Plant-plant competition outcomes are modulated by plant effects on the soil bacterial community

Competition is a key process that determines plant community structure and dynamics, often mediated by nutrients and water availability. However, the role of soil microorganisms on plant competition, and the links between above- and belowground processes, are not well understood. Here we show that the effects of interspecific plant competition on plant performance are mediated by feedbacks between plants and soil bacterial communities. Each plant species selects a singular community of soil microorganisms in its rhizosphere with a specific species composition, abundance and activity. When two plant species interact, the resulting soil bacterial community matches that of the most competitive plant species, suggesting strong competitive interactions between soil bacterial communities as well. We propose a novel mechanism by which changes in belowground bacterial communities promoted by the most competitive plant species influence plant performance and competition outcome. These findings emphasise the strong links between plant and soil communities, paving the way to a better understanding of plant community dynamics and the effects of soil bacterial communities on ecosystem functioning and services

Tracking mycorrhizas and extraradical mycelium of the edible fungus Lactarius deliciosus under field competition with Rhizopogon spp

The objective of this study is to evaluate the field persistence of the edible ectomycorrhizal fungus Lactarius deliciosus in competition with two ubiquitous soil fungi. Couples of plants inoculated with either L. deliciosus, Rhizopogon roseolus, or R. luteolus were transplanted, 10 cm apart, in two different sites at the following combinations: L. deliciosus–R. roseolus, L. deliciosus–R. luteolus, L. deliciosus–control (non-inoculated), control–R. roseolus, control–R. luteolus, and control–control. Eight months after transplantation, root colonization and extraradical soil mycelium for each fungal species were quantified. For mycelium quantification, soil cores equidistant to the two plants in each couple were taken, and total deoxyribonucleic acid (DNA) was extracted. Real-time polymerase chain reaction analysis was performed using specific primers and TaqMan® Minor groove binding (MGB) probes designed in the ribosomal DNA internal transcribed spacer region of each fungal species. Field site significantly influenced persistence of both mycorrhizas and extraradical mycelium of L. deliciosus. Extraradical mycelium quantity was positively correlated with the final percentage of ectomycorrhizas for the three fungal species. Different competitive pressure between the two Rhizopogon species on L. deliciosus persistence was observed, with R. luteolus having no effect on L. deliciosus survival. Negative correlation between the final percentage of mycorrhizas of L. deliciosus and R. roseolus was observed. However, no relationship was determined between extraradical mycelia of both fungal species. The results obtained suggest that competition between L. deliciosus and R. roseolus takes place in the root system, for ectomycorrhiza formation in available roots, rather than in the extraradical phase

Field persistence of the edible ectomycorrhizal fungus Lactarius deliciosus : effects of inoculation strain, initial colonization level, and site characteristics

Pinus pinea plants were inoculated with different strains of the edible ectomycorrhizal fungus Lactarius deliciosus. The inoculated plants were established in six experimental plantations in two sites located in the Mediterranean area to determine the effect of the initial colonization level and the inoculated strain on fungal persistence in the field. Ectomycorrhizal root colonization was determined at transplantation time and monitored at different times from uprooted plants. Extraradical soil mycelium biomass was determined from soil samples by TaqMan® real-time polymerase chain reaction (PCR). The results obtained indicate that the field site played a decisive role in the persistence of L. deliciosus after outplanting. The initial colonization level and the selection of the suitable strain were also significant factors but their effect on the persistence and spread of L. deliciosus was conditioned by the physical–chemical and biotic characteristics of the plantation soil and, possibly, by their influence in root growth. Molecular techniques based on real-time PCR allowed a precise quantification of extraradical mycelium of L. deliciosus in the field. The technique is promising for non-destructive assessment of fungal persistence since soil mycelium may be a good indicator of root colonization. However, the accuracy of the technique will ultimately depend on the development of appropriate soil sampling methods because of the high variability observed

Molecular identification of the edible ectomycorrhizal fungus Lactarius deliciosus in the symbiotic and extraradical mycelium stages

Specific rDNAITS amplifications, microsatellite-primedPCRand ITS-SSCP analysis were applied to identify and characterize pre-selected isolates of the edible ectomycorrhizal fungus Lactarius deliciosus in different stages of the life cycle. Sampling was performed from pure cultures, mycorrhizas and soil from experimental plots established with nursery-inoculated pine seedlings. A newly-designed reverse primer (LDITS2R) combined with the universal forward ITS1 allowed to perform specific amplifications of L. deliciosus from all the samples. Microsatellite-primed PCR using the (GTG)5 oligonucleotide as a primer showed clear polymorphisms among the different L. deliciosus isolates. The patterns of mycorrhiza samples showed additional bands corresponding to the plant DNA. Single strand conformation polymorphism (SSCP) analysis of the specific rDNA ITS fragment amplified from 18 L. deliciosus isolates showed nine clearly different patterns. Mycorrhiza and soil samples showed coincident patterns with their respective fungal isolates. Specific rDNA ITS amplifications had not been previously used for SSCP analysis of ectomycorrhizas and extraradical mycelium. This relatively simple and inexpensive technique allows tracking L. deliciosus isolates in different stages of the fungus development. Specific ITS-SSCP analysis is promising in studies of the persistence of inoculated L. deliciosus isolates and their competitiveness with native ectomycorrhizal fungi, especially at the extraradical mycelium stage

Quantitative detection of Lactarius deliciosus extraradical soil mycelium by real-time PCR and its application in the study of fungal persistence and interspecific competition

Real-Time PCR has been applied to quantify extraradical soil mycelium of the edible ectomycorrhizal fungus Lactarius deliciosus in an interspecific competition experiment under greenhouse conditions. Couples of Pinus pinea seedlings inoculated with either L. deliciosus, Rhizopogon roseolus, or non-inoculated (control) were transplanted into pots filled with two types of soil in all the possible combinations. Total DNA was extracted from soil samples at 3 and 6 months after transplantation to perform real-time PCR analysis. DNA extractions from soil mixed with known amounts of mycelium of L. deliciosus were used as standards. Six months after transplantation, the percentage of mycorrhizas of L. deliciosus and seedling growth were significantly affected by the soil type. Extraradical soil mycelium of L. deliciosus was positively correlated with the final percentage of mycorrhizas and significantly affected by the sampling time and soil depth. The competition effect of R. roseolus was not significant for any of the measured parameters, probably due to the sharp decrease of the mycorrhizal colonization by this fungus. We conclude that real-time PCR is a powerful technique for extraradical mycelium quantification in studies aimed at evaluating the persistence of introduced strains of L. deliciosus in field plantations

Complementarity in nurse plant systems : soil drives community composition while microclimate enhances productivity and diversity

Aims Nurse plants facilitate the establishment of other species under their canopies through improvement of microclimate and soil conditions beneath their canopies. We aimed to test whether the effect of nurse soil on understorey communities was independent of the canopy effect, and to what extent soil properties (i.e., nutrients, pH, moisture, temperature, aggregates) can predict understorey biomass and diversity. Methods In a field experiment, we extracted three soil blocks (20 × 20 × 15 cm) under the canopy of eighteen Retama sphaerocarpa shrubs –grouped into small, medium, and large canopy sizes (n = 6). Soil blocks were distributed in a stratified-random design, so that each shrub received back three soil blocks (one from under each Retama canopy size). At the end of the growing season plant biomass, abundance, diversity and soil properties were recorded. Results Canopy and soil independently affected understorey community structure. Biomass and diversity increased under large Retama canopies irrespective of soil origin, whereas biomass was higher -and plant diversity lower- in soils from large Retama shrubs irrespective of canopy size. Biomass and species abundance were influenced more by soil than by canopy. There was, however, complementarity between these factors, as biomass increased in soils with high moisture and low temperature, while diversity was highly influenced by canopy size. Conclusions Soil, rather than microclimate amelioration by the nurse, was the main driver of understorey community structure, soil moisture playing a pivotal role. This is highly relevant as drought is expected to increase in drylands worldwide

Intraspecific variability of Lactarius deliciosus isolates : colonization ability and survival after cold storage

Intraspecific variability in root colonization, extraradical growth pattern, and survival after cold storage of Lactarius deliciosus isolates was determined in pure culture conditions using Pinus pinaster as a host plant. The ectomycorrhizal ability of L. deliciosus at 30, 45, and 60 days from inoculation was highly variable among isolates and was negatively correlated to the age of the culture (time elapsed from isolation). The formation of rhizomorphs was related to colonization ability, but no relationship was found between colonization and formation of extraradical mycelium. The final colonization achieved at 60 days from inoculation was not related to the tree species under which the sporocarps were collected. However, isolates from sporocarps collected under P. pinaster colonized more rapidly the seedlings than those collected under other pine species. The climatic range of the sporocarps from which the isolates were obtained (maritime vs. continental) was not related to the formation of mycorrhizas at 60 days from inoculation. However, isolates from sporocarps collected from a maritime climate area colonized more rapidly the P. pinaster seedlings than those collected from a continental zone. Tolerance to cold water storage of L. deliciosus was also isolate dependent. Growth revival in agar was obtained from most of the isolates after 28 months of cold storage at 4°C, but only 10 out of 29 isolates showed unaffected growth. The ITS rDNA alignment of all the L. deliciosus isolates showed a low variability with identities over 99%. Most of the variation was detected in the ITS1 region and consisted in single nucleotide changes and/or punctual indel mutations. The number of base differences per sequence from averaging over all sequence pairs was 1.329, which is in the low range when compared with other ectomycorrhizal species. No ITS pattern due to geographical origin of the isolates could be discerned

Interactions among soil, plants, and microorganisms drive secondary succession in a dry environment

Secondary succession studies have mainly focused on plants, but little is known about the fate of soil microbial communities and their relationship with plant succession after disturbance, particularly in dry ecosystems. We examined changes in soil properties and of plant and soil microbial communities across a chronosequence of abandoned arable fields that included five successional stages according to time of abandonment stretching near a century. We hypothesized the existence of a parallel secondary succession above- and below-ground and explored the possible linkages between plant and microbial communities as well as the role of changes in soil properties over the successional gradient. Soil microbial communities were characterized by PLFAs analysis, enzymatic activities, and pyrosequencing of the 16S rDNA. We found clear patterns of plant and microbial secondary succession characterized by an increase in organic C, NHþ 4 , and silt content as well as in soil microbial biomass and activity along the successional stages, linked to an increase in plant productivity and diversity. Plant and microbial composition were significantly different among successional stages, although no distinct microbial communities were observed in the two initial stages, suggesting that microbial succession may lag behind plant succession. However, the degree of change in the composition of soil microbial communities and plant communities across our chronosequence evidenced that above- and below-ground secondary succession developed with similar patterns and correlated with changes in multiple ecosystem functions such as increases in above-and below-ground productivity, diversity and nutrient accumulation as plant and microbial succession progressed

Tracking inoculated edible ectomycorrhizal fungi by real-time PCR

The inoculation of different plant species with edible ectomycorrhizal fungi is a promising technology for sustainable production of non-wood forest resources, especially in areas unsuitable for agricultural production. Key species as truffles (Tuber), boletes (Boletus) and saffron-milk caps (Lactarius) are especially valued in local markets but their availability depends, almost exclusively, on the erratic natural production

Competitive interactions among three ectomycorrhizal fungi and their relation to host plant performance

1. Competition strongly influences many species assemblages, but its role in mycorrhizal fungal interactions is not well understood. We examined interactions among three ectomycorrhizal (ECM) species to determine if the structure of competition could be characterized by either competitive networks (where no clear hierarchy exists in the outcome of competition between various species pairs) or competitive hierarchies (where one species out competes all other species). 2. Using a bioassay experiment, we inoculated Pinus muricata seedlings with three Rhizopogon species (R. occidentalis, R. salebrosus, and R. vulgaris) in single-, two-, and three-species treatments. After 7 months, we assessed the relative abundance of each species in each treatment using real-time PCR of internal transcribed spacer rDNA. 3. We found that R. occidentalis was strongly inhibited by R. vulgaris and R. salebrosus in all competition treatments. In contrast, R. vulgaris and R. salebrosus had similar ECM biomasses in the two-species treatment, but R. vulgaris had significantly higher biomass than R. salebrosus in the three-species treatment. 4. In the single-species treatments, seedlings colonized by the competitive dominants had higher shoot biomass and total leaf nitrogen, but also higher percentage ECM biomass. In the multi-species treatments, seedlings had either equivalent or somewhat lower shoot biomass and total leaf nitrogen than their respective single-species treatments. 5. Synthesis. Our results indicate that ECM competition does not appear to be characterized by strict networks or hierarchies. Instead, the outcome is dependent on the conditions of the local environment in which it occurs. There also does not seem to be a clear relationship between ECM competitive ability and plant performance, but competition does appear to negatively affect the ability of ECM fungi to provide benefits to their hosts