Inferring Phytoplankton, Terrestrial Plant and Bacteria Bulk delta C-13 Values from Compound Specific Analyses of Lipids and Fatty Acids

Abstract

Stable isotope mixing models in aquatic ecology require delta C-13 values for food web end members such as phytoplankton and bacteria, however it is rarely possible to measure these directly. Hence there is a critical need for improved methods for estimating the delta C-13 ratios of phytoplankton, bacteria and terrestrial detritus from within mixed seston. We determined the delta C-13 values of lipids, phospholipids and biomarker fatty acids and used these to calculate isotopic differences compared to the whole-cell delta C-13 values for eight phytoplankton classes, five bacterial taxa, and three types of terrestrial organic matter (two trees and one grass). The lipid content was higher amongst the phytoplankton (9.5 +/- 4.0%) than bacteria (7.3 +/- 0.8%) or terrestrial matter (3.9 +/- 1.7%). Our measurements revealed that the delta C-13 values of lipids followed phylogenetic classification among phytoplankton (78.2% of variance was explained by class), bacteria and terrestrial matter, and there was a strong correlation between the delta C-13 values of total lipids, phospholipids and individual fatty acids. Amongst the phytoplankton, the isotopic difference between biomarker fatty acids and bulk biomass averaged -10.7 +/- 1.1 parts per thousand for Chlorophyceae and Cyanophyceae, and -6.1 +/- 1.7 parts per thousand for Cryptophyceae, Chrysophyceae and Diatomophyceae. For heterotrophic bacteria and for type I and type II methane-oxidizing bacteria our results showed a -1.3 +/- 1.3 parts per thousand, -8.0 +/- 4.4 parts per thousand, and -3.4 +/- 1.4 parts per thousand delta C-13 difference, respectively, between biomarker fatty acids and bulk biomass. For terrestrial matter the isotopic difference averaged -6.6 +/- 1.2 parts per thousand. Based on these results, the delta C-13 values of total lipids and biomarker fatty acids can be used to determine the delta C-13 values of bulk phytoplankton, bacteria or terrestrial matter with +/- 1.4 parts per thousand uncertainty (i.e., the pooled SD of the isotopic difference for all samples). We conclude that when compound-specific stable isotope analyses become more widely available, the determination of delta C-13 values for selected biomarker fatty acids coupled with established isotopic differences, offers a promising way to determine taxa-specific bulk delta C-13 values for the phytoplankton, bacteria, and terrestrial detritus embedded within mixed seston.