Ashley Campbell edited Results & Discussion.tex  over 9 years ago

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The \textsuperscript{13}C-incorporation reveals temporal dynamics of C degradation demonstrated by \textsuperscript{13}C-xylose incorporation at days 1, 3, and 7 and \textsuperscript{13}C-cellulose incorporation at days 14 and 30 (\href{https://www.authorea.com/users/3537/articles/3612/master/file/figures/ordination_all1/ordination_all1.png}{Fig. 1B}). In support of this, the bulk community sequencing demonstrates significant (pval) microbial community changes over time. Although within a single time point, the bulk community demonstrated no significant difference between treatments (Fig Sx).   The temporal dynamics reveal the Heavy fraction amplicon pools from samples that received \textsuperscript{13}C-xylose diverged from corresponding controls on days 1 through 7. Furthermore, amplicon pool  composition of varied across these days (Fig x) indicating dynamic changes in  \textsuperscript{13}C-xylose assimilating amplicons assimilation with time. At day 14 and 30 heavy fractions from \textsuperscript{13}C-xylose labeled samples  are different for each no longer differentiated from corresponding controls indicating that \textsuperscript{13}C is no longer detectable in DNA. The decline in \textsuperscript{13}C-labelling  of the days the label DNA  is detected based on their separate distributions for each likely due to isotopic dilution resulting from assimilation  of the time points unlabeled C and/or due to cell turnover resulting from mortality  (\href{https://www.authorea.com/users/3537/articles/3612/master/file/figures/ordination_all1/ordination_all1.png}{Fig. 1B}). The disappearance of the \textsuperscript{13}C-xylose incorporation signature (relative to control) for days 14 and 30 result from loss of \textsuperscript{13}C-label in DNA over time. This occurs by dilution of \textsuperscript{13}C-label out of the DNA when a switch from \textsuperscript{13}C to \textsuperscript{12}C substrate utilization takes place during biomass turnover and/or predation. \textsuperscript{13}C-cellulose incorporation isn't detected until day 14 and amplicon composition is consistent for both days 14 and 30 (\href{https://www.authorea.com/users/3537/articles/3612/master/file/figures/ordination_all1/ordination_all1.png}{Fig. 1B}).The consistency of amplicon composition for cellulose degradation over time compared to xylose suggests a wider array of microorganisms utilize xylose, whereas, cellulose utilization occurs in a select few. This is consistent with long standing notions that more microorganisms are capable of utilizing simple carbohydrates than complex C substrates.  Overall patterns of C degradation observed in this study demonstrate different microbial community members are responsible for the consumption of these two substrates; xylose is consumed quickly, whereas, cellulose decomposition takes longer. This suggests a pattern of microbial community transition accompanying the decomposition process. This is consistent with \cite{Engelking_2007} who observed as much as 75\% of labile C respired or converted into microbial biomass in the first 5 days of decomposition, whereas, cellulose degraders take longer to respond \cite{Hu_1997} with less than 42\% of cellulose metabolized over the first 5 days of incubation \cite{Engelking_2007}. \textbf{Differential C utilization by taxa.} Individual OTUs that assimilated \textsuperscript{13}C-substrates were identified using the DESeq framework \cite{Anders_Huber_2010} to analyze differential representation in heavy fractions (\href{https://www.authorea.com/users/3537/articles/3612/master/file/figures/l2fc_fig1/l2fc_fig.pdf}{Fig. 2}). There were 43 and 35 unique OTUs that significantly (\textit{p}-value \textless 0.10) assimilated \textsuperscript{13}C-xylose and \textsuperscript{13}C-cellulose, respectively; herein called 'responders' (\href{https://www.authorea.com/users/3537/articles/8459/master/file/figures/OTU_screening_schematic/OTU_screening_schematic.pdf}{Fig. S2}, \href{https://www.authorea.com/users/3537/articles/8459/master/file/figures/l2fc_fig_pVal/l2fc_fig_pVal.png}{Fig. S3}). Overall, we found xylose responders were from higher rank abundances than cellulose responders, however, cellulose responders exhibited a greater change in buoyant density (i.e. assimilated more \textsuperscript{13}C) than xylose responders in response to isotope incorporation (Figure 3).