Cells save their energy during nitrogen starvation by selective autophagy of ribosomes and degradation of RNA to ribonucleotides and nucleosides. Nucleosides are hydrolyzed by nucleoside N-ribohydrolases (nucleosidases, NRHs). Subclass I of NRHs preferentially hydrolyzes the purine ribosides while subclass II is more active towards uridine and xanthosine. Here, we performed a crystallographic and kinetic study to shed light on nucleoside preferences among plant NRHs followed by in vivo metabolomic and phenotyping analyses to reveal the consequences of enhanced nucleoside breakdown. We report the crystal structure of Zea mays NRH2b (subclass II) and NRH3 (subclass I) in complexes with the substrate analog forodesine. Purine and pyrimidine catabolism are inseparable because nucleobase binding in the active site of ZmNRH is mediated via a water network and is thus unspecific. Dexamethasone-inducible ZmNRH overexpressor lines of Arabidopsis thaliana, as well as double nrh knockout lines of moss Physcomitrium patents, reveal a fine control of adenosine in contrast to other ribosides. ZmNRH overexpressor lines display an accelerated early vegetative phase including faster root and rosette growth upon nitrogen starvation or osmotic stress. Moreover, the lines enter the bolting and flowering phase much earlier. We observe changes in the pathways related to nitrogen-containing compounds such as β-alanine and several polyamines, which allow plants to reprogram their metabolism to escape stress. Taken together, crop plant breeding targeting enhanced NRH-mediated nitrogen recycling could therefore be a strategy to enhance plant growth tolerance and productivity under adverse growth conditions.

Centre of the Region Haná for Biotechnological and Agricultural Research Department of Genetic Resources for Vegetables Medicinal and Special Plants Crop Research Institute Šlechtitelů 29 78371 Olomouc Czech Republic

Czech Advanced Technology and Research Institute Palacký University Šlechtitelů 27 78371 Olomouc Czech Republic

Department of Experimental Biology Faculty of Science Palacký University Olomouc CZ 78371 Czech Republic

Institute of Plant Science and Microbiology Universität Hamburg Ohnhorststr 18 22609 Hamburg Germany

Laboratory of Growth Regulators Institute of Experimental Botany of the Czech Academy of Sciences and Palacký University Šlechtitelů 11 Olomouc CZ 78371 Czech Republic

Université Paris Saclay CEA CNRS Institute for Integrative Biology of the Cell Gif sur Yvette F 91198 France

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Comprehensive LC-MS/MS analysis of nitrogen-related plant metabolites