Lea Corkidi

NITROGEN ENRICHMENT ALTERS MYCORRHIZAL ALLOCATION AT FIVE MESIC TO SEMIARID GRASSLANDS

Arbuscular mycorrhizal (AM) fungi are integral components of grasslands because most plants are associated with interconnected networks of AM hyphae. Mycorrhizae generally facilitate plant uptake of nutrients from the soil. However, mycorrhizal associations are known to vary in their mutualistic function, and there is currently no metric that links AM functioning with fungal colonization of roots. Mycorrhizal structures differ in their physiological and ecological functioning, so changes in AM allocation to intraradical (inside roots) and extraradical (in soil) structures may signal shifts in mycorrhizal function. We hypothesize that the functional equilibrium model applies to AM fungi and that fertilization should reduce allocation to arbuscules, coils, and extraradical hyphae, the fungal structures that are directly involved in nutrient acquisition and transfer to plants. This study compared AM responses to experimental N enrichment at five grasslands distributed across North America. Samples were collected from replicated N-enriched (and some P-enriched) and control plots throughout the growing season for three years. Intraradical AM structures were measured in over 1400 root samples, extraradical hyphal density was measured in over 590 soil samples, and spore biovolume was analyzed in over 400 soil samples. There were significant site × N interactions for spore biovolume, extraradical hyphae, intraradical hyphae, and vesicles. Nitrogen enrichment strongly decreased AM structures at Cedar Creek, the site with the lowest soil N:P, and it increased AM structures at Konza Prairie, the site with the highest soil N:P. As predicted by the functional equilibrium model, in soils with sufficient P, relative allocation to arbuscules, coils, and extraradical hyphae was generally reduced by N enrichment. Allocation to spores and hyphae was most sensitive to fertilization. At the mesic sites, this response was associated with a shift in the relative abundance of Gigasporaceae within AM fungal communities. This study demonstrates that N enrichment impacts mycorrhizal allocation across a wide range of grassland ecosystems. Such changes are important because they suggest an alteration in mycorrhizal functioning that, in turn, may impact plant community composition and ecosystem function.

Nitrogen fertilization alters the functioning of arbuscular mycorrhizas at two semiarid grasslands

The effects of nitrogen (N) fertilization on arbuscular mycorrhizas were studied at two semiarid grasslands with different soil properties and N-enrichment history (Shortgrass Steppe in Colorado, and Sevilleta National Wildlife Refuge in New Mexico). These sites are part of the National Science Foundations Long-Term Ecological Research Network. The experimental plots at Shortgrass Steppe were fertilized with ammonium nitrate (NH4NO3) from 1971 to 1975, and have not received additional N since then. The experimental plots at Sevilleta were also fertilized with NH4NO3, but were established in 1995, 2 years before the soils were used for this study. Greenhouse experiments were conducted to compare the growth response of local grasses to arbuscular mycorrhizal (AM) fungi from fertilized (FERT) and unfertilized (UNFERT) field soils, at each site. Two species per site were chosen, Bouteloua gracilis and Elymus elymoides from Shortgrass Steppe, and B. gracilis and B. eriopoda from Sevilleta. Plants were grown for 3 months at HIGH N and LOW N levels, with FERT or UNFERT soil inoculum and in a non-mycorrhizal condition. Fertilization with N altered the functioning of AM fungi at both sites. Grasses inoculated with AM fungi from UNFERT soils had the most tillers, greatest biomass and highest relative growth rates. There were no significant differences in the growth response of plants inoculated with AM fungi from FERT soils and the non-mycorrhizal controls. These results were consistent across sites and species except for the plants grown at LOW N in Sevilleta soils. These plants were deficient in N and phosphorus (P) and did not show growth enhancement in response to AM inoculation with either FERT or UNFERT soils. Percent root length colonized by AM fungi was not directly related to plant performance. However, enrichment with N consistently decreased root colonization by AM fungi in the grasses grown in soils from Shortgrass Steppe with high P availability (18.4 mg kg−1), but not in the grasses grown in Sevilleta soils with low P availability (6.6 mg kg−1). Our study supports the hypotheses that (1) fertilization with N alters the balance between costs and benefits in mycorrhizal symbioses and (2) AM fungal communities from N fertilized soils are less beneficial mutualists than those from unfertilized soils.

Impacts of early- and late-seral mycorrhizae during restoration in seasonal tropical forest, Mexico

Disturbance of vegetation and soil may change the species composition of arbuscular mycorrhizal fungi (AMF), which may in turn affect plant species responses to AMF. Seasonal tropical forest in Mexico is undergoing rapid conversion to early-successional forest because of increased wildfire and may require restoration. The responses of six early- and late-successional tree species were tested using early- and late-successional AMF inoculum. The plants were germinated in the shadehouse and received three inoculum treatments: (1) soil from a two-year-old burned site, (2) soil from a mature forest site, or (3) uninoculated controls. They were transplanted as seedlings to a site prepared by burning, and their growth was measured from September 1997 to November 2000. All six species had the greatest growth response to early-seral inoculum, but the response to late-seral inoculum varied. Two tree species, Ceiba pentandra and Guazuma ulmifolia, were smallest with late-seral inoculum, even smaller than the uninocu...

PLANT WINNERS AND LOSERS DURING GRASSLAND N-EUTROPHICATION DIFFER IN BIOMASS ALLOCATION AND MYCORRHIZAS

Human activities release tremendous amounts of nitrogenous compounds into the atmosphere. Wet and dry deposition distributes this airborne nitrogen (N) on otherwise pristine ecosystems. This eutrophication process significantly alters the species composition of native grasslands; generally a few nitrophilic plant species become dominant while many other species disappear. The functional equilibrium model predicts that, compared to species that decline in response to N enrichment, nitrophilic grass species should respond to N enrichment with greater biomass allocation aboveground and reduced allocation to roots and mycorrhizas. The mycorrhizal feedback hypothesis states that the composition of mycorrhizal fungal communities may influence the composition of plant communities, and it predicts that N enrichment may generate reciprocal shifts in the species composition of mycorrhizal fungi and plants. We tested these hypotheses with experiments that compared biomass allocation and mycorrhizal function of four grass ecotypes (three species), two that gained and two that lost biomass and cover in response to long-term N enrichment experiments at Cedar Creek and Konza Long-Term Ecological Research grasslands. Local grass ecotypes were grown in soil from their respective sites and inoculated with whole-soil inoculum collected from either fertilized (FERT) or unfertilized (UNFERT) plots. Our results strongly support the functional equilibrium model. In both grassland systems the nitrophilic grass species grew taller, allocated more biomass to shoots than to roots, and formed fewer mycorrhizas compared to the grass species that it replaced. Our results did not fully support the hypothesis that N-induced changes in the mycorrhizal fungal community were drivers of the plant community shifts that accompany N eutrophication. The FERT and UNFERT soil inoculum influenced the growth of the grasses differently, but this varied with site and grass ecotype in both expected and unexpected ways suggesting that ambient soil fertility or other factors may be interacting with mycorrhizal feedbacks.

Feedbacks of Soil Inoculum of Mycorrhizal Fungi Altered by N Deposition on the Growth of a Native Shrub and an Invasive Annual Grass

Anthropogenic nitrogen (N) deposition causes shifts in vegetation types as well as species composition of arbuscular mycorrhizal (AM) fungi and other soil microorganisms. A greenhouse experiment was done to determine whether there are feedbacks between N-altered soil inoculum and growth of a dominant native shrub and an invasive grass species in southern California. The region is experiencing large-scale loss of Artemisia californica shrublands and replacement by invasive annual grasses under N deposition. Artemisia californica and Bromus madritensis ssp. rubens were grown with soil inoculum from experimental plots in a low N deposition site that had (1) N-fertilized and (2) unfertilized soil used for inoculum, as well as (3) high-N soil inoculum from a site exposed to atmospheric N deposition for four decades. All treatments plus a nonmycorrhizal control were given two levels of N fertilizer solution. A. californica biomass was reduced by each of the three inocula compared to uninoculated controls under at least one of the two N fertilizer solutions. The␣inoculum from the N-deposition site caused the greatest growth depressions. By contrast, B.␣madritensis biomass increased with each of the three inocula under at least one, or both, of the N solutions. The different growth responses of the two plant species may be related to the types of AM fungal colonization. B. madritensis was mainly colonized by a fine mycorrhizal endophyte, while A. californica had primarily coarse endophytes. Furthermore, A. californica had a high level of septate, nonmycorrhizal root endophytes, while B. madritensis overall had low levels of these endophytes. The negative biomass response of A. californica seedlings to high N-deposition inoculum may in part explain its decline; a microbially-mediated negative feedback may occur in this system that causes poor␣seedling growth and establishment of A.␣californica in sites subject to N deposition and B. madritensis invasion.

An Introduction to Propagation of Arbuscular Mycorrhizal Fungi in Pot Cultures for Inoculation of Native Plant Nursery Stock

Arbuscular mycorrhizal (AM) fungi can generally be collected from undisturbed sites and propagated in the nursery following a 7-step process. These fungi form symbiotic associations with most native plants and this relationship provides many benefits to plant health and function. Although specific techniques vary depending on the fungus, the plant, and the nursery, this introduction should help growers begin their own AM program.