CYTOKININ OXIDASE/DEHYDROGENASE3 maintains cytokinin homeostasis during root and nodule development in Lotus japonicus

Dugald Elgin Reid, Anne Birgitte Heckmann, Ondrej Novak, Simon Kelly, Jens Stougaard

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88 Citations (Scopus)

Abstract

Cytokinins are required for symbiotic nodule development in legumes, and cytokinin signaling responses occur locally in nodule primordia and in developing nodules. Here, we show that theLotus japonicus Ckx3cytokinin oxidase/dehydrogenase gene is induced by Nod factor during the early phase of nodule initiation. At the cellular level,pCkx3::YFPreporter-gene studies revealed that theCkx3promoter is active during the first cortical cell divisions of the nodule primordium and in growing nodules. Cytokinin measurements inckx3mutants confirmed that CKX3 activity negatively regulates root cytokinin levels. Particularly, tZ and DHZ type cytokinins in both inoculated and uninoculated roots were elevated inckx3mutants, suggesting that these are targets for degradation by the CKX3 cytokinin oxidase/dehydrogenase. The effect of CKX3 on the positive and negative roles of cytokinin in nodule development, infection and regulation was further clarified usingckx3insertion mutants. Phenotypic analysis indicated thatckx3mutants have reduced nodulation, infection thread formation and root growth. We also identify a role for cytokinin in regulating nodulation and nitrogen fixation in response to nitrate asckx3phenotypes are exaggerated at increased nitrate levels. Together, these findings show that cytokinin accumulation is tightly regulated during nodulation in order to balance the requirement for cell divisions with negative regulatory effects of cytokinin on infection events and root development.

Original languageEnglish
JournalPlant Physiology
Volume170
Issue2
Pages (from-to)1060-1074
Number of pages15
ISSN0032-0889
DOIs
Publication statusPublished - 2016

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