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Antisense expression of a caffeic acidO-methyltransferase ofStylosanthes humilis in transgenic poplar: Effect of expression onO-methyltransferase activity and lignin composition
Authors:Kasim Bajrovic  Kemal Kazan  Zeliha İpekçi  Nermin Gözükırmızı
Affiliation:(1) Research Institute for Genetic Engineering and Biotechnology, TüBITAK, MRC, 41470 Gebze-Kocaeli, Turkey;(2) Cooperative Research Center for Tropical Plant Pathology, University of Queensland Brisbane, 4072, Australia;(3) Biology Department, University of Istanbul, Faculty of Science, 34459 Vezneciler, Istanbul, Turkey
Abstract:
Poplar (Populus tremula) was transformed with a construct carrying an antisense caffeic acidO-methyltransferase (COMT) cDNA (pOMT8) from a tropical pasture legume,Stylosanthes humilis. pOMT8 shows 83% overall homology to the corresponding COMT gene (pPCLA) of poplar. Of the 200 putatively-transformed plants regenerated on selective media after co-cultivation of poplar stem explants withAgrobacterium tumefaciens harbouring a CaMV 35S-antisensepOMT8 construct, a subset of 20 plants were randomly chosen for further analysis. PCR and Southern blot analysis demonstrated the stable integration of T-DNA into the genome of these plants. Antisense expression ofpOMT8 resulted in reductions in total COMT activity in the majority of the transgenic plants with the lowest total COMT activities (61–70% of untransformed control plants) being observed in four transgenic plants. The composition of lignin in transgenic plants was also changed, as detected by reductions in the content of syringyl units using infrared spectroscopy. However, no changes were found in the amount of insoluble lignin in transgenic plants as compared to untransformed control plants. These results indicate the potential of thepOMT8 gene to partially suppress COMT activity and modify the composition of lignin in transgenic poplar. This work was partly supported by General Management of Turkish Pulp and Paper Mills.
Keywords:caffeic acidO-methyltransferase  lignin   Populus tremula    Stylosanthes humilis
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