Defoliation effects on the community structure of arbuscular mycorrhizal fungi based on 18S rDNA sequences

The effects of defoliation on arbuscular mycorrhizal (AM) associations in the field were investigated in terms of the community structure of AM fungi colonizing roots of grassland plants; the carbohydrate balance of the host plants was also determined. We focused on two plant species dominating Japanese native grasslands: the grazing-intolerant species Miscanthus sinensis and the grazing-tolerant species Zoysia japonica. Community structures of AM fungi were determined from 18S rRNA gene sequences. The dominant fungal group in both plant species was the Glomus clade, which was classified into several phylogenetic groups based on genetic distances and topology. In Miscanthus roots, the Glomus-Ab, Glomus-Ac, and Glomus-Ad groups were detected almost equally. In Zoysia roots, the Glomus-Ab group was dominant. Defoliation effects on the community structure of AM fungi differed between the plant species. In Miscanthus roots, the percentage of root length colonized (%RLC) by the Glomus-Ac and Glomus-Ad groups was significantly reduced by defoliation treatment. On the other hand, AM fungal group composition in Zoysia roots was unaffected by defoliation except on the last sampling date. Decreased %RLC by Glomus-Ac and Glomus-Ad coincided with decreased non-structural carbohydrate (NSC) levels in host plants; also, significant positive correlations were found between the %RLC and some NSC levels. On the other hand, the %RLC by Glomus-Ab in both plant species was unaffected by the NSC level. These results suggest that AM fungal groups have different carbohydrate requirements from host plants.Electronic Supplementary Material Supplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s00572-003-0286-x     

Arbuscular Mycorrhizal Fungi (AMF) and Biosolids Enhance the Growth of a Native Australian Grass on Sulphidic Gold Mine Tailings

We tested the effect of the addition of biosolids combined with a native mycorrhizal inoculum (Arbuscular Mycorrhizal Fungi [AMF]) on growth of a native Australian grass, and on trace element stabilization of sulphidic gold mine tailings. A glasshouse trial was established on four substrates: tailings (T); tailings with a layer of 5 cm topsoil (TS); tailings amended with 100 dry t ha^?1 biosolids (LB), and tailings amended with 500 dry t ha^?1 biosolids (HB). Pots of 1.2 L of capacity were established; some were inoculated with a mixture of Glomus sp. (WUM51–9227), Scutelospora aurigloba (WUM51–53), and Acaulospora levis (WUM46) culture mix, and others were uninoculated controls. Seeds of the native Australian grass, Bothriochloa macra were sown in the pots. Root infection, plant biomass production, nutrients and trace element concentrations in shoots were investigated. Addition of biosolids significantly increased AMF infection of roots compared to unamended substrates. No clear qualitative differences in colonization were detected. Addition of biosolids and AMF together clearly improved the establishment and growth of the native grass. Similar trends in nutritional status were shown for biosolids and inoculation with AMF treatments. Mycorrhizal inoculation increased plant biomass production and the effectiveness of nutrient uptake. The combined use of biosolids and mycorrhizal inoculation could be a reliable method for phytostabilization purposes in polluted substrates.     

Phosphorus Acquisition Strategies within Arbuscular Mycorrhizal Fungal Community of a Single Field Site

Diversity in phosphorus (P) acquisition strategies was assessed among eight isolates of arbuscular mycorrhizal fungi (AMF) belonging to three Glomus species, all obtained from the same field site. Maize (Zea mays L. cv. Corso) was used as a test plant. Compartmented cultivation containers coupled with ³³P radioisotope labeling of soil P were employed to estimate (1) the distance from the roots that AMF were able to acquire soil P from, (2) the rate of soil colonization, (3) the efficiency of uptake of soil P by AMF, (4) benefits provided to maize in terms of P acquisition and growth. Glomus mosseae and G. intraradices took up P 10 cm from roots, whereas G. claroideum only up to 6 cm from the roots. G. mosseae most rapidly colonized the available soil volume and transported significant amounts of P to maize from a distance, but provided no net P uptake benefit to the plants. On the other hand, both G. intraradices and three out of four G. claroideum isolates significantly improved net P uptake by maize. These effects seem to be related to variability between and to a limited extent also within AMF species, in mycelium development, efficiency of hyphal P uptake and effects on plant P acquisition via the root pathway. In spite of absence of maize growth responses to inoculation with any of the AMF isolates, this study indicates remarkable functional diversity in the underground component of the studied field site.     

Time-course study and partial characterization of a protein on hyphae of arbuscular mycorrhizal fungi during active colonization of roots

Material on the surface of hyphal walls of arbuscular mycorrhizal fungi (AMF) during active colonization of plant roots was detected by a monoclonal antibody. Pot-cultured isolates of Glomus, Acaulospora, Gigaspora, Scutellospora, and Entrophospora had immunofluorescent material (IM) on younger, thinner, intact hyphae, but IM was scant to absent on thicker, melanized or lysing hyphae. Colonization of corn (Zea mays L.), Sudangrass (Sorghum sudanense (Piper) Staph.) or red clover (Trifolium pratense L.) was examined during 5 months of plant growth by removing cores and performing an indirect immunoassay on roots with attached hyphae. Fresh spores of some Glomus spp. had IM on the outer layer of the spore wall. Abundant IM was seen on root hairs of plants colonized by some isolates, and some IM was detected on root surfaces of all plants examined even during early colonization. After cultures were dried, hyphae, roots and spores had little to no IM. Uninoculated control roots had very rare, small patches of IM. An immunoreactive protein was extracted from hyphae of Gigaspora and Glomus isolates by using 20mM citrate (pH 7.0) at 121°C for 90 min. Gel electrophoresis profiles indicated that all isolates tested had the same banding patterns. Lectin-binding of extracted protein is suggestive of a glycoprotein. The immunofluorescence assay can be used to examine root sections for active colonization by AMF, and the potential use of the protein to quantify AMF activity in soil is discussed.     

A comparison of the development and metabolic activity of mycorrhizas formed by arbuscular mycorrhizal fungi from different genera on two tropical forage legumes

 Two glasshouse experiments were done to assess the development and metabolic activity of mycorrhizas formed by isolates of arbuscular mycorrhizal fungi (AMF) from three different genera, Acaulospora, Gigaspora and Glomus on either Pueraria phaseoloides L. or Desmodium ovalifolium L. plants. The second of the two experiments included three levels of a localised phosphate source in the pots. Alkaline phosphatase (ALP), stained histochemically in the intra-radical mycelium (IRM) of AMF over sequential harvests, did not provide a direct marker for the efficiency of AMF in mobilising phosphorus (P) for plant growth and development. The ability of the extra-radical mycelium (ERM) to scavenge a localised phosphate source, determined by its extraction from buried 35-μm mesh pouches, was dependent on the species of AMF tested. This work indicates that AMF from different genera have unique patterns of mycelial development when forming mycorrhizas with tropical hosts in the presence of a localised phosphate source. AMF also appear to have different mechanisms for the control of P transfer, within the mycelium, to the host. The significance of the architecture of the ERM is discussed as well as the localisation of ALP in the IRM in determining the efficiency of AMF in terms of P accumulation in planta and subsequent growth of plants. Accepted: 19 August 1998     

Aggressiveness spectrum and genetic variability of Fusarium spp. on rice and wheat varieties

Abstract

A total of 106 Fusarium spp. were isolated from infected roots and soil samples of wheat and rice. Of the 106 isolates, 32 from wheat, and 74 from rice, were isolated. Six Fusarium spp. (F. oxysporum, F. moniliforme, F. poae, F. graminearum, F. tricinctum and F. equiseti) were identified at specie level. In aggressiveness tests Fusarium spp. root rot causing fungi were screened out into different aggressiveness classes according to disease severity scales. The aggressiveness of Fusarium spp. was studied on wheat varieties (Inqalab-91 and chakwal-86) and on rice varieties (Basmati-385 and IRRI-6) under controlled conditions. The overall total number of aggressive isolates was higher in rice than in wheat. However, the percentage of severely aggressive isolates was high in wheat, whereas the percentage of moderately and slightly aggressiveness isolates was high in rice. In rice, five isolates were non-aggressive and on wheat 17 were non-aggressive. Random Amplified Polymorphism DNAs (RAPDs) were used to study the polymorphism and genetic variations within the population of Fusarium spp. that established to study correlation between taxonomical and genetical characters of fungi. Five random primers were used P1 (5′-AGGAGGACCC-3′), P2 (5′-ACGAGGGACT-3′), PE7 (5′-AGATGCAGCC-3′), P14 (5′-CCACAGCACG-3′) and PE20 (5′-AACGGTGACC-3′). Each of the 10-mer primers produced results based on the respective banding patterns they generated in present investigations. Primers distinguished the F. oxysporum, F. moniliforme, F. graminearum, F. tricinctum, F. poa and F. equiseti. All the tested primers yielded amplification products, and that were reproducible. Although there was some intraspecific variation with primers, some strains were similar and some were different in banding pattern. In F. oxysporum, F. moniliforme, F. graminearum, F. tricinctum, F. poa and F. equiseti were seen clustered close to one another but each primer separated them unambiguously. All primer (P1, P2, P14, PE7 and PE20) combination produced 62 bands. All primers have shown interspecific and intraspecific variations in banding patterns.     

Arbuscular mycorrhizae in wheat and field pea crops on a low P soil: increased Zn-uptake but no increase in P-uptake or yield

Few field studies have investigated the contribution of arbuscular mycorrhizal fungi (AMF) to agricultural systems. In this study, the role of AMF in nutrition and yield of dryland autumn-sown wheat and field pea was examined through a 2-year crop sequence experiment on a red loam (Kandosol) in SE Australia. The soil was P-deficient and had low levels of root pathogens. In Year 1, levels of AMF were increased by growing subterranean clover or Linola^TM and decreased by growing canola or through maintenance of bare fallow with herbicides or tillage. In Year 2, hosts of AMF (wheat and field pea) and non-mycorrhizal canola were grown with 0 P or 20 kg ha^–1 of P as superphosphate. Yields of all Year 2 crops were increased by P-fertiliser. Year 1 treatment led to 2–3 fold variation in colonisation by AMF at each P-level for Year 2 wheat and field pea. High colonisation did not correspond with greater crop growth, yield, or uptake of P, K, Ca, Cu or S in wheat or field pea. However, total crop Zn-uptake and grain Zn concentration were positively correlated with colonisation by AMF, due to enhanced Zn-uptake after anthesis. For wheat, high colonisation also corresponded with reduced Mn-uptake and lower grain Mn concentrations. In a glasshouse experiment using a second P-deficient Kandosol, inoculation of wheat with Glomus intraradices and Scutellospora calospora enhanced uptake of Zn and P when no P-fertiliser was applied. We conclude that high colonisation by AMF is unimportant for productivity of the major field crops grown on the Kandosol soils that occupy large areas of cropland in temperate SE Australia, even under P-limiting conditions. Investigation of the factors that control functioning of arbuscular mycorrhizae under field conditions, especially temperature, is required.     

The effect of agricultural practices on the development of indigenous arbuscular mycorrhizal fungi. II. Studies in experimental microcosms

Two glasshouse experiments were performed to assess the development and metabolic activity of mycorrhizas formed by isolates of arbuscular mycorrhizal fungi (AMF) from three different genera, Acaulospora, Gigaspora and Glomus on Desmodium ovalifolium L. plants. In the first experiment the effect of disturbance of a pre-established extra-radical mycelium (ERM) was studied. In the second experiment the effect of phosphate addition as either organic matter (OM) or fertiliser was studied. Disturbance of a pre-established ERM reduced the formation of mycorrhizas by Gigaspora rosea (BEG111) and increased that by Glomus manihotis (BEG112) on D. ovalifolium plants. Acaulospora tuberculata (BEG41) failed to form mycorrhizas in the experiment. Either Gi. rosea (BEG111) or G. manihotis (BEG112) appeared to be the major component of the colonisation resulting from treatments with combinations of two or three of the AMF and determined the sensitivity of these treatments to disturbance of a pre-established ERM. The addition of phosphate fertiliser (10 mg P kg^-1) reduced mycorrhiza formation by each species of AMF compared with the addition of OM (10 mg P kg^-1). This work indicates that AMF from different genera respond differently to management by agricultural practices when in association with a tropical legume. Clearly, there is potential to alter the formation of mycorrhizas of AMF from different genera, through the use of agricultural practices. The significance of the development and metabolic activity of mycorrhizas formed by AMF from different genera for plant growth is discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Diversity and efficiency of arbuscular mycorrhizal fungi in soils from organic chili (Capsicum frutescens) farms

No previous studies have been conducted on the diversity and population of arbuscular mycorrhizal fungi (AMF) in relation to organically grown chili (Capsicum frutescens L.) in Thailand. This study was carried out to investigate the diversity and status of AMF populations at four organically managed farms in Ubon Ratchathani and Sisaket provinces. The effects of each AMF species on the growth and nutrient uptake of chili grown in sterile, organically managed soil were determined. Fourteen AM fungal taxa belonging to the genera Acaulospora (4 spp.), Entrophospora (1 sp.), Glomus (7 spp.) and Scutellospora (2 spp.) were found. Among these, Glomus was the dominant genus found at all sites, followed by Acaulospora. The spore density and root colonization of AMF on chili did not vary significantly among the sites. The effects of ten selected AMF species on the growth of chili showed that Gl. clarum RA0305 increased the growth, flowering, and fruit production of chili, and also increased the P uptake significantly, compared to non-mycorrhizal plants. This fungus showed the highest potential as a promoter of growth, flowering and yield in organically managed chili production.     

Accumulation of mansonones E and F in seedlings of Ulmus americana in response to inoculation with Ophiostoma ulmi

Summary The accumulation of mansonones E and F was investigated in Ulmus americana L. seedlings 5 weeks after inoculation with three aggressive and three non-aggressive isolates of Ophiostoma ulmi (Buism.) Nannf. The three non-aggressive isolates stimulated significantly more mansonone E and F accumulation than the three aggressive isolates of O. ulmi. Mansonone induction also varied within both the aggressive and the non-aggressive groups. Aggressive and non-aggressive isolates were recovered in equal frequencies from the inoculation wounds, whereas the aggressive isolates were recovered more frequently than the non-aggressive isolates 15 cm and 25 cm up the seedlings' stem. Vascular browning in the outer xylem of the seedlings correlated with mansonone E and F accumulation. Mansonone accumulation in U. americana seedlings is therefore associated with vascular browning and a reduction in fungal spread.     

A method for assessing arbuscular mycorrhizal fungi group distribution in tree roots by intergenic transcribed sequence variation

We identified five taxonomic groups of arbuscular mycorrhizal fungi (AMF) inside roots of young trees of six species of legumes and six species of non-legumes from a field site in southern Costa Rica using an AMF group-specific PCR assay of the intergenic transcribed sequence and 18S rRNA gene fragment. Assay specificity was verified by cloning and sequencing representatives from four of the five AMF groups. We found no difference in overall AMF diversity levels between legumes and non-legumes or between plant species. Some groups of AMF may associate more frequently with legumes than others, as Glomus Group A (Glomus mosseae/intradices group) representatives were detected more frequently in legumes than non-legumes relative to Glomus Group B (Glomus etunicatum/claroideum) representatives.     

Influence of arbuscular mycorrhizal fungi on growth and selenium uptake by garlic plants

The aim of this study is to investigate the effects of arbuscular mycorrhizal fungi (AMF) on garlic plants growth and the uptake of selenium (Se). Garlic plants were grown in the pots inoculated with Glomus fasciculatum and G. mosseae and maintained in a greenhouse. Three weeks after planting, the pots had received different concentrations of Se (5, 10, 15, 20, 25 mg kg^?1 of soil) in the form of selenium dioxide (SeO₂) at 3 weeks intervals up to 12 weeks. For physiological and biochemical analysis, the samples were randomly collected from five plants of each experiment. Maximum AM infection, spore population and plant biomass were observed in the roots of mycorrhizal-mediated plants without Se, and they were gradually declined in both mycorrhizal and non-mycorrhizal (NM) plants with increasing concentrations of Se. Among the two Glomus species tested, G. fasciculatum-mediated plants showed higher AM infection, spore population and plant biomass than G. mosseae. No differences were observed for the uptake of Se in mycorrhizal plants and NM plants. However, NM plants uptake more Se than mycorrhizal plants. Higher contents of total chlorophyll and sugars were observed in plants inoculated with G. fasciculatum without Se and they were decreased in the presence of Se. In contrast, increased amount of glutathione peroxidase was observed at increasing concentrations of Se up to 20 mg kg^?1. High-performance liquid chromatography data revealed that SeO₂ converted to organic form of Se as γ-glutamyl-Se-methylselenocysteine. These results are basis for further investigations on the role of AMF on plant growth and uptake of Se in crop plants.     

Inheritance of pathogenicity and cultural characters in Ceratocystis ulmi: hybridization of aggressive and non-aggressive strains

When British isolates of Ceratocystis ulmi were surveyed for compatibility type, both A- and B-types were found in the non-aggressive strain, but only the B-type in the aggressive strain. Single ascospore progeny from crosses between compatible aggressive and non-aggressive isolates showed a near-normal growth rate distribution, with a mean lying between the parents. Many grew either faster than the aggressive or slower than the non-aggressive parent. The progeny were highly variable in culture morphology and could not be classified in terms of the parental types. When inoculated into English elm they showed a marked skewness towards low pathogenicity. None approached the aggressive strain in pathogenicity. It is concluded that the above characters are under polygenic control, and that the aggressive strain could not arise from the non-aggressive by a simple mutation. The results suggest that the two strains may be reproductively isolated.     

Fern-associated arbuscular mycorrhizal fungi are represented by multiple Glomus spp.: do environmental factors influence partner identity?

Symbioses involving arbuscular mycorrhizal fungi (AMF) are among the most important ecological associations for many plant species. The diversity of AMF associated with ferns, however, remains poorly studied. Using recently designed Glomus-specific primers, we surveyed the AMF community associated with ferns from deciduous, broad-leaved second-growth forest habitats at the eastern edge of the piedmont region of central Virginia, USA. Results indicate that this molecular approach may be a useful tool for detecting AMF in ferns compared to traditional techniques based on morphology. Over 30 potential fungal ribotypes were identified from eight fern species using denaturing gradient gel electrophoresis. Fungal ribotypes were found to differ widely in terms of (1) the number of fern partners with which they interact and (2) their relative frequency within each fern. Sequence analysis of fungal isolates from three species of fern indicated that the primers were generally highly specific for Glomus species but some non-target DNA was also amplified. Cloned polymerase chain reaction (PCR) products from Polystichum acrostichoides and Osmunda regalis revealed several phylogenetically distinct Glomus species. A single Glomus species was identified in the cloned PCR products from Botrychium virginianum. These findings challenge the hypothesis that the extent or degree of fern–fungal symbiosis is somehow tied to root complexity. Environmental factors appear to influence the suite of AMF that form partnerships with ferns. Some species of fern from similar habitats associated with dissimilar fungal partners (e.g., P. acrostichoides and Athyrium filix-femina var. asplenioides), whereas others harbored uniform fungal communities (e.g., Asplenium platyneuron). The significance of these data in terms of ecological and evolutionary dynamics of the AMF–fern symbiosis is discussed. Brittany West, Jessica Brandt, and Kay Holstien contributed equally to this work.     

Differential effects of abiotic factors and host plant traits on diversity and community composition of root-colonizing arbuscular mycorrhizal fungi in a salt-stressed ecosystem

Arbuscular mycorrhizal fungi (AMF) were investigated in roots of 18 host plant species in a salinized south coastal plain of Laizhou Bay, China. From 18 clone libraries of 18S rRNA genes, all of the 22 AMF phylotypes were identified into Glomus, of which 18 and 4 were classified in group A and B in the phylogenetic tree, respectively. The phylotypes related to morphologically defined Glomus species occurred generally in soil with higher salinity. AMF phylotype richness, Shannon index, and evenness were not significantly different between root samples from halophytes vs. non-halophytes, invades vs. natives, or annuals vs. perennials. However, AMF diversity estimates frequently differed along the saline gradient or among locations, but not among pH gradients. Moreover, UniFrac tests showed that both plant traits (salt tolerance, life style or origin) and abiotic factors (salinity, pH, or location) significantly affected the community composition of AMF colonizers. Redundancy and variation partitioning analyses revealed that soil salinity and pH, which respectively explained 6.9 and 4.2 % of the variation, were the most influential abiotic variables in shaping the AMF community structure. The presented data indicate that salt tolerance, life style, and origin traits of host species may not significantly affect the AMF diversity in roots, but do influence the community composition in this salinized ecosystem. The findings also highlight the importance of soil salinity and pH in driving the distribution of AMF in plant and soil systems.     

Molecular analysis of AMF diversity in aquatic macrophytes: A comparison of oligotrophic and utra-oligotrophic lakes

This study aimed to assess AMF diversity in various plant species in lakes with low and relatively high P concentrations to elucidate possible correlations with environmental factors in order for better understanding the functioning of mycorrhizal fungi in submerged plants. A considerable diversity of AMF communities was observed in the lakes with low dissolved P concentrations, especially in the roots of Littorella uniflora. Glomus group A, Archaeospora and Acaulospora were the most frequent and diverse AMF lineages with eight, seven and four phylotypes at Littorella uniflora in at least six lakes with low dissolved P concentrations. In theses lakes, AMF were for the first time observed in the roots of J. bulbosus, a member of a family previously thought to be non-mycorrhizal. In the lakes with relatively high dissolved P concentrations, the frequency decreased from Acaulospora, found at three locations, to Archaeospora at two locations and Glomus group A and Paraglomus at one location.All chemical parameters of the surface water layer, except pH, revealed significant (p ≤ 0.01) differences between the lakes with low and relatively high dissolved P concentrations. Mean Mg²⁺, Ca²⁺, K⁺, NH₄⁺, CO₂, o-PO₄³⁻ and HCO₃⁻ were 3, 13.5, 15.7, 19.5, 31 and 42.6 times higher, respectively, in the lakes with relatively high dissolved P concentrations compared to the lakes with low dissolved P concentrations. AMF occurred more abundantly with low phosphate and high redox values in the lakes than with high phosphate and low redox values. The pH-value, the total-calcium and total-phosphorus concentrations were strongly correlated with the occurrence of Glomus phylotypes 4 and Archaeospora phylotypes 5 and 8, and a bit less with Acaulospora phylotype 4 and Archaeospora phylotype 3. In such lakes the presence of a diverse AMF community still enables the uptake of sufficient P for isoetid plant species despite the prevailing ‘ultra-oligotrophic’ conditions. As a consequence, macrophyte plant communities in lakes with relatively high dissolved P concentrations are less dependent on AMF colonization for their development.     

Simulated nitrogen deposition affects community structure of arbuscular mycorrhizal fungi in northern hardwood forests

Our previous investigation found elevated nitrogen deposition caused declines in abundance of arbuscular mycorrhizal fungi (AMF) associated with forest trees, but little is known about how nitrogen affects the AMF community composition and structure within forest ecosystems. We hypothesized that N deposition would lead to significant changes in the AMF community structure. We studied the diversity and community structure of AMF in northern hardwood forests after more than 12 years of simulated nitrogen deposition. We performed molecular analyses on maple (Acer spp.) roots targeting the 18S rDNA region using the fungal‐specific primers AM1 and NS31. PCR products were cloned and identified using restriction fragment length polymorphism (RFLP) and sequencing. N addition significantly altered the AMF community structure, and Glomus group A dominated the AMF community. Some Glomus operational taxonomic units (OTUs) responded negatively to N inputs, whereas other Glomus OTUs and an Acaulospora OTU responded positively to N inputs. The observed effect on community structure implies that AMF species associated with maples differ in their response to elevated nitrogen. Given that functional diversity exists among AMF species and that N deposition has been shown to select less beneficial fungi in some ecosystems, this change in community structure could have implications for the functioning of this type of ecosystem.     

Influence of Arbuscular Mycorrhiza and Phosphorus Supply on Polyamine Content, Growth and Photosynthesis of Plantago lanceolata

A greenhouse pot experiment with different phosphorus supply was conducted to study growth, photosynthesis and free polyamine (PA) content in Plantago lanceolata L. plants in relation to arbuscular mycorrhizal (AM) colonization. Inoculum of Glomus fasciculatum (BEG 53) was used. Inoculated plants had high colonization intensities which were related to the P supply. Non-mycorrhizal (NM) plants showed a typical yield response curve for P availability. Dry masses of mycorrhizal (M) plants were higher at the lowest soil P content than those of NM plants, but the opposite was found at the highest P supply. P contents in M plants were always higher. There were no differences in chlorophyll (Chl) concentrations (except the lowest soil P content) and ratios of variable to maximum Chl fluorescence (Fv/Fm) values between M and NM plants, whereas M plants had higher ratios of leaf area to fresh mass (A/f.m.) at low soil P contents and they had significantly higher CO₂ fixation capacities per unit leaf area. Free putrescine (Put), spermidine (Spd) and spermine (Spm) contents in NM plants were usually highest at the lowest P supply. The ratios of Put/(Spd+Spm) were identical in M and NM leaves. They were significantly higher, however, in NM roots at the two low P doses. It is concluded, that a P nutritional status might exist, below which PA concentrations and ratio are increased drastically, possibly indicating P deficiency or a certain state of plant development with a higher demand for AM symbiosis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of long-term fertilization on AM fungal community structure and Glomalin-related soil protein in the Loess Plateau of China

Arbuscular mycorrhizal fungi (AMF) are crucial for ecosystem functioning, and thus have potential use for sustainable agriculture. In this study, we investigated the impact of organic and mineral fertilizers on the AMF community composition and content of Glomalin-related soil protein (GRSP) in a field experimental station which was established in 1979, in the Loess Plateau of China. Roots and soils were sampled three times during the growing period of winter wheat in 2008. The treatments including: N (inorganic N), NP (inorganic N and P), SNP (straw, inorganic N and P), M (farmyard manure), MNP (farmyard manure, inorganic N and P), and CK (no fertilization). AMF communities of root and soil samples were analyzed using PCR-DGGE, cloning and sequencing techniques; and GRSP content was determined by Bradford assay. Our results indicated that spore density, GRSP, and AMF community varied significantly in soils of long-term fertilization plots at three different wheat growing stages. The effects of wheat growing period on AMF community in roots were much more evident than fertilization regimes. However, the diversity of AMF was low in our study field. Up to five AMF phylotypes appeared in each sample, with the overwhelming dominance of a Glomus-like phylotype affiliated to G. mosseae. GRSP content was correlated positively with organic carbon, total phosphorus, available phosphorus, soil pH, and spore densities, but correlated negatively with soil C/N (P?<?0.05). The results of our study highlight that the richness of AMF in Loess Plateau agricultural region is low, and long-term fertilization, especially amendments with manure and straw, has beneficial effects on accumulation of soil organic carbon, spore density, GRSP content, and AMF diversity. Host phenology, edaphic factors (influenced by long-term fertilization), and habitats interacted to affect the AMF community and agoecosystem functioning. Additionally, soil moisture and pH make a greater contribution than other determined soil parameters to the AMF community dynamics in such a special semi-arid agroecosystem where crops rely greatly on rainfall.     

Wheat Responses to Arbuscular Mycorrhizal Fungi in a Highly Calcareous Soil Differ from those of Clover, and Change with Plant Development and P supply

We evaluated the roles of arbuscular mycorrhizal (AM) fungi in growth and phosphorus (P) nutrition of wheat (Triticum aestivum L.) in a highly calcareous soil and compared the responses of wheat with those of clover (Trifolium subterraneum L). In the first experiment wheat (cv. Brookton) was harvested at 6 wk. Colonisation by four AM fungi was low (<20%). Clover was harvested at 8 wk. Colonisation varied with different fungi, with the highest value (52%) obtained with Glomus intraradices. Although suffering from P deficiency, non-mycorrhizal (NM) wheat grew relatively well with no added P (P0) and application of P at 100 mg kg⁻¹ (P100) increased the dry weight (DW). Shoot P concentrations increased with P application and there were positive effects of all AM fungi at P100. In contrast, NM clover grew very poorly at P0 and did not respond to P application. Clover responded positively to all AM fungi at both P levels, associated with increases in P uptake. In the second experiment colonisation by a single AM fungus (G. intraradices) of two wheat cultivars (Brookton and Krichauff) was well established at 6 wk (~50% in P0 plants) and continued to increase up to maturity (~70%), but decreased greatly at both harvests as P supply was increased (up to 150 mg P kg⁻¹: P150). Addition of P significantly increased plant growth, grain yield and P uptake irrespective of cultivar and harvest time, and the optimum soil P for grain yield was P100. In both cultivars, a growth depression in AM plants occurred at 6 wk at all P levels, but disappeared at 19 wk with added P. At P0, AM plants also produced lower grain yield (weight) per plant, but with higher P, AM plants produced higher grain yields than NM plants. There was a significant positive effect of AM on grain P concentration at P0, but not at other P levels. Brookton was somewhat more P efficient than Krichauff, and the latter responded more to AM fungi. This study showed that responses of wheat to AM inoculation and P supply were quite different from those of clover, and changed during development. Results are discussed in relation to the underlying soil properties.