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Does NO participate in cytokinin signaling?

BioMed Central
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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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