Research Centre for Biosystems, Land Use, and Nutrition, Justus Liebig University, 35392 Giessen, Germany.
Proc Natl Acad Sci U S A. 2010 Apr 6;107(14):6198-203. doi: 10.1073/pnas.1001945107. Epub 2010 Mar 22.
The aim of the present study was to assess possible adverse effects of transgene expression in leaves of field-grown barley relative to the influence of genetic background and the effect of plant interaction with arbuscular mycorrhizal fungi. We conducted transcript profiling, metabolome profiling, and metabolic fingerprinting of wild-type accessions and barley transgenics with seed-specific expression of (1,3-1, 4)-beta-glucanase (GluB) in Baronesse (B) as well as of transgenics in Golden Promise (GP) background with ubiquitous expression of codon-optimized Trichoderma harzianum endochitinase (ChGP). We found more than 1,600 differential transcripts between varieties GP and B, with defense genes being strongly overrepresented in B, indicating a divergent response to subclinical pathogen challenge in the field. In contrast, no statistically significant differences between ChGP and GP could be detected based on transcriptome or metabolome analysis, although 22 genes and 4 metabolites were differentially abundant when comparing GluB and B, leading to the distinction of these two genotypes in principle component analysis. The coregulation of most of these genes in GluB and GP, as well as simple sequence repeat-marker analysis, suggests that the distinctive alleles in GluB are inherited from GP. Thus, the effect of the two investigated transgenes on the global transcript profile is substantially lower than the effect of a minor number of alleles that differ as a consequence of crop breeding. Exposing roots to the spores of the mycorrhizal Glomus sp. had little effect on the leaf transcriptome, but central leaf metabolism was consistently altered in all genotypes.
本研究的目的是评估转基因在田间生长的大麦叶片中表达的可能不良反应,相对于遗传背景的影响和植物与丛枝菌根真菌相互作用的影响。我们对野生型和具有种子特异性表达(1,3-1,4)-β-葡聚糖酶(GluB)的转基因大麦 Baronesse(B)以及在 Golden Promise(GP)背景下具有普遍表达的优化密码子 Trichoderma harzianum 内切几丁质酶(ChGP)的转基因进行了转录谱分析、代谢组分析和代谢指纹分析。我们发现,在品种 GP 和 B 之间有超过 1600 个差异转录本,其中防御基因在 B 中强烈过表达,表明在田间对亚临床病原体挑战有不同的反应。相比之下,基于转录组或代谢组分析,无法检测到 ChGP 和 GP 之间存在统计学上的显著差异,尽管当比较 GluB 和 B 时,有 22 个基因和 4 个代谢物差异丰富,这导致了这两种基因型在主成分分析中的区分。在 GluB 和 GP 中大多数这些基因的共调控,以及简单序列重复标记分析,表明 GluB 中的特有等位基因是从 GP 遗传而来的。因此,两个研究的转基因对全局转录谱的影响远低于由于作物育种而导致的少数等位基因差异的影响。将根暴露于菌根 Glomus sp 的孢子中对叶片转录组几乎没有影响,但在所有基因型中,中心叶片代谢都发生了一致的改变。