Secondary active transport was studied as one of the most important mechanism controlling cytokinin distribution between shoots and roots enabling adaptive growth reaction of barley. It is shown that under phosphate deficit maintenance of root growth, inhibition of their branching and decline in shoot mass of barley plants (Prairie cultivar) was due to decline in shoot cytokinins and their accumulation in the roots. Application of protonophore carbonyl cyanide-m-chlorophenylhydrazone (CCCP) as an inhibitor of secondary active transport leveled off hormonal reaction to phosphate deficit: prevented accumulation of cytokinins in the roots and increased their content in the shoots and also changed percentage of zeatin and its derivative in the total cytokinin content in the roots and shoots. These data analysis showed that in the roots the content of ribosides and nucleotides of zeatin was not change significantly by protonophore, while free zeatin, whose uptake by cells, unlike its riboside, is mostly affected by protonophore, decreased to the greatest extent. Important role of secondary active trans-membrane transfer is discussed in the context of cytokinins transport from roots to shoots under phosphate deficit.