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White lupin is very often used as a model plant for root exudation studies due to its capability to release huge amounts of organic acids and flavonoids. The complex nature of these organic compounds makes not only their analytical determination difficult but also their extraction from soil samples. For these reasons simplified approaches, as hydroponic-based systems are widely used to study the root exudation. Therefore, the composition of a trap solution is crucial to limit artefacts causing over/underestimation of exudation rates and/or a biased molecular composition of the collected compounds. The present study was aimed at assessing the influence of different trap solutions and collection times on the quali- and quantitative root exudation pattern of white lupin ( Lupinus albus L.) grown under phosphorus (P) and iron (Fe) deficiency. Our results suggest that, in works aimed at studying root exudation processes, water is the most effective trap solution to collect the exudates like organic acids and flavonoids, especially in short time (e.g. 2 h). For longer times, low concentrations of Ca could be helpful to limit osmotic stress and possible passive leakage and/or diffusion. The use of bacteriostatic compounds as NaN and Micropur bias the results, due to interferences either with the metabolism or inhibition of the exudation processes, especially in the case of flavonoids such as quercetin. Also, the use of a pH buffer solution like 2-(N-morpholino)ethanesulfonic acid (MES) should be avoided for its undesired interferences with the release. [ABSTRACT FROM AUTHOR]

The low bioavailability of nutrients, especially nitrogen (N) and phosphorus (P), is one of the most limiting factors for crop production. In this study, under N- and P-free nutrient solution (-N-P), nodulating white lupin plants developed some nodules and analogous cluster root structures characterized by different morphological, physiological, and molecular responses than those observed upon single nutrient deficiency (strong acidification of external media, a better nutritional status than -N+P and +N-P plants). The multi-elemental analysis highlighted that the concentrations of nutrients in white lupin plants were mainly affected by P availability. Gene-expression analyses provided evidence of interconnections between N and P nutritional pathways that are active to promote N and P balance in plants. The root exudome was mainly characterized by N availability in nutrient solution, and, in particular, the absence of N and P in the nutrient solution triggered a high release of phenolic compounds, nucleosides monophosphate and saponines by roots. These morphological, physiological, and molecular responses result from a close interplay between N and P nutritional pathways. They contribute to the good development of nodulating white lupin plants when grown on N- and P-free media. This study provides evidence that limited N and P availability in the nutrient solution can promote white lupin-Bradyrhizobium symbiosis, which is favourable for the sustainability of legume production. [ABSTRACT FROM AUTHOR]