A metabolomic study of substantial equivalence of field-grown genetically modified wheat
A metabolomic study of substantial equivalence of field-grown genetically modified wheat
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DOI：
10.1111/j.1467-7652.2006.00197.x
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发表时间：
2006-07-01
影响因子：
13.8
通讯作者：
Beale, Michael H.
中科院分区：
文献类型：
--
作者：
Baker, John M.;Hawkins, Nathaniel D.;Beale, Michael H.
通过对英国两个地点（赫特福德郡的罗萨姆斯特德和布里斯托尔附近的朗阿什顿）重复田间试验中生长的样品进行为期 3 年的代谢物分析，检查了表达额外高分子量亚基基因的三种转基因小麦和相应亲本系（两个系加上无效转化体）的“实质等同性”。氘化甲醇-水提取的极性代谢物的质子核磁共振谱的多变量比较表明，地点和年份的影响比基因型的影响更强。然而，观察到转基因系和亲本系之间存在一些分离，特别是转基因系B73-6-1（其具有最高水平的转基因表达）与其亲本系L88-6之间。光谱比较表明，这种分离是由于该转基因品系中麦芽糖和/或蔗糖水平增加造成的，并且游离氨基酸的差异也很明显。使用气相色谱-质谱法对 2000 年生长的材料的氨基酸组成进行了更详细的研究。最显着的差异是，在洛桑培养的样品始终含有大量酸性氨基酸（谷氨酸、天冬氨酸）及其酰胺（谷氨酰胺、天冬酰胺）。此外，相关品系 L88-6 和 B73-6-1 在 Long Ashton 生长时都含有比在 Rothamsted 生长时更多的脯氨酸和 γ-氨基丁酸。结果清楚地表明环境影响代谢组，并且对照品系和转基因品系之间的任何差异通常与在不同地点和不同年份生长的对照品系之间观察到的差异在同一范围内。
The 'substantial equivalence' of three transgenic wheats expressing additional high-molecular-weight subunit genes and the corresponding parental lines (two lines plus a null transformant) was examined using metabolite profiling of samples grown in replicate field trials on two UK sites (Rothamsted, Hertfordshire and Long Ashton, near Bristol) for 3 years. Multivariate comparison of the proton nuclear magnetic resonance spectra of polar metabolites extracted with deuterated methanol-water showed a stronger influence of site and year than of genotype. Nevertheless, some separation between the transgenic and parental lines was observed, notably between the transgenic line B73-6-1 (which had the highest level of transgene expression) and its parental line L88-6. Comparison of the spectra showed that this separation resulted from increased levels of maltose and/or sucrose in this transgenic line, and that differences in free amino acids were also apparent. More detailed studies of the amino acid composition of material grown in 2000 were carried out using gas chromatography-mass spectrometry. The most noticeable difference was that the samples grown at Rothamsted consistently contained larger amounts of acidic amino acids (glutamic, aspartic) and their amides (glutamine, asparagine). In addition, the related lines, L88-6 and B73-6-1, both contained larger amounts of proline and gamma-aminobutyric acid when grown at Long Ashton than at Rothamsted. The results clearly demonstrate that the environment affects the metabolome and that any differences between the control and transgenic lines are generally within the same range as the differences observed between the control lines grown on different sites and in different years.