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Plant Sunscreen: The Formation of Sinapoylmalate by O -Methyl Transferases. Beth Kauffman Under the Guidance of: Nick Anderson and Clint Chapple. Outline. Why are methyltransferases important? What is the phenotype of an atomt1 mutant

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plant sunscreen the formation of sinapoylmalate by o methyl transferases

Plant Sunscreen: The Formation of Sinapoylmalate by O-Methyl Transferases

BethKauffman

Under the Guidance of:

Nick Anderson and Clint Chapple

outline
Outline
  • Why are methyltransferases important?
  • What is the phenotype of an atomt1 mutant
  • How is a candidate COMT tested for redundant ATOMT1 activity?
  • Do any of the candidate COMT’s have redundant activity?
  • Future Directions?
methylation is a common reaction in nature
Methylation is a common reaction in nature

Methyl Donor

Methyltransferase

Methyl Donor

Methyltransferase

a short life cycle and small genome make arabidopsis a model organism1
A short Life cycle and small genome make Arabidopsis a model organism
  • Model organism
  • Small genome which has been sequenced
  • Short life cycle compared to other plants
  • Relatively easy to make a mutation
a caffeic acid o methyltransferase comt atomt1 participates in the phenylpropanoid pathway
A Caffeic Acid O-Methyltransferase (COMT), ATOMT1 participates in the phenylpropanoid pathway.

sinapoylmalate

atomt1

ATOMT1

sinapylaldehyde

phenylalanine

5-OH coniferaldehyde

5-OH feruloylmalate

S-lignin

o methyltransferases may be responsible sinapoylmalate formation in an atomt1 mutant
O-methyltransferases may be responsible Sinapoylmalate formation in an atomt1 mutant
  • S-lignin falls below level of detection
  • Sinapoylmalate is still formed.
  • Arabidopsis has many candidate COMT genes
candidate comt s are investigated using a reverse genetic approach
Candidate COMT’s are investigated using a reverse genetic approach

Disruption

Gene

comt1

atomt1

comt8

atomt1

comt2

atomt1

atomt1

comt9

comt3

atomt1

atomt1

comt10

atomt1

comt4

comt5

atomt1

comt11

atomt1

atomt1

atomt1

comt12

comt6

comt13

atomt1

comt7

atomt1

hplc analysis is used to quantify soluble metabolite levels
HPLC analysis is used to quantify soluble metabolite levels
  • Determined metabolic Profile of HPLC Analysis
  • If an COMT was responsible, then the mutant’s metabolic profile would contain no sinapoylmalate.
hplc analysis is used to quantify soluble metabolite levels1
HPLC analysis is used to quantify soluble metabolite levels
  • Determined metabolic Profile of HPLC Analysis
  • If an COMT was responsible, then the mutant’s metabolic profile would contain no sinapoylmalate.

5-OH feruloylmalate

sinapoylmalate

future directions
Future Directions
  • It may be that a combination of several COMT’s are responsible for the redundant activity
  • Triple mutants generated
  • Same method of study conducted with triple mutants. If a triple mutant does not form sinapoylmalate, then combination is responsible

comt2

atomt1

comt1

acknowledgements

Chapple Lab

Nick Anderson

Clint Chapple

Jo Cusumano

Nick Bonawitz

Jeong-Im Kim

Peng Wang

Yi Li

Whitney Dolan

WenjieZeng

Marwa El-Hindawy

Alexandra Smith

Nicholas Pogranichniy

Misha Remy

Acknowledgements

references
References
  • Besse, P., Da Silva, D., Bory, S., Noirot, M., and Grissoni, M.(2009). Variation in intron length in caffeic acid O-methyltransferase (COMT) in Vanilla species (Orchidaceae). Plant Sci. 176, 452-460.
  • Goujon, T., Sibout, R., Pollet, B., Maba, B., Nussaume, L., et. al. (2003). A new Arabidopsis thaliana mutant deficient in the expression of O-methyltransferase impacts lignins and sinapoylesters.Plant. Mol. Bio.51, 973-989.
  • Do, C., Pollet, B., Thevenin, J., Sibout, R., Denoue, D., et. al. (2007). Both caffeoul Coenzyme A 3-O-methyltransferase 1 and caffeic acid O-methyltransferase 1 are involved in redundant functions for lignin, flavonoids and sinapoylmalate biosynthesis in Arabidopsis. Planta.226, 1117-1129.
  • Dang, T. T., and Facchini, P. J. (2012). Characterization of Three O-Methyltransferases Involved in Noscapine Biosynthesis in Opium Poppy. Plant Phys.159, 618-631.
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