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Volatile fatty acids (VFAs) are increasingly viewed as platform substances for the production of sustainable biofuels and green chemicals. Optimising the acidogenic fermentation requires determining which environmental parameters (temperature, pH, substrate, inoculum, etc) have the largest impact on the metabolite profile. The present study investigates the influence of substrate and mixed inoculum origin. Nine combinations of three complex lignocellulosic substrates and three different mixed inocula were monitored under batch conditions (35 °C; pH 6.5; triplicates, monitoring of pH, soluble COD and VFAs) during 30 days. All nine combinations led to a significantly different fermentation profile, and each combination represented a unique situation. The metabolite profiles were highly reproducible within each substrate–inoculum combination. The substrate COD conversion yield to VFAs was up to 64% (average 31%). VFAs represented most (between 61 and 98%) of the soluble COD. Acetic, propionic and butyric acids were always the major VFAs. [ABSTRACT FROM AUTHOR]

Abstract: Several ectomycorrhizal fungi, including Hebeloma cylindrosporum, actively release large quantities of phosphatase enzymes into their growth medium. We fractionated the phosphatase activity of the ectomycorrhizal association between H. cylindrosporum and its host plant, Pinus pinaster, with the aim to quantify its spatial and temporal variation in response to contrasting soil phosphorus conditions. Seedlings were grown in mini-rhizoboxes and the phosphomonoesterase activity of rhizosphere soil, released by roots, surface-bound to roots or mycelium was determined spectrophotometrically with the p-nitrophenyl phosphate method or microscopically with the ELF-method as a function of culture time. We showed that acid phosphatase activity of the soil and the root increased with mycorrhizal association. We also observed that the phosphatase activity associated with ectomycorrhizal plants was related to soil type. All phosphatase fractions decreased over culture time, except the proportion of hyphae exhibiting phosphatase activity in the extramatrical mycelium, which increased over time. The specific fractions of phosphatase activity associated with the mycorrhizal plants were clearly related to the soil phosphorus type and content. Soils showed an increase in acid phosphomonoesterase activity with mycorrhizal association, supporting a role for this enzyme in the degradation of soil bound phosphorus. The gradually increasing proportion of hyphae in the extramatrical mycelium exhibiting alkaline phosphatase activity, particularly under low phosphorus conditions, indicates an induction of alkaline phosphatase activity by phosphorus limitation. [Copyright &y& Elsevier]

Pyrrolnitrin (PRN) is a tryptophan-derived secondary metabolite produced by a narrow range of Gram-negative bacteria. The PRN biosynthesis by rhizobacteria presumably has a key role in their life strategies and in the biocontrol of plant diseases. The biosynthetic operon that encodes the pathway that converts tryptophan to PRN is composed of four genes, prnA through D, whose diversity, genomic context and spread over bacterial genomes are poorly understood. Therefore, we launched an endeavour aimed at retrieving, by in vitro and in silico means, diverse bacteria carrying the prnABCD biosynthetic loci in their genomes. Analysis of polymorphisms of the prnD gene sequences revealed a high level of conservation between Burkholderia, Pseudomonas and Serratia spp. derived sequences. Whole-operon- and prnD-based phylogeny resulted in tree topologies that are incongruent with the taxonomic status of the evaluated strains as predicted by 16S rRNA gene phylogeny. The genomic composition of c. 20 kb DNA fragments containg the PRN operon varied in different strains. Highly conserved and distinct transposase-encoding genes surrounding the PRN biosynthetic operons of Burkholderia pseudomallei strains were found. A prnABCD-deprived genomic region in B. pseudomallei strain K96243 contained the same gene composition as, and shared high homology with, the flanking regions of the PRN operon in B. pseudomallei strains 668, 1106a and 1710b. Our results strongly suggest that the PRN biosynthetic operon is mobile. The extent, frequency and promiscuity of this mobility remain to be understood. [ABSTRACT FROM AUTHOR]

• Colonization of two plant species byGlomus intraradiceswas studied to investigate the two morphological types (ArumandParis), their symbiotic interfaces and metabolic activities.• Root pieces and sections were stained to observe the colonization and metabolic activity of all mycorrhizal structures.• There were no growth responses observed in the plants caused by mycorrhizal symbiosis. The two morphological types had a similar percentage of root colonized, but theArum-type had higher metabolic activity. Most of the mycorrhizal structures (88%) showed succinate dehydrogenase activity; about half showed acid phosphatase activity; and a small percentage showed alkaline phosphatase activity. Phosphatase activity was highest in arbuscules and low in intercellular hyphae in theArum-type colonization. In theParis-type, hyphal coils and arbusculate coils showed a similar intermediate percentage of phosphatase activity.• We conclude that acid phosphatase is more important than alkaline phosphatase in both colonization types. We discuss the possibility that, whereas arbuscules inArum-type are the main site for phosphorus release to the host plant, both the hyphal and arbusculate coils may be involved in theParis-type.New Phytologist(2005)doi: 10.1111/j.1469-8137.2005.01340.x© New Phytologist(2005) [ABSTRACT FROM AUTHOR]

The ubiquitous arbuscular mycorrhizal fungi consume significant amounts of plant assimilated C, but this C flow has been difficult to quantify. The neutral lipid fatty acid 16:1ω5 is a quantitative signature for most arbuscular mycorrhizal fungi in roots and soil. We measured carbon transfer from four plant species to the arbuscular mycorrhizal fungus Glomus intraradices by estimating 13C enrichment of 16:1ω5 and compared it with 13C enrichment of total root and mycelial C. Carbon allocation to mycelia was detected within 1 day in monoxenic arbuscular mycorrhizal root cultures labeled with [13C] glucose. The 13C enrichment of neutral lipid fatty acid 16:1ω5 extracted from roots increased from 0.14% 1 day after labeling to 2.2% 7 days after labeling. The colonized roots usually were more enriched for 13C in the arbuscular mycorrhizal fungal neutral lipid fatty acid 16:1ω5 than for the root specific neutral lipid fatty acid 18:2ω6,9. We labeled plant assimilates by using 13CO2 in whole-plant experiments. The extraradical mycelium often was more enriched for 13C than was the intraradical mycelium, suggesting rapid translocation of carbon to and more active growth by the extraradical mycelium. Since there was a good correlation between 13C enrichment in neutral lipid fatty acid 16:1ω5 and total 13C in extraradical mycelia in different systems (r2 = 0.94), we propose that the total amount of labeled C in intraradical and extraradical mycelium can be calculated from the 13C enrichment of 16:1ω5. The method described enables evaluation of C flow from plants to arbuscular mycorrhizal fungi to be made without extraction, purification and identification of fungal mycelia. [ABSTRACT FROM AUTHOR]

Arbuscular mycorrhizal (AM) development in different soil types, and the influence of AM fungal hyphae on their original soil were investigated. Plantago lanceolata, which can grow in soils of a very wide pH range, was grown in two closely related limestone soils and an acid soil from rock habitats. Plants were colonised by the indigenous AM fungal community. The use of compartmented systems allowed us to compare soil with and without mycorrhizal hyphae. Root colonisation of P. lanceolata was markedly higher in the limestone soils (30–60%) than in the acid soil (5–20%), both in the original habitat and in the experimental study. Growth of extraradical AM fungal hyphae was detected in the limestone soils, but not in the acid soil, using the signature fatty acid 16:1ω5 as biomass indicator. Analysis of signature fatty acids demonstrated an increased microbial biomass in the presence of AM fungal hyphae as judged for example from an increased amount of NLFA 16:0 with 30 nmol g−1 in one of the limestone soils. Bacterial activity, but not soil phosphatase activity, was increased by around 25% in the presence of mycorrhizal hyphae in the first harvest of limestone soils. AM fungal hyphae can thus stimulate microorganisms. However, no effect of AM hyphae was observed on the soil pH or organic matter content in the limestone soils and the available P was not depleted. [Copyright &y& Elsevier]

We monitored the development of intraradical and extraradical mycelia of the arbuscular mycorrhizal (AM) fungi Scutellospora calospora and Glomus intraradices when colonizing Piantago lanceolata. The occurrence of arbuscules (branched hyphal structures) and vesicles (lipid storage organs) was compared with the amounts of signature fatty acids. The fatty acid 16:1ω5 was used as a signature for both AM fungal phospholipids (membrane constituents) and neutral lipids (energy storage) in roots (intraradical mycelium) and in soil (extraradical mycelium). The formation of arbuscules and the accumulation of AM fungal phospholipids in intraradical mycelium followed each other closely in both fungal species. In contrast, the neutral lipids of G. intraradices increased continuously in the intraradical mycelium, while vesicle occurrence decreased after initial rapid root colonization by the fungus. S. calospora does not form vesicles and accumulated more neutral lipids in extraradical than in intraradical mycelium, while the opposite pattern was found for G. intraradices. G. intraradices allocated more of its lipids to storage than did S. calospora. Thus, within a species, the fatty acid 16:1ω5 is a good indicator for AM fungal development. The phospholipid fatty acid 16:1ω5 is especially suitable for indicating the frequency of arbuscules in the symbiosis. We propose that the ratio of neutral lipids to phospholipids is more important than is the presence of vesicles in determining the storage status of AM fungi. [ABSTRACT FROM AUTHOR]

To establish relationships between soil phosphorus (P) fractions and leaf P, a mycorrhizal species ( Plantago lanceolata L.) was compared with a typically non-mycorrhizal species (Rumex acetosella L.) in a glasshouse experiment. The plants were grown in 40 soils from non-fertilised, abandoned pastures or abandoned arable fields and leaf P concentration were found to be related to various soil P fractions after six weeks of growth. The differences in the P fractions in soil can account for a large share of the variation in leaf P concentration in both species, but the two species differed in their utilisation of P fractions. Leaf P concentration of R. acetosella was more related to extractable soil P than that of P. lanceolata. Rumex acetosella showed a higher maximum P concentration. The P fractions accounting for the largest share of the variation in leaf P concentration was the Bray 1 extractable and the weak oxalate (1 mM) extractable P, and for P. lanceolata also the Na2SO4+NaF extractable P fraction. P extracted with these methods accounted for up to 80% of the variation in P concentration in leaves of R. acetosella and 65% of the variation in leaves of P. lanceolata. More P extractable with weak oxalate, Na2SO4+NaF and strong oxalate (50 mM) was released from the soil than was taken up by the plants during the experimental period. The Bray 1 extractable P fraction, however, decreased in both unplanted and planted soils. Phosphatase release was not induced in any of the plants during the experimental period, indicating that they were not mobilising soil organic P. However, some of the methods extracted a large share of the organic P and still explained much of the variation in leaf P concentration. Mycorrhizal colonisation of P. lanceolata was inversely related to the extractable soil P. The consistently fast P uptake of R. acetosella indicates that this species have a high demand for P. The differences in P utilisation between R. acetosella and P. lanceolata could be caused by their different mycorrhizal status. [ABSTRACT FROM AUTHOR]

Summary • To test the response of arbuscular mycorrhizal (AM) fungi to a difference in soil pH, the extraradical mycelium of Scutellospora calospora or Glomus intraradices , in association with Plantago lanceolata , was exposed to two different pH treatments, while the root substrate pH was left unchanged. • Seedlings of P. lanceolata , colonized by one or other of the fungal symbionts, and nonmycorrhizal controls, were grown in mesh bags placed in pots containing pH-buffered sand (pH around 5 or 6). The systems were harvested at approximately 2-wk intervals between 20 and 80 d. • Both fungi formed more extraradical mycelium at the higher pH. Glomus intraradices formed almost no detectable extraradical mycelium at lower pH. The extraradical mycelium of S. calospora had higher acid phosphatase activity than that of G. intraradices . Total AM root colonization decreased for both fungi at the higher pH, and high pH also reduced arbuscule and vesicle formation in G. intraradices . • In conclusion, soil pH influences AM root colonization as well as the growth and phosphatase activities of extraradical mycelium, although the two fungi responded differently. [ABSTRACT FROM AUTHOR]

• The influence of external nitrogen (N) on carbon (C) allocation and processes related to phosphorus (P) metabolism were studied in monoxenic arbuscular mycorrhiza (AM) cultures of Daucus carota. • Fungal hyphae of Glomus intraradices proliferated from colonized roots growing on solid medium into C-free liquid minimal medium with two different N and P levels. Furthermore, we exposed the colonized roots to high or low N availability and then studied the mycelial development. Roots were provided with 13C-glucose in order to follow the C allocation. The mycelium was analysed for phosphatase activity and transcription levels of two nutrient regulated genes. • High N availability to the monoxenic AM root reduced the C allocation to the AM fungus while N availability to the mycelium was important for the upregulation of the fungal inorganic phosphorus (Pi)-transporter GiPT. • We found that N availability can regulate nutritional processes in arbuscular mycorrhiza. We conclude that negative impacts of N on AM abundance are caused by reduced C allocation from the plant. Upregulation of the fungal Pi-transporter GiPT indicated that increased N availability might induce P limitation in the mycelium. [ABSTRACT FROM AUTHOR]

Reciprocal effects between arbuscular mycorrhizal fungi (AMF) and plant communities are essential to study the complexity of interactions in a grassland ecosystem. Here, we investigated the effects of plant community density and composition on AMF colonization, plant growth, and reproduction investment. We developed an experimental system with three compartments, each containing either three or six Medicago truncatula Gaertn. plants, or three M. truncatula plants associated with three Silene vulgaris (Moench) Garcke plants. All three compartments shared the same common mycorrhizal network built either by Rhizophagus irregularis MUCL 43194, by Rhizophagus clarus MUCL 46238, or by both AMF in association grown in a central compartment on Plantago lanceolata L. Our results demonstrate an absence of effect of plant density but a positive influence of mixed cultures on AMF root colonization compared with monocultures. This higher AMF development resulted in a positive feedback on shoot biomass and number of flowers and fruits produced by M. truncatula. Although both fungal strains were present in root systems, co-inoculation did not generate a synergistic effect on plant development. These results highlight the importance of plant associations on AMF dynamics, which requires further investigation at the community scale to improve our understanding of the intricate AMF - host plant relationships. [ABSTRACT FROM AUTHOR]

Arbuscular mycorrhizal fungi ( AMF) are key determinants of plant interactions in ecosystems. Through their effects on competition, they are regulators of the structure of communities. Conversely, the composition of plant assemblages may also influence the AMF colonization dynamics of plant species. Here, we tested under in vitro culture conditions the effects of M edicago truncatula, a highly mycorrhizal plant species, and S ilene vulgaris, a weakly mycorrhizal plant species, grown single (monospecies treatments) or in combination (bispecies treatment) on the colonization dynamics of the AMF R hizophagus irregularis MUCL 43194. The seedlings were placed in a pre-established hyphal network developing from a mature M . truncatula mycorrhizal donor plant. Extraradical mycelium ( ERM) and root colonization parameters as well as root morphology were measured over a period of 12 days. An increased ERM length, total root colonization and proportion of arbuscules were noted in the bispecies treatment. Conversely, the bispecies treatment seemed to have no effect on root growth. This study also demonstrated the suitability of the in vitro culture system for studying the interactions between AMF and host plants grown as mono- and bispecies combinations. [ABSTRACT FROM AUTHOR]

Abstract: We investigated element accumulation in vesicles of the arbuscular mycorrhizal (AM) fungus Glomus intraradices, extracted from the roots of inoculated leek plants. The elemental composition (elements heavier than Mg) was quantified using particle-induced X-ray emission (PIXE), in combination with scanning transmission ion microscopy (STIM). In vesicles, P was the most abundant of the elements analysed, followed by Ca, S, Si and K. We analysed 12 vesicles from two root systems and found that the variation between vesicles was particularly high for P and Si. The P content related positively to Si, Zn and K, while its relation to Cl fitted to a negative power function. Vesicle transects showed that P and K were present in central parts, while Ca was present mainly near the vesicle surfaces. The results showed that P is an important part (0.5% of the dry weight) of the vesicle content and that the distribution of some elements, within mycelia, may be strongly correlated. [Copyright &y& Elsevier]

Ectomycorrhizal symbiosis markedly improves plant phosphate uptake, but the molecular mechanisms underlying this benefit are still poorly understood. We identified two ESTs in a cDNA library prepared from the ectomycorrhizal basidiomycete Hebeloma cylindrosporum with significant similarities to phosphate transporters from the endomycorrhizal fungus Glomus versiforme and from non-mycorrhizal fungi. The full-length cDNAs corresponding to these two ESTs complemented a yeast phosphate transport mutant (Δ pho84). Measurements of 33P-phosphate influx into yeast expressing either cDNA demonstrated that the encoded proteins, named HcPT1 and HcPT2, were able to mediate Pi:H+ symport with different affinities for Pi ( Km values of 55 and 4 μm, respectively). Real-time RT-PCR showed that Pi starvation increased the levels of HcPT1 transcripts in H. cylindrosporum hyphae grown in pure culture. Transcript levels of HcPT2 were less dependent on Pi availability. The two transporters were expressed in H. cylindrosporum associated with its natural host plant, Pinus pinaster, grown under low or high P conditions. The presence of ectomycorrhizae increased net Pi uptake rates into intact Pinus pinaster roots at low or high soil P levels. The expression patterns of HcPT1 and HcPT2 indicate that the two fungal phosphate transporters may be involved in uptake of phosphate from the soil solution under the two soil P availability conditions used. [ABSTRACT FROM AUTHOR]

Focuses on the use of ergosterol and fatty acids in biomass estimation of mycorrhizal fungi. Biomass indicators of microorganisms; High performance liquid chromatography; Tandem mass spectrometry.