To overcome nitrogen deficiency, legume roots establish symbiotic interactions with nitrogen-fixing rhizobia that are fostered in specialized organs (nodules). Similar to other organs, nodule formation is determined by a local maximum of the phytohormone auxin at the primordium site. However, how auxin regulates nodule development remains poorly understood. Here, we found that in soybean, (Glycine max), dynamic auxin transport driven by PIN-FORMED (PIN) transporter GmPIN1 is involved in nodule primordium formation. GmPIN1 was specifically expressed in nodule primordium cells and GmPIN1 was polarly localized in these cells. Two nodulation regulators, (iso)flavonoids trigger expanded distribution of GmPIN1b to root cortical cells, and cytokinin rearranges GmPIN1b polarity. Gmpin1abc triple mutants generated with CRISPR-Cas9 showed the impaired establishment of auxin maxima in nodule meristems and aberrant divisions in the nodule primordium cells. Moreover, overexpression of GmPIN1 suppressed nodule primordium initiation. GmPIN9d, an ortholog of Arabidopsis thaliana PIN2, acts together with GmPIN1 later in nodule development to acropetally transport auxin in vascular bundles, fine-tuning the auxin supply for nodule enlargement. Our findings reveal how PIN-dependent auxin transport modulates different aspects of soybean nodule development and suggest that the establishment of auxin gradient is a prerequisite for the proper interaction between legumes and rhizobia.

College of Advanced Agricultural Sciences University of Chinese Academy of Sciences Beijing 100049 China

College of Life Science Fujian Agriculture and Forestry University Fuzhou Fujian 350002 China

College of Resources and Environment Fujian Agriculture and Forestry University Fuzhou Fujian 350002 China

Department of Experimental Plant Biology Faculty of Science Charles University Viničná 5 128 43 Prague 2 Czech Republic

Haixia Institute of Science and Technology Horticultural Plant Biology and Metabolomics Center Fujian Agriculture and Forestry University Fuzhou Fujian 350002 China

Institute of Science and Technology Austria Am Campus 1 3400 Klosterneuburg Austria

Photosynthesis Research Center Key Laboratory of Photobiology Institute of Botany Chinese Academy of Sciences Beijing 100093 China

State Key Laboratory of Plant Genomics and National Center for Plant Gene Research Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing 100101 China

State Key Laboratory of Plant Physiology and Biochemistry College of Biological Sciences China Agricultural University Beijing 100193 China

The Czech Academy of Sciences Institute of Experimental Botany Rozvojová 263 165 02 Prague 6 Czech Republic

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