Cytokinin profiling of long-term in vitro pea (Pisum sativum L.) shoot cultures
Forty-seven endogenous cytokinin metabolites were determined in samples of long-term in vitro multiple shoot cultures of pea (Pisum sativum L.). Significant differences were found in the levels of all cytokinin groups except cis-zeatin and some of its derivatives. Old cultures (30 or 10 years old) contained larger total and active cytokinin pools than freshly prepared cultures. Long-term shoot cultures maintained on cytokinin-supplemented media initially exhibit gradually increasing concentrations of endogenous cytokinins that subsequently stabilise at a high level, which can make it more difficult to induce rooting later on. This is consistent with previous findings concerning the growth and development of plantlets derived from long-term cultures and the generally accepted role of cytokinins during root development. The results presented herein demonstrate the utility of long-term pea cultures as model systems for studying the influence of long-term cultivation on genetic stability, and suggest ways of circumventing the rooting problems associated with such cultures.
KeywordsCytokinins Pea Quantification UHPLC–MS/MS Long-term culture in vitro
Ultra high-performance liquid chromatography–electrospray tandem mass spectrometry
This work was supported by the Ministry of Education Youth and Sports, Czech Republic (Grant LO1204 from the National Program of Sustainability), by IGA of Palacký University (IGA_PrF_2014006) and by the Ministry of Agriculture of the Czech Republic project No. RO 1014. The authors would like to thank to Dave Richardson and Sees-editing Ltd. for careful reading and editing of this manuscript.
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