Durand Mickaël, Morin Amélie, Porcheron Benoît, Pourtau Nathalie
Écologie et Biologie des Interactions (EBI), Université de Poitiers, CNRS, EBI, Poitiers, France.
EA2106 "Biomolécules et Biotechnologies Végétales", Université de Tours, Tours, France.
Methods Mol Biol. 2023;2642:375-386. doi: 10.1007/978-1-0716-3044-0_20.
The study of root growth and plasticity in situ is rendered difficult by the opacity and mechanical barrier of soil substrates. Therefore, for the analysis of developmental processes and abiotic stress and development relationships, it is essential to set up cultivation systems that overcome these hindrances in a non-invasive and non-destructive manner. For this purpose, we have developed a useful and powerful rhizobox culture system, where the roots are separated from the soil substrate by a porous membrane with a mesh of such width that allows the exchange of water and solutes without allowing the roots to penetrate the soil. This system provides direct, easy, and quick access to the roots and allows to follow root growth and development, root system architecture, and root system plasticity at different stages of plant development and under various environmental conditions. Moreover, these rhizoboxes provide clean and intact roots that can be easily harvested to perform further physiological, biochemical, and molecular analyses at different stages of development and in response to various environmental constraints. This rhizobox method was validated by assessing root response plasticity of drought-stressed Arabidopsis and pea plants grown in soil displaying water content alterations. This rhizobox system is suitable for many types of abiotic stress-development studies, including the comparison of different stress intensities or of various mutants and genotypes.
土壤基质的不透明性和机械屏障使得原位研究根系生长和可塑性变得困难。因此，为了分析发育过程以及非生物胁迫与发育的关系，建立以非侵入性和非破坏性方式克服这些障碍的培养系统至关重要。为此，我们开发了一种实用且强大的根箱培养系统，在该系统中，根系通过具有特定宽度网孔的多孔膜与土壤基质分离，这种宽度的网孔允许水和溶质交换，但不允许根系穿透土壤。该系统提供了直接、简便且快速观察根系的途径，并能追踪植物发育不同阶段以及各种环境条件下的根系生长与发育、根系结构和根系可塑性。此外，这些根箱提供了干净且完整的根系，便于在发育的不同阶段以及应对各种环境限制时进行进一步的生理、生化和分子分析。通过评估在土壤水分含量发生变化的情况下生长的干旱胁迫拟南芥和豌豆植株的根系响应可塑性，验证了这种根箱方法。这种根箱系统适用于许多类型的非生物胁迫 - 发育研究，包括不同胁迫强度或各种突变体和基因型的比较。
