Saiz-Fernández Iñigo, Černý Martin, Skalák Jan, Brzobohatý Břetislav
Phytophthora Research Centre, Department of Molecular Biology and Radiobiology, Faculty of AgriSciences, Mendel University in Brno, Zemědělská 3, 613 00, Brno, Czech Republic.
Department of Molecular Biology and Radiobiology, Faculty of AgriSciences, Mendel University in Brno, Zemědělská 3, 613 00, Brno, Czech Republic.
Plant Methods. 2021 Jan 9;17(1):7. doi: 10.1186/s13007-020-00706-1.
Split-root systems (SRS) have many applications in plant sciences, but their implementation, depending on the experimental design, can be difficult and time-consuming. Additionally, the system is not exempt from limitations, since the time required for the establishment of the SRS imposes a limit to how early in plant development experiments can be performed. Here, we optimized and explained in detail a method for establishing a SRS in young Arabidopsis thaliana seedlings, both in vitro and in soil.
We found that the partial de-rooting minimized the recovery time compared to total de-rooting, thus allowing the establishment of the split-root system in younger plants. Analysis of changes in the Arabidopsis leaf proteome following the de-rooting procedure highlighted the distinct metabolic alterations that totally and partially de-rooted plants undergo during the healing process. This system was also validated for its use in drought experiments, as it offers a way to apply water-soluble compounds to plants subjected to drought stress. By growing plants in a split-root system with both halves being water-deprived, it is possible to apply the required compound to one half of the root system, which can be cut from the main plant once the compound has been absorbed, thus minimizing rehydration and maintaining drought conditions.
Partial de-rooting is the suggested method for obtaining split-root systems in small plants like Arabidopsis thaliana, as growth parameters, survival rate, and proteomic analysis suggest that is a less stressful procedure than total de-rooting, leading to a final rosette area much closer to that of uncut plants. Additionally, we provide evidence that split root-systems can be used in drought experiments where water-soluble compounds are applied with minimal effects of rehydration.
分根系统（SRS）在植物科学中有许多应用，但根据实验设计，其实施可能困难且耗时。此外，该系统也有局限性，因为建立分根系统所需的时间限制了植物发育实验能够开展的最早时间。在此，我们优化并详细解释了一种在拟南芥幼苗早期建立分根系统的方法，该方法可在体外和土壤中进行。
我们发现，与完全去根相比，部分去根能使恢复时间最短，从而能够在更幼小的植株中建立分根系统。对去根处理后拟南芥叶片蛋白质组变化的分析突出了完全去根和部分去根的植株在愈合过程中所经历的不同代谢改变。该系统在干旱实验中的应用也得到了验证，因为它提供了一种向遭受干旱胁迫的植物施加水溶性化合物的方法。通过在两半根系均缺水的分根系统中种植植物，可以将所需化合物施加到一半根系上，一旦化合物被吸收，就可以将其从主植株上切断，从而最大限度地减少再水化并维持干旱条件。
对于像拟南芥这样的小型植物，建议采用部分去根的方法来获得分根系统，因为生长参数、存活率和蛋白质组分析表明，这一过程比分根更不易产生压力，最终莲座叶面积更接近未切割植株。此外，我们提供的证据表明，分根系统可用于干旱实验，在该实验中施加水溶性化合物时再水化的影响最小。
