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Warner Orozco-Obando, Gwen N. Hirsch and Hazel Y. Wetzstein

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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Yi He, Hazel Y. Wetzstein and Barrv A. Palevitz

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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Hazel Y. Wetzstein, Elizabeth A. Richardson and Yi He

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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Hazel Y. Wetzstein, Weiguang Yi, Justin A. Porter and Nadav Ravid

Pomegranate trees (Punica granatum) produce large numbers of both hermaphroditic (bisexual) flowers that produce fruit and functionally male flowers that characteristically abort. Excessive production of male flowers can result in decreased yields resulting from their inability to set fruit. Within hermaphroditic flowers, sex expression appears to follow a spectrum ranging from those exhibiting strong to weak pistil development. Unknown is the scope that flower quality plays in influencing fruit production. A description of floral characteristics and how they vary with flowers of different sizes and positions is lacking in pomegranate and was the focus of this study. Furthermore, the effects of flower size and position on fruit set and fruit size were evaluated. This study documents that flower size characteristics and ovule development can be quite variable and are related to flower type and position. Single and terminal flowers within a cluster were larger than lateral flowers. In addition, lateral flowers exhibited a high frequency of flowers with poor ovule development sufficient to negatively impact fruiting in that flower type. Ovule numbers per flower were significantly influenced by flower size with more ovules in larger flowers. Pollination studies verified significantly higher fruit set and fruit weight, and larger commercial size distributions were obtained with larger vs. smaller flowers. Thus, flower quality is an important issue in pomegranate. Cultural and environmental factors that influence flower size and vigor may have a direct consequence on fruit production and yield.

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Hazel Y. Wetzstein*, Allan M. Armitage, Gwen N. Hirsch and Stephanie L. Anderson

Tissue culture is a useful means to clonally propagate new ornamental plant selections, particularly when plant material is limited and/or conventional propagation methods are ineffective. An efficient in vitro multiplication protocol was established to propagate a new goatsbeard hybrid (Aruncus dioicus, × A. aethusifolia). The hybrid is of interest because it exhibits a dwarf habit, delicate white flower panicles and fern-like leaves, yet is tolerant to heat and humidity. Experiments were conducted to evaluate explant type (nodes, stems, leaves, and floral parts), disinfestation procedures, and media formulations including varying concentrations of 6-benzylaminopurine (BAP) and naphthalene acetic acid (NAA). Rapid plant regeneration was obtained with a shoot organogenesis system using a half strength Murashige and Skoog medium supplemented with 4.4 μmol BAP, 0.54 μmol NAA, 30 g·L-1 sucrose, and 3.0 g·L-1 GelGro. Studies compared the performance and yield of plants rooted using different in vitro and ex vitro methods. Ex vitro rooting of shoots during greenhouse acclimatization under mist was most effective. Regenerated plants exhibited uniform and rapid growth, and performed well in greenhouse and field evaluations.

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Kimberly A. Pickens, Jan Wolf, James M. Affolter and Hazel Y. Wetzstein

Many bromeliad species indigenous to the rain forests of Central and South America are threatened because of over-collection and habitat destruction. Studies were conducted to develop propagation protocols for Tillandsia eizii, a rare ornamental bromeliad of ceremonial significance to the Highland Maya communities in Chiapas, Mexico. We anticipate using in vitro propagation for the conservation of this species with the potential of utilizing bromeliads as an alternative and sustainable forest resource. Protocols were developed for the sterilization and germination of axenic seed. Seedling growth in vitro was assessed and outplanting studies were conducted. Media were evaluated to promote adventitious bud production in experiments using the plant growth regulators naphthaleneacetic acid and benzylaminopurine. Pulse time and duration, as well as the stage of seed development, had a marked effect on bud production. The effects of various potting media on plant growth and survival were assessed. A pure pine bark medium elicited over 95 percent survival. Plants exhibited a “tank-like” morphology characteristic of plants in the wild.

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Benjamin Jeyaretnam, Hazel Y. Wetzstein, Sharad C. Phatak and Russel W. Carlson

Changes in lipid and total protein content of somatic embryos of pecan (Carya illinoinensis) were estimated during maturation, cold treatment alone (3, 5, or 8 weeks) or cold followed by dessication (3, 5, or 7 days). Triglyceride was estimated colorimetrically and methyl esters of fatty acids were analyzed by GC-MS. Total protein was extracted from the same tissue with 2% SDS in Tris·HCL buffer. Triglyceride content of enlarged somatic embryos was significantly lower than zygotic embryos and further declined after 5 weeks cold treatment. An even greater decline was observed during the desiccation treatment. The most abundant fatty acids in small and enlarged somatic embryos are linolenic > palmitic > oleic > stearic acid. However, the molar ratio of linolenic to oleic reached 1:1 after 5 weeks of cold treatment. During enlargement, protein content increased to levels found in zygotic embryos, with desiccation resulting in further elevation.

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Justin A. Porter, Hazel Y. Wetzstein, David Berle, Phillip A. Wadl and Robert N. Trigiano

Georgia plume, Elliottia racemosa (Ericaceae), is a small tree endemic only to the state of Georgia, where it is listed as a threatened species. Information about genetic relatedness is critical for establishing approaches for safeguarding, reintroduction, and conservation of this rare species. The genetic relationships among and within selected georgia plume populations were evaluated using random amplified polymorphic DNA (RAPD) in conjunction with site visits at which time a census and GPS survey were conducted. Populations ranged from those containing eight to over 1000 individuals with most populations containing few plants (less than 50 individuals). With one exception, small populations with less than 50 individuals had more genetic similarity than populations with greater numbers of plants. Two protected populations containing large numbers of individuals were sampled extensively. Genetic similarity of individuals was not associated with plant proximity within a population. The small number of individuals and geographic isolation characteristic of many populations were associated with high within-population genetic similarity. Conservation priorities should be given to preserving as many different populations as possible to retain the genetic diversity of the species. Whether the narrow genetic variation found in some populations may be contributing to lack of sexual reproduction in the wild is an area for further study.

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Kimberly A. Pickens, James M. Affolter, Hazel Y. Wetzstein and Jan H.D. Wolf

Tillandsia eizii is an epiphytic bromeliad that due to over-collection, habitat destruction, and physiological constraints has declined to near threatened status. This species exhibits high mortality in the wild, and seed are characterized by low percentages of germination. As a means to conserve this species, in vitro culture protocols were developed to enhance seed germination and seedling growth. A sterilization protocol using 70% ethanol for 2 minutes followed by 2.6% NaOCl for 40 minutes disinfested seed and promoted seedling growth. Sucrose incorporated into the culture medium had no effect on germination or growth, while NAA inhibited growth, but not germination. Cultures maintained under a 16-hour photoperiod at 22 °C exhibited greater growth than those grown at 30 °C. Seed that germinated in the dark remained etiolated and failed to develop even after transfer to light conditions. Plants grown in vitro were successfully acclimatized and transferred to the greenhouse. Over 86% survival and rapid growth were obtained with either an all-pine-bark medium, or a mixture of 2 redwood bark: 2 fir bark: 2 potting mix: 1 perlite. This demonstrated that in vitro culture of seed may be used to rapidly produce large numbers of T. eizii, and thus can be used for the conservation and reintroduction of this species.

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Patricio A. Brevis, D. Scott NeSmith, Hazel Y. Wetzstein and Dorothy B. Hausman

Fruit set of rabbiteye blueberries (Vaccinium ashei Reade) can be pollen-limited under certain conditions. The objective of this study was to determine production, release, and viability of pollen, as well as pollen-ovule ratios in the rabbiteye blueberry cultivars Austin, Brightwell, Climax, and Tifblue. In vitro tetrad germination varied among genotypes, although, values were high (≥80%) in all cultivars. Pollen viability does not seem to contribute to reproductive failure in the cultivars studied. Total pollen production per flower averaged 8434 tetrads across all cultivars. On a per ovule basis, pollen production was very low relative to other xenogamous species. The low pollen-ovule ratio of rabbiteye blueberry (≈400) may be an indicator of the high efficiency of its pollen dispensing mechanism. Total pollen production varied among cultivars. Furthermore, a significant difference in pollen release was found between two cultivars with similar total pollen production per flower. The possible mechanism regulating pollen release in these cultivars is discussed.