Profiles of Cytokinins Metabolic Genes and Endogenous Cytokinins Dynamics during Shoot Multiplication In Vitro of .

Shoot multiplication induced by exogenous cytokinins (CKs) has been commonly used in micropropagation for commercial production. Despite this, mechanisms of CKs action on shoot multiplication remain unclear in . In this study, we first identified key CKs metabolic genes, including six isopentenyltransferase (), six cytokinin riboside 5' monophosphate phosphoribohydrolase (), and six cytokinin dehydrogenase (), from the genome. Then, we investigated expression profiles of these CKs metabolic genes and endogenous CKs dynamics in shoot proliferation by thidiazuron (TDZ) treatments (an artificial plant growth regulator with strong cytokinin-like activity). Our data showed that these CKs metabolic genes have organ-specific expression patterns. The shoot proliferation in vitro was effectively promoted with increased TDZ concentrations. Following TDZ treatments, the highly expressed CKs metabolic genes in micropropagated shoots were , and . By 30 days of culture, TDZ treatments significantly induced CK-ribosides levels in micropropagated shoots, such as ZR and iPR (2000-fold and 200-fold, respectively) as compared to the controls, whereas ZR showed only a 10-fold increase. Overexpression of and by agroinfiltration assays resulted in increased CK-ribosides levels in tobacco leaves, while overexpression of resulted in decreased CK-ribosides levels. These findings suggest de novo biosynthesis of CKs induced by TDZ, primarily in elevation of ZR and iPR levels. Our results provide a better understanding of CKs metabolism in micropropagation.