Does NO participate in cytokinin signaling?

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Does NO participate in cytokinin signaling?

BioMed Central
Publication Date
May 31, 2005

Abstract ral ss BioMed CentBMC Plant Biology Open AcceMeeting abstract Does NO participate in cytokinin signaling? Georgy Romanov*1, Natalia Rakova1 and Thomas Schmülling2 Address: 1Institute of Plant Physiology, Russian Academy of Sciences, 127276 Moscow, Russia and 2Freie Universität Berlin, Institute of Biology, Applied Genetics, D-14195 Berlin, Germany Email: Georgy Romanov* - [email protected] * Corresponding author Background Nitric oxide (NO) is a unique ubiquitous molecule in ani- mals and plants [1]. It has been demonstrated that NO mediates ABA-induced stomatal closure and interferes with ethylene during the maturation and senescence of plant tissues. Recently published data [2,3] suggested that NO is also involved in cytokinin signaling. However the above-mentioned data did not provide defenitive proof of a role of NO in cytokinin signaling. In order to obtain more conclusive evidence we have performed a study using Parr5::GUS transgenic Arabidopsis plants which express the GUS reporter gene under the transcriptional control of the cytokinin-responsive ARR5 promoter. Addi- tional experiments were also performed using Amaranthus seedlings. Materials and methods The Arabidopsis and Amaranthus assay systems were described in detail earlier [4,5]. The cytokinin effects were measured quantitatively in both systems. For RNA blot analysis, 3-day-old Arabidopsis seedlings were incubated with BA (with oor without tested compounds) for 35 min. Total RNA was isolated and separated in a 1.5% agarose- formaldehyde gel, transferred to nylon membrane (Amer- sham) and hybrydized with radioactive-labeled DNA probe. For radioautography blots were exposed with BioMax MS films (Kodak) for 1–3 days. Quantification of signal was done on an integrative densitometer CD-50 (Desaga). As a control for loading, the blot was rehybrid- ized with an actin 2 gene probe. Results In our experiments, a strong competitive inhibitor of all Arabidopsis harboring the GUS gene driv

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