1. Enzyme capacities to decompose plant-biomass in microbial symbionts associated with fungus-growing termites
Rafael R. da Costa, Haofu Hu, Bo Pilgaard, Julia Schükel, Stjepan K. Kračun, Sabine Vreeburg, Lene Lange, Duur Aanen, Michael Poulsen
MSc Rafael Rodrigues da Costa - PhD Student at Københavns Universitet
University of Copenhagen, Department of Biology, 
Section for Ecology and Evolution - Centre for Social Evolution (CSE)
Universitetsparken 15, Building 3, 1st floor, 2100 Copenhagen East
EU IUSSI 2016 Conference
10/08/2016

2. Fungus-farming termites
Section for Ecology and Evolution
Fungus-farming termites
Supports vegetation
Supports large grazers
Important for nutrient cycling: Decompose 90% of plant material in African savannahs
1.85m
Bonachela et al. Science 2015
Daniel Kapijimpanga

3. Fungus-growing termites and their microbial symbionts
Co-dependent mutualism with fungi and gut bacteria
@Aanen et al., 2009

4. Macrotermes natalensis
Decomposition in fungus-growing termites
Enzymes to break down nearly any plant polysaccharide.
Mainly enzymes to break down fungus material and sugars (also presence of a gene expressing a cellulase)
Mainly enzymes to break the fungus cell-wall and to utilize simple sugars
Macrotermes natalensis
Poulsen et al. 2014, PNAS

5. Section for Ecology and Evolution
How do Termitomyces and gut bacteria work synergistically to decompose plant-biomass?
Chromogenic substrates
AZCL
Termite guts and fungus combs
Julia Schückel
Jesper Harholt
CoMPP
Haofu Hu
Kristine Pedersen
Sabine
Vreeburg
Acid hydrolyses (HPCL)
Enzyme assays
Content analyses
RNAseq
Sabine Vreeburg
Duur Aanen
RNAseq and PPR
Illumina HiSeq
Characterise decomposition enzymes
Plant component content
What enzymes are expressed?
Lene Lange
Bo Pilgaard
Duur Aanen

6. AZCL enzyme activities
Section for Ecology and Evolution
AZCL enzyme activities
Fresh
Nodules
Old
AZCL
Guts
Odontotermes badius
Odontotermes sp.
Macrotermes natalensis
17 different substrates presence of halo = positive result
Closed symbols: colonies sampled in Open symbols: colonies sampled in 2016

7. Enzyme capacities in components involved in the decomposition process
Fresh comb
Old comb
Nodules
Old major worker guts
AZCL
RAFA – SOMETHING OFF WITH THE MANIPULATIONS.

8. CoMPP suggests decomposition of polysaccharides from fresh comb>old comb>guts
Termite guts

9. Content analysis – Plant components
Objective: measure up to 90% of sample composition (plant components + fungal biomass) What has been measured: Lignin Xylose Glucose Mannose Galactose Fucose Galacturonic acid Glucuronic acid What is ongoing: Cellulose Xylan Fungal biomass
General decrease in the polymers amount;
Consumption polymers throughout decomposition process;
Decomposition could possibly start in the guts;
Fresh comb
Old comb
Old major worker guts
Lignin
Xylose
Glucose
Arabinose
Lignin
Xylose
Glucose
Arabinose
Lignin
Xylose
Glucose
Arabinose
Macrotermes
natalensis
* To be done: Soluble sugars measurements Cellulose measurements Lignin measurements Fungal biomass measurements
Odontotermes sp.

10. Plant-biomass decomposition in Macrotermes natalensis symbionts
Section for Ecology and Evolution
AZCL activity
CAZy transcript levels
RNAseq from guts ongoing…
Illumina HiSeq

11. Section for Ecology and Evolution
Summary
Production and optimal distribution of plant-degrading enzymes in Macrotermes natalensis
 Enzymes produced in comb  transported to nodules  transported via guts to the fresh plant substrate?
1- Decomposition process is initiated and finalized in the gut
2- Different processes take place in different locations (i.e. nodules, guts and comb)
3- Different species are likely to have specific enzyme capacities

12. Section for Ecology and Evolution
Acknowledgements