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Genomic Science Program

2006 Awardee

Biochemical Genomics of Wood Formation:O-Acyltransferases for Alteration of LignocellulosicProperty and Enhancement of Carbon Deposition inPoplar

INVESTIGATOR(S): Liu, C.-J.

INSTITUTION: Brookhaven NationalLaboratory

NON-TECHNICAL SUMMARY: The goal of GTL isto use newly acquired genomic data to better understandfundamental biological processes and enhance thetranslation of that scientific knowledge into newtechnologies for energy and environmental applications. Ourproject is going to characterize the O -acylationreactions participated in lignocellulosic biosynthesesusing poplar genomics resources. O -acylation isinvolved in the cell-wall component polysaccharide andlignin biosyntheses. It affects the cell-wall sugar'ssolubility and the lignocellulosic bio-digestibility. O -acylation is also responsible for the structuralmodification and sequestration of a variety of polyphenolicmetabolites required for wood formation. Understanding themechanism of O-acylation at molecular level implicatesbiotechnological applications in genetic modification oflignocellulosic structures to facilitate biomass tobioethanol conversion, and in improvement of feedstockbiomass production.

OBJECTIVES: 1) genome-wide identifyacyl-CoA dependent acyltransferase genes from poplargenomics database; 2) systemically explore the tissuespecific and stress-responsible expression patterns of O -acyltransferase genes to identify the enzymesspecifically involved in lignocellulosic biosynthesis; 3)systemically characterize the biochemical functions ofacyltransferases responsible for polysaccharideacetylation, lignol biosynthesis and phenolic compoundmodification.

APPROACH: Obj. 1. tblastn algorithm willbe applied to search poplar genomics resources ( P.trichocarpa V1.0, http://genome.jgi-psf.org )by using the highly conserved sequence motifs (HXXXD andDFGWG) of acy-CoA dependent acyltransferases. In order todistinguish the potential soluble and membrane boundproteins, the encoded polypeptides of the identified genecandidates (at least ~50 gene models) will be subjected tothe computational topology and post-translationalmodification analyses by using PSORT and SignalP webservices to predict the protein sorting signal, subcellularlocalization site and the location of signal peptidecleavage sites in amino acid sequences. Obj. 2. Thetranscriptional profiling of putative acyltransferases willbe analyzed both by " in silico " northern, basedupon the high resolution poplar EST/microarray databases,and by QRT-PCR against mRNAs from different types oftissues (leaf, shoot, root, stem etc.) and the tissuesections representing different stages of developing wood,including the early expansion, late expansion, secondarycell wall formation, and programmed cell death(sapwood/heartwood) from poplars under normal growingconditions, the drought, salt stresses, and insect damageor physical wounding. Obj. 3. The recombinant proteins ofthe gene candidates that are highly expressed inwood-forming tissues will be produced using either E.coli , yeast, or Drosophila Gatewayexpression systems. Subsequently the combinatorial invitro assays will be conducted by using differentacyl-CoA donors and potential substrates including sugars,oligosaccharides, hydrolyzed and pre-deacylated pectin andxyloglucan, lignols, and other phenolics. Productsdetection and identification will be performed byLC-UV-ESI-MS n , HPAEC-MS and MALDI-TOF-MSanalyses.

PROJECT CONTACT:

Name: Liu, C.-J.
Phone: 631-344-2966
Fax: 631-344-3407
Email: cliu@bnl.gov

 

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