Cytokinins, a group of mobile phytohormones, play an important role in plant growth and development, and their activity is finely controlled by environmental factors in the control of morphogenic and metabolic adaptations. Inorganic nitrogen sources, such as nitrate, are a major factor regulating gene expression of adenosine phosphate-isopentenyltransferase (IPT), a key enzyme of cytokinin biosynthesis. Modulation of IPT and macronutrient transporter gene expression in response to nitrate, sulphate and phosphate, and cytokinin-dependent repression of the transporter genes suggest that cytokinins play a critical role in balancing acquisition and distribution of macronutrients. Biased distribution of trans-zeatin (tZ)-type cytokinins in xylem and N(6)-(Delta(2)-isopentenyl)adenine (iP)-type cytokinins in phloem saps suggest that, in addition to acting as local signals, cytokinins communicate acropetal and systemic long-distance signals, and that structural side chain variations mediate different biological messages. The compartmentalization of tZ- and iP-type cytokinins implies the involvement of a selective transport system. Recent studies have raised the possibility of subsets of the purine permease family as a transporter of cytokinin nucleobases and equilibrative nucleoside transporters (ENT) for cytokinin nucleosides. These biochemical and transgenic data suggest that AtENT6, an Arabidopsis ENT, could also participate in cytokinin nucleoside transport with a preference for iP riboside in vascular tissue.

No abstract

Genome-wide analyses of rice (Oryza sativa L.) cytokinin (CK)-responsive genes using the Affymetrix GeneChip(R) rice genome array were conducted to define the spectrum of genes subject to regulation by CK in monocotyledonous plants. Application of trans-zeatin modulated the expression of a wide variety of genes including those involved in hormone signaling and metabolism, transcriptional regulation, macronutrient transport and protein synthesis. To understand further the function of CK in rice plants, we examined the effects of in planta manipulation of a putative CK signaling factor on morphology, CK metabolism and expression of CK-responsive genes. Overexpression of the CK-inducible type-A response regulator OsRR6 abolished shoot regeneration, suggesting that OsRR6 acts as a negative regulator of CK signaling. Transgenic lines overexpressing OsRR6 (OsRR6-ox) had dwarf phenotypes with poorly developed root systems and panicles. Increased content of trans-zeatin-type CKs in OsRR6-ox lines indicates that homeostatic control of CK levels is regulated by OsRR6 signaling. Expression of genes encoding CK oxidase/dehydrogenase decreased in OsRR6-ox plants, possibly accounting for elevated CK levels in transgenic lines. Expression of a number of stress response genes was also altered in OsRR6-ox plants.

We identified four genes for potential equilibrative nucleoside transporters (ENTs) from rice (Oryza sativa; designated OsENT1 through OsENT4). Growth analysis of budding yeast (Saccharomyces cerevisiae) cells expressing OsENTs showed that OsENT2 transported adenosine and uridine with high affinity (adenosine, K m 5 3.0 mM; uridine, K m 5 0.7 mM). Purine or pyrimidine nucleosides and 2#-deoxynucleosides strongly inhibited adenosine transport via OsENT2, suggesting that OsENT2 possesses broad substrate specificity. OsENT2-mediated adenosine transport was resistant to the typical inhibitors of mammalian ENTs, nitrobenzylmercaptopurine ribonucleoside, dilazep, and dipyridamole. The transport activity was maximal at pH 5.0 and decreased slightly at lower as well as higher pH. In competition experiments with various cytokinins, adenosine transport by OsENT2 was inhibited by isopentenyladenine riboside (iPR). Direct measurements with radiolabeled cytokinins demonstrated that OsENT2 mediated uptake of iPR (K m 5 32 mM) and trans-zeatin riboside (K m 5 660 mM), suggesting that OsENT2 participates in iPR transport in planta. In mature plants, OsENT2 was predominantly expressed in roots. The OsENT2 promoter drove the expression of the b-glucuronidase reporter gene in the scutellum during germination and in vascular tissues in germinated plants, suggesting a participation of OsENT2 in the retrieval of endosperm-derived nucleosides by the germinating embryo and in the long-distance transport of nucleosides in growing plants, respectively.