Effects of soil moisture, needle age and leaf morphology on carbon and oxygen uptake, incorporation and allocation: a dual labeling approach with 13CO2 and H218O in foliage of a coniferous forest

The carbon and oxygen isotopic composition of water and assimilates in plants reveals valuable information on plant responses to climatic conditions. Yet, the carbon and oxygen uptake, incorporation and allocation processes determining isotopic compositions are not fully understood. We carried out a dual-isotope labeling experiment at high humidity with 18O-enriched water (H218O) and 13C-enriched CO2 (13CO2) with attached Scots pine (Pinus sylvestris L.) branches and detached twigs of hemiparasitic mistletoes (Viscum album ssp. austriacum) in a naturally dry coniferous forest, where also a long-term irrigation takes place. After 4 h of label exposure, we sampled previous- and recent-year leaves, twig phloem and twig xylem over 192 h for the analysis of isotope ratios in water and assimilates. For both species, the uptake into leaf water and the incorporation of the 18O-label into leaf assimilates was not influenced by soil moisture, while the 13C-label incorporation into assimilates was significantly higher under irrigation compared with control dry conditions. Species-specific differences in leaf morphology or needle age did not affect 18O-label uptake into leaf water, but the incorporation of both tracers into assimilates was two times lower in mistletoe than in pine. The 18O-label allocation in water from pine needles to twig tissues was two times higher for phloem than for xylem under both soil moisture conditions. In contrast, the allocation of both tracers in pine assimilates were similar and not affected by soil moisture, twig tissue or needle age. Soil moisture effects on 13C-label but not on 18O-label incorporation into assimilates can be explained by the stomatal responses at high humidity, non-stomatal pathways for water and isotope exchange reactions. Our results suggest that non-photosynthetic 18O-incorporation processes may have masked prevalent photosynthetic processes. Thus, isotopic variation in leaf water could also be imprinted on assimilates when photosynthetic assimilation rates are low.

Limited vertical CO2 transport in stems of mature boreal Pinus sylvestris trees

Several studies have suggested that CO2 transport in the transpiration stream can considerably bias estimates of root and stem respiration in ring-porous and diffuse-porous tree species. Whether this also happens in species with tracheid xylem anatomy and lower sap flow rates, such as conifers, is currently unclear. We infused 13C-labelled solution into the xylem near the base of two 90-year-old Pinus sylvestris L. trees. A custom-built gas exchange system and an online isotopic analyser were used to sample the CO2 efflux and its isotopic composition continuously from four positions along the bole and one upper canopy shoot in each tree. Phloem and needle tissue 13C enrichment was also evaluated at these positions. Most of the 13C label was lost by diffusion within a few metres of the infusion point indicating rapid CO2 loss during vertical xylem transport. No 13C enrichment was detected in the upper bole needle tissues. Furthermore, mass balance calculations showed that c. 97% of the locally respired CO2 diffused radially to the atmosphere. Our results support the notion that xylem CO2 transport is of limited magnitude in conifers. This implies that the concerns that stem transport of CO2 derived from root respiration biases chamber-based estimates of forest carbon cycling may be unwarranted for mature conifer stands.

A Lowly Digestible-Starch Diet after Weaning Enhances Exogenous Glucose Oxidation Rate in Female, but Not in Male, Mice

Starches of low digestibility are associated with improved glucose metabolism. We hypothesise that a lowly digestible-starch diet (LDD) versus a highly digestible-starch diet (HDD) improves the capacity to oxidise starch, and that this is sex-dependent. Mice were fed a LDD or a HDD for 3 weeks directly after weaning. Body weight (BW), body composition (BC), and digestible energy intake (dEI) were determined weekly. At the end of the intervention period, whole-body energy expenditure (EE), respiratory exchange ratio (RER), hydrogen production, and the oxidation of an oral 13C-labelled starch bolus were measured by extended indirect calorimetry. Pancreatic amylase activity and total 13C hepatic enrichment were determined in females immediately before and 4 h after administration of the starch bolus. For both sexes, BW, BC, and basal EE and RER were not affected by the type of starch, but dEI and hydrogen production were increased by the LDD. Only in females, total carbohydrate oxidation and starch-derived glucose oxidation in response to the starch bolus were higher in LDD versus HDD mice; this was not accompanied by differences in amylase activity or hepatic partitioning of the 13C label. These results show that starch digestibility impacts glucose metabolism differently in females versus males.

Lack of ¹³C-label incorporation suggests low turnover rates of thaumarchaeal intact polar tetraether lipids in sediments from the Iceland shelf

Thaumarchaeota are amongst the most abundant microorganisms in aquatic environments, however, their metabolism in marine sediments is still debated. Labeling studies in marine sediments have previously been undertaken, but focused on complex organic carbon substrates which Thaumarchaeota have not yet been shown to take up. In this study, we investigated the activity of Thaumarchaeota in sediments by supplying different 13C-labeled substrates which have previously been shown to be incorporated into archaeal cells in water incubations and/or enrichment cultures. We determined the incorporation of 13C-label from bicarbonate, pyruvate, glucose and amino acids into thaumarchaeal intact polar lipid-glycerol dibiphytanyl glycerol tetraethers (IPL-GDGTs) during 4–6 day incubations of marine sediment cores from three sites on the Iceland shelf. Thaumarchaeal intact polar lipids, in particular crenarchaeol, were detected at all stations and concentrations remained constant or decreased slightly upon incubation. No 13C incorporation in any IPL-GDGT was observed at stations 2 (clay-rich sediment) and 3 (organic-rich sediment). In bacterial/eukaryotic IPL-derived fatty acids at station 3, contrastingly, a large uptake of 13C label (up to + 80‰ ) was found. 13C was also respired during the experiment as shown by a substantial increase in the 13C content of the dissolved inorganic carbon. In IPL-GDGTs recovered from the sandy sediments at station 1, however, some enrichment in δ13C (1–4‰ ) was detected after incubation with bicarbonate and pyruvate. The low incorporation rates suggest a low activity of Thaumarchaeota in marine sediments and/or a low turnover rate of thaumarchaeal IPL-GDGTs due to their low degradation rates. Cell numbers and activity of sedimentary Thaumarchaeota based on IPL-GDGT measurements may thus have previously been overestimated.

Lack of ¹³C-label incorporation suggests low turnover rates of thaumarchaeal intact polar tetraether lipids in sediments from the Iceland Shelf

Thaumarchaeota are amongst the most abundant microorganisms in aquatic environments, however, their metabolism in marine sediments is still debated. Labeling studies in marine sediments have previously been undertaken, but focused on complex organic carbon substrates which Thaumarchaeota have not yet been shown to take up. In this study, we investigated the activity of Thaumarchaeota in sediments by supplying different 13C-labeled substrates which have previously been shown to be incorporated into archaeal cells in water incubations and/or enrichment cultures. We determined the incorporation of 13C-label from bicarbonate, pyruvate, glucose and amino acids into thaumarchaeal intact polar lipid-glycerol dibiphytanyl glycerol tetraethers (IPL-GDGTs) during 4–6 day incubations of marine sediment cores from three different sites on the Iceland Shelf. Thaumarchaeal intact polar lipids were detected at all stations and concentrations remained constant or decreased slightly upon incubation. No 13C incorporation in any IPL-GDGT was observed at stations 2 (clay-rich sediment) and 3 (organic-rich sediment). In bacterial/eukaryotic IPL-derived fatty acids at station 3, contrastingly, a large uptake of 13C label (up to +80‰) was found. 13C was also respired during the experiment as shown by a substantial increase in the 13C content of the dissolved inorganic carbon. In IPL-GDGTs recovered from the sandy sediments at station 1, however, some enrichment in 13C (1–4‰) was detected after incubation with bicarbonate and pyruvate. The low incorporation rates suggest a low activity of Thaumarchaeota in marine sediments and/or a low turnover rate of thaumarchaeal IPL-GDGTs due to their low degradation rates. Cell numbers and activity of sedimentary Thaumarchaeota based on IPL-GDGT measurements may thus have previously been overestimated.

Sediment community responses to marine vs. terrigenous organic matter in a submarine canyon

The Whittard Canyon is a branching submarine canyon on the Celtic continental margin, which may act as a conduit for sediment and organic matter (OM) transport from the European continental slope to the abyssal sea floor. In situ stable-isotope labelling experiments were conducted in the eastern and western branches of the Whittard Canyon, testing short-term (3–7 days) responses of sediment communities to deposition of nitrogen-rich marine (Thalassiosira weissflogii) and nitrogen-poor terrigenous (Triticum aestivum) phytodetritus. 13C and 15N labels were traced into faunal biomass and bulk sediments, and the 13C label traced into bacterial polar lipid fatty acids (PLFAs). Isotopic labels penetrated to 5 cm sediment depth, with no differences between stations or experimental treatments (substrate or time). Macrofaunal assemblage structure differed between the eastern and western canyon branches. Following deposition of marine phytodetritus, no changes in macrofaunal feeding activity were observed between the eastern and western branches, with little change between 3 and 7 days. Macrofaunal C and N uptake was substantially lower following deposition of terrigenous phytodetritus with feeding activity governed by a strong N demand. Bacterial C uptake was greatest in the western branch of the Whittard Canyon, but feeding activity decreased between 3 and 7 days. Bacterial processing of marine and terrigenous OM were similar to the macrofauna in surficial (0–1 cm) sediments. However, in deeper sediments bacteria utilised greater proportions of terrigenous OM. Bacterial biomass decreased following phytodetritus deposition and was negatively correlated to macrofaunal feeding activity. Consequently, this study suggests that macrofaunal–bacterial interactions influence benthic C cycling in the Whittard Canyon, resulting in differential fates for marine and terrigenous OM.

Food quality determines sediment community responses to marine vs. terrigenous organic matter in a submarine canyon

The Whittard canyon is a branching submarine canyon on the Celtic continental margin, which may act as a conduit for sediment and organic matter (OM) transport from the European continental slope to the abyssal sea floor. In situ stable-isotope labelling experiments were conducted in the eastern and western branches of the Whittard canyon testing short term (3–7 day) responses of sediment communities to deposition of nitrogen-rich marine (Thallassiosira weissflogii) and nitrogen-poor terrigenous (Triticum aestivum) phytodetritus. 13C and 15N labels were traced into faunal biomass and bulk sediments, and the 13C label traced into bacterial polar lipid fatty acids (PLFAs). Isotopic labels penetrated to 5 cm sediment depth, with no differences between stations or experimental treatments (substrate or time). Macrofaunal assemblage structure differed between the eastern and western canyon branches. Following deposition of marine phytodetritus, no changes in macrofaunal feeding activity were observed between the eastern and western branches, with little change between 3 and 7 days. Macrofaunal C and N uptake was substantially lower following deposition of terrigenous phytodetritus with feeding activity governed by a strong N demand. Bacterial C uptake was greatest, in the western branch of the Whittard canyon, but feeding activity decreased between 3 and 7 days. Bacterial processing of marine and terrigenous OM were similar to the macrofauna in surficial (0–1 cm) sediments. However, in deeper sediments bacteria utilised greater proportions of terrigenous OM. Bacterial biomass decreased following phytodetritus deposition and was negatively correlated to macrofaunal feeding activity. Consequently, this study suggests that macrofaunal-bacterial interactions influence benthic C cycling in the Whittard canyon, resulting in differential fates for marine and terrigenous OM.