TY - JOUR

T1 - Enzymes and cellular interplay required for flux of fixed nitrogen to ureides in bean nodules

AU - Voß, Luisa

AU - Heinemann, Katharina J.

AU - Herde, Marco

AU - Medina-Escobar, Nieves

AU - Witte, Claus Peter

N1 - Funding Information: We thank André Specht and Hildegard Thölke for technical assistance and Markus Niehaus for help with the design and construction of CRISPR vectors as well as Jana Streubel for donating the MoClo compatible shuttle vector. We also thank Marta Santalla (CSIC Pontevedra, Spain) and Javier Ollero (University of Sevilla, Spain) for the Negro Jamapa seeds and the Rhizobium tropici CIAT 899 strain, respectively. Jannis Rinne we thank for his assistance in harvesting nodules and technical expertise in AFLP analysis. We also are grateful to Leonie Fischer for contributing to the cloning of the pY vector series, Melina Wehrspohn for her help in preparing cross sections and Helge Küster for access to the vibratome. This work was supported by the Deutsche Forschungsgemeinschaft (DFG) grants WI3411/7-1, WI3411/8-1 and INST 187/741-1 FUGG to C.-P.W.

PY - 2022/9/10

Y1 - 2022/9/10

N2 - Tropical legumes transport fixed nitrogen in form of ureides (allantoin and allantoate) over long distances from the nodules to the shoot. Ureides are formed in nodules from purine mononucleotides by a partially unknown reaction network that involves bacteroid-infected and uninfected cells. Here, we demonstrate by metabolic analysis of CRISPR mutant nodules of Phaseolus vulgaris defective in either xanthosine monophosphate phosphatase (XMPP), guanosine deaminase (GSDA), the nucleoside hydrolases 1 and 2 (NSH1, NSH2) or xanthine dehydrogenase (XDH) that nodule ureide biosynthesis involves these enzymes and requires xanthosine and guanosine but not inosine monophosphate catabolism. Interestingly, promoter reporter analyses revealed that XMPP, GSDA and XDH are expressed in infected cells, whereas NSH1, NSH2 and the promoters of the downstream enzymes urate oxidase (UOX) and allantoinase (ALN) are active in uninfected cells. The data suggest a complex cellular organization of ureide biosynthesis with three transitions between infected and uninfected cells.

AB - Tropical legumes transport fixed nitrogen in form of ureides (allantoin and allantoate) over long distances from the nodules to the shoot. Ureides are formed in nodules from purine mononucleotides by a partially unknown reaction network that involves bacteroid-infected and uninfected cells. Here, we demonstrate by metabolic analysis of CRISPR mutant nodules of Phaseolus vulgaris defective in either xanthosine monophosphate phosphatase (XMPP), guanosine deaminase (GSDA), the nucleoside hydrolases 1 and 2 (NSH1, NSH2) or xanthine dehydrogenase (XDH) that nodule ureide biosynthesis involves these enzymes and requires xanthosine and guanosine but not inosine monophosphate catabolism. Interestingly, promoter reporter analyses revealed that XMPP, GSDA and XDH are expressed in infected cells, whereas NSH1, NSH2 and the promoters of the downstream enzymes urate oxidase (UOX) and allantoinase (ALN) are active in uninfected cells. The data suggest a complex cellular organization of ureide biosynthesis with three transitions between infected and uninfected cells.

UR - http://www.scopus.com/inward/record.url?scp=85138127851&partnerID=8YFLogxK

U2 - 10.1038/s41467-022-33005-5

DO - 10.1038/s41467-022-33005-5

M3 - Article

C2 - 36088455

AN - SCOPUS:85138127851

VL - 13

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

IS - 1

M1 - 5331

ER -