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Isolation and characterization of hybrid poplar galactinol synthases Unda, Faride; Mansfield, Shawn Sep 13, 2011

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POSTER PRESENTATION Open AccessIsolation and characterization of hybrid poplargalactinol synthasesFaride Unda1*, Shawn Mansfield2From IUFRO Tree Biotechnology Conference 2011: From Genomes to Integration and DeliveryArraial d Ajuda, Bahia, Brazil. 26 June - 2 July 2011The raffinose family of oligosaccharides (RFOs) likelyfulfill at least few major physiological roles in plants,translocation of carbon in the phloem, storage in sinktissues, and as putative biological agents to combat bothabiotic and biotic stress [1-3]. The synthesis of galacti-nol from myo-inositol + UDP-galactose, by galactinolsynthase (GolS), is considered a key regulatory step inRFO synthesis [4].To investigate the functional roles ofthis class of compounds in trees, two cDNAs thatencode galactinol synthase (GolS), were identified andcloned from hybrid poplar (P. alba × grandidentata).Phylogenetic analyses of the Populus GolS isoforms withother known galactinol synthases suggested a putativerole for these enzymes during biotic or abiotic stress inhybrid poplar. The predicted protein sequences of bothisoforms (PaxgGolSI and PaxgGolSII) showed character-istics of galactinol synthases from other species, includ-ing a serine phosphorylation site at position 266 and thepentapeptide hydrophobic domain ASAAP [5]. Kineticanalyses of recombinant Pa×gGolSI and PaxgGolSIIresulted in Km values for UPD-galactose of 0.79 and0.65 mM and Vmax values of 660.4 and 1245 nM min-1,respectively. PaxgGolSI inherently possessed broader pHrange and temperature sensitivity when compared toPa×gGolSII. Interestingly, spatial and temporal expres-sion analyses revealed that PaxgGolSII transcript levelsvaried seasonally, while PaxgGolSI did not, therebyimplying a temperature-regulated transcriptional controlof this gene in addition to the observed thermosensitiv-ity of the respective enzyme. Based on this evidence, wesuggest that Pa×gGolSI may be involved in basic meta-bolic activities (i.e.storage), while Pa×gGolSII is likelyinvolved in seasonal mobilization of carbohydrates.Author details1Wood Science Department -University of British Columbia, Canada. 2WoodScience Department- University of British Columbia, Canada.Published: 13 September 2011References1. Taji T, Ohsumi C, Iuchi S, Seki M, Kasuga M, Kobayashi M, et al: Importantroles of drought- and cold-inducible genes for galactinol synthase instress tolerance in Arabidopsis thaliana. Plant Journal 2002, 29(4):417-426.2. Panikulangara TJ, Eggers-Schumacher G, Wunderlich M, Stransky H,Schoffl F: Galactinol synthase1. A novel heat shock factor target generesponsible for heat-induced synthesis of raffinose familyoligosaccharides in Arabidopsis. Plant Physiology 2004, 136(2):3148-3158.3. Philippe RN, Ralph SG, Mansfield SD, Bohlmann J: Transcriptome profiles ofhybrid poplar (Populus trichocarpa x deltoides) reveal rapid changes inundamaged, systemic sink leaves after simulated feeding by forest tentcaterpillar (Malacosoma disstria). New Phytologist 2010, 188(3):787-802.4. Keller F, Pharr DM: Metabolism of carbohydrates in sinks and sources:Galactosyl-sucrose oligosaccharides. In Photoassimilate distribution inplants and crops. New York: Marcel Dekker;Zamski E, & Schaffer AA1996:115-184.5. Sprenger N, Keller F: Allocation of raffinose family oligosaccharides totransport and storage pools in Ajuga reptans: The roles of two distinctgalactinol synthases. Plant Journal 2000, 21(3):249-258.doi:10.1186/1753-6561-5-S7-P77Cite this article as: Unda and Mansfield: Isolation and characterization ofhybrid poplar galactinol synthases. BMC Proceedings 2011 5(Suppl 7):P77.Submit your next manuscript to BioMed Centraland take full advantage of: • Convenient online submission• Thorough peer review• No space constraints or color figure charges• Immediate publication on acceptance• Inclusion in PubMed, CAS, Scopus and Google Scholar• Research which is freely available for redistributionSubmit your manuscript at www.biomedcentral.com/submit* Correspondence: farideu@interchange.ubc.ca1Wood Science Department -University of British Columbia, CanadaFull list of author information is available at the end of the articleUnda and Mansfield BMC Proceedings 2011, 5(Suppl 7):P77http://www.biomedcentral.com/1753-6561/5/S7/P77© 2011 Unda and Mansfield; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the CreativeCommons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, andreproduction in any medium, provided the original work is properly cited.

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