Anna Liisa Ruotsalainen

Seasonality of root fungal colonization in low-alpine herbs

Abstract. Arbuscular mycorrhizal (AM) and dark septate endophytic (DSE) fungal colonization of Alchemilla glomerulans, Carex vaginata, Ranunculus acris ssp. pumilus and Trollius europaeus growing in low-alpine meadows in the Finnish subarctic were studied at different times during the growing season. Fungal colonization was correlated to soil soluble phosphorus (P) concentration. The influence of flower bud removal on fungal colonization was investigated in A. glomerulans, C. vaginata and R. acris and the correlation between AM and DSE colonization was studied. The fungal colonization patterns were found to be species-specific. R. acris maintained a relatively high rate of fungal colonization throughout the summer, while the rates of colonization of T. europaeus were lower and decreased towards the end of the season. A. glomerulans had constant arbuscular and vesicular colonization throughout the summer, but hyphal and DSE colonization declined towards the end of the season. C. vaginata did not form arbuscular mycorrhiza, but was colonized by DSE fungi and hyaline septate hyphae throughout the season. The soil soluble P concentration showed some seasonal variation, but was also highly variable between the study sites. Bud removal decreased arbuscular colonization of R. acris, but no unique effects were seen in any other parameters or the other species studied. The root fungal parameters correlated with soil P in some species at some sites, but no consistent trend was found. DSE colonization was positively correlated with root vesicular and hyphal colonization in some cases. The differences in fungal colonization parameters may be related to species-specific phenologies.

Root endophytes along a primary succession gradient in northern Finland

Primary successional vegetation gradients are characterized by changes in the soil microbial communities. However, information on possible shifts of the root endophytes along these gradients is scarce. The objective of the current study was to identify root endophytic fungi from a primary successional gradient on land uplift seashore of a geographically isolated island area. We applied a sequencing approach by amplifying the ITS region with fungal specific primers. We used mainly an isolate-based method, and to compare the abundance of culturable and unculturable endophytes, direct sequencing of one representative plant specimen Deschampsia flexuosa was also carried out. A total of 38 cultured endophytic strains were sequenced from Empetrum nigrum (Empetraceae), Vaccinium vitis-idaea (Ericaceae) and Deschampsia flexuosa (Poaceae). Out of these, 27 were identified as Phialocephala fortinii, three as Mollisia minutella, four as Phialophora sp., one as Ascomycetes sp. and three remained unidentified. The strains clustered into five clades in the phylogram, mostly irrespective of the successional stages and hosts from which they had been isolated. The early successional seashore dune ridge plants however, seemed to host a distinct fungal taxon, Phialophora sp. Culture-independent methods were applied on a root sample of a mid-successional Deschampsia flexuosa specimen and a total of 16 clones were randomly selected and sequenced. Out of 16 sequences, 13 were identified as unculturable strains and three showed closest similarity to a basidiomycete Cortinarius callisteus. The unculturable sequences were grouped into two main clades and were different from any culturable isolate in this study. Our results suggest that (i) P. fortinii dominates the isolate data at mid to late successional stages, (ii) roots of the ericaceous plants and the grass Deschampsia flexuosa are colonized by the same endophytic fungi in this ecosystem, and (iii) unculturable endophytes are common and potentially more abundant than the culturables. To our knowledge, this is the first report of the molecular phylogenies of the DSE in the mid-boreal zone and also the first report of the unculturable root endophytes of D. flexuosa.

Mycorrhizal benefit in two low arctic herbs increases with increasing temperature

Climate change may influence the relationship between arctic plants and their symbiotic mycorrhizal fungi. The benefit of the symbiosis for the host plant affects vegetation succession and may be a key parameter in predicting vegetation responses to warming. We investigated the mycorrhizal benefit in the low arctic perennial herbs Potentilla crantzii and Ranunculus acris in symbiosis with the arbuscular mycorrhizal fungus Glomus claroideum. Temperature response in the mycorrhiza-mediated acquisition of nitrogen (N) and phosphorus (P), growth, and photosynthetic nutrient-use efficiency were determined. Near the average natural soil temperature (12°C), mycorrhiza did not improve plant nutrient capture but significantly enhanced plant P capture at 17°C. Photosynthetic nitrogen-use efficiency was higher at 17°C than at 12°C and was further increased by mycorrhiza at 17°C. Photosynthetic phosphorus-use efficiency was not affected by temperature or mycorrhiza. Increasing the growing temperature by 5°C increased the relative shoot growth rate by 15%. Mycorrhizal symbiosis did not enhance plant growth rate, but the plants gained between 20% and 90% more mycorrhiza-mediated P when grown at higher temperature. The results suggest that these low arctic species have good potential to respond positively to increasing temperatures.

Mycorrhiza does not alter low temperature impact on Gnaphalium norvegicum

Extreme arctic-alpine vegetation has relatively low affinity to form mycorrhizal symbiosis. We asked whether the mycorrhizal growth benefit for the host plant is lower at low temperatures. We investigated the role of two root-associated fungi and temperature in growth, carbon–nitrogen relations and germination of an arctic-alpine herb. Seeds of Gnaphalium norvegicum were germinated at 8° or 15°C with or without arbuscular mycorrhizal (AM, Glomus claroideum) and dark septate endophytic (DSE, Phialocephala fortinii) inocula in a climate chamber. We found that germination percentage, shoot and root biomass, shoot N% and root AM colonization were lower at 8°C than at 15°C. P. fortinii inoculation had a positive impact on germination at both temperatures, whereas G. claroideum produced no effect. N% was lower in AM plants at both temperatures. Plant biomass and shoot N content were higher in AM plants than in control plants at 15°C, but not at 8°C. DSE inoculation tended also to have positive effects on plant biomass and N content at 15°C. At 15°C, rate of photosynthesis, photosynthetic nutrient use efficiency and specific leaf area were positively affected by G. claroideum, which suggests that G. claroideum formed a carbon sink and possibly enhanced the seedling water economy. The positive effects of P. fortinii were probably due to its saprotrophic function in the substrate because it did not colonize the roots. These results suggest that the effects of AM and DSE on plant growth are affected by temperature and that the mycorrhizal benefit for the host plant was lower at the lower temperature. Low saprotrophic activity and decreased mycorrhiza-mediated nutrient acquisition may thus constrain plant nutrient acquisition in cold environments. Decreased mycorrhizal benefit may be related to the comparatively low mycotrophy of cold environment vegetation.

Root fungal symbionts interact with mammalian herbivory, soil nutrient availability and specific habitat conditions

Herbivory, competition and soil fertility interactively shape plant communities and exhibit an important role in modifying conditions for host-dependent fungal symbionts. However, field studies on the combined impacts of natural herbivory, competition and soil fertility on root fungal symbionts are rare. We asked how mammalian herbivory, fertilization, liming and plant–plant competition affect the root colonization of arbuscular mycorrhizal fungi (AMF) and dark septate endophytic (DSE) fungi of the dicot herb, Solidago virgaurea. The 2-year full-factorial experiment was conducted in two contrasting habitats: non-acidic and acidic mountain tundra. We found that herbivory increased arbuscular colonization (i.e. the site of resource exchange) at fertile non-acidic sites, where vegetation was rich in species having AMF symbionts, whereas at infertile acidic sites, where plants having AMF symbiont are scarce, the response was the opposite. Herbivory of the host plant negatively affected DSE hyphal and sclerotial colonization in unfertilized plots, possibly due to reduced carbon flow from the host plant while there was no effect of herbivory in fertilized plots. DSE colonization was highest in unfertilized exclosures where soil nutrient concentrations were also lowest. Liming had a negative effect on DSE hyphal colonization, and its effect also interacted with herbivory and the habitat. Biomass removal of the neighboring plants did not affect the root colonization percent of either arbuscules or DSE. Our results show that the impacts of aboveground mammalian herbivory, soil nutrient availability and specific habitat conditions on belowground root fungal symbionts are highly dependent on each other. Arbuscule response to herbivory appeared to be regulated by specific habitat conditions possibly caused by differences in the AMF availability in the soil while DSE response was associated with availability of host-derived carbon. Our result of the relationship between herbivory and soil nutrients suggests an important role of DSE in ecosystem processes.

Moth herbivory enhances resource turnover in subarctic mountain birch forests

Massive moth outbreaks cause large-scale damage in subarctic mountain birch forests with a concomitant decrease in carbon flux to mycorrhizal fungi and an increased deposition of dissolved carbon and nutrients as moth frass into soil. We investigated impacts of moth herbivory along three replicated gradients with three levels of moth herbivory (undamaged, once damaged, repeatedly damaged) on soil nutrient levels and biological parameters. We found an increase in soil nutrients and in the biomass of enchytraeid worms, which are key faunal decomposers. Fungi bacteria ratio and C:N ratio decreased in humus with increasing severity of herbivory. Our findings suggest enhanced resource turnover in mountain birch forests due to massive moth herbivory. This may provide a shortcut for carbon and nutrient input to subarctic soils, which largely bypasses the main routes of carbon from plants to soil via mycorrhizal and litter-decomposing fungi. Moreover, a temporal shift occurs in carbon allocation to soil, providing decomposers an opportunity to use an early-season peak in resource availability. Our results suggest a hitherto unappreciated role of massive insect herbivore attacks on resource dynamics in subarctic ecosystems.

Contrasting impacts of defoliation on root colonization by arbuscular mycorrhizal and dark septate endophytic fungi of Medicago sativa

Individual plants typically interact with multiple mutualists and enemies simultaneously. Plant roots encounter both arbuscular mycorrhizal (AM) and dark septate endophytic (DSE) fungi, while the leaves are exposed to herbivores. AMF are usually beneficial symbionts, while the functional role of DSE is largely unknown. Leaf herbivory may have a negative effect on root symbiotic fungi due to decreased carbon availability. However, evidence for this is ambiguous and no inoculation-based experiment on joint effects of herbivory on AM and DSE has been done to date. We investigated how artificial defoliation impacts root colonization by AM (Glomus intraradices) and DSE (Phialocephala fortinii) fungi and growth of Medicago sativa host in a factorial laboratory experiment. Defoliation affected fungi differentially, causing a decrease in arbuscular colonization and a slight increase in DSE-type colonization. However, the presence of one fungal species had no effect on colonization by the other or on plant growth. Defoliation reduced plant biomass, with this effect independent of the fungal treatments. Inoculation by either fungal species reduced root/shoot ratios, with this effect independent of the defoliation treatments. These results suggest AM colonization is limited by host carbon availability, while DSE may benefit from root dieback or exudation associated with defoliation. Reductions in root allocation associated with fungal inoculation combined with a lack of effect of fungi on plant biomass suggest DSE and AMF may be functional equivalent to the plant within this study. Combined, our results indicate different controls of colonization, but no apparent functional consequences between AM and DSE association in plant roots in this experimental setup.

The modelled growth of mycorrhizal and non-mycorrhizal plants under constant versus variable soil nutrient concentration

Abstract. We studied the response of mycorrhizal and non-mycorrhizal plants to variation in soil nutrient concentration. A model for the relative growth rate (RGR) of plant biomass was constructed with soil nutrients as an explanatory variable. A literature survey was carried out to find the relative magnitudes of parameter values for mycorrhizal and non-mycorrhizal plants. Mycorrhizal plants had higher RGR at low nutrient concentrations and non-mycorrhizal plants at high nutrient concentrations. The RGR of mycorrhizal and non-mycorrhizal plants at constant versus log-normally distributed soil nutrient concentration were compared to see the effect of mycorrhizal status on responses to variation. Variation in nutrient concentration generally reduced RGR, especially in mycorrhizal plants. The RGR of a non-mycorrhizal plant may increase with variation where a growth function threshold exists, i.e. a soil nutrient concentration that must be exceeded to allow growth. Mycorrhizal plants appeared more sensitive to variation in nutrient concentration than non-mycorrhizal plants due to the higher affinity of mycorrhizal roots at low nutrient levels. However, this prediction may be reversed if mycorrhizal symbiosis considerably stabilises flow of nutrients to plant physiological processes, such that mycorrhizal plants experience less variation in soil nutrient concentration than non-mycorrhizal plants. Our results also attain broader significance by suggesting a general trade-off between competitive ability in a constant versus variable resource availability.

Contrasting preferences of arbuscular mycorrhizal and dark septate fungi colonizing boreal and subarctic Avenella flexuosa

Arbuscular mycorrhizal (AM) and dark septate endophytic (DSE) fungi are ubiquitous in grass roots, but their colonizations may vary according to latitudinal gradient and site conditions. We investigated how vegetation zone (boreal vs. subarctic), humus thickness, and site openness affect root fungal colonizations of the grass Avenella flexuosa. More precisely, we hypothesized that AM and DSE fungal colonizations would have different responses to environmental conditions such that AM fungi could be more common in boreal zone, whereas we expected DSE fungi to be more affected by the amount of humus. We found site openness to affect AM and DSE fungi in a contrasting manner, in interaction with the vegetation zone. AM colonization was high at open coastal dunes, whereas DSE fungi were more common at forested sites, in the boreal zone. Humus thickness affected AM fungi negatively and DSE fungi positively. To conclude, the observed AM and DSE fungal colonization patterns were largely contrasting. AM fungi were favored in seashore conditions characterized by thin humus layer, whereas DSE fungi were favored in conditions of higher humus availability.

Root fungal colonisations of the understory grass Deschampsia flexuosa after top-canopy harvesting

AimsRoot fungal relationships in forest understory may be affected by tree harvesting. Deschampsia flexuosa forms a mutualistic symbiosis with arbuscular mycorrhizal (AM) fungi functioning in nutrient uptake, and a more loose association with dark septate endophytic (DSE) fungi. We asked how harvesting affects fungal colonisations and whether DSE is more prone to change than AM.MethodsDeschampsia flexuosa plants were sampled close to a control or a cut tree after top-canopy harvesting in a primary successional site. Colonisations were studied using light microscopy. Shoot N%, vegetation cover and soil nutrients were determined.ResultsTree harvesting did not affect vegetation and soil parameters, except potassium (K+) increasing near cut trees. AM colonisation did not change, while DSE increased. Shoot N% increased with increasing DSE near cut trees. Hyaline septate (HSE) hyphae and soil K+ and magnesium (Mg2+) were positively correlated near control trees. Lichen cover and HSE correlated negatively.ConclusionsDSE colonisation increased but AM did not change after harvesting. Positive correlation of DSE with shoot N% near cut trees may suggest a role for DSE in favouring plant nitrogen uptake after disturbance in an open microsite. HSE may play a role in K+ and Mg2+ uptake.