Resilience of soil aggregation and exocellular enzymatic functions associated with arbuscular mycorrhizal fungal communities along a successional gradient in a tropical dry forest.

Arbuscular mycorrhizal (AM) fungi are well-known contributors to soil aggregation and nutrient cycling functions, but we still know little about their capacity to resist or recover from persistent disturbance. Rangeland management may deteriorate these functions by affecting the activity of soil biota, including AM fungi, among other consequences. If affected, some soil properties show recovery when management stops and natural regeneration is allowed. We conducted an experiment to evaluate if the functions related to soil aggregation and promotion of exocellular enzymatic activities associated with AM fungal communities had been affected by rangeland management and, if they had, whether they recovered with successional time when management stopped. AM fungal communities from ten sites with different successional ages in a tropical dry forest region were inoculated to the same host growing in pots divided by mesh into a plant compartment and an AM mycelium compartment. We examined soil stable aggregates fractions and enzymatic activities produced or promoted by AM fungi. Soil aggregation changed significantly only after the study had run for 3 years, was higher in the hyphosphere than in the root compartment, and showed a low but positive relation with the successional age of the communities. The activity of phosphatase, but not casein-protease and beta-glucosidase, increased with successional age. Therefore, soil aggregation and enzyme activities associated with AM fungal communities seemed resilient because casein-protease and beta-glucosidase were unchanged, and aggregation and phosphatase were reduced by rangeland management but recovered with successional time.

Direct evidence for modulation of photosynthesis by an arbuscular mycorrhiza-induced carbon sink strength.

It has been suggested that plant carbon (C) use by symbiotic arbuscular mycorrhizal fungi (AMF) may be compensated by higher photosynthetic rates because fungal metabolism creates a strong C sink that prevents photosynthate accumulation and downregulation of photosynthesis. This mechanism remains largely unexplored and lacks experimental evidence. We report here two experiments showing that the experimental manipulation of the mycorrhizal C sink significantly affected the photosynthetic rates of cucumber host plants. We expected that a sudden reduction in sink strength would cause a significant reduction in photosynthetic rates, at least temporarily. Excision of part of the extraradical mycorrhizal mycelium from roots, and causing no disturbance to the plant, induced a sustained (10-40%) decline in photosynthetic rates that lasted from 30 min to several hours in plants that were well-nourished and hydrated, and in the absence of growth or photosynthesis promotion by mycorrhizal inoculation. This effect was though minor in plants growing at high (700 ppm) atmospheric CO2 . This is the first direct experimental evidence for the C sink strength effects exerted by arbuscular mycorrhizal symbionts on plant photosynthesis. It encourages further experimentation on mycorrhizal source-sink relations, and may have strong implications in large-scale assessments and modelling of plant photosynthesis.

Morphological and physiological responses of the external mycelium of Rhizophagus intraradices to water stress.

Most studies dealing with mycorrhizal associations and drought have focused on the plants, not on the fungi, and tolerance and adaptations of arbuscular mycorrhizal (AM) fungi to cope with water stress are virtually unknown. This study was conducted to assess how water stress directly affects an AM fungus isolate, particularly through morphological and physiological changes in the external mycelium. We used two-compartment pots separated by mesh and an air gap that allowed us to apply water stress treatments only to the external mycelium. Clover (Trifolium subterraneum L.) plants inoculated with Rhizophagus intraradices grew at high humidity until external mycorrhizal mycelium developed in the mycelium compartment. Then, we started three watering treatments: high (H, 70% of soil water holding capacity), low (L, 10%), and mixed watering (HLHL, 70-10-70-10%) only in the hyphal compartment. The HLHL treatment was rewetted once to 70% after 42 days. We measured total mycelium length, hyphal length in diameter categories, respiration activity, and protoplasm fragmentation 42 and 76 days after starting the treatments. Rhizophagus intraradices mycelium responded to water stress by reducing its length, maintaining larger diameter hyphae, and concentrating protoplasm activity in fragments in the HLHL and L treatments. In both water stress treatments, changes suggested a trade-off between avoiding desiccation and storing resources, and maintaining soil exploration and water uptake capacity.

Explaining long-term inter-individual growth variation in plant populations: persistence of abiotic factors matters.

An unanswered question in ecology is whether the environmental factors driving short-term performance also determine the often observed long-term performance differences among individuals. Here, we analyze the extent to which temporal persistence of spatial heterogeneity in environmental factors can contribute to long-term inter-individual variation in stem length growth. For a natural population of a long-lived understorey palm, we first quantified the effect of several environmental factors on stem length growth and survival. We then performed individual-based simulations of growth trajectories, in which we varied, for two environmental factors: (1) the strength of the effect on stem length growth and (2) the temporal persistence. Short-term variation in stem length growth was strongly driven by light availability. Auto-correlation in light availability and soil pH increased simulated variation in stem length growth among 20-year-old palms to levels similar to the observed variation. Analyses in which we varied both the strength of the effect on stem length growth and the temporal persistence of the environmental factors revealed that a large fraction of observed long-term growth differences can be explained, as long as one of these effects is strong. This implies that environmental factors that are relatively unimportant for short-term performance can still drive long-term performance differences when the environmental variation is sufficiently persistent over time.

Soil resistance and recovery during neotropical forest succession.

The recovery of soil conditions is crucial for successful ecosystem restoration and, hence, for achieving the goals of the UN Decade on Ecosystem Restoration. Here, we assess how soils resist forest conversion and agricultural land use, and how soils recover during subsequent tropical forest succession on abandoned agricultural fields. Our overarching question is how soil resistance and recovery depend on local conditions such as climate, soil type and land-use history. For 300 plots in 21 sites across the Neotropics, we used a chronosequence approach in which we sampled soils from two depths in old-growth forests, agricultural fields (i.e. crop fields and pastures), and secondary forests that differ in age (1-95 years) since abandonment. We measured six soil properties using a standardized sampling design and laboratory analyses. Soil resistance strongly depended on local conditions. Croplands and sites on high-activity clay (i.e. high fertility) show strong increases in bulk density and decreases in pH, carbon (C) and nitrogen (N) during deforestation and subsequent agricultural use. Resistance is lower in such sites probably because of a sharp decline in fine root biomass in croplands in the upper soil layers, and a decline in litter input from formerly productive old-growth forest (on high-activity clays). Soil recovery also strongly depended on local conditions. During forest succession, high-activity clays and croplands decreased most strongly in bulk density and increased in C and N, possibly because of strongly compacted soils with low C and N after cropland abandonment, and because of rapid vegetation recovery in high-activity clays leading to greater fine root growth and litter input. Furthermore, sites at low precipitation decreased in pH, whereas sites at high precipitation increased in N and decreased in C : N ratio. Extractable phosphorus (P) did not recover during succession, suggesting increased P limitation as forests age. These results indicate that no single solution exists for effective soil restoration and that local site conditions should determine the restoration strategies. This article is part of the theme issue 'Understanding forest landscape restoration: reinforcing scientific foundations for the UN Decade on Ecosystem Restoration'.

Isotope Labeling to Study Phosphorus Uptake in the Arbuscular Mycorrhizal Symbiosis.

Isotope labeling enables the detection and quantification of nutrient fluxes between soil and plants through arbuscular mycorrhizal (AM) fungi. Here we describe the use of radioactive isotopes, 33P and 32P, to study the uptake of P from soil by AM fungal mycelium and its transfer to the host plant through the mycorrhizal pathway.

Phosphorus uptake by arbuscular mycorrhizal hyphae does not increase when the host plant grows under atmospheric CO2 enrichment.

• We conducted an experiment to test whether phosphorus (P) uptake by mycorrhizal hyphae could be enhanced by growing the host plant under [CO2 ] enrichment and whether any response to [CO2 ] was dependent on C source-sink relationships. • Plant C assimilation, mass allocation, growth and P uptake were measured in pea (Pisum sativum) plants inoculated with 0, 1 or 5% of a mixture of three Glomus spp. Intra- and extra-radical mycorrhizal development was followed and hyphal 33 P uptake from a root-exclusion compartment was measured. • Total P and 33 P content measurements indicated that root, not hyphal, P uptake was increased by elevated [CO2 ] in the mycorrhizal treatments and that hyphal P uptake was actually reduced by elevated [CO2 ] after 57 d. Neither intra- nor extraradical mycorrhizal development was related to this response. • Plant and fungal measurements suggested positive interactions in plant growth and P uptake only when C source-sink relationships were balanced; high C source (enhanced assimilation at elevated [CO2 ]) and high C sink (increasing mycorrhizal development). The results also indicated that enhanced plant C supply does not alter growth or function of arbuscular mycorrhizal fungi.

Neither mycorrhizal inoculation nor atmospheric CO2 concentration has strong effects on pea root production and root loss.

• Root responses to elevated CO2 concentrations, where nutrient demand was expected to be higher than at ambient CO2 , and possible interactions with mycorrhizal symbionts are reported for pea (Pisum sativum). These are important below-ground components affecting carbon flow into the soil. • A video-minirhizotron system was used to study root growth in pot-grown mycorrhizal (inoculated with Glomus caledonium) and nonmycorrhizal pea plants at ambient or elevated CO2 concentrations over 9 wk. Analyses were made of root length changes, cohort size and survivorship. • Root length production at ambient, but not at elevated CO2 , was higher in nonmycorrhizal than in mycorrhizal plants from week 4-7. Root loss began at week 5, peaking 2 wk later with 40-50% loss of the root length produced by week 8. The decline in root production and increase in root loss coincided with the onset of flowering. • Neither mycorrhizal inoculation nor CO2 concentration has a strong effect on pea root production and root loss, although mycorrhizal infection has a greater effect than CO2 .

Short-term consequences of slash-and-burn practices on the arbuscular mycorrhizal fungi of a tropical dry forest.

Rates of land conversion from forest to cultivated land by slash-and-burn practices are higher in tropical dry forest (TDF) than any other Neotropical forest type. This study examined the short-term consequences of the slash-and-burn process on arbuscular mycorrhizal fungi (AMF). We expected that slash-and-burn would reduce mycorrhizal colonization and propagules and change species richness and composition. Soil and root samples were taken from TDF control and pasture plots originated after slash-and-burn at four dates during the year of conversion to examine species composition, spore abundance, and infective propagules. Additionally, spore abundance and viability and viable intraradical colonization were measured twice during the second year after conversion. Forest and pasture plots maintained similar species richness and an overall 84% similarity during the first year after conversion. Infective propagules were reduced in pasture plots during the first year after slash-and-burn, whereas spore abundance and intraradical colonization remained similar in TDF and pasture plots both years of the study. Our results suggest, contrary to the expected, that forest conversion by means of slash-and-burn followed by cultivation resulted in few immediate changes in the AMF communities, likely because of the low heat conductivity of the soil and rapid combustion of plant residues.

High compatibility between arbuscular mycorrhizal fungal communities and seedlings of different land use types in a tropical dry ecosystem.

We conducted this study to explore limitations for the establishment of mycorrhizal associations in disturbed areas of the tropical dry ecosystem in the Chamela region of Jalisco, Mexico. Specifically, we: (1) assessed the diversity and composition of arbuscular mycorrhizal fungal (AMF) communities through spore morphospecies identification in three common land uses (primary forest, secondary forest, and pasture), (2) tested the inoculum potential of the AMF communities and the effect of water stress on the establishment of mycorrhizal associations in seedlings of various plant species, and (3) explored the importance of AMF community composition on early seedling development. Soil and root samples were taken from 15 random points in each of three plots established in two primary forests, two 26-year-old secondary forests, and two 26-year-old pastures. We expected that because of soil degradation and management, pastures would have the lowest and primary forests the highest AMF species richness. We found evidence for changes in AMF species composition due to land use and for higher morphospecies richness in primary forests than in secondary forests and pastures. We expected also that water stress limited plant and mycorrhizal development and that plants and AMF communities from secondary forests and pastures would be less affected by (better adapted to) water stress than those from the primary forest. We found that although all plant species showed biomass reductions under water stress, only some of the plant species had lower mycorrhizal development under water stress, and this was regardless of the AMF community inoculated. The third hypothesis was that plant species common to all land use types would respond similarly to all AMF communities, whereas plant species found mainly in one land use type would grow better when inoculated with the AMF community of that specific land use type. All plant species were however equally responsive to the three AMF communities inoculated, indicating that all plants established functionally compatible AMF in each community, with no preferences. The results suggest that early seedling growth and mycorrhizal development in secondary forests and pastures is not likely limited by diversity, quantity, or quality of mycorrhizal propagules but by the high temperature and water stress conditions prevailing at those sites.

13C incorporation into signature fatty acids as an assay for carbon allocation in arbuscular mycorrhiza.

The ubiquitous arbuscular mycorrhizal fungi consume significant amounts of plant assimilated C, but this C flow has been difficult to quantify. The neutral lipid fatty acid 16:1omega5 is a quantitative signature for most arbuscular mycorrhizal fungi in roots and soil. We measured carbon transfer from four plant species to the arbuscular mycorrhizal fungus Glomus intraradices by estimating (13)C enrichment of 16:1omega5 and compared it with (13)C enrichment of total root and mycelial C. Carbon allocation to mycelia was detected within 1 day in monoxenic arbuscular mycorrhizal root cultures labeled with [(13)C]glucose. The (13)C enrichment of neutral lipid fatty acid 16:1omega5 extracted from roots increased from 0.14% 1 day after labeling to 2.2% 7 days after labeling. The colonized roots usually were more enriched for (13)C in the arbuscular mycorrhizal fungal neutral lipid fatty acid 16:1omega5 than for the root specific neutral lipid fatty acid 18:2omega6,9. We labeled plant assimilates by using (13)CO(2) in whole-plant experiments. The extraradical mycelium often was more enriched for (13)C than was the intraradical mycelium, suggesting rapid translocation of carbon to and more active growth by the extraradical mycelium. Since there was a good correlation between (13)C enrichment in neutral lipid fatty acid 16:1omega5 and total (13)C in extraradical mycelia in different systems (r(2) = 0.94), we propose that the total amount of labeled C in intraradical and extraradical mycelium can be calculated from the (13)C enrichment of 16:1omega5. The method described enables evaluation of C flow from plants to arbuscular mycorrhizal fungi to be made without extraction, purification and identification of fungal mycelia.

Temperature constraints on the growth and functioning of root organ cultures with arbuscular mycorrhizal fungi.

In this study we investigated the effects of temperature on fungal growth and tested whether the differences in fungal growth were related to the effects of temperature on carbon movement to, or within, the fungus. Growth curves and C uptake-transfer-translocation measurements were obtained for three arbuscular mycorrhizal fungi (AMF) isolates cultured within a 6-30 degrees C temperature range. A series of experiments with a model fungal isolate, Glomus intraradices, was used to examine the effects of temperature on lipid body and 33P movement, and to investigate the role of acclimation and incubation time. Temperature effects on AMF growth were both direct and indirect because, despite clear independent root and AMF growth responses in some cases, the uptake and translocation of 13C was also affected within the temperature range tested. Root C uptake and, to a lesser extent, C translocation in the fungus, were reduced by low temperatures (< 18 degrees C). Uptake and translocation of 33P by fungal hyphae were, by contrast, similar between 10 and 25 degrees C. We conclude that temperature, between 6 and 18 degrees C, reduces AMF growth, and that C movement to the fungus is involved in this response.

Allocation of plant carbon to foraging and storage in arbuscular mycorrhizal fungi.

Abstract Foraging strategies, the cost-benefit associated with the search for new resources, have only begun to be explored in arbuscular mycorrhizal fungi (AMF). We show the use of (13)C-labelling, via shoot photosynthesis, of the 16:1omega5 fatty acid biomarker (the dominant and rather specific fatty acid in AMF storage lipids) to study the immediate patterns of carbon allocation to fungal lipids in response to inorganic and organic nutrient amendments. Signature fatty acid measurements, the incorporation of the label and complementary hyphal length density measurements showed that the extraradical mycelium of AMF proliferated in response to all the amendments provided whereas its development into unamended sand was minor in all treatments. We demonstrate the foraging capacity of AMF, linked to plant carbon, through their hyphal proliferation and accumulation of energy reserves.