Glucosyltransferase UGT76C1 finely modulates cytokinin responses via cytokinin N-glucosylation in Arabidopsis thaliana

Cytokinins are master regulators of plant growth and development. The glucosyltransferase UGT76C1 capable of N-glucosylation of different cytokinins at the N7- and N9-position was previously identified in Arabidopsis thaliana, but its physiological relevance in plants remains unclear. In the present work, we investigated the physiological characteristics of UGT76C1 mutant (ugt76c1) and its overexpressors. Under normal growth conditions, although ugt76c1 plants and UGT76C1 overexpressors did not display obvious phenotypic alteration, ugt76c1 plants significantly reduced the accumulation of cytokinin N-glucosides, whereas UGT76C1 overexpressors increased cytokinin N-glucosides. Unexpectedly, the concentrations of free forms of cytokinins (mainly trans-zeatin and N6-isopentenyladenine) were comparable to those of the wild type. Upon application of exogenous cytokinin, the mutant showed the same tendency of more sensitive cytokinin response in primary root elongation, chlorophyll retention and anthocyanin accumulation. In contrast, overexpressors showed a tendency of less sensitive cytokinin response in most tests. Furthermore, cytokinin-related genes were investigated for their expression; and the expression levels of AHK3, ARR1, CYP735A2 and LOG2 noticeably changed in ugt76c1 plants, suggesting that plants employ a set of cytokinin regulation mechanisms to coordinate the loss-of-function of UGT76C1. Tissue-specific expression of UGT76C1 showed a high level of expression in germinating seeds and young seedlings. Taken together, our data suggest that the glucosyltransferase UGT76C1 could finely modulate cytokinin responses in planta via N-glucosylation of cytokinins.

► The physiological relevance of glucosyltransferase UGT76C1 was studied.
► UGT76C1 is involved in N-glucosylation of cytokinins in plants.
►  UGT76C1 finely modulates cytokinin responses of plants via N-glucosylation.
► UGT76C1 is expressed mainly in germinating seeds and young seedlings.