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TECHNICAL REPORTS

Ecological Risk Assessment

Molecular Composition of Leaves and Stems of Genetically Modified Bt and Near-Isogenic Non-Bt Maize—Characterization of Lignin Patterns

^a UFZ-Center for Environmental Research Leipzig-Halle Ltd., D-04318 Leipzig, Germany
^b RWTH Aachen, Institute of Environmental Research (Biology V), Chair of Ecology, Ecotoxicology, Ecochemistry, D-52076 Aachen, Germany
^c Leibniz-Center for Agricultural Landscape and Land Use Research, D-15374 Muncheberg, Germany
^d Department of Chemistry, University of Waterloo, Waterloo, ON, N2L 3G1 Canada

^* Corresponding author (juergen.poerschmann{at}ufz.de)

Received for publication February 25, 2005. Transformation of crops, including maize (Zea mays L.), with the cry1Ab gene from Bacillus thuringiensis to combat lepidopteran pests results in pleiotropic effects regarding lignin biosynthesis. Lignin patterns in stems and leaves of two genetically modified Bt-maize varieties (Novelis T and Valmont T) were studied along with their non-Bt near-isolines (Nobilis and Prelude, respectively). Molecular-level based thermochemolysis using tetramethylammonium hydroxide (TMAH) in combination with gas chromatography–mass spectrometry (GC–MS) was used to quantitate the total lignin contents and to identify monomeric lignin subunits including p-hydroxyphenyl (P), guaiacyl (G), and syringyl (S) moieties. The results were supplemented and confirmed by cupric oxide oxidation. The stems of the transgenic lines had higher concentrations of total lignin than the respective isogenic lines: Valmont T/Prelude by 18% and Novelis T/Nobilis by 28%. In contrast, differences in the total lignin concentration of leaves between the transgenic and the respective near-isogenic lines were marginal. There were significant modifications in the ratio of p-hydroxyphenyl/guaiacyl/syringyl molecular marker units of stem lignin between transgenic and isogenic lines. The guaiacyl units (in particular the G18 marker) accounted chiefly for the higher total lignin contents in the transgenic lines. The leaf lignin patterns did not show significant differences in molecular markers between isogenic and transgenic lines. TMAH-induced thermochemolysis—conducted in both the on-line and off-line modes—provided detailed information on the molecular composition of lignin, thus proving superior to the established "wet chemistry" methods of lignin determination.

Abbreviations: amu, atomic mass unit • FFAP, free fatty acid phase • G, guaiacyl unit • GC–MS, gas chromatography–mass spectrometry • P, p-hydroxyphenyl unit • RSD, relative standard deviation • S, syringyl unit • TMAH, tetramethylammonium hydroxide

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