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Ann C. Smigocki

Cytokinins were first recognized as a class of phytohormones for their ability to promote cytokinesis in cultured plant cells and have since been shown to be involved in a wide range of physiological processes. Most recently, the availability of phytohorm one-specifying genes from Agrobacterium tumefaciens has allowed for direct in planta manipulation of cytokinin levels. Overexpression of the isopentenyl transferase (ipt) gene by constitutive promoters led to enhanced ability of plant cells to undergo shoot organogenesis but the high endogenous cytokinin levels almost completely suppressed root development. Transient overproduction of cytokinins using promoters regulated by environmental and/or developmental factors did not inhibit regeneration of rooted plants. Transgenic plants in which cytokinin levels can be modulated are being used to characterize the participation of cytokinins in fundamental regulatory mechanisms of morphogenesis, delayed senescence, disease resistance and directed nutrient transport. The potential for using reconstructed cytokinin biosynthesis genes in economically important crops is of tremendous agronomic significance.

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Yan Zhang, Cuiyue Liang, Yan Xu, Thomas Gianfagna and Bingru Huang

Leaf senescence is a natural process of leaf maturation, but it can be induced prematurely by environmental stresses. Cytokinins (CKs) have been well known for their function in delaying leaf senescence, and even in reversing the process of leaf

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Lisa J. Rowland and Elizabeth L. Ogden

Conditions for improving the efficiency of shoot regeneration from leaf sections of highbush blueberry (Vaccinium corymbosum L.) were investigated. Effectiveness of tissue culture medium supplemented with the cytokinin conjugate zeatin riboside or the cytokinin zeatin at 10, 20, or 30 μm was compared with medium supplemented with the optimum 2iP concentration of 15 μm. Use of 20 μm zeatin riboside resulted in the most shoots per leaf section, » 6-fold higher than the number of shoots produced on 2iP medium. The number of shoots produced on medium supplemented with zeatin was not significantly higher than the number of shoots produced on 2iP medium. Consequently, we concluded that the cytokinin conjugate zeatin riboside was more effective than either of the free cytokinins, 2iP or zeatin, in promoting shoot regeneration from leaf sections of highbush blueberry. Chemical names used: 6-(y,y-dimethylallylamino)-purine (2iP); 6-(4-hydroxy-3-methyl-but-2-enylamino)purine (zeatin).

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Eric W. Mercure, Carol A. Auer and Mark H. Brand

Tissue proliferation (TP) is characterized primarily by the formation of galls or tumors at the crown of container-grown rhododendrons propagated in vitro. However, TP of Rhododendron `Montego' is observed initially in in vitro shoot cultures and it is characterized by the formation of multiple shoots with small leaves and nodal tumors. The formation of shoots in `Montego' TP (TP+) shoot cultures occurs without the presence of exogenous cytokinin in the medium, unlike normal `Montego' (TP–) shoot cultures, which require cytokinin for shoot growth. Structural studies have shown that tumors are composed of many adventitious buds and parenchyma cells, suggesting that TP is a result of abnormal cytokinin regulation that is controlling tumor and shoot formation. Two approaches are being used to determine if differences in cytokinin concentration and/or metabolism exist between TP+ and TP– shoot cultures. In the first approach, shoot cultures are grown in vitro for 1 week in the presence of tritiated isopentenyladenine (iP). Cytokinin uptake and metabolism are analyzed using HPLC and other analytical methods. Experiments suggest that extensive degradation and N-glucoside conjugation occur in TP+ and TP– shoots, resulting in the removal of most of the exogenous iP. In the second approach, the levels of endogenous cytokinins such as iP, isopentenyladenosine, zeatin, and zeatin riboside, are being measured in TP+ tumors and shoots and in TP– shoots by an ELISA method.

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Sandra B. Wilson and Dennis R. Decoteau

Similarities exist between the effects of phytochrome and cytokinins on plant growth and development (e.g., chloroplast development, amaranthin synthesis, seed germination). It is unclear, however, if and how these two systems interact. The coaction between phytochrome and cytokinins was investigated by using Nicotiana plumbaginifolia plants transformed with the isopentenyl transferase (ipt) cytokinin gene and treated with end-of-day (EOD) red (R) and far-red (FR) light. The ipt gene was under control of either a constitutive cauliflower mosaic virus promoter (35S-plants) or an inducible, heat shock promoter (HS-plants). When treated with EOD FR light, whole plants were characterized by decreased chlorophyll concentrations and increased fresh weights. When treated with EOD R light, 35S-plants contained high concentrations of zeatin riboside (ZR) compared to plants treated with EOD FR light. When treated with EOD FR light, HS-plants contained high concentrations of ZR compared to plants treated with EOD R light. Both cytokinin responses were photoreversible. The reasons for the differences between the 35S- and HS-plant responses are not known. Results appear to implicate interactions between phytochrome and cytokinins in plant growth and development.

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James W. Rushing

Broccoli (Brassica oleracea L. var. italica `Citation') florets were treated postharvest with either benzyladenine or trans-zeatin at either 10 or 50 ppm before packaging in perforated polyethylene bags and storage at 16C. The most pronounced effects were observed with benzyladenine at 50 ppm. Compared to controls, respiration rate was reduced 50% and ethylene production increased 40% throughout the first 4 days of storage. Total chlorophyll content had dropped 60% in controls, but was unchanged in cytokinin-treated florets, which had a 90% longer shelf life than controls. These effects depended on the amount of cytokinin applied and were of greater magnitude with benzyladenine than with zeatin.

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Stuart R. Reitz and John T. Trumble

We examined two aspects of treating plants with a cytokinin-containing seaweed extract (SWE). In the first series of experiments, we tested the hypothesis that immature lima bean (Phaseolus lunatus L.) and tomato (Lycopersicon esculentum Mill.) plants provided with exogenous cytokinins could recover from defoliation by a generalist insect herbivore, Spodoptera exigua (Hübner), more rapidly than plants without cytokinin supplements. However, the SWE inhibited growth of lima beans at all levels of herbivore damage. The SWE neither inhibited nor stimulated growth of tomatoes following defoliation. Because SWE effects largely were neutral for tomato growth, we conducted a second series of experiments to test the hypothesis that SWE treatments alter the attractiveness of tomato foliage to S. exigua larvae. In these experiments, we determined consumption of, and preference for, SWE-treated tomato foliage by S. exigua larvae. Repeated root applications of SWE led to increased consumption and preference by S. exigua. Repeated foliar applications did not alter consumption or preference compared with controls. Spodoptera exigua larvae gained significantly more mass when feeding on SWE-treated foliage compared with controls. While these data indicate that plant responses to exogenous cytokinin-containing materials depend on taxa and application method, the practical uses of SWE appear limited given the negative effects on plant growth and increased attractiveness of treated foliage to herbivores.

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Servet Kefi, M.M. Meagher, P.E. Read and A.D. Pavlista

The effects of different cytokinin-like compounds on invertase activities at different tuberization stages of potato (Solanum tuberosum L. `Atlantic') were examined. Single nodal segments were cultured on MS medium plus 6% sucrose and supplemented with either 2 mg kinetin/L, 0.1 mg thidiazuron (TDZ)/L, 1.0 mg AC 243,654/L, 0.1 mg AC 239,604/L, or no cytokinin. Tissue samples for determining invertase activity were taken at three stages of tuberization: stage 1, the “hook stage”; stage 2, the “swelling stage”; and stage 3, “tuber initials.” Invertase activity was significantly affected by the interaction between cytokinin-like compounds and tuberization between cytokinin-like compounds and tuberization stages. The highest invertase activities in the stolons at stage 1 were found in kinetin and TDZ treatments. Invertase activity in the stolons on the control medium significantly increased from stage 1 to 2 and decreased at stage 3. Invertase might play a role in either stolon elongation or carbohydrate utilization by increasing the pool of reducing sugars.

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Georgia Vlachou, Maria Papafotiou and Konstantinos F. Bertsouklis

given in Vlachou et al. (2016b) . The cultures were maintained with a number of subcultures on the initiation medium followed by one subculture on Hf-MS medium. Effect of cytokinin type and concentration on shoot multiplication Adult plant- or seedling

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Fang Geng, Renae Moran, Michael Day, William Halteman and Donglin Zhang

sour cherry, 6 to 8 weeks was optimum for promoting shoot growth, but longer than 8 weeks was detrimental ( Borkowska, 1986 ). Cytokinin in the media is necessary for proliferation, but can inhibit shoot elongation at concentrations that promote rapid