Shrubs influence arbuscular mycorrhizal fungi communities in a semi-arid environment

Interactions between arbuscular mycorrhizal fungi (AMF) and plants are essential components of ecosystem functioning; however, they remain poorly known in dry ecosystems. We examined the relationship between seven shrub species and their associated AMF community in a semi-arid plant community in southern Spain. Soil characteristics and plant physiological status were measured and related to AMF community composition and genetic diversity by multivariate statistics. We found differences in AMF communities in soils under shrubs and in gaps among them, whereas no differences were detected among AMF communities colonizing roots. Soil nutrients content drove most of the spatial variations in the AMF community and genetic diversity. AMF communities were more heterogeneous in fertile islands with low nitrogen-to-phosphorus ratio and vice versa. AMF genetic diversity increased in soils limited by phosphorus and with high soil organic matter content, while AMF genetic diversity increased in roots growing in soil not limited by phosphorus. Overall, we could not find a clear link between plant performance and the associated AMF community. Our findings show that different shrub species generate islands of fertility which differ in nutrient content and, therefore, support different AMF communities, increasing AMF diversity at the landscape level.     

Methyl-mercury affects microbial activity and biomass,bacterial community structure but rarely the fungal community structure

Monomethyl-mercury is one of the most toxic compounds. Methylation of Hg usually appears under anoxic conditions. In Swiss forest soils, methyl-Hg concentrations of up to 3 μg kg⁻¹ soil dw have been observed, but the impact of methyl-Hg on soil microorganisms have rarely been examined so far. In this study, we investigated the effect of increasing concentrations of methyl-Hg (0, 5, 20, 90 μg kg⁻¹ soil dw) on the microbial communities in various forest soils differing in their physico-chemical properties. Experiments were conducted in microcosms under controlled conditions and the basal respiration (BR), the microbial biomass carbon (MBC) and the bacterial and fungal community structures using T-RFLP-profiling were investigated. BR was significantly affected by methyl-Hg. In general, the BR increased with increasing methyl-Hg concentrations, whereas the MBC was significantly reduced. Bacterial communities were more sensitive to methyl-Hg than fungal communities. In five out of seven soils, the bacterial community structures differed significantly between the treatments whereas the fungal communities did not. The impact of methyl-Hg on the soil bacterial communities was site specific. In one soil, a methyl-Hg concentration of already 5 μg kg⁻¹ soil dw significantly affected the relative abundance of 13% bacterial operational taxonomic units (OTU), whereas in other soils concentrations of even 90 μg kg⁻¹ soil dw rarely affected the abundance of OTUs. In this study, for the first time, the impact of methyl-Hg on soil bacterial and fungal communities in forest soils was assessed. We showed that its impact strongly depends on the physico-chemical conditions of the soil and that bacterial communities were more sensitive to methyl-Hg than fungi.     

Evaluating the physical capture method of terminal restriction fragment length polymorphism for comparison of soil microbial communities

Terminal restriction fragment length polymorphism (T-RFLP) is a popular method of comparative microbial community analysis which is normally accomplished by tagging terminal restriction fragments (T-RFs) with a fluorescent primer. Here, we evaluate an alternative method of T-RFLP where T-RFs are physically captured using a biotinylated primer and streptavidin-coated beads. This eliminates one of the primary criticisms of T-RFLP, namely that T-RFs cannot be identified by sequence analysis, and also represents an alternative method for collecting T-RFLP profiles. Microbial communities from forest, agricultural, and turf soils were investigated using several sets of primers specific for different microbial groups. The physical capture method of T-RFLP resulted in similar profiles to those generated by fluorescent T-RFLP. The relationships among ecosystem types captured by both methods and revealed by ordination were virtually identical. The total variance in the profiles that was attributed to ecosystem type was approximately equal, or greater, when generated by the physical capture method, depending on the primers used. However, physical capture T-RFLP resolved fewer T-RFs than fluorescent T-RFLP, and this may reduce the sensitivity to changes in non-dominant populations within the community. Direct cloning and sequencing of physical capture T-RFs revealed that most bands were not comprised of sequences related to those in the database that would generate T-RFs of similar size. T-RFs should therefore be identified by sequencing, rather than by comparing the sizes of T-RFs to computer digests of database sequences. Physical capture T-RFLP should be a useful tool to identify T-RFs by sequencing, and for laboratories without economical access to equipment required to perform fluorescent T-RFLP.     

No evidence that nitrogen enrichment affect fungal communities of Vaccinium roots in two contrasting boreal forest types

In boreal forests ericaceous shrubs often dominate the forest floor vegetation. Nitrogen enrichment has been shown to decrease shrub abundance and in this study we explored whether it also affects the root associated fungal communities. Fine roots of Vaccinium myrtillus were collected in a Norway spruce dominated forest and of Vaccinium vitis-idaea in a Scots pine dominated forest. In both forests, nitrogen enrichment was experimentally induced by adding 12.5 and 50 kg N ha⁻¹ yr⁻¹ for 12 (spruce forest) and four (pine forest) years. Based on terminal restriction fragment length polymorphisms, subcloning and sequencing analyses, the root associated fungal communities were examined. We found 93 fungal species including Asco-, Basidio- and Zygo-mycota. In general, the Rhizoscyphus ericae aggregate was the most dominant and this was followed by Herpotrichiellaceae and Sebacina. Ordination analysis revealed that nitrogen enrichment did not change species composition of the fungal communities in neither the spruce nor the pine forest, while fungal community structures were clearly discriminated between the dominant shrub species in each forest. Similarly, no fungal species showed a significant response to nitrogen enrichment. Therefore, nitrogen enrichment appears to have no effect on root associated fungi of understorey dwarf shrubs in boreal forests, while it is clear that spruce and pine forests harbor distinctive communities of these fungi.     

Woodland trees modulate soil resources and conserve fungal diversity in fragmented landscapes

Resource islands around woody plants are thought to define the structure and function of many semiarid and arid ecosystems, but their role in patterning of soil microbial communities remains largely unexamined in dry environments. This study examined soil resource distribution and associated fungal communities in two Allocasuarina luehmannii (buloke) remnants of semiarid north-western Victoria, Australia. These savannah-like woodlands are listed as endangered due to extensive clearing for agriculture. We used the DNA-based profiling technique T-RFLP and ordination-based statistical methods to compare fungal community compositions in surface soils from two remnants (located 1.6 km apart) and three sampling positions (beneath individual buloke canopies; grassy inter-canopy areas; and adjoining cleared paddocks). Resource island formation beneath buloke trees was clearly evident in soil physicochemical properties (e.g. threefold concentrations of total carbon and nitrogen in canopy versus non-canopy soils). This heterogeneity of resources was moderately correlated with soil fungal community compositions, which were distinct for each sampling position. We argue that fungal composition patterns reflected multiple roles of fungi in dryland ecosystems, namely: responses of saprotrophic fungi to tree organic matter inputs; specificity of ectomycorrhizal fungi to tree rooting zones; and fungal involvement in biological soil crusts that variably covered non-canopy soils. Our data did not indicate that buloke canopy areas were particular hotspots of soil fungal diversity, but that they increased landscape-level diversity by supporting a distinct suite of fungi. In addition, we provide evidence of phylogenetic differentiation of soil fungal communities between our two remnants, which adds to growing evidence of fungal genetic structure at localised scales. These findings highlight the importance of remnant trees in conserving both soil resources and microbial genetic diversity. In addition, evidence of differentiation of soil fungal phylogenetics between nearby but isolated remnants suggests that conserving soil fungal diversity requires conservation of host habitats over their entire (remaining) range, and indicates previously unseen consequences of tree loss from extensively cleared landscapes.     

Response of soil microbial communities to compost amendments

Soil organic matter is considered as a major component of soil quality because it contributes directly or indirectly to many physical, chemical and biological properties. Thus, soil amendment with composts is an agricultural practice commonly used to improve soil quality and also to manage organic wastes. We evaluated in laboratory scale experiments the response of the soilborne microflora to the newly created soil environments resulting from the addition of three different composts in two different agricultural soils under controlled conditions. At a global level, total microbial densities were determined by classical plate count methods and global microbial activities were assessed by measuring basal respiration and substrate induced respiration (SIR). Soil suppressiveness to Rhizoctonia solani diseases was measured through bioassays performed in greenhouses. At a community level, the modifications of the metabolic and molecular structures of bacterial and fungal communities were assessed. Bacterial community level physiological profiles (CLPP) were determined using Biolog™ GN microtiter plates. Bacterial and fungal community structures were investigated using terminal restriction fragment length polymorphism (T-RFLP) fingerprinting. Data sets were analyzed using analysis of variance and ordination methods of multivariate data. The impact of organic amendments on soil characteristics differed with the nature of the composts and the soil types. French and English spent mushroom composts altered all the biological parameters evaluated in the clayey soil and/or in the sandy silty clay soil, while green waste compost did not modify either bacterial and fungal densities, SIR values nor soil suppressiveness in any of the soils. The changes in bacterial T-RFLP fingerprints caused by compost amendments were not related to the changes in CLPP, suggesting the functional redundancy of soil microorganisms. Assessing the density, the activity and the structure of the soil microflora allowed us not only to detect the impact of compost amendment on soil microorganisms, but also to evaluate its effect at a functional level through the variation of soil disease suppressiveness. Differences in disease suppressiveness were related to differences in chemical composition, in availability of nutrients at short term and in microbial composition due to both incorporation and stimulation of microorganisms by the compost amendments.     

秸秆腐熟剂对水稻秧苗素质及育秧基质微生物的影响

为探究接种腐熟剂堆沤秸秆作为育秧基质培育水稻秧苗的可行性及其在生态学方面的机制,本研究结合理化测定并采用末端限制性片段长度多态性(T-RFLP)分析,探究了浅盘试验条件下的水稻秧苗素质、基质矿质成分及基质微生物多样性。结果表明,与对照组相比,接种腐熟剂堆沤秸秆作为育秧基质使得水稻鲜重、水稻株高、水稻假茎宽、水稻根等形态学指标有所提高;基质中有机质、全氮、有效磷含量增加显著;基质中细菌和霉菌的生物量分别增加了3.61、70.67倍;细菌中参与C循环、S循环、N循环、抗致病微生物、有益微生物、未知功能微生物、营养代谢相关细菌的生物量分别提高了0.80、1.33、4.86、2.62、6.27、3.24、2.69倍;基质中亚硝化细菌、反硝化细菌、好气性纤维素分解菌、硅酸盐细菌、硝化细菌、好气性自身固氮菌、硅酸盐细菌等功能微生物的数量分别提高了136.50、17.61、8.26、5.65、3.99、2.63倍。研究表明,秸秆腐熟剂改变了原始环境的微生物群落结构并有益于水稻生长。本研究为合理利用堆沤秸秆基质育秧提供了科学依据。