1. Lignin Degradation From Plants to Energy: Efficient Biofuels
Margarette Francis, Maham Hijazi, Reeda Mahmood,
Riza Unabia & Manaum Zain

2. Fossil fuels: usage and concerns
“Fossil Fuels” in today’s world
Green house gasses, climate change and global warming
Why still use them?
Economically, it is more beneficial
Hybrid electric car
inefficent

3. Alternative options… Biofuels  Ethanol
Increasingly common option in agricultural and forestry industry in Canada
DISADVANTAGE  presence of lignin
Genomics to overcome this problem
An exemplary model  SOYBEAN

4. Why SOYBEAn? Soybean (Glycine max) Abundant all over the world
Grown in Southern and Eastern Ontario
Breeding advances (E.O)
Significant source of protein and oil

5. Ethanol as an energy source…
Reduces greenhouse gases
Does not contribute to carbon dioxide
emissions (34% reduction)
Input of energy required to produce
ethanol has dropped by 36%
Gives more energy compared to other renewable sources

6. Why Is lignin a Barrier?
Barrier for cellulose and hemicellulose extraction
Lignin removal improve the digestibility of the biomass
Mechanical and chemical treatments are used to breakdown lignin
rate of reaction is slow
Altering the pathway in order to reduce lignin is more effective
rate of reaction is comparatively faster
Wood Composition. (2007). Retrieved from
Yarris, L. (2013, November 13). Lignin-Feasting Microbe Holds Promise for Biofuels. States News Service. Retrieved from

7. The effect of lignin reduction on the structure of plants
Removing too much lignin from a plant affects its phenotype
Research conducted on the plant Arabidopsis thaliana
Reduction of 64% of lignin, compensated by the increase in matrix polysaccharides or cellulose
Afingen technique was used for lignin reduction,
producing functionally and structurally normal plants
Stems & Vascular Tissue. (2012). Retrieved from

8. The Phenylpropanoid Pathway
Enzymes and transcription factors of the phenylpropanoid pathway 
Source of metabolites in plants, being required for the biosynthesis of lignin, and serving as a starting point for the production of many other important compounds, such as the flavonoids, coumarins, and lignans.
Reinprecht, Y., Yadegari, Z., Perry, G., Siddiqua, M., Wright, L., Mcclean, P., & Pauls, K. (2013). In silico comparison of genomic regions containing genes coding for enzymes and transcription factors for the phenylpropanoid pathway in Phaseolus vulgaris L. and Glycine max L. Merr. Frontiers in Plant Science. Retrieved March 19, 2015, from

9. A closer look….
Cinnamyl alcohol dehydrogenase (CAD) catalyzes the reaction that is involved in the formation of the monolignols that make up lignin.
CAD
CAD
CAD
P-Hydoxyphenol lignin
Guaiacyl lignin
Syringyl lignin
Baldoni, A., Von Pinho, E. V. R., Fernands, J. S., Abreu, V. M., & Carvalho, M. L. M. (2013). Gene Expression in the lignin biosynthesis pathway during soybean seed development. Genet. Mol. Res 12 (3)

10. METHODS for GENOME EDITING
Finger nucleases
RNA interference
Transcription activator-like effector nucleases
Expensive and time-consuming
A recent and more innovative process..

11. Clustered regularly interspaced short palindromic repeats (CRISPRs) and CRISPR-associated (Cas) proteins
CRISPR/Cas9 System

12. What is it?
derived from a natural process found in bacteria to protect themselves from pathogens
targets genes for editing and regulating
comparable to Photoshop
Horizon Licenses Harvard University Gene-Editing Technology. (2013). Drug Discovery & Development.

13. To initiate gene modification,
sgRNA
(single guide RNA)
Cas9 complex
Cas9 nuclease

14. Protospacer Adjacent Motif (PAM)
Target Sequence
Gene of Interest

15. Non-Homologous End Joining (NHEJ) DNA repair pathway
Stop Codon

16. Summation: crispr/Cas system
sgRNA/Cas9 complex binds to gene
Cas breaks gene (double strand breaks)
Induced mutation in gene sequence
altered gene sequence  dysfunctional CAD gene

17. What makes a good biofuel?
Greenhouse Gas Reduction Potential
Cost-Effectiveness
Energy Allocations for Crops

18. Abundance in Canada - Ontario
Crop Production Data in Metric Tonnes
Soybean is one of the most widely used feedstock for biofuel production.
Canada’s second leading crop.
Crops
2012
2013
2014
Barley
165,500
134,600
151,300
Corn
8,598,300
9,007,300
7,600,000
Soybean
3,401,900
3,238,600
3,791,100
Canola
61,200
49,900
31,300
Wheat, winter
1,750,000
2,277,900
1,627,500
Statistics Canada. CANSIM table 

19. Energy Allocations
WCBA. Canada.
Soybean has a much higher net energy balance as compared to various other feedstocks

20. GREENHOUSE GAS EMISSIONS
According to the Canadian Natural Resources model, GHGenius, soybean biodiesel and corn ethanol were found to reduce GHG emissions by 50% and 21%, respectively.
According to the Greenhouse gases, regulated Emissions and Energy in Transportation (GREET) model for life cycle analysis, all soybean-derived fuels achieve a significant reduction (52–107%) in fossil energy use and in petroleum use (more than 85%)

21. Why is it good for Canada?
Cost-effective
Eco-friendly
Sustainable
Renewable