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在可控环境条件下进行快速且灵敏的体内研究,以替代对转基因植物进行的长期田间试验。

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

作者信息

Horn Patricia, Schlichting André, Baum Christel, Hammesfahr Ute, Thiele-Bruhn Sören, Leinweber Peter, Broer Inge

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 Feb 10;243:48-60. doi: 10.1016/j.jbiotec.2016.12.014. Epub 2016 Dec 21.

DOI:10.1016/j.jbiotec.2016.12.014
PMID:28011129
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

我们介绍一种简便、快速且有效的方法,用于在实验室分析转基因植物对土壤和模式生物的影响,以替代费力且耗时的田间试验。对于此处所述的研究,我们聚焦于所谓第三代的两种转基因植物:生产药物的转基因植物(PMP)和植物源工业产品(PMI)。选择藻青素合成酶(cphA)作为PMI的模型,霍乱毒素B(CTB)作为PMP的模型。模型基因在培养皿中半无菌生长的复合硬毛野豌豆植物的转基因根中表达,或在装有非无菌土壤的小容器中表达。未检测到模型基因表达对根诱导、生长、生物量、与根瘤菌等共生体的相互作用(根瘤的数量、大小和功能、结瘤菌株的选择)或丛枝菌根真菌有显著影响。在体外,而非田间原位条件下,通过基于磷脂脂肪酸(PLFA)图谱得出的细菌与真菌比例以及革兰氏阳性菌与革兰氏阴性菌比例,确定转基因和非转基因品种之间的土壤微生物群落结构多样性。热解场离子质谱(Py-FIMS)分析表明,PLFA比例的显著差异与根际沉积物的数量和分子组成差异相关。与田间试验不同,在田间试验中基于转基因表达的微小影响可能会被各种环境因素的巨大影响所掩盖,而我们的体外系统能够检测到即使是微小的影响,并以更少的空间、时间和劳动力将其与转基因表达相关联。

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