Study of Synthesis Pathways of the Essential Polyunsaturated Fatty Acid 20:5n-3 in the Diatom Chaetoceros Muelleri Using 13C-Isotope Labeling

The present study sought to characterize the synthesis pathways producing the essential polyunsaturated fatty acid (PUFA) 20:5n-3 (EPA). For this, the incorporation of 13C was experimentally monitored into 10 fatty acids (FA) during the growth of the diatom Chaetoceros muelleri for 24 h. Chaetoceros muelleri preferentially and quickly incorporated 13C into C18 PUFAs such as 18:2n-6 and 18:3n-6 as well as 16:0 and 16:1n-7, which were thus highly 13C-enriched. During the experiment, 20:5n-3 and 16:3n-4 were among the least-enriched fatty acids. The calculation of the enrichment percentage ratio of a fatty acid B over its suspected precursor A allowed us to suggest that the diatom produced 20:5n-3 (EPA) by a combination between the n-3 (via 18:4n-3) and n-6 (via 18:3n-6 and 20:4n-6) synthesis pathways as well as the alternative ω-3 desaturase pathway (via 20:4n-6). In addition, as FA from polar lipids were generally more enriched in 13C than FA from neutral lipids, particularly for 18:1n-9, 18:2n-6 and 18:3n-6, the existence of acyl-editing mechanisms and connectivity between polar and neutral lipid fatty acid pools were also hypothesized. Because 16:3n-4 and 20:5n-3 presented the same concentration and enrichment dynamics, a structural and metabolic link was proposed between these two PUFAs in C. muelleri.

Fertilization effects on the fungal biomass in grasslands

<p>Fertilisation is a common practise in grass production systems performed to increase primary production, a supporting ecosystem service essential for other services. However, different fungal groups, like saprothropic fungi (SF) and the obligate symbionts arbuscular mycorrhizal fungi (AMF), have potential differential response to the fertilizer concentration and composition. Three controlled field experiments were utilised in our study, two medium-term (6 years) in the south of Sweden (SE) and one long-term experiment (46 year) in Switzerland (CH), all sampled in 2018. The Swedish sites included the same two factor treatment, i.e. four different plant mixtures and two (SE-Lanna) or three (SE-Alnarp) nitrogen fertilization levels (0, 60, 120 kg ha<sup>-1</sup> yr<sup>-1</sup>); while the Swiss experiment  included different proportions of N, P and K fertilization under different cutting regimes (CH-Bremgarten). The PLFA and NLFA (phospholipid- and neutral lipid fatty acid) analysis was used to estimate the fungal biomass (SF+AMF). The application of N was associated with a decrease in the AMF biomass, with significant effects with the application of 60 and 120 kg N ha<sup>-1</sup> in SE-Alnarp, and 75 and 150 kg N ha<sup>-1</sup> in CH-Bremgarten. On the other hand, the SF biomass was only negatively affected by the N fertilization in SE-Lanna (60 kg N ha<sup>-1</sup>) under the plant mixture that showed the biggest SF biomass in the unfertilized plot; and by the highest application of N in CH-Bremgarten. Our findings indicate that nitrogen fertilization influences microbial community structure and reduces the abundance of AMF, with these being more sensitive than SF to fertilizer application.</p>

Non-symbiotic soil microbes are more strongly influenced by altered tree biodiversity than arbuscular mycorrhizal fungi during initial forest establishment

ABSTRACT While the relationship between plant and microbial diversity has been well studied in grasslands, less is known about similar relationships in forests, especially for obligately symbiotic arbuscular mycorrhizal (AM) fungi. To assess the effect of varying tree diversity on microbial alpha- and beta-diversity, we sampled soil from plots in a high-density tree diversity experiment in Minnesota, USA, 3 years after establishment. About 3 of 12 tree species are AM hosts; the other 9 primarily associate with ectomycorrhizal fungi. We used phospho- and neutral lipid fatty acid analysis to characterize the biomass and functional identity of the whole soil bacterial and fungal community and high throughput sequencing to identify the species-level richness and composition of the AM fungal community. We found that plots of differing tree composition had different bacterial and fungal communities; plots with conifers, and especially Juniperus virginiana, had lower densities of several bacterial groups. In contrast, plots with a higher density or diversity of AM hosts showed no sign of greater AM fungal abundance or diversity. Our results indicate that early responses to plant diversity vary considerably across microbial groups, with AM fungal communities potentially requiring longer timescales to respond to changes in host tree diversity.

Neutral lipid fatty acid analysis is a sensitive marker for quantitative estimation of arbuscular mycorrhizal fungi in agricultural soil with crops of different mycotrophy

The impact of host mycotrophy on arbuscular mycorrhizal fungal (AMF) markers was studied in a temperate agricultural soil cropped with mycorrhizal barley, flax, reed canary-grass, timothy, caraway and quinoa and non-mycorrhizal buckwheat, dyer's woad, nettle and false flax. The percentage of AMF root colonization, the numbers of infective propagules by the Most Probable Number (MPN) method, and the amounts of signature Phospholipid Fatty Acid (PLFA) 16:1ω5 and Neutral Lipid Fatty Acid (NLFA) 16:1ω5 were measured as AMF markers. Crop had a significant impact on MPN levels of AMF, on NLFA 16:1ω5 levels in bulk and rhizosphere soil and on PLFA 16:1ω5 levels in rhizosphere soil. Reed canary-grass induced the highest levels of AMF markers. Mycorrhizal markers were at low levels in all non-mycorrhizal crops. NLFA 16:1ω5 and the ratio of NLFA to PLFA 16:1ω5 from bulk soil are adequate methods as indicators of AMF biomass in soil.

Use of the Signature Fatty Acid 16:1ω5 as a Tool to Determine the Distribution of Arbuscular Mycorrhizal Fungi in Soil

Biomass estimation of arbuscular mycorrhiza (AM) fungi, widespread plant root symbionts, commonly employs lipid biomarkers, predominantly the fatty acid 16:1ω5. We briefly reviewed the application of this signature fatty acid, followed by a case study comparing biochemical markers with microscopic techniques in an arable soil following a change to AM non-host plants after 27 years of continuous host crops, that is, two successive cropping seasons with wheat followed by amaranth. After switching to the non-host amaranth, spore biomass estimated by the neutral lipid fatty acid (NLFA) 16:1ω5 decreased to almost nil, whereas microscopic spore counts decreased by about 50% only. In contrast, AM hyphal biomass assessed by the phospholipid (PLFA) 16:1ω5 was greater under amaranth than wheat. The application of PLFA 16:1ω5 as biomarker was hampered by background level derived from bacteria, and further enhanced by its incorporation from degrading spores used as microbial resource. Meanwhile, biochemical and morphological assessments showed negative correlation for spores and none for hyphal biomass. In conclusion, the NLFA 16:1ω5 appears to be a feasible indicator for AM fungi of the Glomales group in the complex field soils, whereas the use of PLFA 16:1ω5 for hyphae is unsuitable and should be restricted to controlled laboratory studies.