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Anil P. Ranwala and William B. Miller

Three soluble invertase isoforms from Lilium longiflorum flower buds that had been separated by DEAE-Sephacel chromatography were purified to near homogeneity by further chromatography on hydroxylapetite, Con-A sepharose, phenyl agarose, and Sephacryl S-200 gel filtration. Nondenaturing polyacrylamide gel electrophoresis (PAGE) gave a single band in all three invertases that corresponded to a band of invertase activity in a duplicate gel. The SDS-PAGE of the purified invertase I resulted in a single band with apparent relative molecular mass of 78 kDa. Invertase II and III were resolved to a similar polypeptide pattern by SDS-PAGE with three bands of 54, 52, and 24 kDa. Antiserum of tomato acid invertase cross-reacted with all three invertase protein bands. Antiserum of wheat coleoptile acid invertase cross-reacted only with 54 and 52 kDa bands of invertase II and III but did not recognize invertase I protein. Con-A peroxidase was bound to invertase I protein and all three protein bands of invertase II and III, suggesting that all proteins were glycosylated. Invertase I protein could be completely deglycosylated by incubating with peptide-N-glycosidase F to result in a peptide of 75 kDa. Invertase II and III were partially deglycosylated by peptide-N-glycosidase F resulting proteins bands of 53, 51, 50, and 22 kDa.

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Anil P. Ranwala and William B. Miller

Easter lily flower buds at five stages of development (stage 1, 3–4 cm in length; stage 2, 6–7 cm; stage 3, 9–10 cm; stage 4, unopened buds, 13–14 cm; and stage 5, open flower 1 day after anthesis) were harvested, and flower organs were dissected for invertase assay. On a fresh weight (FW) basis, anthers had the highest soluble invertase activity (about 10-fold greater) than all other organs reaching to 15 units/g FW by the stage 2. The activity dropped to about 3 units/g FW at stage 3 and 4, and then increased up to 10 units/g FW in open flowers. Specific activity (units per mg of protein) also showed the same trend. On a specific activity basis, sepal invertase activity steadily increased during bud development, but was relatively constant on a fresh weight basis. stigma, style, and ovary, soluble invertase activity expressed on a FW and specific activity basis steadily increased as bud development. Filament soluble invertase activity on FW basis dropped at the stage 2 and 3, while specific activity steadily increased during bud development. Cell wall-bound invertase activity (released with 1 m NaCl) was present in all flower organs. However, soluble activity accounted for the most of total activity in sepal, ovary and filament (about 90%). About 75% of total activity was soluble in anther and style, whereas nearly equal amounts of soluble and cell wall activities were present in the stigma. The cell wall bound invertase activity increased throughout the bud development in sepal, stigma, style, and ovary parallel to soluble activity. Anther cell wall-bound activity fluctuated in a similar pattern as the soluble activity.

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Maxim J. Schlossberg and William P. Miller

Coal combustion by-products (CCB) are produced nationwide, generating 108 Mg of waste annually. Though varied, the majority of CCB are crystalline alumino-silicate minerals. Both disposal costs of CCB and interest in alternative horticultural/agricultural production systems have increased recently. Field studies assessed the benefit of CCB and organic waste/product mixtures as supplemental soil/growth media for production of hybrid bermudagrass [Cynodon dactylon (L.) Pers. × C. transvaalensis Burtt-Davy] sod. Growth media were applied at depths of 2 to 4 cm (200 to 400 m3·ha-1) and vegetatively established by sprigging. Cultural practices typical of commercial methods were employed over 99- or 114-day growth periods. Sod was monitored during these propagation cycles, then harvested, evaluated, and installed offsite in a typical lawn-establishment method. Results showed mixtures of CCB and biosolids as growth media increased yield of biomass, with both media and tissue having greater nutrient content than the control media. Volumetric water content of CCB-containing media significantly exceeded that of control media and soil included with a purchased bermudagrass sod. Once installed, sod grown on CCB-media did not differ in rooting strength from control or purchased sod. When applied as described, physicochemical characteristics of CCB-media are favorable and pose little environmental risk to soil or water resources.

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Joseph P. Albano and William B. Miller

Irradiation of FeDTPA-containing nutrient solutions by a fluorescent plus incandescent light source resulted in the loss of both Fe-chelate and soluble Fe, the formation of a precipitate that was composed mostly of Fe, and a rise in pH. The rate of Fe-chelate photodegradation in solution increased with irradiance intensity and with solution temperature under irradiation, but irradiance had the greater effect. Fe-chelates absorb in the blue and UV regions of the spectrum. Removal of these wavelengths with a spectral filter eliminated photodegradation. Chemical name used: ferric diethylenetriaminepentaacetic acid (FeDTPA).

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Joseph P. Albano and William B. Miller

Marigold (Tagetes erecta L.) grown hydroponically in an irradiated nutrient solution containing FeDTPA had root ferric reductase activity 120% greater, foliar Fe level 33% less, and foliar Mn level 90% greater than did plants grown in an identical, nonirradiated solution, indicating that the plants growing in the irradiated solution were responding to Fe-deficiency stress with physiological reactions associated with Fe efficiency. The youngest leaves of plants grown in the irradiated solution had symptoms of Mn toxicity (interveinal chlorosis, shiny-bronze necrotic spots, and leaf deformation). Plants grown in irradiated solution in which the precipitated Fe was replaced with fresh Fechelate were, in general, no different from those grown in the nonirradiated solution. Chemical name used: ferric diethylenetriaminepentaacetic acid (FeDTPA).

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Joseph P. Albano and William B. Miller

Irradiating a ferric ethylenediaminetetraacetic acid (FeEDTA)-containing commercially available soluble fertilizer with ultraviolet (UV) and blue radiation from high intensity discharge (HID) lamps caused the photooxidation of the FeEDTA complex, resulting in the loss of 98% of soluble iron. The loss of soluble iron coincided with the development of a precipitate that was mostly composed of iron. The effects of using an irradiated FeEDTA-containing fertilizer solution on plant growth and nutrition under commercial conditions were studied. Application of the irradiated fertilizer solutions to greenhouse grown tomato plants (Lycopersicon esculentum) resulted in lower levels of iron (6%) and zinc (9%), and higher levels of manganese (8%) and copper (25%) in leaf tissue compared to control plants that received a nonirradiated fertilizer solution. Leaf macronutrient levels (phosphorous, potassium, calcium, and magnesium), leaf dry weight, leaf number, and plant height was not affected by application of the irradiated fertilizer solution.

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Joseph P. Albano and William B. Miller

Irradiation of, ferric ethylenediaminetetraacetic acid (FeEDTA, iron chelate)-containing commercial fertilizer solutions by fluorescent plus incandescent lamps resulted in the loss of both FeEDTA and soluble iron (Fe), and the formation of a yellow-tan precipitate that was mostly composed of Fe. The ratio of soluble Fe:manganese (Mn) was altered due to FeEDTA photodegradation from 2:1 in the nonirradiated solutions to 1:4 in the irradiated solutions, respectively. Storing fertilizer solutions in containers that were impervious to light prevented FeEDTA photodegradation.

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Joseph P. Albano and William B. Miller

The susceptibility of seven African marigold (Tagetes erecta L.) cultivars to iron toxicity was assessed. Plants were grown in a greenhouse in a soilless medium and Fe-DTPA was incorporated into the nutrient solution at either 0.018 mmol·L-1 (low) or 0.36 mmol·L-1 (high). Symptoms of Fe toxicity (bronze speckle disorder in marigold characterized by chlorotic and necrotic speckling and downward leaf cupping and curling) developed only in the high-Fe treatment. The concentration of Fe in leaves in the high-Fe treatment was 5.6 and 1.7 times as great as in the low-Fe treatment for `Orange Jubilee' and `Discovery Orange', respectively. Based upon the percentage of plants affected and leaf symptom severity, relative cultivar susceptibility to Fe toxicity was Orange Jubilee > First Lady > Orange Lady > Yellow Galore > Gold Lady > Marvel Gold > Discovery Orange. Chemical names used: ferric diethylenetriaminepentaacetic acid, disodium salt dihydrate (Fe-DTPA).

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Anil P. Ranwala and William B. Miller

Amylolytic activities extracted from scales of tulip (Tulipa gesneriana L. cv. Apeldoorn) bulbs stored at 4 °C for 6 weeks under moist conditions were characterized. Anion exchange chromatography of enzyme extract on DEAE-Sephacel revealed three peaks of amylolytic activity. Three enzymes showed different electrophoretic mobilties on nondenaturing polyacrylamide gels. The most abundant amylase activity was purified extensively with phenyl-agarose chromatography, gel filtration on Sephacryl S-200, and chromatofocusing on polybuffer exchanger PBE 94. The purified amylase was determined to be an endoamylase based on substrate specificity and end product analysis. The enzyme had a pH optimum of 6.0 and a temperature optimum of 55 °C when soluble starch was used as the substrate. The apparent Km value for soluble starch was 1.28 mg/ml. The inclusion of 2 mM CaCl2 in the reaction mixture resulted in a 1.4-fold increase in the enzyme activity. The presence of calcium ions also enhanced the thermo-stability of the enzyme at higher temperatures. The enzyme was able to hydrolyze soluble starch, amylose, amylopectin, and beta-limit dextrin, but it had no activity against pullulan, inulin, maltose, or p-nitrophenyl alpha-glucopyranoside. Only maltooligosaccharides, having a degree of polymerization of 7 or more, were hydrolyzed to a significant extent by the enzyme. Exhaustive hydrolysis of soluble starch with the enzyme yielded a mixture of maltose and matlooligosaccharides. This amylase activity was not inhibited by alpha- or beta-cyclodextrin upto a concentration of 10 mM. Maltose at a 50 mM concentration partially inhibited the enzyme activity, whereas glucose had no effect at that concentration.

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Joseph P. Albano and William B. Miller

Our objective was to determine the effects on plant growth and physiology that a photodegraded Fe-chelate containing lab-prepared nutrient solution would have when used in plant culture. Plants grown hydroponically in the irradiated Fe-DTPA containing nutrient solution had ferric reductase activity 2.2 times greater, foliar Fe level 0.77 times less, and foliar Mn level 1.9 times greater than in plants grown in an identical but non-irradiated solution, indicating that plants growing in the irradiated solution were responding to Fe deficiency stress with physiological reactions associated with Fe efficiency. The youngest leaves of plants that were grown in the irradiated solution had symptoms of Mn toxicity. Restoration of the irradiated solution by removing the precipitated Fe by centrifugation and adding fresh Fe-chelate resulted in plants that were, in general, not different from those grown in the non-irradiated solution (control).