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499 - October 18

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by

Tina Wong

on 16 October 2012

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Transcript of 499 - October 18

Tobacco Cell walls VIGS Can be coppiced – multiple harvests per year
Does not flower at high latitudes – helps with transgenic containment Adrianov V, Borisjuk N, Pogrebnyak N, Brinker A, Dixon J, Spitsin S, Flynn J, Matyszczuk P, Adryszak K, Laurelli M, Golovkin M, Koprowski H (2010) Tobacco as a production platform for biofuel: overexpression of Arabidopsis DGAT and LEC2 genes increases accumulation and shifts the composition of lipids in green biomass. Plant Biotech J 8: 277-287 http://3.bp.blogspot.com/-9dzyEvRnEpI/TnbKzKRg9gI/
AAAAAAAAAGw/1kWEBlSSS-c/s1600/tobacco+flower.jpg Nicotiana tabacum often overlooked due to smoking industry
Broad growth range
Easy to grow & manipulate
Large amount of biomass: 170 tons/hectare Tobacco for bioproducts Adrianov V, Borisjuk N, Pogrebnyak N, Brinker A, Dixon J, Spitsin S, Flynn J, Matyszczuk P, Adryszak K, Laurelli M, Golovkin M, Koprowski H (2010) Tobacco as a production platform for biofuel: overexpression of Arabidopsis DGAT and LEC2 genes increases accumulation and shifts the composition of lipids in green biomass. Plant Biotech J 8: 277-287 Tobacco can reach up to 40% seed weight in oil, but has lower yield than traditional biofuel oil crops
Primary oil synthesis occurs in the leaves of many plant species
Tobacco: 1.7%-4% oil per dry weight
This oil is extracted as fatty acid esters – primary component of biofuel oil
Transformation of tobacco with upregulated Arabidopsis DGAT gave 20-fold increase in TAG accumulation and shifted lipid composition to favour biodiesel production Tobacco for oil production http://micro.magnet.fsu.edu/cells/plants/images/cellwallfigure1.jpg Raven PH, Evert RF, Eichhorn SE (2005) Biology of plants.7th ed. Freeman, New York, pp 53-57 Functions:
Retains protoplast
Absorption
Transport
Secretion
Defense
Communication
Composition:
Cellulose
Hemi-cellulose
Pectin
Glycoprotein Target: primary cell walls Raven PH, Evert RF, Eichhorn SE (2005) Biology of plants.7th ed. Freeman, New York, pp 53-57 http://upload.wikimedia.org/wikipedia/commons/thumb/c/c3/Cellulose_strand.jpg/260px-Cellulose_strand.jpg Linear chain of β(1→4) ᴅ-glucose molecules Cellulose Raven PH, Evert RF, Eichhorn SE (2005) Biology of plants.7th ed. Freeman, New York, pp 53-57 http://nutrition.jbpub.com/resources/chemistryreview9.cfm Cellulose Raven PH, Evert RF, Eichhorn SE (2005) Biology of plants.7th ed. Freeman, New York, pp 53-57 http://doctortee.com/dsu/tiftickjian/cse-img/botany/plant-anat/cell/rosette.jpg Exists as a rosette-shaped complex in the plasma membrane
Synthesizes cellulose microfibrils using UDP-glucose
Extrudes microfibril – walls grow from outside in
Complexes organized in Golgi and carried to PM via dictyosomes Cellulose synthase Lu R, Martin-Hernandez AM, Peart JR, Malcuit I, Baulcombe DC (2003) Virus-induced gene silencing in plants. Methods 30: 296-303 Exploits intrinsic RNA silencing defense mechanism in plants
ssRNA virus  dsRNA via RNA-dependent RNA Polymerase
dsRNA processed to make siRNAs
Endogenous RNase complex is guided by pairing of siRNAs to target RNA for degradation
VIGS: vector contains insert corresponding to host gene  posttranscriptional gene silencing
Very specific and rapid
Silencing occurs in 6-10 days Virus-induced gene silencing (VIGS) http://www.sassykandle.com/tobacco-flower/ Dr. Stacy Singer
Dr. Mike Deyholos
Dr. Randall Weselake
Dr. Rodrigo Siloto
Robin Miles
Scott Greer Acknowledgements The Big Picture Adrianov V, Borisjuk N, Pogrebnyak N, Brinker A, Dixon J, Spitsin S, Flynn J, Matyszczuk P, Adryszak K, Laurelli M, Golovkin M, Koprowski H (2010) Tobacco as a production platform for biofuel: overexpression of Arabidopsis DGAT and LEC2 genes increases accumulation and shifts the composition of lipids in green biomass. Plant Biotech J 8: 277-287 http://www.asergeev.com/pictures/archives/2008/651/jpeg/09.jpg Down-regulate cellulose synthase in N. benthamiana using VIGS
Resultant carbon from suppressed cell wall synthesis re-allocated into oil in leaves
Bioproduct applications
Proof of concept for increased oil content in oilseeds
Attain diminished cell wall formation without introducing a lethal or very severe mutation Putting it all together http://www.sassykandle.com/tobacco-flower/ Dr. Stacy Singer
Dr. Mike Deyholos
Dr. Randall Weselake
Dr. Rodrigo Siloto
Robin Miles
Scott Greer Acknowledgements Ideas for hand-sectioning Brassica napus seeds? Question for the lab Tobacco as a platform for bioproducts
Oil in leaves instead of seeds
Target: primary cell walls
Function & composition
Cellulose & cellulose synthase
VIGS
Putting it all together
Wanted: smart ideas from smart people! Outline
Full transcript