Hans Ek

Extramatrical mycelial growth, biomass allocation and nitrogen uptake in ectomycorrhizal systems in response to collembolan grazing

The effects of Collembola grazing activities on the ectomycorrhizal symbiosis were studied in laboratory microcosms. Pinus contorta seedlings in association with Paxillus involutus were grown in a sandy soil and the Collembola Onychiurus armatus was added in different densities. To study effects on nutrient uptake by the extramatrical mycorrhizal mycelium, cups containing 15NH4+ and phytin amended soil were evenly distributed in the microcosms. These cups were covered with a net that allowed the mycelium to penetrate but neither Collembola nor plant roots. Extramatrical hyphal growth was impeded at a high density of O. Armatus. Low densities of O. armatus increased the extramatrical hyphal growth, the colonization rate of side plants and the biomass of P. involutus. However, the amount of P. involutus on/in the plant roots was not affected. Thus, low densities of collembolans induced a shift towards a larger proportion of P. involutus growing extramatrically. The presence of O. armatus in low numbers enhanced the uptake and transfer of 15N by P. involvus to the plants by up to 76%. The Collembola population growth was not higher in mycorrhizal compared with non-mycorrhizal microcosms while nematode population size was reduced in mycorrhizal compared with non-mycorrhizal microcosms. (Less)

Carbon and nitrogen flow in silver birch and Norway spruce connected by a common mycorrhizal mycelium

Abstract Spruce and birch seedlings were grown together in boxes filled with unsterile peat. Both seedlings were colonized by the ectomycorrhizal fungus Scleroderma citrinum. The two plants thus shared a common external mycelium. 15N-labelled ammonium was supplied exclusively to the fungus, while the birch or the spruce plant was continuously fed with 13C-labelled CO2 for 72 h. The carbon and nitrogen transfer rates were strikingly different for birch and spruce seedlings. The mycorrhizal mycelium received carbohydrates mainly from the birch plant and the nitrogen transfer by the fungus to the plants was largely directed towards the birch. Carbon assimilates were also transferred in both directions between birch and spruce; however, there was no conclusive evidence for a net transfer of carbon between the plants.

Determination of 13C-enrichment in bacterial fatty acids using chemical ionization mass spectrometry with negative ion detection

Saturated, monoenoic and β-hydroxysubstituted fatty acids, 13C-labelled at the carboxyl group, were prepared from natural or synthetic unlabelled analogues. The synthetic route involves decarboxylation of the unlabelled fatty acid to the next lower iodide, displacement of iodide for [13C]cyanide and hydrolysis. The fatty acids were converted to their pentafluorobenzyl esters and analysed by selected ion monitorint using chemical ionization and negative ion detection. Measurements of the signal ratios for the negative carboxylate ion (m) and the (m + 1) ion showed that at 95% confidence level and n = 5, mean values differing by 1.0 atom% 13C will be significantly resolved. The calculated standard deviation was the same for the studied bacterial acids including the phospholipid ester-linked palmitoleic acid, β-hydroxymyristic acid in the lipopolysaccharides and β-hydroxybutyric acid in the storage polymer poly-β-hydroxyalkanoate. Sodium [1-13C]acetate or D-[13C6]glucose were pulse administered to a Gram-negative marine bacterium isolate. Phospholipid ester-linked fatty acids and β-hydroxybutyric acid showed extensive 13C-incorporation within 15 min after the pulse. After approximately 60 min a maximum of 10 atom% excess of 13C was reached for palmitoleic acid. The method provides the potential to measure the metabolic activity of bacterial communities by measuring the incorporation of 13C-labelled substrates into specific fatty acids that can be utilized as biomarkers for biomass, community structure and nutritional status. (Less)

Determination of 15N-labelled ammonium and total nitrogen in plant and fungal systems using mass spectrometry.

A selected ion-monitoring method to measure 15N-labelled ammonia in biological samples was improved to simplify sample handling, to obviate interference from ammonia due to the decomposition of glutamine and to allow the determination of total N. Ammonia is derivatized with pentafluorobenzoylchloride to yield pentafluorobenzamide which is analysed by gas chromatography/mass spectrometry after clean-up using disposable silicic acid columns. The sensitivity achieved when operating in the negative ion-chemical ionization mode was somewhat higher than when using electron-impact ionization. Use of methyl amine as an internal standard improved the accuracy and precision of the measurements. The method was applied to samples taken from an intact ectomycorrhizal system fed with ISN-labelled ammonium and used to determine patterns of N assimilation into ammonium, free amino acids and macromolecular compounds. (Less)

Mycelial uptake, translocation and assimilation of N-15-labeled nitrogen by ectomycorrhizal Pinus sylvestris plants

Mycolial uptake, translocation and assimilation of 15N-labelled ammonium was followed in Pinus sylvestris plants infected with the ectomycorrhizal fungus. Paxillus involutus. The distribution of labelled compounds within the systems was examined using gas chromatography / mass spectrometry. Labelled nitrogen was incorporated into a range of free amino acids in the fungal mycelium, the principal sinks being alanine and glutamatelglutamine. Levels of 15 N enrichment declined throughout the transport pathway from 30-45% in the mycelium a~d mycorrhlzal root lips to 3-11% in the plant shoots. A significant proportion of the total assimilated nitrogen (27%) was incorporated into proteinaceous material and the proportion of label present in this fraction increased from 27% in the mycelium to 32% and 58% in the plant roots and needles respectively. The protein-incorporated amino acids showed a wider spectrum of labelling with significant amounts of 15N incorporated into valine, isoleucine, phenylalanine and lysine. (Less)