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Yao-Chien Chang and William B. Miller

Upper leaf necrosis (ULN) on Lilium `Star Gazer' has been recently demonstrated to be a calcium (Ca) deficiency disorder. In the current studies, we confirmed this by using a Ca-free nutrient regime to reproduce ULN symptoms. The ability of a bulbous storage organ to supply calcium to a growing shoot is poorly understood. Therefore, we conducted experiments to determine Ca partitioning during early growth stages, and under suboptimal Ca levels to determine how the bulb affects the symptomatology. The results indicated that ULN is originally caused by an insufficient Ca supply from the bulb. In the most susceptible period, bulb dry matter decreased dramatically and Ca concentrations in immature folded leaves dropped to very low levels. Consequently, necrosis began to appear on the upper, young leaves. The bulb was able to supply Ca to other organs, but only to a limited extent since Ca concentration in bulbs was low (0.04% w/w). To confirm this result, we cultivated lilies with low-Ca or Ca-free nutrient solution and obtained bulbs with extremely low internal Ca concentrations. Upon forcing these low-Ca bulbs, we found, as expected, prominent necrosis symptoms on the lower and middle leaves. Data suggested the lower and middle leaves relied more on Ca supplied from the bulb, while upper leaves and flowers relied more on Ca uptake from the roots. Different organs have different Ca requirements, and tissue sensitivity to Ca deficiency varies according to the growth stage.

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Yao-Chien Chang and William B. Miller

A necrotic disorder occurs on upper leaves of many oriental hybrid lily (Lilium L.) cultivars, including the most-widely-grown `Star Gazer'. We term this disorder “upper leaf necrosis” (ULN) and hypothesize that it is a calcium (Ca) deficiency. We demonstrated that Ca concentration in necrosed tissues was nearly six-fold below that of normal leaves (0.10% vs. 0.57% dry weight), and that Ca concentration was negatively associated with percentage necrosed leaf area. It was concluded that ULN is a Ca deficiency disorder. When the symptoms were slight, early ULN symptoms appeared as tiny depressed spots on the lower surface of the leaf, or as water-soaked areas when the disorder was severe. Most commonly, ULN began on the leaf margin. The injured areas turned brown, leading to leaf curling, distortion, or tip death. ULN occurred on leaves associated with flower buds and leaves immediately below the flower buds. For the plants grown from 16-18 cm circumference bulbs, the five leaves directly below the flower buds and larger leaves associated with the 1st and the 2nd flower buds were most susceptible. In general, flower buds were not affected by ULN, and continued to develop and flower normally, even though they were associated with subtending, highly distorted leaves. Eighty-five percent of plants began to exhibit ULN symptoms 30-40 days after planting (i.e., 24-34 days after shoot emergence). This was the stage when the 6th or 7th leaf under the bottom flower bud was just unfolded. Light was not the main factor that initiated ULN, however, ULN severity was greatly increased by light reduction, as leaf transpiration was reduced.

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William B. Miller and Robert W. Langhans

Easter liliy (Lilium longiflorum Thunb. `Nellie White') bulbs were stored in moist peatmoss for up to 85 days at – 1.0 or 4.5C. Bulbs were periodically removed from storage and analyzed to determine levels of soluble carbohydrates and starch. Storage at – 1.0C induced large accumulations of sucrose, mannose, fructose, and oligosaccharide in both mother and daughter scales. Starch concentration declined substantially during this period. Storage at 4.5C resulted in less dramatic alterations in bulb carbohydrates, although trends toward increased soluble carbohydrates and reduced starch levels were seen. The accumulation of mannose suggests that glucomannan, a secondary storage carbohydrate, was also degraded during – 1.0C storage.

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Anil P. Ranwala and William B. Miller

In mature Lilium longiflorum flower buds, anther and stigma had the highest soluble acid invertase activity [3.29 and 2.31 μmol of reducing sugars (RS)/min per gram of fresh weight (FW), respectively] compared to style, ovary, petal, and filament with activities of 1.52, 1.08, 0.99 and 0.98 μmol RS/min per gram of FW, respectively. DEAE-sephacel chromatography revealed that invertase activity in petal, ovary, style, and stigma was composed exclusively of invertase II and III isoforms. Anther invertase was mainly invertase I with small amounts of invertase II and III. Filament tissue mainly had invertase II and III isoforms with a small amount of invertase I. Wall-bound invertases were extracted with 1.0 m NaCl. Anthers had the highest wall-bound invertase activity (4.42 μmol RS/min per gram of FW) followed by stigma (0.42 μmol RS/min per gram of FW). Other tissues had low wall-bound invertase activity (<0.1 μmol RS/min per gram FW). For further purification, the binding of soluble invertases to nine different reactive dyes was investigated. Invertase I was bound to Reactive Green 5, Reactive Green 19, and Reactive Red 120 columns and was eluted with 0.5 m NaCl, resulting in increase in specific activity ≈10-fold with ≈70% recovery. Invertase II and III bound only to Reactive Red 120. Elution with 0.5 m NaCl resulted in an ≈6-fold increase in specific activity.

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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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Joseph P. Albano and William B. Miller

Iron chelate photodegradation is a problem in tissue culture where limited soluble Fe in agar reduces callus tissue growth. Our objectives were to determine if Fe chelate photodegradation occurs in commercial fertilizers used in greenhouse plant production and, if so, the effects on plant Fe acquisition. Commercial 20N–10P–20K soluble fertilizers containing Fe-EDTA were prepared as 100x stocks based on a 100 mg N/liter (1x) concentration. A modified Hoagland's solution with Fe-DTPA was prepared as a 10x stock based on a 200 mg N/liter (1x) concentration. Samples then were kept in darkness or were irradiated with 500 μmol·m–2·s–1 from fluorescent and incandescent sources for ≤240 hours. Soluble Fe in the irradiated commercial fertilizer solutions decreased 85% in 240 h. Soluble Fe in the Hoagland's solution, prepared in the lab, decreased 97% in 72 h. There was no loss in soluble Fe in any dark-stored treatment; demonstrating photodegradation of Fe-chelates under commercial settings. Excised roots of marigold (Tagetes erecta L.), grown hydroponically in the irradiated solutions, had Fe(III)-DTPA reductase activity 2 to 6 times greater than roots of plants grown in solutions kept in darkness. Plants growing in irradiated solutions acidified the rhizosphere more than plants growing in solutions kept dark. The increase in Fe reductase activity and rhizosphere acidification are Fe-efficiency reactions of marigold responding to the photodegradation of Fe-chelates and subsequent decrease in soluble Fe in both commercial fertilizers and lab-prepared nutrient solution.

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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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Jeff S. Kuehny and William B. Miller

The majority of Hippeastrum bulbs sold in the U.S. market are shipped from other countries. The shipping time and temperature varies by the country that the bulbs are shipped from and the storage time and temperature also varies by the company that packages the bulbs for retail sale. These packaged bulbs then sit on a shelf until they are purchased and forced by the consumer. These various storage times and temperatures can affect the longevity after packaging (premature emergence) and quality of the finished plant. The objectives of this research were to determine the effects of various storage temperatures and durations on emergence and forcing of Hippeastrum hybrids. Bulbs were stored at temperatures of 5, 9, 13, 21, and 29 °C for 6, 9, 12, and 15 weeks after which time one set was stored at 21 °C (packaged display temperature) and the other set forced in the greenhouse. Emergence of leaves and buds when stored at the 21 °C display temperature and during greenhouse forcing varied by specific hybrid according to storage duration at 5, 9 and 13 °C. Storage at 21 and 29 °C resulted in only leaf emergence and no flower bud emergence during the 21 °C display temperature and greenhouse forcing. Storage at 5 and 9 °C generally resulted in slower leaf emergence and quicker bud emergence. Results from this research can be used to help determine the best storage times and temperatures for preventing premature emergence of Hippeastrum based upon previous shipping times and temperatures.

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Joseph P. Albano and William B. Miller

We have shown previously that Fe-chelates incorporated into soluble fertilizers are vulnerable to photodegradation, and that such solutions can cause modifications in root reductase activity. The objective of this research was to determine the effects of Fe-chelate photodegradation under commercial production conditions. Marigolds were grown in a greenhouse and transplanted stepwise from #200 plug trays to 804 packs to 11.4-cm (4.5-inch) pots. Plants were harvested at the end of each stage, and treatments consisted of either irradiated (complete loss of soluble Fe) or non-irradiated fertilizer solutions ranging from 100-400 mg/L N (0.5–2 mg/L Fe). In the plug and pack stages, foliar Fe was significantly lower and Mn significantly higher in plants treated with the irradiated than nonirradiated fertilizer solutions, averaging 97 μg·g–1 and 115 μg·g–1 Fe, and 217 μg·g–1 and 176 μg·g–1 Mn, respectively. Fe(III)-DTPA reductase activity of roots of plugs treated with the irradiated fertilizer solution was 1.4-times greater than for roots treated with the non-irradiated fertilizer solution. Leaf dry weight in the plug and pack stages was not affected by treatment, and averaged 0.1 g and 1.2 g per plant, respectively.

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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).