Insights into nitrogen and carbon dynamics of ectomycorrhizal and saprotrophic fungi from isotopic evidence

The successful use of natural abundances of carbon (C) and nitrogen (N) isotopes in the study of ecosystem dynamics suggests that isotopic measurements could yield new insights into the role of fungi in nitrogen and carbon cycling. Sporocarps of mycorrhizal and saprotrophic fungi, vegetation, and soils were collected in young, deciduous-dominated sites and older, coniferous-dominated sites along a successional sequence at Glacier Bay National Park, Alaska. Mycorrhizal fungi had consistently higher δ¹⁵N and lower δ¹³C values than saprotrophic fungi. Foliar δ¹³C values were always isotopically depleted relative to both fungal types. Foliar δ¹⁵N values were usually, but not always, more depleted than those in saprotrophic fungi, and were consistently more depleted than in mycorrhizal fungi. We hypothesize that an apparent isotopic fractionation by mycorrhizal fungi during the transfer of nitrogen to plants may be attributed to enzymatic reactions within the fungi producing isotopically depleted amino acids, which are subsequently passed on to plant symbionts. An increasing difference between soil mineral nitrogen δ¹⁵N and foliar δ¹⁵N in later succession might therefore be a consequence of greater reliance on mycorrhizal symbionts for nitrogen supply under nitrogen-limited conditions. Carbon signatures of mycorrhizal fungi may be more enriched than those of foliage because the fungi use isotopically enriched photosynthate such as simple sugars, in contrast to the mixture of compounds present in leaves. In addition, some ¹³C fractionation may occur during transport processes from leaves to roots, and during fungal chitin biosynthesis. Stable isotopes have the potential to help clarify the role of fungi in ecosystem processes. Received: 7 January 1998 / Accepted: 9 November 1998     

Fertilization alters nitrogen isotopes and concentrations in ectomycorrhizal fungi and soil in pine forests

To assess how nitrogen (N) availability affected ectomycorrhizal functioning and to test a theoretical model of ectomycorrhizal ¹⁵N partitioning, we measured C/N and δ¹⁵N in soils and nine fungal taxa in two Swedish N addition experiments. Sporocarp C/N and soil C/N decreased with fertilization, implying that N uptake per unit fungal growth increased. The S horizon was more responsive than the F and H horizons to changes in N addition, with N turnover for these horizons of 24, 57, and 57 y, respectively. Fungal and soil δ¹⁵N patterns identified fungal N sources, with N acquisition primarily from the S, F, or H horizon for two, five, and two taxa, respectively. With increasing N availability, sporocarp ¹⁵N enrichment increased in five taxa, in agreement with our model of fungal-plant N partitioning. However, it decreased in Lactarius rufus and Russula aeruginea, perhaps indicating shifts towards greater inorganic N uptake in these two taxa. This may relate to the generally lower sensitivity of these taxa to N deposition compared to the Cortinarius and Suillus taxa that fit our model of ¹⁵N partitioning.     

Carbon and nitrogen acquisition strategies by wood decay fungi influence their isotopic signatures in Picea abies forests

We examined whether sporocarp carbon and nitrogen isotope ratios (δ¹³C and δ¹⁵N values) reflected different functional strategies in 15 species of wood decay fungi. In Finnish Picea abies forests, we compared sporocarp δ¹³C and δ¹⁵N against log diameter, proximity to ground, and three wood decay types, specifically brown rot, nonselective white rot, and selective white rot (targeting hemicellulose and lignin preferentially). In regression analysis (adjusted r² = 0.576), species accounted for 31% of variability in δ¹³C, with factors influencing wood δ¹³C accounting for the remainder. Brown rot fungi and three white rot fungi that selectively attacked hemicellulose (Heterobasidion parviporum, Phellopilus nigrolimitatus, and Trichaptum abietinum) were higher in δ¹³C than nonselective white rot fungi. This was attributed to greater assimilation of ¹³C-enriched pentoses from hemicellulose by these fungi. The pathogenic white rot fungus Heterobasidion parviporum had higher δ¹⁵N with proximity to ground and increasing log diameter. This suggested that ¹⁵N-enriched soil N contributed to decomposing logs and that Heterobasidion growing from a bigger resource base had increased access to soil N. These isotopic patterns accordingly reflected both functional diversity of wood decay fungi and site-specific factors.     

The stable carbon and nitrogen isotopic composition of vegetation in tropical forests of the Amazon Basin, Brazil

Here we present the within-site, seasonal, and interannual variations of the carbon (δ¹³C) and nitrogen (δ¹⁵N) isotope ratios of leaves, wood, bark and litter from four sites in the Amazon region, Brazil. Samples were collected in Manaus (3° 06′07′′ S; 60°01′30′′ W), Ji-Paraná (10°53′07′′ S; 61°57′06′′ W), and Santarém (2°26′35′′ S; 54°42′30′′ W) with mean annual precipitation of 2207, 2040 and 1909 mm respectively. The overall average for all leaf samples was for δ¹³C and for δ¹⁵N (n=756). The leaf δ values at these sites were often but not always statistically distinct from each other. The δ¹³C values varied from to . Pronounced differences in δ¹³C values occurred with height associated with differences in forest structure. The δ¹³C of leaf dry matter showed seasonal variations associated with the length of the dry season, despite the fact that total annual precipitation was similar among the studied sites. Leaf δ¹⁵N values ranged from to a maximum value of , and the Santarém sites showed more enriched values than Manaus and Ji-Paraná sites. No seasonal variation was detected in the δ¹⁵N of leaves, but significant differences were observed among sites and with changes in canopy height. The isotope ratio data are consistent with our current understanding of the roles of light, water availability, and recycling of soil-respired CO₂ influences on δ¹³C and consistent with our understanding that an open nitrogen cycle can lead to high δ¹⁵N values despite a significant number of legumes in the vegetation.     

Saprotrophic and ectomycorrhizal fungi exhibit contrasting richness patterns along elevational gradients in cool-temperate montane forests

Increasing evidence suggests that elevational gradients of soil fungal richness are highly variable, but few studies have examined how diversity components of each guild contribute to overall fungal diversity. Here, we aimed to disentangle the relationships between total, saprotrophic, and ectomycorrhizal fungal richness, and environmental factors across elevational gradients in cool-temperate montane forests. We observed that total and saprotrophic richness decreased but ectomycorrhizal richness increased with increase in elevation. Elevational range size and nestedness analyses illustrated that saprotrophic and ectomycorrhizal source communities were located at lower and higher elevations, respectively. The observed total and saprotrophic richness were directly influenced by soil properties and indirectly influenced by climate and plant communities. Ectomycorrhizal fungal richness was affected by climate and the dominance of ectomycorrhizal trees. We highlight that two directional source–sink dynamics lead to opposite elevational patterns between saprotrophic and ectomycorrhizal fungal richness, shaping the variation in elevational richness gradients.     

长期氮添加对亚热带森林土壤微生物碳源代谢多样性的影响

土壤微生物功能多样性对维持生态系统功能和稳定性具有非常重要的意义。人类活动造成全球氮沉降量激增,会引起土壤微生物群落结构和功能的改变,但是目前有关亚热带森林生态系统土壤微生物碳源利用多样性对模拟N沉降的响应还不是很清楚。依托位于鼎湖山的长达15年的长期野外模拟氮沉降试验样地平台,借助Biolog-Eco微平板技术,分析探讨了不同N添加量对季风常绿阔叶林中土壤微生物群落碳代谢功能多样性的影响差异及机制。研究结果表明:(1)与对照相比,长期低N、中N和高N量添加使土壤微生物碳源代谢活性(AWCD)分别显著降低了15.3%、32.9%和38.0%;并且显著降低了微生物碳源利用Shannon多样性指数和丰富度指数;(2)微生物对糖类、羧酸、氨基酸、胺类和酚酸类的利用随着施N水平的提高而显著降低;其中胺类最为敏感,长期高N添加下其利用强度与对照相比显著降低了80.2%;(3)主成分分析表明,不同N添加水平显著影响了土壤微生物碳源利用(P=0.001);(4)分类变异分析表明,土壤和植物因素可解释不同N水平下碳源利用差异的90.7%;(5)典型对应分析发现,土壤pH(P=0.009)是解释不同N水平添加处理间土壤微生物碳源代谢多样性差异的主要环境因子;而植被丰富度和凋落物量没有显著影响。综上所述,长期不同N添加显著影响了亚热带森林土壤微生物碳源代谢多样性,并且结果表明土壤pH是影响碳源利用多样性的主要影响因素。研究结果对于揭示全球变化影响下热带森林生态系统地下生物多样性维持机制提供了重要理论依据。     

The carbon and nitrogen isotopic values of particulate organic material from the Florida Keys: a temporal and spatial study

The δ¹⁵N and δ¹³C values of particulate organic material (POM) were analyzed from 35 sites in the Florida Keys over the time interval 2000 to 2002. The sites within the study area were delineated into nine transects stretching from Key West to Key Largo. Each transect consisted of three to five sites extending from close to the Keys to the edge of the reef tract. The POM had mean δ¹⁵N and δ¹³C values of +3.6‰ (σ = ±3.2‰) and −19.9‰ (σ = ±0.6‰) respectively. Over the study period there were no statistically significant changes in δ¹⁵N, δ¹³C, or C:N. For the majority of the sampling dates, the δ¹³C values showed a distinct inshore (δ¹³C = −18.3‰, σ = ±1.0‰) to offshore gradient (δ¹³C = −21.4, σ = ±0.9‰). In contrast, the δ¹⁵N values showed no consistent patterns related to the distance from land. The more positive δ¹³C values of the nearshore samples suggest that the source of the carbon and the nitrogen in the POM in the nearshore was mainly derived from the degradation of seagrass detritus and not from the input of anthropogenically derived material from the Florida Keys. In contrast, the POM on the outer reef was dominated by marine plankton. As mineralization and nitrification of the organic nitrogen pool are major contributors to the dissolved inorganic nitrogen in the water column, it is unlikely that variations in the δ¹⁵N of the algae and other benthic organisms reported in the Florida Keys are related to the input of sewage.     

Epigeous fruiting bodies of ectomycorrhizal fungi as indicators of soil fertility and associated nitrogen status of boreal forests

Soil fertility and associated nitrogen (N) status was a key ecosystem attribute, and surveys of ectomycorrhizal fungal (EMF) communities via epigeous fruiting bodies could provide an effective biotic indicator of forest soil productivity. We explored the utility of aboveground EMF communities in this regard by surveying sporocarps over a 3-year period from contrasting plant associations of southern old-growth boreal forests of British Columbia (Canada). Cumulative richness ranged from 39 to 89 EMF species per plot (0.15 ha) and followed a skewed parabolic correlation with foliar N concentrations and soil N availability. EMF species composition was consistently distinct in ordinations and strongly correlated to the increasing rates of N mineralization aligned with soil productivity. Approximately 40 EMF species were specialists, as they collectively indicated oligotrophic, mesotrophic, and eutrophic nutrient regimes, while the remaining species were categorized as broadly tolerant (distributed over 100% of the N gradient), partially intolerant (approximately 70%), or satellites (rare). The functional organization of EMF communities reflected by distribution classes could help define the ecological integrity of forests, which was characterized in this boreal landscape by an average allotment of 20 broadly tolerant, 25 partially intolerant, 15 specialist, and ten satellite species per plot. Epigeous fruiting bodies provided a disparate yet complementary view to the belowground assessment of EMF communities that was valuable in identifying indicators for ecosystem monitoring.     

Effects of fixation on freshwater invertebrate carbon and nitrogen isotope composition and its arithmetic correction

Retrospective investigations using carbon and nitrogen stable isotope composition of archived material have a great potential for describing past effects of anthropogenic ecosystem alterations or natural shifts in ecosystems. In this study, we examined the effects of two commonly used preservation substances of freshwater invertebrates, ethanol and lugol, on δ¹³C and δ¹⁵N of various planktonic and benthic taxa. For both isotopes, the average effect of fixation in ethanol was stronger than in lugol, and the effects on δ¹³C were stronger than on δ¹⁵N (average ± SD: 1.18 ± 0.94 and −0.47 ± 0.99 for δ¹³C ethanol and lugol fixed samples, respectively, and 0.39 ± 0.68 and 0.17 ± 0.77 for δ¹⁵N, respectively). The changes in the isotopic composition were not dependent on the initial isotopic composition of each taxon, but were related with concomitant changes in the carbon or nitrogen content. Application of a mass balance correction equation to the fixed samples resulted in a significantly lower average effect of fixation in ethanol (0.01 ± 0.59 and 0.44 ± 0.65 for δ¹⁵N and δ¹³C, respectively), while corrections had little effect for lugol fixed samples (0.24 ± 0.53 and −0.39 ± 0.85, respectively). For both isotopes and fixatives, corrections resulted in linear relationships between fixed vs. control samples, with slopes and intercepts not significantly different from 1 and 0, respectively. Therefore, mass balance correction of stable isotopes in fixed invertebrates is recommended for minimising the effects of fixation. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Handling editor: M. Power     

Diversity and abundance of resupinate thelephoroid fungi as ectomycorrhizal symbionts in Swedish boreal forests

Resupinate thelephoroid fungi (hereafter called tomentelloid fungi) have a world-wide distribution and comprise approximately 70 basidiomycete species with inconspicuous, resupinate sporocarps. It is only recently that their ability to form ectomycorrhizas (EM) has been realized, so their distribution, abundance and significance as mycobionts in forest ecosystems is still largely unexplored. In order to provide baseline data for future ecological studies of tomentelloid fungi, we explored their presence and abundance in nine Swedish boreal forests in which the EM communities had been analysed. Phylogenetic analyses were used to compare the internal transcribed spacer of nuclear ribosomal DNA (ITS rDNA) sequence data obtained from mycobionts on single ectomycorrhizal tips with that obtained from sporocarps of identified tomentelloid fungi. Five species of Tomentella and one species of Pseudotomentella were identified as ectomycorrhizal fungi. The symbiotic nature of Tomentella bryophila, T. stuposa, T. badia and T. atramentaria is demonstrated for the first time. T. stuposa and Pseudotomentella tristis were the most commonly encountered tomentelloid fungi, with the other species, including T. sublilacina, only being recorded from single stands. Overall, tomentelloid fungi were found in five of the studies, colonizing between 1 and 8% of the mycorrhizal root tips. Two of the five sites supported several tomentelloid species. Tomentelloid fungi appear to be relatively common ectomycorrhizal symbionts with a wide distribution in Swedish coniferous forests. The results are in accordance with accumulating data that fungal species which lack conspicuous sporocarps may be of considerable importance in EM communities.     

Contrasting carbon metabolism in saprotrophic and pathogenic microascalean fungi from Protea trees

Protea-associated Knoxdaviesia species grow on decaying inflorescences, yet are closely related to plant pathogens such as Ceratocystis albifundus. C. albifundus also infects Protea, but occupies a distinct niche. We investigated substrate utilization in two Knoxdaviesia saprotrophs, a generalist and a specialist, and the pathogen C. albifundus by integrating phenome and whole-genome data. On shared substrates, the generalist grew slightly better than its specialist counterpart, alluding to how it has maintained its Protea host range. C. albifundus grew on few substrates and had limited cell wall-degrading enzymes. It did not utilize sucrose, but may prefer soluble oligosaccharides. Nectar monosaccharides are likely important carbon sources for early colonizing Knoxdaviesia species. Once the inflorescence ages, they could switch to degrading cell wall components. C. albifundus likely uses its limited cell wall-degrading arsenal to gain access to plant cells and exploit internal resources. Overall, carbon metabolism and gene content in three related fungi reflected their ecological adaptations.     

Carbon and nitrogen cycling in North American boreal forests

A model of boreal forest dynamics was adapted to examine the factors controlling carbon and nitrogen cycling in the boreal forests of interior Alaska. Empirical relationships were used to simulate decomposition and nitrogen availability as a function of either substrate quality, the soil thermal regime, or their interactive effects. Test comparisons included black spruce forests growing on permafrost soils and black spruce, birch, and white spruce forests growing on permafrost-free soils. For each forest, simulated above-ground tree biomass, basal area, density, litterfall, moss biomass, and forest floor mass, turnover, thickness, and nitrogen concentration were compared to observed data. No one decay equation simulated forests entirely consistent with observed data, but over the range of upland forest types in interior Alaska, the equation that combined the effects of litter quality and the soil thermal regime simulated forests that were most consistent with observed data. For black spruce growing on permafrost soils, long-term simulated forest dynamics in the absence of fire resulted in unproductive forests with a thick forest floor and low nitrogen mineralization. Fires were an important means to interrupt this sequence and to restart forest succession.     

Recovery of the ectomycorrhizal community after termination of long-term nitrogen fertilisation of a boreal Norway spruce forest

Ectomycorrhizal fungi (ECM) are a fundamental component of boreal forests promoting tree growth and participating in soil nutrient cycling. Increased nitrogen (N) input is known to largely influence ECM communities but their potential recovery is not well understood. Therefore, we studied the effects of long-term N-fertilisation on ECM communities, and their recovery after termination of N treatment. Fungal ITS sequencing data indicated that N-fertilisation (34 kg N ha⁻¹ y⁻¹) for 46 y decreased the relative abundance of ECM species in the fungal community and suppressed originally dominating medium-distance fringe exploration types adapted to N-limited conditions, while the ECM diversity remained unaffected. In other plots, 23 y after termination of fertilisation at 73 kg N ha⁻¹ y⁻¹ for 23 y, the relative abundance of ECM species shifted closer to, but did not reach, control levels. These observations indicate only slow recovery of ECM community, likely due to a high soil N retention capacity.     

Understorey plant and soil responses to disturbance and increased nitrogen in boreal forests

Question: How do N fertilization and disturbance affect the understorey vegetation, microbial properties and soil nutrient concentration in boreal forests? Location: Kuusamo (66°22′N; 29°18′E) and Oulu (65°02′N; 25°47′E) in northern Finland. Methods: We conducted a fully factorial experiment with three factors: site (two levels), N fertilization (four levels) and disturbance (two levels). We measured treatment effects on understorey biomass, vegetation structure, and plant, soil and microbial N and C concentrations. Results: The understorey biomass was not affected by fertilization either in the control or in the disturbance treatment. Fertilization reduced the biomass of deciduous Vaccinium myrtillus. Disturbance had a negative effect on the biomass of V. myrtillus and evergreen Vaccinium vitis‐idaea and decreased the relative proportion of evergreen species. Fertilization and disturbance increased the biomass of grass Deschampsia flexuosa and the relative proportion of graminoids. The amount of NH₄⁺ increased in soil after fertilization, and microbial C decreased after disturbance. Conclusions: Our results suggest that the growth of slow‐growing Vaccinium species and soil microbes in boreal forests are not limited by N availability. However, significant changes in the proportion of dwarf shrubs to graminoids and a decrease in the biomass of V. myrtillus demonstrate the susceptibility of understorey vegetation to N enrichment. N enrichment and disturbance seem to have similar effects on understorey vegetation. Consequently, increasing N does not affect the rate or the direction of recovery after disturbance. Moreover, our study demonstrates the importance of understorey vegetation as a C source for soil microbes in boreal forests.