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探究叶片生长和分子表型的可重复性:在十个实验室中培养的三个拟南芥品系的比较。

Probing the reproducibility of leaf growth and molecular phenotypes: a comparison of three Arabidopsis accessions cultivated in ten laboratories.

机构信息

INRA, SUPAGRO-UMR LEPSE, 34060 Montpellier cedex, France.

出版信息

Plant Physiol. 2010 Apr;152(4):2142-57. doi: 10.1104/pp.109.148338. Epub 2010 Mar 3.

Abstract

A major goal of the life sciences is to understand how molecular processes control phenotypes. Because understanding biological systems relies on the work of multiple laboratories, biologists implicitly assume that organisms with the same genotype will display similar phenotypes when grown in comparable conditions. We investigated to what extent this holds true for leaf growth variables and metabolite and transcriptome profiles of three Arabidopsis (Arabidopsis thaliana) genotypes grown in 10 laboratories using a standardized and detailed protocol. A core group of four laboratories generated similar leaf growth phenotypes, demonstrating that standardization is possible. But some laboratories presented significant differences in some leaf growth variables, sometimes changing the genotype ranking. Metabolite profiles derived from the same leaf displayed a strong genotype x environment (laboratory) component. Genotypes could be separated on the basis of their metabolic signature, but only when the analysis was limited to samples derived from one laboratory. Transcriptome data revealed considerable plant-to-plant variation, but the standardization ensured that interlaboratory variation was not considerably larger than intralaboratory variation. The different impacts of the standardization on phenotypes and molecular profiles could result from differences of temporal scale between processes involved at these organizational levels. Our findings underscore the challenge of describing, monitoring, and precisely controlling environmental conditions but also demonstrate that dedicated efforts can result in reproducible data across multiple laboratories. Finally, our comparative analysis revealed that small variations in growing conditions (light quality principally) and handling of plants can account for significant differences in phenotypes and molecular profiles obtained in independent laboratories.

摘要

生命科学的主要目标之一是了解分子过程如何控制表型。由于理解生物系统依赖于多个实验室的工作,生物学家默认具有相同基因型的生物体在可比条件下生长时会表现出相似的表型。我们研究了在使用标准化和详细方案在 10 个实验室中生长的三种拟南芥(Arabidopsis thaliana)基因型的叶片生长变量以及代谢物和转录组谱中,这种情况在多大程度上是正确的。一个由四个实验室组成的核心小组产生了相似的叶片生长表型,证明了标准化是可行的。但是,一些实验室在某些叶片生长变量上存在显著差异,有时会改变基因型排名。来自同一叶片的代谢物图谱显示出强烈的基因型 x 环境(实验室)成分。可以根据其代谢特征对基因型进行分离,但前提是分析仅限于来自一个实验室的样本。转录组数据显示出相当大的植物间变异,但标准化确保了实验室间的变异并不比实验室内部的变异大得多。标准化对表型和分子图谱的不同影响可能是由于这些组织水平上涉及的过程的时间尺度不同造成的。我们的研究结果强调了描述、监测和精确控制环境条件的挑战,但也证明了专门的努力可以在多个实验室中产生可重复的数据。最后,我们的比较分析表明,生长条件(主要是光照质量)和植物处理的微小变化可以解释在独立实验室中获得的表型和分子图谱的显著差异。

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