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Gibberellin (GA) plays a controversial role in the legume-rhizobium symbiosis. Recent studies have shown that the GA level in legumes must be precisely controlled for successful rhizobial infection and nodule organogenesis. However, regulation of the GA level via catabolism in legume roots has not been reported to date. Here, we investigate a novel GA inactivating C20-GA2-oxidase gene  MtGA2ox10  in  Medicago truncatula . RNA sequencing analysis and quantitative polymerase chain reaction revealed that  MtGA2ox10  was induced as early as 6 h post-inoculation (hpi) of rhizobia and reached peak transcript abundance at 12 hpi. Promoter::β-glucuronidase fusion showed that the promoter activity was localized in the root infection/differentiation zone during the early stage of rhizobial infection and in the vascular bundle of the mature nodule. The CRISPR/Cas9-mediated deletion mutation of  MtGA2ox10  suppressed infection thread formation, which resulted in reduced development and retarded growth of nodules on the  Agrobacterium rhizogenes -transformed roots. Over-expression of  MtGA2ox10  in the stable transgenic plants caused dwarfism, which was rescued by GA 3  application, and increased infection thread formation but inhibition of nodule development. We conclude that  MtGA2ox10  plays an important role in the rhizobial infection and the development of root nodules through fine catabolic tuning of GA in  M. truncatula .

MtGA2ox10 encoding C20-GA2-oxidase regulates rhizobial infection and nodule development in Medicago truncatula