Responses of soil microbial diversity, network complexity and multifunctionality to three land-use changes.

Land-use change is one of the greatest challenges for natural ecosystem services. Soil microbiomes are essential for modulating multiple ecosystem functions. However, little is known about the impact of land-use changes on soil microbial communities and their associated soil functions. In this study, 150 alpine soil samples representing conversion of forests to shrublands or grasslands, and of shrublands to grasslands were investigated for bacterial, fungal and protistan community diversity, co-occurrence network, as well as their relationships with soil multifunctionality via a sampling strategy of space-for-time substitution. The conversion of forest to grassland increased the diversity of fungi and bacteria, and altered the microbial community structures of bacteria, fungi and protists, resulting a greater impact on soil microbiome than other land-use conversions. Cross-trophic interaction analyses demonstrated this conversion increased microbial network complexity and robustness, whereas forest to shrubland had the opposite trend. The land-use induced changes in soil multifunctionality were related with microbial network modules, but were not always associated with variations of microbial diversity. Random forest modeling further suggested the significant role of microbial modules in explaining soil multifunctionality, together with environmental factors. These findings indicate divergent responses of belowground multitrophic organisms to land-use changes, and the potential role of microbial module in forecasting soil multifunctionality.

Effects of Amendments and Indigenous Microorganisms on the Growth and Cd and Pb Uptake of Coriander (Coriandrum sativum L.) in Heavy Metal-Contaminated Soils.

Heavy metal (HM) contamination of soils is a worldwide problem with adverse consequences to the environment and human health. For the safe production of vegetables in contaminated soil, efficient soil amendments need to be applied such as nano-hydroxyapatite (n-HAP) and poly γ-glutamic acid (γ-PGA), which can mitigate heavy metal uptake and enhance crop yield. However, the combined effects of soil amendments and indigenous microorganisms (IMOs) on HMs immobilisation and accumulation by crops have received little attention. We established a pot experiment to investigate the effects of IMOs combined with n-HAP and γ-PGA on coriander (Coriandrum sativum L.) growth and its Cd and Pb uptake in two acidic soils contaminated with HMs. The study demonstrated that applying n-HAP, with and without IMOs, significantly increased shoot dry biomass and reduced plant Cd and Pb uptake and diethylenetriaminepentaacetic acid (DTPA) extractable Cd and Pb concentrations in most cases. However, γ-PGA, with and without IMOs, only reduced soil DTPA-extractable Pb concentrations in slightly contaminated soil with 0.29 mg/kg Cd and 50.9 mg/kg Pb. Regardless of amendments, IMOs independently increased shoot dry biomass and soil DTPA-extractable Cd concentrations in moderately contaminated soil with 1.08 mg/kg Cd and 100.0 mg/kg Pb. A synergistic effect was observed with a combined IMOs and n-HAP treatment, where DTPA-extractable Cd and Pb concentrations decreased in slightly contaminated soil compared with the independent IMOs and n-HAP treatments. The combined treatment of γ-PGA and IMOs substantially increased shoot dry biomass in moderately contaminated soil. These results indicate that solo n-HAP enhanced plant growth and soil Cd and Pb immobilisation, and mitigated Cd and Pb accumulation in shoots. However, the combination of n-HAP and IMOs was optimal for stabilising and reducing HMs' uptake and promoting plant growth in contaminated soil, suggesting its potential for safe crop production.

Diversity and distribution of CO2-fixing microbial community along elevation gradients in meadow soils on the Tibetan Plateau.

Soil CO2-fixing microbes play a significant role in CO2-fixation in the terrestrial ecosystems, particularly in the Tibetan Plateau. To understand carbon sequestration by soil CO2-fixing microbes and the carbon cycling in alpine meadow soils, microbial diversity and their driving environmental factors were explored along an elevation gradient from 3900 to 5100 m, on both east and west slopes of Mila Mountain region on the Tibetan Plateau. The CO2-fixing microbial communities were characterized by high-throughput sequencing targeting the cbbL gene, encoding the large subunit for the CO2-fixing protein ribulose 1, 5-bisphosphate carboxylase/oxygenase. The overall OTU (Operational Taxonomic Unit) abundance is concentrated at an altitude between 4300 and 4900 m. The diversity of CO2-fixing microbes is the highest in the middle altitude area, and on the east slope is higher than those on the west slope. In terms of microbial community composition, Proteobacteria is dominant, and the most abundant genera are Cupriavidus, Rhodobacter, Sulfurifustis and Thiobacillus. Altitude has the greatest influence on the structural characteristics of CO2-fixing microbes, and other environmental factors are significantly correlated with altitude. Therefore, altitude influences the structural characteristics of CO2-fixing microbes by driving environmental factors. Our results are helpful to understand the variation in soil microbial community and its role in soil carbon cycling along elevation gradients.

Elevational distribution and occurrence of arbuscular mycorrhizal fungi in non-host Carex capillacea.

Little is known about Arbuscular mycorrhizal (AM) fungal colonization and community composition in non-mycorrhizal (NM) plants, especially along elevational gradients. This study explores this question using a NM plant, Carex capillacea, at Mount Segrila, Tibet. Here, C. capillacea, its rhizosphere soil, and the neighboring mycotrophic plant Poa annua were sampled at four elevations to evaluate and compare their AM fungi colonization and communities. The results showed that AM fungal colonization density of C. capillacea was negatively correlated with elevation and biomass of total NM plants per quadrat. AM fungal diversity and community composition between C. capillacea and P. annua showed a similar pattern. In addition, elevation and soil did not significantly influence the AM community in C. capillacea, while they were important abiotic factors for assemblages in rhizosphere soil and P. annua. These findings support that a broad array of AM fungi colonize the root of C. capillacea, and a mycelial network from a co-occurring host plant might shape the AM fungal communities in C. capillacea along the elevation gradient. The co-occurrence patterns of AM fungi associated with non-mycotrophic species and adjacent mycotrophic species have important implications for understanding AM fungal distribution patterns and plant-AM interactions.

Interactions between arbuscular mycorrhizal fungi and non-host Carex capillacea.

A topic of confusion over the interactions between arbuscular mycorrhizal (AM) fungi and plants is the mycorrhizal status of some plant families such as Cyperaceae, which is generally considered to be non-mycorrhizal. Here, we conducted experiments to explore how the abiotic environmental conditions and AM network influence the interactions between AM fungi and Carex capillacea. We grew Carex capillacea alone or together with a mycorrhizal host species Medicago sativa in the presence or absence of AM fungi (soil inoculum from Mount Segrila and Rhizophagus intraradices from the Chinese Bank of the Glomeromycota, BGC). Plants were grown in a growth chamber and at two elevational sites of Mount Segrila, respectively. The results indicate that mycorrhizal host plants ensured the presence of an active AM fungal network whether under growth chamber or alpine conditions. The AM fungal network significantly depressed the growth of C. capillacea, especially when native inocula were used and the plants grew under alpine site conditions, although root colonization of C. capillacea increased in most cases. Moreover, the colonization level of C. capillacea was much higher (≤ 30%) when growing under alpine conditions compared with growth chamber conditions (< 8.5%). Up to 20% root colonization by Rhizophagus intraradices was observed in monocultures under alpine conditions. A significant negative relationship was found between shoot phosphorus concentrations in M. sativa and shoot dry mass of C. capillacea. These results indicate that growing conditions, AM network, and inoculum source are all important factors affecting the susceptibility of C. capillacea to AM fungi, and growing conditions might be a key driver of the interactions between AM fungi and C. capillacea.

Temperature-mediated phylogenetic assemblage of fungal communities and local adaptation in mycorrhizal symbioses.

Recent work demonstrates that habitat conditions exert striking effects on symbiont performance by mediating trade-offs in plants, AM fungi and environmental interactions. However, how local temperature conditions influence the functional diversity of mycorrhizal symbioses and the genetics of coexisting AM fungi at the local scale remain unclear. In the present study, we conducted a reciprocal inoculation experiment to explore the performance of sympatric associations against allopatric associations under contrasting temperatures and the AM fungal community in colonized roots. No local adaptation of plant biomass was found under both temperature conditions investigated, but a consistent local versus foreign effect was found in AM fungal performance. The temperature and the origin of the inoculum relative to the plant origin were important in explaining symbiotic function. Correspondingly, the community structure and Nearest Relatedness Index of the AM fungal community of the root symbiont varied with inoculum source, and assemblages with more closely related taxa led to a decline in plant biomass and stronger disequilibrium among AM fungi in roots. Our findings suggest that functional divergence exists in naturally coexisting communities of AM fungi from contrasting climatic origins, and fungal relatedness is an important driver of plant growth.

Temperature-mediated local adaptation alters the symbiotic function in arbuscular mycorrhiza.

Variation in the symbiotic function of arbuscular mycorrhizal fungi (AM fungi) has been demonstrated among distinct biotic and abiotic interactions. However, there is little knowledge on how local temperature conditions influence the functional divergence of AM symbionts in alpine ecosystems. Here, we conduct a reciprocal inoculation experiment to explore the three-way interactions among plants, AM fungal inoculum and temperature at sites of contrasting elevation. Evidence of local adaptation of plant growth was found only under low temperature conditions, with no consistent local versus foreign effect found in AM fungal performance. The origin of either the plant or the inoculum relative to the temperature was important in explaining symbiotic function. Specifically, when inoculum and temperature were sympatric but allopatric to the plant, poor adaptation by the plant to the novel environment was clearly found under both temperature conditions. Further analysis found that the symbiotic function was inversely related to fungal diversity under high temperature conditions. These results suggest that local adaptation represents a powerful factor in the establishment of novel combinations of plant, inoculum and temperature, and confirms the importance of taking into account both biotic and abiotic interactions in the prediction of the response of symbionts to global environmental change.

Responses of arbuscular mycorrhizal symbionts to contrasting environments: field evidence along a Tibetan elevation gradient.

Plant adaptation to alpine ecosystems is not fully explained by plant physiological and morphological traits. Arbuscular mycorrhizal (AM) associations may be involved in mediating plant performance in response to environmental differences. Little is known, however, as to whether or not a close relationship exists between plant performance and arbuscular mycorrhizal fungus status across environmental gradients. We conducted a field investigation of the performance of six plant species and their associated AM fungi along higher and lower elevation gradients on Mount Segrila in Tibet. In most of our species, we observed higher shoot and inflorescence biomass production and a lower root-to-shoot ratio in the populations at those sites where the species was dominant (intermediate elevation sites) than in populations sampled at the limits of the distribution. The elevation pattern of root colonization differed with plant species on both gradients, and the extraradical hypha development of most species showed a unimodal pattern as did plant growth. The relationship between plant and fungus traits shows that AM fungus development generally matched host plant performance on the lower elevation gradient but not on the higher elevation gradient. This study provides evidence that plant distribution and productivity were significantly related to root and soil colonization by AM fungi, especially under less physically stressful conditions.

Altitudinal distribution patterns of AM fungal assemblages in a Tibetan alpine grassland.

A better understanding of biogeography of Glomeromycota is essential for the conservation of arbuscular mycorrhizal (AM) fungal species and the ecosystem services that they provide worldwide. We examined the spatial dynamics of AM fungi along two slopes (4149 m a.s.l. to the summit at 5033 m a.s.l.) of Mount Mila on the Tibetan Plateau. Our hypothesis was that AM fungal communities at higher elevation would show distinct assemblages with lower diversity in conditions of increasing environmental harshness. A total of 52 operational taxonomic units (OTUs) spanning all four orders were detected and some OTUs were habitat specific. Nearly 30% of the OTUs were new phylotypes, including two family-like clades. Distinct communities of AM fungi were found at the higher elevation, demonstrating potential niche differentiation along the elevation gradient. Elevation patterns of taxon richness/diversity differed between the two transects, decreasing with increasing elevation on the eastern slope and being unimodal (or lacking a pattern) on the western slope. Taken together, our findings provide evidence of a significant spatial structure of AM fungi across the elevation gradient, with the distribution patterns of these fungi regulated simultaneously by the plant communities, soil properties and climatic conditions in this plateau montane ecosystem.

Contribution of arbuscular mycorrhizal fungi of sedges to soil aggregation along an altitudinal alpine grassland gradient on the Tibetan Plateau.

The diversity of arbuscular mycorrhizal fungi (AMF) in sedges on the Tibetan Plateau remains largely unexplored, and their contribution to soil aggregation can be important in understanding the ecological function of AMF in alpine ecosystems. Roots of Kobresia pygmaea C.B. Clarke and Carex pseudofoetida Kük. in alpine Kobresia pastures along an elevational transect (4149-5033 m) on Mount Mila were analysed for AMF diversity. A structural equation model was built to explore the contribution of biotic factors to soil aggregation. Sedges harboured abundant AMF communities covering seven families and some operational taxonomic units are habitat specific. The two plant species hosted similar AMF communities at most altitudes. The relative abundance of the two sedges contributed largely to soil macroaggregates, followed by extraradical mycorrhizal hyphae (EMH) and total glomalin-related soil protein (T-GRSP). The influence of plant richness was mainly due to its indirect influence on T-GRSP and EMH. There was a strong positive correlation between GRSP and soil total carbon and nitrogen. Our results indicate that mycorrhization might not be a major trait leading to niche differentiation of the two co-occurring sedge species. However, AMF contribute to soil aggregation and thus may have the potential to greatly influence C and N cycling in alpine grasslands.

Molecular diversity of arbuscular mycorrhizal fungi associated with two co-occurring perennial plant species on a Tibetan altitudinal gradient.

Plant communities on Mount Segrila on the Tibetan Plateau show distinct changes at different altitudes, but little information is available on belowground communities of arbuscular mycorrhizal fungi (AMF). Root samples of two co-occurring species, Pennisetum centrasiaticum and Kobresia sp., growing in open grasslands at eight altitudes (3,446-4,556 m) were analyzed for diversity of AMF by PCR, cloning, and sequencing. Dominant plants were well colonized by AMF even at higher altitudes where spore density in rhizospheres decreased dramatically. A total of 29 operational taxonomic units (OTUs) of AMF were detected, and some novel sequence types were found. Acaulosporaceae and Glomeraceae were the dominant families. There was no significant difference in OTU richness along elevational gradients in Kobresia sp., but OTU richness in P. centrasiaticum was higher at intermediate elevations. Elevation, host plant species, and soil variables (pH, soil organic matter, and available P and N) were found to have significant effects on the overall AMF community across all elevations. Fungal community composition differed significantly between the two plant species at each elevation, and the similarity was generally higher at the intermediate elevations. No significant difference in compositional similarity was observed for Kobresia sp. with increasing elevation, but the dissimilarity increased significantly for P. centrasiaticum. These results suggest that host identity is an important determinant for the structure of the AMF communities along the elevational gradients in high altitude environments.

[Ecological distribution of arbuscular mycorrhizal fungi in alpine grasslands of Tibet Plateau].

Seventy soil samples with the roots of 37 dominant or common plant species on the grasslands in south and north Tibet Plateau were collected to study the ecological distribution of arbuscular mycorrhizal (AM) fungi in the investigation area. A total of 35 AM fungi species belonging to 5 genera were isolated, among which, 18 species belonged to Glomus, 9 species belonged to Acaulospora, 6 species belonged to Scutellospora, 1 species belonged to Entrophospora, and 1 species belonged to Paraglomus. There were 23 AM fungi species belonging to 4 genera isolated from south Tibet, and 22 species belonging to 4 genera from north Tibet. The Shannon diversity index of AM fungi in south and north Tibet Plateau was 2.31 and 2.75, respectively, and the spore density and species richness were significantly higher in north Tibet than in south Tibet. In different ecological zones, lesser AM fungi common species were found, species distribution was more site-specific, and different dominant species were observed. In alpine grassland, mountain meadow, and alpine meadow, the Shannon index of AM fungi was 1.91, 1.83, and 1.80, respectively; while in severely degraded temperate grassland, this index was only 1.64. The highest species richness of AM fungi occurred at the altitude of 4000-4600 m, but the highest Shannon index and species evenness occurred at the altitude of 4600-5220 m, with the values being 2.42 and 0.79, respectively. At all altitudes, Glomus was the dominant genus, and its relative abundance was higher when the altitude was below 4000 m. Acaulospora was mainly observed at the altitudes higher than 4000 m, Scutellospora was mainly distributed at the altitude 3500-5220 m, Paraglomus mainly occurred in the north alpine meadow with an altitude of 4000-5220 m and occasionally in the alpine steppe, whereas Entrophospora was only found in the south temperate grassland with an altitude of 3500-3700 m.

[Field inoculation effect of AM fungi on Tibet plateau Stipa bungeana grassland].

A field experiment was conducted on the Tibet plateau Stipa bungeana grassland to investigate the effects of arbuscular mycorrhizal fungi (AMF) inoculation on S. bungeana growth and its phosphorus uptake, and on soil microbial communities under unsterilized condition. The results showed that AMF inoculation increased the AMF spore density, infection rate and infection intensity in rhizosphere soil significantly, but had less effect on arbuscule richness. The infection rate of AMF increased with its increasing spore density, and the plant dry weight and P uptake of S. bungeana increased significantly with AMF infection rate. Similar results were observed in S. bungeana root dry weight and its P content. AMF inoculation increased the acid and alkaline phosphatase activities and bacterial biomass in rhizosphere soil, but had less effect on fungal and actinomyces communities. The infection effect of AMF varied with inoculation treatments, being in the sequence of Glomus mosseae + G. intraradices + Scutellospora calospora > G. mosseae + G. aggregatum > Glomus sp. > G. mosseae > G. mosseae + G. etunicatum + G. intraradices + S. erythropa > G. geosporum.

[Effects of environmental factors on AM fungi around steppe plant roots in Tibet Plateau].

The study on the representative steppe plant species in Tibet Plateau showed that the density of AM fungi spores in host plant rhizosphere did not correlate with the infection rate of AM fungi. The big changes in air temperature and rainfall at different altitudes played an important role in determining the growth and infection of AM fungi specific to steppe plants, and steppe type and soil texture also had obvious effects on AM fungi's growth and infection. Within a certain range, the spore density increased significantly with increasing soil pH (r = 0.5319, n = 20), but showed a declining trend with the improvement of soil organic matter (r = - 0.1973, n = 20). In contrast, the infection rates of AM fungi to host plants were to some extent negatively and positively correlated with soil pH and soil organic matter, respectively. Phosphorus (P) enrichment in soil environment led to the inhibition of the reproduction and infection of AM fungi. The suitable soil pH, OM and Olsen P contents for the growth and reproduction of AM fungi ranged from 8.0-8.7, 3.8-4.8 g.kg(-l) and 7.8-10.1 mg.kg(-1), respectively. Moderate and serious degradation of steppe (especially the serious degradation) had negative or detrimental impacts on the reproduction and infection of AM fungi. Reasonable grazing was helpful to the conservation of critical species of AM fungi. AM fungi also showed a relatively high infection rate on the roots of sedge species such as Carex praecpara, Kobresia humilis and Cyperus compressus.

[Field distribution pattern and metabolic activity of AM fungi and their effects on Ipomoea batatas growth].

The study showed that after eight weeks of inoculating sweet potato seedlings with AM fungus Glomus intraradices BEG141 under field condition, the shoot dry weight, individuals of tuber and their fresh weight, as well as the P uptake by the plant increased significantly. Compared with the control, the root colonization rate, extraradical hyphal density and intraradical alkaline phosphatase (ALP) activity were greatly increased. After inoculation with BEG141, the total and succinate dehydrogenase (SDH)- active hyphae paralleling with ridge were much more than those in underside direction, while there was no significant difference in different directions in the control. The increase of intra- and extraradical active hyphae density and the change of AM fungus distribution pattern after inoculation with mycorrhizal fungi may be the main reasons for the growth improvement of sweet potato.

[Effects of soil factors on arbuscular mycorrhizae (AM) fungi around roots of wild plants].

150 rhizospheric soil samples were collected from 45 wild plants distributed in Shandong Province during 1995-1997. More than forty species of AM fungi were isolated, and the effects of some soil factors on AM fungi were also investigated. It was proved that soil conditions were important factors to the colonization, growth, and distribution of AM fungi. Spore numbers were highest in brown earth, and lowest in alkali-saline soil. Glomus occurred in all types of soil. The occurrence frequency of Gigaspora and Scutellospora was much higher in brown earth. The distribution of AM fungi was also affected by soil pH. Glomus occurred in soil with a wide pH range. The greater of soil alkalinity, the more Glomus were found, while the greater of soil acidity, the more Acaulospora were isolated. Scutellospora occurred mostly in soil with pH of 6.0-7.0, and Gigaspora distributed mainly in acid soil.