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Mary W. George and Robert R. Tripepi

Plant Preservative Mixture™ (PPM), a relatively new, broad-spectrum preservative and biocide for use in plant tissue culture, was evaluated as an alternative to the use of conventional antibiotics and fungicides in plant tissue culture. Concentrations of 0.5 to 4.0 mL·L-1 were tested with leaf explants of chrysanthemum (Dendranthem×grandiflora Kitam), European birch (Betula pendula Roth), and rhododendron (Rhododendron catawbiense Michx.). PPM had little effect on the percentage of explants forming shoots and the number of shoots formed per explant in birch and rhododendron, but dramatically reduced both responses in chrysanthemum. Therefore, the effects of PPM must be evaluated for each species of interest prior to use.

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Joseph J. King and Dennis P. Stimat

Changes in quantity and form of nitrogen used were evaluated during development of Dendranthema × grandiflora `Iridon', `Sequoia' and `Sequest' grown in hydroponic solutions containing 28 ppm NO 3 - (as Ca(NO3)2) and 28 ppm NH4 + (as(NH4)2SO4). Four plants of each cultivar were harvested every ten days over ninety days. Data were collected on plant height, leaf number, flower bud diameter, and root and shoot dry weights. Nutrient solutions were changed every ten days and sampled at days 1, 5, and 10 of each cycle. Concentrations of NO3 - and NH 4 + of nutrient solutions and dried plant tissues were determined by micro-Kjeldahl. Total tissue nitrogen was determined for `Sequoia' by macro-Kjeldahl. Depletion of nitrogen from solutions approximated a 1:1 ratio for NH4 +:NO3 - throughout the crop cycle (r = 0.96). Although the sample date × cultivar interaction was significant for both forms of nitrogen, overall patterns of nitrogen uptake were similar among cultivars. Nitrogen uptake expressed per gram of tissue dry weight was greatest in the first month of development. However, total nitrogen uptake was greatest from days 40 to 60. Developmental stages corresponding to this period are a cessation of new leaf development and flower bud diameters of approximately 1.0 cm. Nitrogen uptake decreased rapidly as flowers expanded. Correlations between morphological changes and nitrogen demands could maximize the efficiency of nitrogen applications.

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Joseph J. King and Dennis P. Stimat

Changes in quantity and form of nitrogen used were evaluated during development of Dendranthema × grandiflora `Iridon', `Sequoia' and `Sequest' grown in hydroponic solutions containing 28 ppm NO 3 - (as Ca(NO3)2) and 28 ppm NH4 + (as(NH4)2SO4). Four plants of each cultivar were harvested every ten days over ninety days. Data were collected on plant height, leaf number, flower bud diameter, and root and shoot dry weights. Nutrient solutions were changed every ten days and sampled at days 1, 5, and 10 of each cycle. Concentrations of NO3 - and NH 4 + of nutrient solutions and dried plant tissues were determined by micro-Kjeldahl. Total tissue nitrogen was determined for `Sequoia' by macro-Kjeldahl. Depletion of nitrogen from solutions approximated a 1:1 ratio for NH4 +:NO3 - throughout the crop cycle (r = 0.96). Although the sample date × cultivar interaction was significant for both forms of nitrogen, overall patterns of nitrogen uptake were similar among cultivars. Nitrogen uptake expressed per gram of tissue dry weight was greatest in the first month of development. However, total nitrogen uptake was greatest from days 40 to 60. Developmental stages corresponding to this period are a cessation of new leaf development and flower bud diameters of approximately 1.0 cm. Nitrogen uptake decreased rapidly as flowers expanded. Correlations between morphological changes and nitrogen demands could maximize the efficiency of nitrogen applications.

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M.W. George and R.R. Tripepi

Plant preservative mixture (PPM) is a new broad-spectrum biocide that may be useful for plant tissue culture. The objective of this study was to determine if PPM interfered with adventitious shoot regeneration on leaf explants from several plant species. Leaf explants from Dendranthema grandiflora `Iridon', Betula pendula, Rhododendron catawbiense var. album and R.c. `America' were made from the top two apical leaves on the microshoots. In the first experiment, 0, 0.5, 1, 2, or 4 mL·L-1 PPM were added to species-appropriate regeneration media. In the second experiment, only mum leaf explants were placed on regeneration media containing 0, 0.1, 0.2, 0.3, or 0.4 mL·L-1 PPM. The percentage of explants forming shoots and the number of shoots per regenerating explant were recorded after 4, 6, and 10 weeks, for mum, birch, and rhododendron leaves, respectively. The percentages of shoot regeneration from birch and rhododendron leaf explants were unaffected by up to 4 mL·L-1 PPM, and the number of shoots formed per R.c. album explant were also unaffected by the tested concentrations of PPM. In contrast, the numbers of shoots formed on birch and `America' explants were reduced 48% and 25%, respectively, when 4 mL·L-1 PPM was used in the media. The percentages of shoot regeneration and number of shoots per explant were drastically reduced on mum explants when only 0.5 mL·L-1 PPM was used in the medium. In fact, 0.3 mL·L-1 PPM or higher reduced shoot formation by more than 5-fold. This study demonstrates that the effects of PPM on shoot regeneration from leaf explants are species specific.