Cytokinins (CKs) play a crucial role in various phases of plant growth and development, but the basic molecular mechanisms of their biosynthesis and signal transduction only recently became clear. The progress was achieved by identifying a series of key genes encoding enzymes and proteins controlling critical steps in biosynthesis, translocation, and signaling. Basic schemes for CK homeostasis and root/shoot communication at the whole-plant level can now be devised. This review summarizes recent findings on the relationship between CK structural variation and activity, distinct features in CK biosynthesis between higher plants and Agrobacterium infected plants, CK translocation at whole-plant and cellular levels, and CKs as signaling molecules for nutrient status via root-shoot communication.

CYTOKININS: Activity, Biosynthesis, and Translocation

■ Abstract Cytokinins are structurally diverse and biologically versatile. The chemistry and physiology of cytokinin have been studied extensively, but the regulation of cytokinin biosynthesis, metabolism, and signal transduction is still largely undefined. Recent advances in cloning metabolic genes and identifying putative receptors portend more rapid progress based on molecular techniques. This review centers on cytokinin metabolism with connecting discussions on biosynthesis and signal transduction. Important findings are summarized with emphasis on metabolic enzymes and genes. Based on the information generated to date, implications and future research directions are presented.

Cytokinin metabolism and action

Thidizuron (TDZ) is among the most active cytokinin-like substances for woody plant tissue culture. It facilitates efficient micropropagation of many recalcitrant woody species. Low concentrations (<1 µM) can induce greater axillary proliferation than many other cytokinins; however, TDZ may inhibit shoot elongation. In some cases it is necessary to transfer shoots to an elongation medium containing a lower level of TDZ and/or a less active cytokinin. At concentrations higher than 1 µM, TDZ can stimulate the formation of callus, adventitious shoots or somatic embryos. Subsequent rooting of microshoots may be unaffected or slightly inhibited by prior exposure to TDZ. The main undesirable side effect of TDZ is that cultures of some species occasionally form fasciated shoots. The high cytokinin activity and positive response of woody species to TDZ have established it as among the most active cytokinins forin vitro manipulation of many woody species.

Thidiazuron: a potent cytokinin for woody plant tissue culture

TDZ (N-phenyl-N’-1,2,3-thidiazol-5-ylurea) is a substituted phenylurea compound which was developed for mechanized harvesting of cotton bolls and has now emerged as a highly efficacious bioregulant of morphogenesis in the tissue culture of many plant species. Application of TDZ induces a diverse array of cultural responses ranging from induction of callus to formation of somatic embryos. TDZ exhibits the unique property of mimicking both auxin and cytokinin effects on growth and differentiation of cultured explants, although structurally it is different from either auxins or purine-based cytokinins. A number of physiological and biochemical events in cells are likely to be influenced by TDZ, but these may or may not be directly related to the induction of morphogenic responses, and hence, the mode of action of TDZ is unknown. However, the recent approaches applied to study the morphogenic events initiated by TDZ are clearly beginning to reveal the details of a variety of underlying mechanisms. Various reports indicate that TDZ may act through modulation of the endogenous plant growth regulators, either directly or as a result of induced stress. The other possibilities include the modification in cell membranes, energy levels, nutrient uptake, or nutrient assimilation. In this review, several of these possiblities are presented and discussed in light of recently published studies on characterization of TDZ-induced morphogenic effects.

Thidiazuron : A potent regulator of in vitro plant morphogenesis

Leaf or stem explants of a hybrid poplar clone (Populus tremula X Populus alba), sensitive to Agrobacterium tumefaciens, were co-cultivated either by an octopine or a nopaline disarmed A. tumefaciens modified strain. Transformed poplar shoots were readily regenerated from explants. The protocol was improved using the nopaline disarmed strain C58/pMP90 with the binary vector pBI121. This protocol was then used to test three other vectors. The first one, possessing a nptII gene fused to the CaMV 19S promoter, permitted regeneration of transformed shoots in presence of 50 to 100 mg/l kanamycin. The two other vectors carried an additional nptII gene under the control of the CaMV 35S or CaMV 35S promoter with a double enhancer sequence (CaMV 70). CaMV 70 promoter provided consistently higher level of gene expression than the other promoters in both callus and leaf tissues.

M.C. Mok

Papers

Cytokinin activity of N-phenyl-N′-1, 2,3-thiadiazol-5-ylurea (thidiazuron)