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“在受控环境条件下进行快速和敏感的体内研究,以替代具有转基因植物的长期田间试验”的重印本。

Reprint of "Fast and sensitive in vivo studies under controlled environmental conditions to substitute long-term field trials with genetically modified plants".

机构信息

Agrobiotechnology, Faculty of Agricultural and Environmental Sciences, University of Rostock, Germany.

Soil Science, Faculty of Agricultural and Environmental Sciences, University of Rostock, Germany.

出版信息

J Biotechnol. 2017 Sep 10;257:22-34. doi: 10.1016/j.jbiotec.2017.07.012. Epub 2017 Jul 27.

Abstract

We introduce an easy, fast and effective method to analyze the influence of genetically modified (GM) plants on soil and model organisms in the laboratory to substitute laborious and time consuming field trials. For the studies described here we focused on two GM plants of the so-called 3rd generation: GM plants producing pharmaceuticals (PMP) and plant made industrials (PMI). Cyanophycin synthetase (cphA) was chosen as model for PMI and Choleratoxin B (CTB) as model for PMP. The model genes are expressed in transgenic roots of composite Vicia hirsuta plants grown in petri dishes for semi-sterile growth or small containers filled with non-sterile soil. No significant influence of the model gene expression on root induction, growth, biomass, interaction with symbionts such as rhizobia (number, size and functionality of nodules, selection of nodulating strains) or arbuscular mycorrhizal fungi could be detected. In vitro, but not in situ under field conditions, structural diversity of the bulk soil microbial community between transgenic and non-transgenic cultivars was determined by PLFA pattern-derived ratios of bacteria: fungi and of gram: gram bacteria. Significant differences in PLFA ratios were associated with dissimilarities in the quantity and molecular composition of rhizodeposits as revealed by Py-FIMS analyses. Contrary to field trials, where small effects based on the transgene expression might be hidden by the immense influence of various environmental factors, our in vitro system can detect even minor effects and correlates them to transgene expression with less space, time and labour.

摘要

我们介绍了一种简单、快速和有效的方法,用于分析基因改良(GM)植物对实验室土壤和模式生物的影响,以替代费力且耗时的田间试验。在本文描述的研究中,我们专注于两种所谓的第三代 GM 植物:生产药物的 GM 植物(PMP)和植物制造的工业产品(PMI)。我们选择蓝藻藻蓝蛋白合酶(cphA)作为 PMI 的模型,霍乱毒素 B(CTB)作为 PMP 的模型。该模型基因在复合 Vicia hirsuta 植物的转基因根中表达,这些植物在培养皿中进行半无菌生长或在装满非无菌土壤的小容器中生长。未发现模型基因表达对根诱导、生长、生物量、与共生体(如根瘤菌(根瘤的数量、大小和功能,根瘤菌的选择)或丛枝菌根真菌)的相互作用有显著影响。在体外,但不在田间条件下的原位,通过 PLFA 模式衍生的细菌:真菌和革兰氏阳性菌:革兰氏阴性菌的比值来确定转基因此外和非转基因品种之间的土壤微生物群落的结构多样性。PLFA 比值的显著差异与根分泌物的数量和分子组成的差异有关,这是通过 Py-FIMS 分析揭示的。与田间试验不同,基于转基因表达的微小影响可能被各种环境因素的巨大影响所掩盖,我们的体外系统即使在转基因表达的微小影响也可以检测到,并通过更少的空间、时间和劳动力将其与转基因表达相关联。

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