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  • Author or Editor: Hazel Y. Wetzstein x
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Propiconazole, a triazole fungicide, has been reported to inhibit leaf expansion in pecan [Carya illinoensis (Wangenh.) K. Koch] trees when applied under field conditions. This study was conducted to determine the effect of propiconazole on pecan leaf morphology and structure using light and transmission electron microscopy. Mature pecan trees were sprayed once or three times per week from budbreak to pollen maturity. Fungicide sprays resulted in significantly reduced leaf area. Compared to controls, leaves from propiconazole-treated shoots had alterations in cell arrangement characterized by more tightly packed palisade parenchyma cells with fewer intercellular spaces; neither leaf thickness nor palisade or spongy layer thickness were affected. Propiconazole caused modifications in the chloroplasts, with a tendency for internal membranes to be less defined, and for thylakiods to exhibit less stacking. The extent of structural changes was related to fungicide dosage. Results show that propiconazole applications during leaf development can inhibit leaf expansion and modify cellular organization of the mesophyll cells. Chemical name used: 1-[[2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolan-2-yl] methyl]-1H-1,2,4-triazole (propiconazole).

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Fungicides have been shown to negatively affect pollen germination, tube growth, and fruit set in important crops. However, little is known regarding possible modes of action in higher plant cells. To address this, the effects of propiconazole or benomyl on pollen germination and tube growth were evaluated in Tradescantia virginiana using light microscopy and immunocytochemistry. Concentrations were selected at levels that had inhibitory effects, but did not totally arrest germination and tube elongation, i.e., propiconazole and benomyl were added at 0, 102, 136, or 170 μl·liter–1, and 0, 480, 600, or 720 mg·liter–1, respectively. Both fungicides inhibited germination, cytoplasmic streaming, tube elongation, and induced abnormal tube morphology and cytoskeletal distribution. Propiconazole-treated tubes had weaker microfilament signals, with amorphous staining. Microtubule (Mt) distribution was severely affected. In benomyl-treated tubes, Mts were fewer in number, fragmented, sinuous, and increasingly disorganized. Possible mechanism(s) will be discussed.

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Zinc deficiency is a nutrient disorder that is observed in pecan production areas. In the field it is characterized by a rosette shoot habit and interveinal leaf chlorosis. Up to now, the induction of zinc deficiency has not been accomplishable in the field or greenhouse. Thus any critical evaluations of effects of zinc nutrition on tree growth and development have been lacking. A hydroponic culture system was developed where zinc deficiency was induced. Seedstocks collected from `Stuart', `Curtis', and `Wichita' trees were grown with and without zinc supply. Biomass, leaf area, node number, and visual symptoms were assessed. Foliar deficiency symptoms were rated 4 and structural evaluations were conducted using light and electron microscopy. Significant differences in visual symptoms were observed between treatments and among cultivars. Leaf area significantly decreased in `Stuart' and `Curtis' under zinc deficient conditions. Zinc had no significant effect on biomass and internodal length. Foliar nutrient contents were compared between cultivars. Our data suggest that genotypic differences in sensitivity to zinc deficiency exists and improving pecan production through genetic selection for zinc efficiency appears promising.

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Georgia plume (Elliottia racemosa, Ericaceae) is a threatened, woody plant endemic to Georgia's Coastal Plain region in the southeastern United States. Populations of the plant have a fragmented distribution within a restricted range and are characterized by low genetic diversity and a lack of sexual recruitment. Georgia plume cannot be effectively propagated using conventional methods. We have developed an in vitro shoot regeneration system that is effective with explants obtained from mature plants in the wild. The objective of this study was to determine the efficacy of using this in vitro protocol to regenerate proliferating shoot cultures from 34 georgia plume genotypes obtained from divergent populations. Young expanding leaves were cultured on Gamborg's media supplemented with 10 μM thidiazuron and 5 μM indole-3-acetic acid. After 8 weeks, tissues were transferred to a shoot elongation medium with 25 μM 2-isopentenyl adenine. Of the 34 genotypes tested, 91% formed shoot primordia and 85% regenerated shoots within 6 months of inoculation. This study verifies that tissue culture can be used to produce adventitious shoots from a wide range of georgia plume genotypes. Within a coordinated conservation program, tissue culture is a feasible system to use for safeguarding and reintroduction of genetically diverse plant material, which may be critical to the survival of this rare species.

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As a plant nutrient, nitrogen is the element in highest demand in terms of quantity and makes up about 2% to 3% of plant dry matter. In this study, we evaluated the effect of nitrogen source on plant growth and nutrient uptake in pecan (Carya illinoensis). Seedlings were hydroponically grown under three nitrogen nutrient regimes where the ratio of nitrate: ammonium was varied, i.e., 3:1, 1:1, and 1:3. High ammonium nutrition had an inhibiting effect on seedling growth. Plants grown under 1:3 (nitrate: ammonium) exhibited significantly lower biomass, decreased root/shoot ratio, and lower specific leaf weight than other treatments. Total nitrogen uptake on a dry weight basis was highest in the high ammonium treatment. In the equal molar treatment (1:1 nitrate: ammonium), plants exhibited preferential uptake of ammonium-form nitrogen. Ammonium-form nitrogen is generally used in pecan orchard practice. Our data suggest that further studies evaluating the effects of nitrogen source are warranted to determine if similar detrimental effects on pecan growth occur in the field. Such studies would be useful for optimizing current fertilization practices.

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A simple and efficient protocol is reported for the isolation of RNA from embryos and leaves of pecan [Carya illinoinensis (Wangenh.) K. Koch]. The method relies on suppression of the polyphenols from interaction with the RNA and their rapid removal from the homogenate by chloroform extraction. This method produced abundant amounts of high-quality RNA. This protocol is likely to be useful for Juglandaceous species and other recalcitrant plants with high levels of phenolic compounds.

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Effects of autoclaving volume, gelling agent (Bactoagar, Gel-gro, Phytagar), and basal salts [Murashige and Skoog (MS); Woody Plant Medium (WPM); Gamborg B5 (GB)] on gel strength and pH of tissue culture media were tested. Gel strength was significantly affected by gelling agent and basal medium. MS media were generally softer than comparable WPM or GB media. As the vessel volume during autoclaving decreased, gel strength significantly decreased with Phytagar and Bactoagar gelling agents; Gel-gro had greater gel strength at the intermediate volume of medium autoclave. In all cases, autoclaving resulted in a pH decrease of 0.2 to 0.5 pH units. Lower pH values were associated with softer gels. The type of gelling agent did not greatly affect the postautoclave pH; mean values among gelling agents were within 0.05 pH units. Postautoclave pH of MS medium was lower than that of WPM or GB. This study verifies the need to observe uniform sterilization protocols to maintain consistency in the chemical and physical properties of media.

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The general doctrine of flowering in Hydrangea macrophylla (Thunb.) Ser. is that floral induction occurs during the fall months with the flower appearing the following spring or summer. However, hydrangea cultivars differ widely in their relative abundance and duration of flower production. The objective of this study was to determine how developmental flowering patterns compared among different hydrangea genotypes. Flowering was characterized in 18 cultivars by assessing flower initiation in dormant buds of 1-year-old stems that received natural outdoor inductive conditions. Terminal and lateral buds were dissected and floral developmental stage categorized microscopically. In terminal buds, flower development was very consistent and occurred in 100% of buds for all cultivars except `Ayesha' (33%). In contrast, lateral buds showed a wide variation in flower induction among genotypes. `Ayesha', `Blushing Pink', `Freudenstein', and `Nigra' had 10% or fewer lateral buds with floral initials. `All Summer Beauty', `David Ramsey', `Masja', `Nightingale', and `Penny Mac' showed high levels of floral induction (>92%). Within a cultivar, flower development was more advanced in terminal than lateral buds. In several cultivars, a significant correlation between bud size (length) and floral stage was found. However, low r-square values indicated that flower stage was explained largely due to factors other than bud length. This study shows that floral induction patterns vary markedly among hydrangea cultivars and provides insight into why cultivars differ in the extent and reliability of seasonal blooming. Genotypes that possess floral primordia in lateral buds would be amenable to cultural practices that enhance lateral budbreak and recurrent blooming.

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In almond [Prunis dulcis (Mill.) D.A. Webb.], fungicide sprays are required to prevent blossom blight, which can infect open flowers. Numerous studies have reported detrimental effects of agrochemical sprays on pollination, fruit set, and yield in tree fruit crops. However, effects of fungicides on pollen germination and growth in almond are little known, particularly those from recently developed active ingredients. In this study we evaluated the effects of commercial formulations of 10 fungicides on pollen germination and tube growth in almond using in vitro assays. Assays conducted at 1/100 recommended field rates (RFR) were effective in delineating differences in almond pollen sensitivity to different fungicides. Captan and azoxystrobin were the most inhibitory, with germination percentages of less than 1% of the no-fungicide control. Germination was not significantly affected by propiconazole and benomyl. Intermediate inhibitory effects on pollen germination were observed with ziram, cyprodinil, maneb, thiophanate-methyl, iprodione, and myclobutanil. In contrast to germination, tube growth was less affected by the presence of fungicide. In pollen that germinated, tube elongation was the same as in controls in five of 10 of the fungicides evaluated. Nonetheless, azoxystrobin and captan reduced tube elongation by ≈90%. Some fungicide treatments also influenced tube morphology. In the absence of field evaluation studies, in vitro germination data may provide insight on how specific chemicals may impact pollination processes and further guide in vivo studies, particularly in the case of new chemical formulations.

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