 replicated trials) or (2) in more pristine soils from coffee plantations (CA; 205 replicated trials) were examined. Plant response to inoculation with different AM strains in CA soils planted with coffee was related to soil properties associated with soil types. The strains Glomus fasciculatum-like and Glomus etunicatum-like were particularly performant in soil relatively rich in nutrients and organic matter. Paraglomus occultum and Glomus mosseae-like performed best in relatively poor soils, and G. mosseae and Glomus manihotis did best in soils of medium fertility. Acaulospora scrobiculata, Diversispora spurca, G. mosseae-like, G. mosseae and P. occultum stimulated coffee growth best in Chromic, Eutric Alluvial Cambisol, G. fasciculatum-like and G. etunicatum-like in Calcaric Cambisol and G. manihotis, in Chromic, Eutric Cambisols. Acaulospora scrobiculata and Diversispora spurca strains performed best in Chromic Alisols and Rodic Ferralsols. There was no significant relationship between plant response to AM fungal strains and soil properties in the HIA soil data set, may be due to variation induced by the use of different host plant species and to modification of soil properties by a history of intensive production. Consideration of the performance of AM fungal strains in target soil environments may well be the key for efficient management of the AM symbiosis in plant production.     

Accumulation of reactive oxygen species in arbuscular mycorrhizal roots

We investigated the accumulation of reactive oxygen species (ROS) in arbuscular mycorrhizal (AM) roots from Medicago truncatula, Zea mays and Nicotiana tabacum using three independent staining techniques. Colonized root cortical cells and the symbiotic fungal partner were observed to be involved in the production of ROS. Extraradical hyphae and spores from Glomus intraradices accumulated small levels of ROS within their cell wall and produced ROS within the cytoplasm in response to stress. Within AM roots, we observed a certain correlation of arbuscular senescence and H₂O₂ accumulation after staining by diaminobenzidine (DAB) and a more general accumulation of ROS close to fungal structures when using dihydrorhodamine 123 (DHR 123) for staining. According to electron microscopical analysis of AM roots from Z. mays after staining by CeCl₃, intracellular accumulation of H₂O₂ was observed in the plant cytoplasm close to intact and collapsing fungal structures, whereas intercellular H₂O₂ was located on the surface of fungal hyphae. These characteristics of ROS accumulation in AM roots suggest similarities to ROS accumulation during the senescence of legume root nodules.     

Effects of mycorrhizal fungus isolates on mineral acquisition by Panicum virgatum in acidic soil

 Plant ability to withstand acidic soil mineral deficiencies and toxicities can be enhanced by root-arbuscular mycorrhizal fungus (AMF) symbioses. The AMF benefits to plants may be attributed to enhanced plant acquisition of mineral nutrients essential to plant growth and restricted acquisition of toxic elements. Switchgrass (Panicum virgatum L.) was grown in pH_Ca (soil:10 mM CaCl₂, 1 : 1) 4 and 5 soil (Typic Hapludult) inoculated with Glomus clarum, G. diaphanum, G. etunicatum, G. intraradices, Gigaspora albida, Gi. margarita, Gi. rosea, and Acaulospora morrowiae to determine differences among AMF isolates for mineral acquisition. Shoots of mycorrhizal (AM) plants had 6.2-fold P concentration differences when grown in pH_Ca 4 soil and 2.9-fold in pH_Ca 5 soil. Acquisition trends for the other mineral nutrients essential for plant growth were similar for AM plants grown in pH_Ca 4 and 5 soil, and differences among AMF isolates were generally higher for plants grown in pH_Ca 4 than in pH_Ca 5 soil. Both declines and increases in shoot concentrations of N, S, K, Ca, Mg, Zn, Cu, and Mn relative to nonmycorrhizal (nonAM) plants were noted for many AM plants. Differences among AM plants for N and Mg concentrations were relatively small (<2-fold) and were large (2- to 9-fold) for the other minerals. Shoot concentrations of mineral nutrients did not relate well to dry matter produced or to percentage root colonization. Except for Mn and one AMF isolate, shoot concentrations of Mn, Fe, B, and Al in AM plants were lower than in nonAM plants, and differences among AM plants for these minerals ranged from a low of 1.8-fold for Fe to as high as 6.9-fold for Mn. Some AMF isolates were effective in overcoming acidic soil mineral deficiency and toxicity problems that commonly occur with plants grown in acidic soil. Accepted: 14 June 1999     

A preliminary survey of the arbuscular mycorrhizal status of grassland plants in southern Tibet

We report for the first time the arbuscular mycorrhizal (AM) status of native plant species and AM fungal diversity in the grasslands of southern Tibet. A total of 51 soil samples were collected from the rhizospheres of the dominant plant species, and AM fungal structures were observed in 18 (82%) of 22 plant species examined. Vesicles and aseptate hyphae were the structures most frequently observed in the plant roots. After trap culture for 5 months, 25 AM fungal taxa were identified in the soil samples collected, of which nine belonged to Glomus, ten to Acaulospora, one to Entrophospora and five to Scutellospora. The frequency of occurrence of different genera and species varied greatly. Glomus was the dominant genus, and the most frequent and abundant species was Glomus mosseae. Over the whole sampling area, spore density in the rhizosphere soil of different host plant species ranged from 2 to 66 per 20 g air-dried soil. Overall AM fungal species richness was 2.10 and species diversity was 2.35. AM fungal diversity was also compared among the four different land use types (farmland and normal, disturbed and highly disturbed montane scrub grassland). Spore densities in the farmland and normal grassland were much higher than in the grasslands that had been degraded to varying degrees. The species richness in normal grassland was the highest of the four land use types examined. Species diversity varied from 1.99 to 0.94 and was highest in normal grassland, intermediate in degraded grassland and farmland, and lowest in the highly disturbed grassland.     

Effects of the mycorrhizal fungus Glomus intraradices on uranium uptake and accumulation by Medicago truncatula L. from uranium-contaminated soil

Phytostabilization strategies may be suitable to reduce the dispersion of uranium (U) and the overall environmental risks of U-contaminated soils. The role of Glomus intraradices, an arbuscular mycorrhizal (AM) fungus, in such phytostabilization of U was investigated with a compartmented plant cultivation system facilitating the specific measurement of U uptake by roots, AM roots and extraradical hyphae of AM fungi and the measurement of U partitioning between root and shoot. A soil-filled plastic pot constituted the main root compartment (C_A) which contained a plastic vial filled with U-contaminated soil amended with 0, 50 or 200 mg KH₂PO₄−P kg^–1soil (C_B). The vial was sealed by coarse or fine nylon mesh, permitting the penetration of both roots and hyphae or of just hyphae. Medicago truncatula plants grown in C_A were inoculated with G. intraradices or remained uninoculated. Dry weight of shoots and roots in C_A was significantly increased by G. intraradices, but was unaffected by mesh size or by P application in C_B. The P amendments decreased root colonization in C_B, and increased P content and dry weight of those roots. Glomus intraradices increased root U concentration and content in C_A, but decreased shoot U concentrations. Root U concentrations and contents were significantly higher when only hyphae could access U inside C_B than when roots could also directly access this U pool. The proportion of plant U content partitioned to shoots was decreased by root exclusion from C_B and by mycorrhizas (M) in the order: no M, roots in C_B > no M, no roots in C_B > M, roots in C_B > M, no roots in C_B. Such mycorrhiza-induced retention of U in plant roots may contribute to the phytostabilization of U contaminated environments.     

Influence of two legume species on hyphal production and activity of two arbuscular mycorrhizal fungi

 Two arbuscular mycorrhizal (AM) fungi (Glomus mosseae and G. intraradices) were compared for abundance of intraradical and soil-borne hyphae in association with Astragalus sinicum, a small-seeded, and Glycine max, a large-seeded legume. A. sinicum was more responsive than G. max to mycorrhizal formation, especially at early growth stages. Biomass allocation was greater in roots than shoots for mycorrhizal A. sinicum, while the opposite was true for G. max. Hyphal development in root and soil compartments was estimated by trypan blue staining and after staining for succinate dehydrogenase (SDH) or alkaline phosphatase (ALP) activity. Total fungal abundance increased steadily in roots and soil with time to a maximum 8 weeks after planting. SDH- and ALP-active AM hyphae increased in roots during plant growth but decreased in soil at later harvests. Mycorrhizal root mass in A. sinicum and G. max increased about 14-fold and 2.5-fold, respectively, but total length of soil hyphae produced per plant differed little, so that the pattern of AM soil to root abundance of the two fungi varied considerably with the host plant. Accepted: 23 July 1997     

Depletion of soil mineral N by roots of Cucumis sativus L. colonized or not by arbuscular mycorrhizal fungi

Two experiments were conducted where Cucumis sativus were grown in uncompartmented pots either alone or in symbiosis with Glomus intraradices Schenck and Smith (Experiment 1) or Glomus sp. (Experiment 2) in order to investigate if root colonization by arbuscular mycorrhizal (AM) fungi has an effect on depletion of the soil mineral N pool. All pots were gradually supplied with 31 mg NH₄NO₃-N kg^-1 dry soil from 12–19 days after planting and an additional 50 mg (NH₄)₂SO₄-N kg^-1 dry soil (¹⁵N-labelled in Experiment 1) was supplied at 21 or 22 days after planting in Experiments 1 and 2, respectively. Dry weight of plant parts, total root length, mycorrhizal colonization rate and soil concentration of NH ₄ ⁺ and NO ₃ ^- were recorded at five sequential harvest events: 21, 24, 30, 35 and 42 days (Experiment 1) and 22, 25, 28, 31 and 35 days (Experiment 2) after planting. In Experiment 1, plants were also analysed for total content of N and ¹⁵N. The mycorrhizal colonization rate increased during time: from 25 to 40% in Experiment 1 and from 50 to 60% in Experiment 2. Plant dry matter accumulation was unaffected by mycorrhizal colonization, except in Experiment 1 where shoot dry weights were slightly increased and in Experiment 2 where root dry weights were slightly decreased compared to non-mycorrhizal control plants. The total root length was similar in the control and mycorrhizal treatments in Experiment 1, while it was decreased (20–30%) by mycorrhizal colonization in the last two harvest in Experiment 2. Mycorrhizal colonization affected the rate of depletion of soil mineral N in Experiment 1, where both NH ₄ ⁺ and NO ₃ ^- concentrations were markedly lower in the first two harvests, when plants were mycorrhizal. As the root length was similar in mycorrhizal and control treatments, this may indicate that the external AM hyphae contributed to the depletion of the soil mineral N pool. A similar pattern was observed in Experiment 2, although the effect was less pronounced. The ¹⁵N enrichment in mycorrhizal plants (Experiment 1) also indicated a faster NH ₄ ⁺ uptake than in the non-mycorrhizal controls in the first two harvests after application of the ¹⁵N-labelled N source. However, the external hyphae and roots seemed to have access to the same N sources as the ¹⁵N enrichment and total N content were similar in mycorrhizal and control plants at the end of the experiment. This revised version was published online in June 2006 with corrections to the Cover Date.     

Changes in arbuscular mycorrhizal fungus community along an exotic plant Eupatorium adenophorum invasion in a chinese secondary forest

Knowledge of the changes in arbuscular mycorrhizal (AM) fungi is fundamental for understanding the success of exotic plant invasions in natural ecosystems. In this study, AM fungal colonization and spore community were examined along an invasive gradient of the exotic plant Eupatorium adenophorum in a secondary forest in southwestern China. With increasing E. adenophorum invasion, the density of arbuscules in the roots of E. adenophorum significantly increased, but the AM root colonization rate and the densities of vesicles and hyphal coils in roots of E. adenophorum were not significantly different. A total of 29 AM fungi belonging to nine genera were identified based on spore morphology. Claroideoglomus etunicatum, Funneliformis geosporus, and Glomus aggregatum were the most common AM fungal species. The E. adenophorum invasion significantly decreased the AM fungal spore density in the soil. Furthermore, with increasing of E. adenophorum invasion the spore densities of C. etunicatum, G. aggregatum, and G. arenarium significantly decreased, whereas F. geosporus significantly increased. Nonmetric multidimensional scaling demonstrated that the AM fungus community composition was significantly different (P=0.003) in the different invasive levels of E. adenophorum, and significantly correlated with plant species richness, soil total P, and soil NO₃ ^?-N. The results suggest that the alteration in AM fungus community might be caused by E. adenophorum invasion via changing the local plant community and soil properties in a Chinese secondary forest ecosystem.     

Contribution by two arbuscular mycorrhizal fungi to P uptake by cucumber (Cucumis sativus L.) from32P-labelled organic matter during mineralization in soil

An experiment was set up to investigate the role of arbuscular mycorrhiza (AM) in utilization of P from organic matter during mineralization in soil. Cucumber (Cucumis sativus L.) inoculated with one of two AM fungi or left uninoculated were grown for 30 days in cross-shaped PVC pots. One of two horizontal compartments contained 100 g soil (quartz sand: clay loam, 1:1) with 0.5 g ground clover leaves labelled with³²P. The labelled soil received microbial inoculum without AM fungi to ensure mineralization of the added organic matter. The labelling compartment was separated from a central root compartment by either 37 m or 700 m nylon mesh giving only hyphae or both roots and hyphae, respectively, access to the labelled soil. The recovery of³²P from the hyphal compartment was 5.5 and 8.6% for plants colonized withGlomus sp. andG. caledonium, respectively, but only 0.6 % for the non-mycorrhizal controls. Interfungal differences were not related to root colonization or hyphal length densities, which were lowest forG. caledonium. Both fungi depleted the labelled soil of NaHCO₃-extractable P and³²P compared to controls. A 15–25% recovery of³²P by roots was not enhanced in the presence of mycorrhizas, probably due to high root densities in the labelled soil. The experiment confirms that AM fungi differ in P uptake characteristics, and that mycorrhizal hyphae can intercept some P immobilization by other microorganisms and P-sorbing clay minerals.     

Hyphal N transport by a vesicular-arbuscular mycorrhizal fungus associated with cucumber grown at three nitrogen levels

Cucumis sativus L. cv. Aminex (F1 hybrid) was grown alone or in symbiosis with Glomus intraradices Schenck and Smith in containers with two hyphal compartments (HC_A and HC_B) on either side of a root compartment (RC) separated by fine nylon mesh. Plants received a total of either 100, 200 or 400 mg N which were applied gradually to the RC during the experiment. ¹⁵N was supplied to HC_A 42 d after plating, at 50 mg ¹⁵NH₄ ⁺-N kg^–1 soil. Lateral movement of the applied ¹⁵N towards the roots was minimized by using a nitrification inhibitor and a hyphal buffer compartment.Non-mycorrhizal controls contained only traces of ¹⁵N after a 27 d labelling period irrespective of the amount of N supplied to the RC. In contrast, 49, 48 and 27% of the applied ¹⁵N was recovered in mycorrhizal plants supplied with 100, 200 and 400 mg N, respectively. The plant dry weight was increased by mycorrhizal colonization at all three levels of N supply, but this effect was strongest in plants of low N status. The results indicated that this increase was due partly to the improved inflow of N via the external hyphae. Root colonization by G. intraradices was unaffected by the amount of N supplied to the RC, while hyphal length increased in HC_A compared to HC_B. Although a considerable ¹⁵N content was detected in mycorrhizal roots adjacent to HC_B, only insignificant amounts of ¹⁵N were found in the external hyphae in HC_B. The external hyphae depleted the soil of inorganic N in both HC_A and HC_B, while the concentration of soil mineral N was still high in non-mycorrhizal containers at harvest. An exception was plants supplied with 400 mg N, where some inorganic N was present at 5 cm distance from the RC in HC_A. The possibility of a regulation mechanism for hyphal transport of N is discussed.     

Effect of the saprophytic fungus Fusarium oxysporum on arbuscular mycorrhizal colonization and growth of plants in greenhouse and field trials

Effects of the saprophytic fungus Fusarium oxysporum on arbuscular mycorrhizal (AM) colonization and plant dry matter were studied in greenhouse and field experiments. Host plants: maize (Zea mays L.), sorghum (Sorghum vulgare L.), lettuce (Lactuca sativa L.), tomato (Lycopersicum esculentum L.), wheat (Triticum vulgare L), lentil (Ervum lens L.) and pea (Pisum sativum L.), the AM fungi: Glomus mosseae, G. fasciculatum, G. intraradices, G. clarum, and G. deserticola and the carriers for F. oxysporum inoculum: aqueous solution, thin agar slices, and pellets of agar and alginate were tested under greenhouse conditions. Greatest plant growth and AM colonization responses in sterilized and unsterilized soils were observed with pea, Glomus deserticola and sodium alginate pellets as the carrier for F. oxysporum inoculum. Under field conditions, adding F. oxysporum increased the survival of transplanted pea, possibly through a beneficial effect on AM fungi. Application of F. oxysporum increased shoot dry matter, N and P concentrations of pea and sorghum plants, and the level of AM colonization attained by indigenous or introduced AM fungi. These parameters were similar in plants inoculated with either G. deserticola or with the indigenous AM fungi. Application of the saprophytic fungus increased the number of propagules of AM fungi in field plots in which pea was grown, but this increase was not sufficient to increase AM colonization of sorghum after the pea crop. This revised version was published online in June 2006 with corrections to the Cover Date.     

Identification of arbuscular mycorrhizal fungi in soils and roots of plants colonizing zinc wastes in southern Poland

 Analysis of the community of arbuscular mycorrhizal (AM) fungi in roots of Fragaria vesca growing in a heavy metal contaminated site was carried out on a Zn waste site near Chrzanow (southern Poland). The waste substratum was characterized by high contents of Pb, Zn, Cd, Cu and As, and by low levels of N, P and organic matter. Spores of Glomales were isolated by wet sieving and DNA was isolated from individual spores. Nested polymerase chain reaction (PCR) with taxon-specific primers was used to identify the species Glomus mosseae, Glomus intraradices and Glomus claroideum. Spores of other fungi were morphologically characterized and new taxon-discriminating molecular probes were developed for two of them (Glomus sp. HM-CL4 and HM-CL5) based on variations in the large ribosomal subunit (25S rDNA). High sequence similarities were found between Glomus sp. HM-CL4 and Glomus gerdemanii, and between Glomus sp. HM-CL5 and Glomus occultum. The designed primers were used to characterize the population of AM fungi colonizing the roots of F. vesca collected from the Zn waste site. The analysis, carried out on roots stained with trypan blue, showed that the most effective colonizer was closely related to G. gerdemannii. G. claroideum and the G. occultum-like fungus were slightly less common whilst frequencies of G. intraradices and G. mosseae in roots were much lower. The analysis of mycorrhiza stained with rhodizoniate to localize heavy metal accumulation showed that the stain does not influence the PCR reaction. Seventy percent of the root samples containing positively stained fungal hyphae were found to be colonized by G. mosseae. The data obtained demonstrate the usefulness of nested PCR for studies carried out in polluted areas. It will enable selection of AM fungi which are able to colonize plant roots under heavy metal stress conditions, as well as the identification of fungi showing high in situ accumulation of potentially toxic elements. Accepted: 7 July 2000     

Soil nitrogen transformations under Populus tremuloides, Betula papyrifera and Acer saccharum following 3 years exposure to elevated CO2 and O3

Increases in atmospheric CO₂ and tropospheric O₃ may affect forest N cycling by altering plant litter production and the availability of substrates for microbial metabolism. Three years following the establishment of our free‐air CO₂–O₃ enrichment experiment, plant growth has been stimulated by elevated CO₂ resulting in greater substrate input to soil; elevated O₃ has counteracted this effect. We hypothesized that rates of soil N cycling would be enhanced by greater plant productivity under elevated CO₂, and that CO₂ effects would be dampened by O₃. We found that elevated CO₂ did not alter gross N transformation rates. Elevated O₃ significantly reduced gross N mineralization and microbial biomass N, and effects were consistent among species. We also observed significant interactions between CO₂ and O₃: (i) gross N mineralization was greater under elevated CO₂ (1.0 mg N kg^?1 day^?1) than in the presence of both CO₂ and O₃ (0.5 mg N kg^?1 day^?1) and (ii) gross NH₄⁺ immobilization was also greater under elevated CO₂ (0.8 mg N kg^?1 day^?1) than under CO₂ plus O₃ (0.4 mg N kg^?1 day^?1). We used a laboratory ¹⁵N tracer method to quantify transfer of inorganic N to organic pools. Elevated CO₂ led to greater recovery of NH₄⁺‐¹⁵N in microbial biomass and corresponding lower recovery in the extractable NO₃^? pool. Elevated CO₂ resulted in a substantial increase in NO₃^?‐¹⁵N recovery in soil organic matter. We observed no O₃ main effect and no CO₂ by O₃ interaction effect on ¹⁵N recovery in any soil pool. All of the above responses were most pronounced beneath Betula papyrifera and Populus tremuloides, which have grown more rapidly than Acer saccharum. Although elevated CO₂ has increased plant productivity, the resulting increase in plant litter production has yet to overcome the influence of the pre‐existing pool of soil organic matter on soil microbial activity and rates of N cycling. Ozone reduces plant litter inputs and also appears to affect the composition of plant litter in a way that reduces microbial biomass and activity.