Effect of peat on mycorrhizal colonization and effectiveness of the arbuscular mycorrhizal fungus Gigaspora margarita

Abstract

The influence of the addition of Chinese peat and Canadian peat on arbuscular mycorrhizal colonization, mycorrhizal effectiveness and host-plant growth was investigated in a pot experiment. Chinese peat or Canadian peat was mixed with Masa soil (weathered granite soil) at different levels (0, 25, 50, 100, 150 or 200 g kg^?1) into which an arbuscular mycorrhizal fungus (AMF) Gigaspora margarita Becker & Hall was inoculated, and seedlings of Miscanthus sinensis Anderess were planted. There was a significant increase in plant growth with increasing amounts of Chinese peat. The growth-promoting effect of the AMF on the host was enhanced when the addition of Chinese peat was increased from 25 to 100 g kg^?1. Root colonization and the number of spores proliferating increased with increases at low levels of Chinese peat (from 25 to 100 g kg^?1), and decreased gradually with higher Chinese peat increments. Although plant growth and root colonization with the addition of Canadian peat increased slightly, Canadian peat suppressed mycorrhizal effectiveness. In contrast to Canadian peat, the addition of Chinese peat improved considerably the physical and chemical properties of the soil, which might result in the promotion of AM formation and mycorrhizal effectiveness.     

Diversity of arbuscular mycorrhizal fungi in soils of yam (Dioscorea spp.) cropping systems in four agroecologies of Nigeria

The diversity of arbuscular mycorrhizal (AM) fungi in soils under a yam cropping system in four agroecologies of Nigeria was investigated. Soil samples were collected from yam fields at Onne (humid forest, high rainfall area), Ibadan (derived savanna), Abuja (Guinea savanna) and Ubiaja (humid forest, medium rainfall area). Soil characteristics, AM fungi species, spore abundance, Shannon diversity index, species richness and evenness were determined. A total of 31 AM fungi species was isolated from the four agroecologies with a range of 14–20 species found in a single location. Glomus species were the most abundant among AM fungi species with G. geosporum, G. intraradices and G. mosseae occurring in large populations in all locations. Ubiaja, which had a cassava/natural vegetation sequence before yam, had significantly higher spore abundance and species richness than the other locations, which had a yam/legumes or a maize/legume sequence before yam. However, diversity was significantly higher at Abuja, which had a maize/legume sequence with yam, than Ibadan, which had only a yam/legume sequence. The study revealed significant diversity in AM fungal species across agroecologies in yam-growing regions. Further research on the functional consequences of changing composition of AM fungi species across the region is recommended.     

Response of a wild-type and modern cowpea cultivars to arbuscular mycorrhizal inoculation in sterilized and non-sterilized soil

Cowpea is an important crop that serves as a legume and vegetable source to many smallholder farmers in sub-Saharan Africa. Soil fertility is a significant limitation to its production thus; inoculation with beneficial soil biota such as arbuscular mycorrhizal fungi (AMF) could improve its performance. However, plant–AMF interaction could vary based on crop cultivar hence affecting overall crop production. The present study aimed at determining the effect of AMF inoculation and soil sterilization on root colonization and growth of a wild-type and three modern cowpea cultivars grown by smallholder farmers in Kenya. Potted cowpea plants were inoculated with a commercial AMF inoculum comprising of Rhizophagus irregularis, Funneliformis mosseae, Glomus aggregatum and Glomus etunicatum and maintained in a greenhouse for 40 days. After harvesting, mycorrhizal colonization, nodule number and dry weight, root and shoot dry weights, nitrogen (N,) phosphorus (P) and potassium (K) content were determined. Interestingly, the modern cultivars showed significantly (p < 0.001) higher root colonization, nodulation, shoot P and N compared to the wild-type cultivar. Moreover, a strong positive correlation between AMF root colonization and shoot P (r² = 0.73, 0.90, p < 0.001), AMF root colonization and shoot N (r² = 0.78; 0.89, p < 0.001) was observed in both sterilized and non-sterilized soil, respectively. Soil sterilization affected root colonization and growth parameters with plants grown in non-sterilized soil performing better than those grown in sterilized soil. This study provides major evidence that modern cowpea cultivars are still responsive to mycorrhizal inoculation suggesting that modern breeding programs are not deleterious AMF symbiosis.     

Global diversity and distribution of arbuscular mycorrhizal fungi

Arbuscular mycorrhizal (AM) fungi form associations with most land plants and can control carbon, nitrogen, and phosphorus cycling between above- and belowground components of ecosystems. Current estimates of AM fungal distributions are mainly inferred from the individual distributions of plant biomes, and climatic factors. However, dispersal limitation, local environmental conditions,and interactions among AM fungal taxa may also determine local diversity and global distributions. We assessed the relative importance of these potential controls by collecting 14,961 DNA sequences from 111 published studies and testing for relationships between AM fungal community composition and geography, environment, and plant biomes. Our results indicated that the global species richness of AM fungi was up to six times higher than previously estimated, largely owing to high beta diversity among sampling sites. Geographic distance, soil temperature and moisture, and plant community type were each significantly related to AM fungal community structure, but explained only a small amount of the observed variance. AM fungal species also tended to be phylogenetically clustered within sites, further suggesting that habitat filtering or dispersal limitation is a driver of AM fungal community assembly. Therefore, predicted shifts in climate and plant species distributions under global change may alter AM fungal communities.     

Effects of avocado (Persea americana Mill.) scion on arbuscular mycorrhizal and root hair development in rootstock

Grafting is an important process to propagate horticulture plants; however, the mechanism through which the scion affects the absorption of rootstock remains poorly understood. The effects of the scion on AM fungi types in the rhizosphere soil of rootstock and the absorption of mycorrhizal root were determined in this study. Composition of arbuscular mycorrhizal (AM) fungi, soil assessment, spore density, hyphal length density, glomalin-related soil protein (GRSP) content in rhizosphere soil, root hair morphology and AM colonisation rate were measured among ‘Kampong’ avocado rootstocks grafted with five scions and ‘Kampong’ seedling trees. Results showed the main types of AM fungi in avocado seedling trees and trees grafted with five scions were nearly identical. However, the proportion of main genera exhibited differences. In addition, alkali-hydrolysable N, alkali-hydrolysable P and available K in rhizosphere soil, root hair density, AM colonization, spore density, hyphal length and GRSP content suggested the absorption of ‘Kampong’ rootstocks grafted with ‘Monroe’, ‘Wilson seedless’, ‘Hass’ and ‘Tonnage’ possessed stronger absorption than ‘Kampong’ seedling trees because of high AM colonisation and root hair density. This study suggested scions regulated both the AM and root hair development systematically and laid the foundation for future research of AM-enhancing avocado production.     

Changes in soil organic carbon,total nitrogen,and abundance of arbuscular mycorrhizal fungi along a large-scale aridity gradient

Changes in soil organic carbon, total nitrogen, pH, and the abundance of arbuscular mycorrhizal fungi are examined along a large-scale aridity gradient from southeast to northwest in China. Soil organic carbon and total nitrogen decreased but pH increased with increased aridity. Aboveground plant biomass, spore abundance, and colonization of roots by arbuscular mycorrhizal fungi also declined as the aridity increased. Soil organic carbon and total nitrogen were positively correlated with aboveground plant biomass, and arbuscular mycorrhizal fungal spore number and root colonization were positively correlated with soil organic carbon, total nitrogen, and aboveground plant biomass but were negatively correlated with soil pH. A structural equation model suggested that aridity affected soil organic carbon and total nitrogen by limiting aboveground plant biomass. Aridity exerted a large direct effect and smaller indirect effects (via changes in aboveground plant biomass) on the abundance of arbuscular mycorrhizal fungi. Soil pH also directly influenced arbuscular mycorrhizal fungal abundance. These results suggest that aboveground plant biomass could be a key factor driving the changes of soil organic carbon, total nitrogen, and arbuscular mycorrhizal fungal abundance along this aridity gradient in China.     

Shrubs influence arbuscular mycorrhizal fungi communities in a semi-arid environment

Interactions between arbuscular mycorrhizal fungi (AMF) and plants are essential components of ecosystem functioning; however, they remain poorly known in dry ecosystems. We examined the relationship between seven shrub species and their associated AMF community in a semi-arid plant community in southern Spain. Soil characteristics and plant physiological status were measured and related to AMF community composition and genetic diversity by multivariate statistics. We found differences in AMF communities in soils under shrubs and in gaps among them, whereas no differences were detected among AMF communities colonizing roots. Soil nutrients content drove most of the spatial variations in the AMF community and genetic diversity. AMF communities were more heterogeneous in fertile islands with low nitrogen-to-phosphorus ratio and vice versa. AMF genetic diversity increased in soils limited by phosphorus and with high soil organic matter content, while AMF genetic diversity increased in roots growing in soil not limited by phosphorus. Overall, we could not find a clear link between plant performance and the associated AMF community. Our findings show that different shrub species generate islands of fertility which differ in nutrient content and, therefore, support different AMF communities, increasing AMF diversity at the landscape level.     

Soil tillage and herbicide applications in pea: arbuscular mycorrhizal fungi,plant growth and nutrient concentration respond differently

ABSTRACT

We conducted a field- and pot experiment with peas to investigate the impact of soil tillage and herbicide applications on arbuscular mycorrhizal fungi (AMF), plant growth, phosphorus concentrations, C:N ratio in plants and yield. The field study was carried out in a long-term soil tillage experiment where four tillage treatments have been compared. Field soil from the experimental plots were used for the pot experiment. AMF were not affected by herbicide (MCPB) application, neither in the field nor in the pot experiments. However, AMF root colonization was enhanced by reduced tillage, minimum tillage and no-tillage practices, compared to conventional tillage. In the pot experiment, plant growth and nodulation of pea roots was negatively affected by the high herbicide dosage. In the field experiment neither tillage nor herbicide treatment exert specific effects on root growth parameters, phosphorus concentrations, C:N ratio and plant dry matter. This work demonstrates that an appropriate herbicide usage coupled with conservation soil tillage techniques can favour AMF root colonization and benefit plant growth.

Abbreviations: AMF: arbuscular mycorrhizal fungi; CT: conventional tillage; RT: reduced tillage; MT: minimum tillage; NT: no tillage; P: Phosphorus; C:N ratio: carbon:nitrogen ratio     

Specificity and selectivity of arbuscular mycorrhizal fungal polymerase chain reaction primers in soil samples by clone library analyses

An open question with regard to the community ecology of arbuscular mycorrhizal fungi (AMF) concerns how to best amplify AMF in the soil, which contains a large proportion of DNA from AM extra-radical mycelium and spores. However, to date, a direct comparison of AMF primers for soil samples, which would systematically assess their amplification efficiency, is still missing. In our present study, we compared and characterized four widely used primer sets targeting AMF 18S rDNA or SSU-ITS-LSU rDNA from three soil samples as follows: (1) SSUmAf/LSUmAr?+?SSUmCf/LSUmBr, (2) GeoA2/Geo11?+?NS31/AM1, (3) AML1/AML2?+?NS31/AM1 and (4) AMV4.5NF/AMDGR. These primer sets were compared in terms of the proportion of Glomeromycota detected, AMF diversity and community composition. Our data revealed that the newly combined primer set 3 was the most suitable one for amplifying AMF from soil samples. It yielded the highest AMF alpha diversity, and was very specific to Glomeromycota. Primer set 2 was unable to amplify Claroideoglomus from soil 1, which was the dominant AMF clade as proved by other three primer sets. Primer set 4 demonstrated its instability among different soil samples, since the proportion of AMF in total sequences varied from 5% to 83%. Although primer set 1 showed the highest proportion of AMF (95–100%) in the soil samples, it captured the lowest AMF diversity, and the operational taxonomic units obtained by this primer set were only 36.4% of that by primer set 4. Taken together, our data suggested that AMF diversity in soil samples could be underestimated by primer set 1, 2 and 4. Our result confirmed the important role of the choice of AMF primers for analyzing AMF communities in soil and explored the most suitable one for amplifying AMF from soil samples.     

Mycelium of arbuscular mycorrhizal fungi increases soil water repellency and is sufficient to maintain water-stable soil aggregates

Using an in vitro bioreactor system in which the arbuscular mycorrhizal (AM) fungus Glomus intraradices was grown in a soil devoid of detectable living microbes, we could show that the mycelium of this fungus contributed to the maintenance of water-stable soil aggregates and increased soil water repellency, as measured by water drop penetration time. This is to our knowledge the first demonstration of a causal link between AM fungal growth and water repellency of soil aggregates. Our results also place AM fungal contributions to soil aggregation on a firm mechanistic footing by showing that hyphae are sufficient to produce effects, in the absence of other soil biota, which have always been included in previous studies.     

Effects of soil application of olive mill wastewaters on the structure and function of the community of arbuscular mycorrhizal fungi

Recycling of olive mill wastewaters (OMW) into agricultural soils is a controversial issue since benefits to soil fertility should counterbalance potential short-term toxicity effects. We investigated the short-term effects of OMW on the soil-plant system, regarding the diversity, structure and root colonization capacity of arbuscular mycorrhizal (AM) fungi and the respective growth response of Vicia faba L, commonly used as green manure in olive-tree plantations. A compartmentalized pot system was used that allowed the establishment of an AM fungal community in one compartment (feeder) and the application of three OMW dose levels in an adjacent second compartment (receiver). At 0, 10, and 30 days after OMW treatment (DAT), V. faba pre-germinated seeds were seeded in the receiver compartment. At harvest, shoot and root dry weights, AM fungal root colonization, soil hyphal length and P availability were recorded in the receiver compartment. In addition, OMW effects on AM fungal diversity in plant roots were studied by DGGE. A transient effect of OMW application was observed; plant growth and AM fungal colonization were initially inhibited, whereas soil hyphal length was stimulated, but in most cases differences were absent when seeding was performed 30 DAT. Similarly, changes induced in the structure of the root AM fungal community were of transient nature. Cloning and sequencing of all the major DGGE bands showed that roots were colonized by Glomus spp. The transient effects of OMW on the structure and function of AM fungi could be attributed to OMW-derived phytoxicity to V. faba plants or to an indirect effect via alteration of soil nutritional status. The high OMW dose significantly increased soil P availability in the presence of AM fungi, suggesting efficient involvement of AM fungi in organic-P minerilization. Overall our results indicate that soil application of OMW would cause transient changes in the AM fungal colonization of V. faba plants, which, would not impair their long-term plant growth promoting ability.     

A PCR-DGGE method for detecting arbuscular mycorrhizal fungi in cultivated soils

Arbuscular mycorrhizal (AM) fungi (AMF) are important components of agro-ecosystems and are especially significant for productive low-input agriculture. Molecular techniques are used to investigate fungal community composition in uncultivated, disturbed, or contaminated soils, but this approach to community analysis of AMF in agricultural soils has not been reported. In this study, a polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) procedure for the detection of fungal 18S ribosomal RNA gene was developed with reference cultures of seven isolates (representing five AMF species). These reference cultures were chosen because isolates of their species were putatively identified in a previous survey of farm field soils in the province of Saskatchewan, Canada. A reference PCR-DGGE profile was generated using DNA extracted and amplified from the spores of these cultures. The effectiveness of the procedure was tested by its application to soil samples from 38 farms. Prominent bands from the PCR-DGGE profiles of these samples were excised for sequence analysis. The total number of species recovered was low in comparison to other AMF community surveys of temperate climate locations. The majority of the sequences recovered were Glomus species. Scutellospora calospora, a previously undetected AM fungus in Saskatchewan was found. Though not without its drawbacks, this approach to community composition analysis of AMF was faster than conventional trap cultivation methods.     

Effectiveness of arbuscular mycorrhizal colonization at nursery-stage on growth and nutrition in wetland rice (Oryza sativa L.) after transplanting under different soil fertility and water regimes

In order to analyze the effectiveness of colonization by arbuscular mycorrhizal fungi (AMF) at the nursery stage on the growth and nutrient concentration of wetland rice after transplanting, the experiments were conducted under glasshouse conditions using two types of soil, namely (i) sterilized paddy soil (PS) and (ii) sterilized paddy soil diluted with sterilized Andosol subsoil 5 times (DS) under two water regimes, (i) flooded conditions changed to non-flooded conditions 30 d before harvest (F-NF) and (ii) continuous flooding (CF) up to harvest. Treatments consisting of mycorrhizal inoculation (+AMF) and non-inoculation ( — AMF) were applied only at the nursery stage when the seedlings were produced under dry nursery (60% moisture of maximum water holding capacity) conditions.

Seedlings grown in PS showed a significantly higher biomass yield and nutrient concentrations than in DS. At 90 and 105 d after transplanting, the mycorrhizal plants showed a higher biomass than non-mycorrhizal plants in PS whereas there were no differences in DS except for roots. Mycorrhizal colonization at the transplanting stage was higher in DS than in PS. However, after transplanting opposite results were obtained, the level in PS being relatively higher than in DS. Grain yield and P concentration of unhulled grain and shoots in PS were higher in the +AMF treatments than in the -AMF treatments under both water regimes. Contents of micronutrients (Zn, Cu, Fe, and Mn) were higher in the +AMF plants than in the -AMF ones at all growth stages up to maturation irrespective of soil fertility and water regimes. These results suggest that AMF inoculation at the nursery-stage was beneficial for wetland rice after transplanting to flooded conditions in terms of growth promotion and increase of nutrient concentrations.     

The bacterial community of tomato rhizosphere is modified by inoculation with arbuscular mycorrhizal fungi but unaffected by soil enrichment with mycorrhizal root exudates or inoculation with Phytophthora nicotianae

Arbuscular mycorrhizal (AM) fungi have been shown to induce the biocontrol of soilborne diseases, to change the composition of root exudates and to modify the bacterial community structure of the rhizosphere, leading to the formation of the mycorrhizosphere. Tomato plants were grown in a compartmentalized soil system and were either submitted to direct mycorrhizal colonization or to enrichment of the soil with exudates collected from mycorrhizal tomato plants, with the corresponding negative controls. Three weeks after planting, the plants were inoculated or not with the soilborne pathogen Phytophthora nicotianae growing through a membrane from an adjacent infected compartment. At harvest, a PCR-Denaturing gradient gel electrophoresis analysis of 16S rRNA gene fragments amplified from the total DNA extracted from each plant rhizosphere was performed. Root colonization with the AM fungi Glomus intraradices or Glomus mosseae induced significant changes in the bacterial community structure of tomato rhizosphere, compared to non-mycorrhizal plants, while enrichment with root exudates collected from mycorrhizal or non-mycorrhizal plants had no effect. Our results support that the effect of AM fungi on rhizosphere bacteria would not be mediated by compounds present in root exudates of mycorrhizal plants but rather by physical or chemical factors associated with the mycelium, volatiles and/or root surface bound substrates. Moreover, infection of mycorrhizal or non-mycorrhizal plants with P. nicotianae did not significantly affect the bacterial community structure suggesting that rhizosphere bacteria would be less sensitive to the pathogen invasion than to mycorrhizal colonization. Of 96 unique sequences detected in the tomato rhizosphere, eight were specific to mycorrhizal fungi, including two Pseudomonas, a Bacillus simplex, an Herbaspirilium and an Acidobacterium. One Verrucomicrobium was common to rhizospheres of mycorrhizal plants and of plants watered with mycorrhizal root exudates.     

Effects of arbuscular mycorrhizal fungi on the growth and zinc uptake of trifoliate orange (Poncirus trifoliata) seedlings grown in low-zinc soil

The effects of the arbuscular mycorrhizal (AM) fungi, Glomus intraradices and G. versiforme, on growth and zinc (Zn) uptake were investigated in trifoliate orange (Poncirus trifoliata) seedlings exposed to low-Zn soil. Low-Zn decreased growth, levels of leaf chlorophyll, soluble protein and sugar, and soil enzymatic activities, and pH in 0–2 cm rhizosphere soil. Low-Zn soil also decreased mineral nutrients (including Zn) concentrations in the shoots and roots. Glomus intraradices especially, significantly enhanced plant biomass, leaf soluble protein and sugar concentrations, root viability, acid phosphatase, catalase, invertase and urease activities, and easily extractable glomalin content in 0–2 cm and 2–4 cm rhizosphere soil. It also increased concentrations of Zn, phosphorus, potassium and magnesium in the shoots and roots, while decreased the soil pH. Arbuscular mycorrhizal fungi, especially G. intraradices, has the potential to improve growth and Zn uptake of triofoliate orange seedlings grown in low-Zn soil.     

Arbuscular mycorrhizal fungi reduce decomposition of woody plant litter while increasing soil aggregation

The decomposition of plant organic matter and the stability of soil aggregates are important components of soil carbon cycling, and the relationship between decomposition rate and arbuscular mycorrhizal fungi (AMF) has recently received considerable attention. The interaction of AMF with their associated microorganisms and the consequences for litter decomposition and soil aggregation still remain fairly unclear. In a laboratory pot experiment we simultaneously tested the single and combined effects of one AMF species (Rhizophagus irregularis) and a natural non-AMF microbial community on the decomposition of small wooden sticks and on soil aggregation. To disentangle effects of hyphae and roots we placed mesh bags as root exclusion compartments in the soil. The decomposition of the wooden sticks in this compartment was significantly reduced in the presence of AMF, but not with the non-AMF microbial community only, compared to the control, while aggregation was increased in all treatments compared to the control. We suggest that AMF directly (via localized nutrient removal or altered moisture conditions) or indirectly (by providing an alternative carbon source) inhibited the activity of decomposers, leading to different levels of plant litter degradation under our experimental settings. Reduced decomposition of woody litter in presence of AMF can be important for nutrient cycling in AMF-dominated forests and in the case of woody plants and perennials that develop lignified roots in grasslands.     

Direct and indirect influence of arbuscular mycorrhizal fungi on abundance and community structure of ammonia oxidizing bacteria and archaea in soil microcosms

Both arbuscular mycorrhizal (AM) fungi and ammonia oxidizers are important soil microbial groups in regulating soil N cycling. However, knowledge of their interactions, especially the direct influences of AM fungi on ammonia oxidizers is very limited to date. In the present study, a controlled microcosm experiment was established to examine the effects of AM fungi and N supply level on the abundance and community structure of ammonia oxidizing bacteria (AOB) and archaea (AOA) in the rhizosphere of alfalfa plants (Medicago sativa L.) inoculated with AM fungus Glomus intraradices. Effects were studied using combined approaches of quantitative polymerase chain reaction (qPCR) and terminal-restriction fragment length polymorphism (T-RFLP). The results showed that inoculation with AM fungi significantly increased the plant dry weights, total N and P uptake. Concomitantly, AM fungi significantly decreased the amoA gene copy numbers of AOA and AOB in the root compartment (RC) but not in the hyphal compartment (HC). Moreover, AM fungi induced some changes in AOA community structure in HC and RC, while only marginal variations in AOA composition were observed to respond to N supply level in HC. Neither RC nor HC showed significant differences in AOB composition irrespective of experimental treatments. The experimental results suggested that AM fungi could directly shape AOA composition, but more likely exerted indirect influences on AOA and AOB abundance via the plant pathway. In general, AM fungi may play an important role in mediating ammonia oxidizers, but the AOA community appeared to be more sensitive than the AOB community to AM fungi.     

The tripartite symbiosis between legumes, rhizobia and indigenous mycorrhizal fungi is more efficient in undisturbed soil

We investigated how the rate of colonization by indigenous arbuscular mycorrhizal fungi (AMF) affects the interaction between AMF, Sinorrhizobium meliloti and Medicago truncatula Gaertn. To generate a differential inoculum potential of indigenous AMF, five cycles of wheat, each of 1 month, were grown in sieved or undisturbed soil before M. truncatula was sown. The early colonization of M. truncatula roots by indigenous AMF was faster in undisturbed soil compared with sieved soil, but by pod-fill the frequency of hyphae, arbuscules and vesicles was similar in both treatments. At this latter stage, M. truncatula grown in undisturbed soil had accumulated a greater biomass in aboveground tissues, had a greater P concentration and derived more N from the atmosphere than plants grown in disturbed soil, although soil compaction resulted in plants having a smaller root system than those from disturbed soil. The difference in plant P content could not be explained by modifications in hydrolytic soil enzymes related to the P cycle as the activity of acid phosphatase was greater in sieved than in undisturbed soil, and the activity of alkaline phosphatase was unaffected by the treatment. Thus, the results observed were a consequence of the different rates of AMF colonization caused by soil disturbance. Together with earlier results for soybean, this study confirms that soil disturbance modifies the interaction between indigenous AMF, rhizobia and legumes leading to a reduced efficacy of the bacterial symbiont.