Sara F. Wright

A survey of soils for aggregate stability and glomalin, a glycoprotein produced by hyphae of arbuscular mycorrhizal fungi

Understanding the contributions of soil microorganisms to soil stabilization at the molecular level will lead to ways to enhance inputs for sustainable agricultural systems. Recent discoveries of copious production of glycoprotein (glomalin) by arbuscular mycorrhizal (AM) fungi and the apparent recalcitrance of this material in soils led to the comparison between concentration of glomalin and aggregate stability. Stability was measured on air-dried aggregates rewetted by capillary action and then subjected to wet sieving for 10 min. Thirty-seven samples from four geographic areas of the U.S. and one area of Scotland were tested. The monoclonal antibody used to discover glomalin on AM hyphae was employed to assess immunoreactive glomalin on aggregate surfaces by immunofluorescence and in extracts from aggregates by enzyme-linked immunosorbent assay (ELISA). Immunofluorescence was observed on at least some surfaces of aggregates from all soils examined, but was most evident on aggregates with high glomalin concentrations. Easily extractable glomalin (EEG) was solubilized by 20 mM citrate, pH 7.0 at 121 °C for 30 min, and total glomalin (TG) was solubilized with 50 mM citrate, pH 8.0 at 121 °C for 90 to 450 min. Some soils required up to seven sequential extractions to remove all of the glomalin. Aggregate stability was linearly correlated (p < 0.001) with all measures of glomalin (mg/g of aggregates) in these soils. The best predictor of aggregate stability (AS) was immunoreactive easily extractable glomalin (IREEG) according to the following relationship: AS = 42.7 +61.3 × log10 IREEG (r2 = 0.86; p <0.001, n = 37).

Extraction of an abundant and unusual protein from soil and comparison with hyphal protein of arbuscular mycorrhizal fungi

It has been speculated that arbuscular mycorrhizal fungi (AMF) produce extracellular compounds and that these are involved in soil stabilization. An unusual and abundant protein was found on hyphae of AMF, and it was hypothesized that the hyphal protein could be found in soil. The purpose of this

Large contribution of arbuscular mycorrhizal fungi to soil carbon pools in tropical forest soils

The origins and composition of soil organic matter (SOM) are still largely uncertain. Arbuscular mycorrhizal fungi (AMF) are recognized as indirect contributors through their influence on soil aggregation, plant physiology, and plant community composition. Here we present evidence that AMF can also make large, direct contributions to SOM. Glomalin, a recently discovered glycoprotein produced by AMF hyphae, was detected in tropical soils in concentrations of over 60 mg cm−3. Along a chronosequence of soils spanning ages from 300 to 4.1 Mio years, a pattern of glomalin concentrations is consistent with the hypothesis that this protein accumulates in soil. Carbon dating of glomalin indicated turnover at time scales of several years to decades, much longer than the turnover of AMF hyphae (which is assumed to be on the order of days to weeks). This suggests that contributions of mycorrhizae to soil carbon storage based on hyphal biomass in soil and roots may be an underestimate. The amount of C and N in glomalin represented a sizeable amount (ca. 4–5%) of total soil C and N in the oldest soils. Our results thus indicate that microbial (fungal) carbon that is not derived from above- or below-ground litter can make a significant contribution to soil carbon and nitrogen pools and can far exceed the contributions of soil microbial biomass (ranging from 0.08 to 0.2% of total C for the oldest soils).

The role of arbuscular mycorrhizal fungi and glomalin in soil aggregation: comparing effects of five plant species

Soil aggregation and soil structure are fundamental properties of natural and managed ecosystems. However, most of our knowledge on the role of plant species in soil aggregation is derived from work in agroecosystems or with agriculturally important plants. Here we examined the effects of five plant species on soil aggregate water stability. The five species (three grasses, one forb, and a legume) were from the same natural grassland, and were grown in monoculture plots in the field. Our first goal was to test if productivity-related or species-specific factors would prevail in determining soil aggregation. We also tested what the relative importance of the soil protein glomalin (produced by arbuscular mycorrhizal fungi, AMF) in soil aggregation is, compared to other factors, including AMF hyphal and root length and percent plant cover. We found significant differences in soil aggregate water stability (1–2 mm size class) for the five plant species examined, and corresponding differences in plant cover, root weight and length, AMF soil hyphal length, and glomalin concentrations. A structural equation modeling approach (path analysis) was used to distinguish direct from indirect effects of factors on soil aggregation based on covariance structures. Root length, soil glomalin, and percent cover contributed equally strong paths to water-stable aggregation. The direct effect of glomalin was much stronger than the direct effect of AMF hyphae themselves, suggesting that this protein is involved in a very important hypha-mediated mechanism of soil aggregate stabilization, at least for the 1–2-mm size class of aggregates.

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.

Aggregate stability and glomalin in alternative crop rotations for the central Great Plains.

Abstract Land productivity, along with improvement or maintenance of soil health, must be evaluated together to achieve sustainable agricultural practices. Winter wheat-fallow (W-F) has been the prevalent cropping system in the central Great Plains for 60 years where moisture is a limitation to crop production. Alternative cropping systems show that producers can crop more frequently if residue management and minimum tillage are used. The impact of different crops, crop rotations and tillage management practices on soil quality was assessed by measuring aggregate stability and glomalin production by arbuscular mycorrhizal (AM) fungi in soil from cropping trials established in 1990. Crops were wheat (W), corn (C), proso millet (M), and sunflower (S). Rotations sampled were W-F, W-C-M, W-C-M-F, W-C-F, and W-S-F. In the same area as the cropping trials, soils were taken from a perennial grass (crested wheatgrass) and from a buffer area that had been planted to Triticale for the past 2 years but prior to that had been extensively plowed for weed control. We found that aggregate stability and glomalin were linearly correlated (r=0.73, n=54, P<0.001) across all treatments sampled. Highest and lowest aggregate stability and glomalin values were seen in perennial grass and Triticale soils, respectively. Aggregate stability in W-S-F was significantly lower than in the other crop rotations (P≤0.03), while W-C-M had significantly higher glomalin than the other rotations (P<0.05). Differences between crop rotations and the perennial grass indicate that selected comparisons should be studied in greater detail to determine ways to manage AM fungi to increase glomalin and aggregate stability in these soils.

Rise in carbon dioxide changes soil structure

Carbon in soil affects the formation and stabilization of aggregates (groups of primary particles that adhere to each other more strongly than to surrounding soil particles). Soil aggregation is important for preventing soil loss through wind and water erosion, and the size distribution and abundance of water-stable aggregates influences a range of physical, chemical, biological and agricultural properties of soil. The effects on soil biota and nutrient cycling of increases in soil carbon availability, brought about by increased CO2, are well studied, but the consequences for soil aggregation and structure have not been examined. Here we show for three ecosystems that the water stability and size distribution of aggregates is affected by long-term CO2fumigation, and we propose a mechanism for this that involves the production by fungi of the glycoprotein glomalin.

Spatial heterogeneity of aggregate stability and soil carbon in semi-arid rangeland

To measure and manage for C sequestration in heterogeneous rangeland systems, we need to more fully understand spatial patterns of soil resources. Spatial distributions of aggregate stability and soil carbon were investigated in a semiarid rangeland in New Mexico, USA. Soil was analyzed from plant interspaces, black grama (Bouteloua eriopoda (Torr.) Torr.), and mesquite (Prosopis glandulosa Torr.) in a landscape-replicated study. Aggregate stability at the 250 microm scale, carbonate C, organic C and N, C:N ratio, and glomalin, were all highest under mesquite. Soil C:N ratio was the best predictor of aggregate stability. Estimates of metric tons of C per hectare in the top 10 cm were highly variable at patch and landscape scales, varying from 4.2 to 10.5 under mesquite and from 3.0 to 7.0 in interspaces. High variability of aggregate stability and soil C has important implications for C sequestration. We argue that this multi-scale soil heterogeneity must be considered when measuring and managing for C sequestration.

Evaluation of vesicular-arbuscular mycorrhizal fungi in diverse plants and soils

Summary-A regional study was made to identify vesicular-arbuscular mycorrhizal (VAM) fungi effective in promoting plant growth in diverse plant and soil systems. Eight cooperators in six states of the eastern United States evaluated six VAM fungal isolates on soybean (Glycine mux L. Merr.) and sorghum (Sorghum bicolor L. Moench) in a shared soil and in at least one regional soil from each location. Plants were grown with high VAM inoculum densities (minimum of 20 VAM propagules ml-‘) for 42-57 days in pasturized soils in greenhouses or growth chambers. Shoot and root dry masses, total and colonized root lengths and shoot-P concentrations were determined at harvest. Under the experimental conditions tested, the VAM fungal isolate was more important than the soil or host plant in determining effectiveness. In the shared soil, inoculation with two isolates of Glomus (GE329 and GENPI) resulted in the greatest shoot masses for soybeans, while the same two isolates and GE312 provided maximum response in sorghum. In the regional soils, GE329 and GENPI had the widest range of growth promotion with both soybean and sorghum; however, for both plant species the mycorrhizal response was greatest in soils with less than 10 mg extractable P kg-‘. For soybeans, colonized root length was not related to VAM growth response. For sorghum, there was a positive correlation between colonized root length and plant growth response. We conclude that VAM isolates exist which are effective in promoting plant growth over a range of edaphic and host conditions.

A fluorescent antibody assay for hyphae and glomalin from arbuscular mycorrhizal fungi

Studies on the role of arbuscular mycorrhizal (AM) fungi in soil have been aided by the use of a monoclonal antibody that detects a molecule common to all isolates of these fungi studied to date. The molecule, glomalin, is a glycoprotein that forms on hyphae, but apparently sloughs off and adheres to soil particles or imbedded plastic mesh. An indirect immunofluorescence (IF) assay is described for detection of glomalin on hyphae attached to roots, in roots, on hyphae traps and on the surface of soil aggregates. Small sieves are used to process hyphae attached to roots and soil aggregates. Glomalin on hyphae and glomalin attached to plastic or nylon are assayed on a 1 cm2 section of meshes. Examples of IF assay results are shown and discussed.