BIOFUELSMargarette Francis, Maham Hijazi, Reeda Mahmood, Riza Unabia & Manaum Zain

From Plants to From Plants to

Energy: Energy: Efficient Lignin Degradation

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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

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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

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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

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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

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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

Yarris, L. (2013, November 13). Lignin-Feasting Microbe Holds Promise for Biofuels. States News Service. Retrieved from http://www.highbeam.com/doc/1G1-348985171.html?

Wood Composition. (2007). Retrieved from http://learn.forestbioenergy.net/learning-modules/module-6/unit-1/lesson-1

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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 http://www.sciencepartners.info/?page_id=534 7

The PhenylpropanoidPathway

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 http://journal.frontiersin.org/article/10.3389/fpls.2013.00317/full 8

A CLOSER LOOK….

Cinnamyl alcohol dehydrogenase (CAD) catalyzes the reaction that is involved in the formation of the monolignols that make up lignin.

P-Hydoxyphenol lignin

Guaiacyl lignin

Syringyl lignin

CADCAD CAD

9Baldoni, 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) 2618-2624

METHODS for GENOME EDITING

Finger nucleasesRNA interference Transcription activator-like effector nucleases

Expensive and time-consuming

A recent and more innovative process..

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CLUSTERED REGULARLY INTERSPACED SHORT PALINDROMIC REPEATS

(CRISPRS) AND CRISPR-ASSOCIATED (CAS) PROTEINS

CRISPR/CAS9 SYSTEM

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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. 12

Cas9 nuclease

sgRNA(single guide RNA)

TO INITIATE GENE MODIFICATION,

Cas9 complex

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Gene of Interest

Protospacer Adjacent Motif (PAM) Target

Sequence

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Stop Codon

Non-Homologous End Joining (NHEJ) DNA repair pathway

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SUMMATION: CRISPR/CAS SYSTEM

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altered gene sequence dysfunctional CAD gene

WHAT MAKES A GOOD BIOFUEL?

1.Greenhouse Gas Reduction Potential

2.Cost-Effectiveness

3.Energy Allocations for Crops

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ABUNDANCE IN CANADA - ONTARIO

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

Soybean is one of the most widely used feedstock for biofuel production.

Canada’s second leading crop.

Statistics Canada. CANSIM table  001-0010

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Crop Production Data in Metric Tonnes

ENERGY ALLOCATIONS

WCBA. Canada. http://westerncanadabiodiesel.org/biodiesel/

Soybean has a much higher net energy balance as compared to various other feedstocks

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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%)

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WHY IS IT GOOD FOR CANADA?

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