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  • Author or Editor: Hazel Y. Wetzstein x
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Although somatic embryogenesis in vitro has been carried out successfully in a number of plants, a limiting factor in many somatic embryogenic systems is that plantlet regeneration is not obtainable or restricted to low frequencies. We have developed a repetitive, high frequency somatic embryogenic system in pecan (Carya illinoensis) and have identified effective treatments for improved somatic embryo conversion. A 6 to 10 week cold treatment followed by a 5 day desiccation, promoted enhanced root germination and extension, and epicotyl elongation. Light and transmission electron microscopic evaluations of somatic embryo cotyledon development will be presented and related to conversion enhancing treatments and their possible roles in embryo maturation.

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Although somatic embryogenesis in vitro has been carried out successfully in a number of plants, a limiting factor in many somatic embryogenic systems is that plantlet regeneration is not obtainable or restricted to low frequencies. We have developed a repetitive, high frequency somatic embryogenic system in pecan (Carya illinoensis) and have identified effective treatments for improved somatic embryo conversion. A 6 to 10 week cold treatment followed by a 5 day desiccation, promoted enhanced root germination and extension, and epicotyl elongation. Light and transmission electron microscopic evaluations of somatic embryo cotyledon development will be presented and related to conversion enhancing treatments and their possible roles in embryo maturation.

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Georgia plume, Elliottia racemosa Muhlenb. ex. Elliott, is an extremely rare small tree or shrub endemic to Georgia, which is being severely affected by habitat loss and lack of sexual recruitment. In vitro plant regeneration of Georgia plume has not been previously reported and may be a method for the conservation and propagation of this threatened species. Studies evaluated the effects of sterilization methods, explant types, medium composition, and light environment on plant regeneration. An efficient plant regeneration system was developed in which adventitious shoot buds were induced using young, expanding leaf explants placed on an induction medium supplemented with 10 μm thidiazuron and 5 μm indole-3-acetic acid with Gamborg's B5 salts. Shoot elongation was promoted on a medium with 25 μm (2-isopentenyl) adenine incorporated into Woody Plant Medium. In vitro rooting studies evaluated continuous and pulse auxin treatments and ex vitro rooting trials after KIBA (indole-3-butric acid, potassium salt) dips. A 5-day pulse treatment on 100 or 150 μm indole-3-butyric acid produced ≈90% rooting of shoots with greater shoot and root dry weight than other pulse times. High rooting frequencies were obtained under in vitro and ex vitro conditions with over 85% survival of plantlets transferred to greenhouse conditions. The culture protocol was found to be effective with material collected from mature specimens in the wild from divergent populations. Tissue culture appears to be a promising approach for the propagation and conservation of this rare and threatened plant.

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Georgia plume (Elliottia racemosa Muhlenb. ex. Elliott) is a rare deciduous shrub or small tree. It has sustained severe loss of habitat and its range is now restricted to a limited number of sites in the state of Georgia. Tissue culture protocols have been developed as a means to propagate and conserve this threatened species using leaf explants induced on medium supplemented with 10 μm thidiazuron (TDZ) and 5 μm indole-3-acetic acid (IAA). Bud-like clusters, elongated embryo-like protrusions, and shoot-like structures were produced from the leaf explants. Morphological and histological evaluations of cultures during induction and development were conducted using light microscopy of sectioned material and scanning electron micrography. Histology of explant tissues indicates that plant regeneration of Georgia plume occurs through a shoot organogenesis pathway that involves the formation of actively dividing meristematic regions originating in subepidermal cell layers that proliferate to form protuberances on the explant surface. Numerous well-formed shoot apical meristems with leaf primordia are produced, as well as fused shoot-like structures. Elongated, embryo-like structures had various degrees of shoot apex development. Evaluations of serial sections found that they lacked a defined root apex, and that basal portions were composed of parenchymatous files of cells with a broad point of attachment to the parent tissue. The lack of bipolarity and a root pole signifies that true somatic embryogenesis does not occur.

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Zinc deficiency is a widespread nutritional disorder in plants and occurs in both temperate and tropical climates. In spite of its physiological importance, cytological and ultrastructural changes associated with zinc deficiency are lacking, in part because zinc deficiency is difficult to induce. A method was developed to induce zinc deficiency in pecan (Carya illinoinensis (Wangenh.) C. Koch) using hydroponic culture. Zinc deficiency was evaluated in leaves using light and electron microscopy. Zinc deficiency symptoms varied with severity ranging from interveinal mottling, overall chlorosis, necrosis, and marginal curving. Zinc deficient leaves were thinner, and palisade cells were shorter, wider, and had more intercellular spaces than zinc sufficient leaves. Cells in zinc deficient leaves had limited cytoplasmic content and accumulated phenolic compounds in vacuoles. Extensive starch accumulation was observed in chloroplasts. This work represents the first detailed microscopic evaluations of zinc deficiency in leaves, and provides insight on how zinc deficiency affects leaf structure and function.

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Tomato (Lycopersicon esculentum Mill.) transplants can be affected by an intermittent physiological problem manifested by loss of apical meristem function and retarded growth rates, referred to herein as apical meristem decline (AMD). Production losses associated with this condition can be substantial. Similar abnormal and arrested development of the shoot apex has been observed in a number of other species, and referred to as blindness, budlessness, toplessness, blindwood, and bud abortion. A developmental study using scanning electron microscopy was conducted in `Agriset' tomato during an occurrence of AMD to evaluate and compare normal and afflicted plants. The AMD condition was associated with cessation of leaf primordia development and lack of flower initiation. The shoot apex of plants with AMD remained vegetative compared to normal plants which at the same age had well-differentiated flower primordia. No evidence of abortion, die back, or necrosis of the shoot apex was observed. The effects of mineral nutrient additions on symptom development varied with year. In year 1, N fertilization reduced the incidence of both AMD and retarded bud growth (i.e., the percentage of normal plants increased from 29% to 97% with N applications). Preplant applications of P, alone or in conjunction with CaCO3 and trace elements, also ameliorated AMD. In year 2, AMD was observed only at very low levels, i.e., 4% or less, and mineral nutrition had no apparent effect on AMD or normal plant number.

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Abstract

Differentiation and development of the pistillate flower in pecan, [Carya illinoensis (Wang.) K. Koch] were examined using scanning electron microscopy (SEM). Floral differentiation did not occur until growth resumed in the spring, when the outer bud scales were shed and buds were swollen, but before the inner bud scale was broken. Subsequent floral and inflorescence development were correlated highly with stages of bud and early leaf development. The organogenesis of the pistillate flower was described from inception to the stage of visible inflorescence.

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Plantlets were recovered from axillary bud cultures of muscadine grape (Vitis rotundifolia, `Summit'). Nodal segments 0.5 to 1.0 cm long were cultured in Murashige and Skoog (MS) basal medium supplemented with 5, 10, 20, or 40 μm BA. Best total shoot production was obtained with 10 μm BA; with higher BA levels, shoots were unexpanded and exhibited high mortalities. MS medium supplemented with IBA enhanced rooting by increasing rooting percentage and number per plantlet. Shoots previously proliferated on medium with 5 μm BA rooted significantly better than those multiplied on 10 μM BA. Shoot vigor during rooting was greater in shoots proliferated on 5 vs. 10 μm BA. Root development was not significantly affected by liquid vs. agar-solidifted medium or shoot length. Chemical names used: N-(phenylmethyl) -1H-purin-6-amine (BA), 1H-indole-3-butyric acid (IBA).

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Abstract

The morphology of pecan [Carya illinoensis (Wangenh C. Koch)] pollen from 4 cultivars was examined using light and scanning electron microscopy. Pollen was triporate, paraisopolar and suboblate, with a tectate and microechinate surface. The exine was thickened around pores. Pollen from the 4 cultivars was indistinguishable. Pollen germinated in vitro after 1 hr. Pollen tubes grew from 1 or 2 pores, with one germ tube becoming dominant. Pollen germination decreased dramatically after anther dehiscence. Less than 1% of the pollen germinated 5 days after collection.

Open Access

Abstract

Inflorescence and staminate flower development in pecan [Carya illinoinensis (Wang.) K. Koch] were examined using scanning electron microscopy. Organogenesis was described from inception to pollen dehiscence. The order of organ initiation was: a single bract, rounded floral apex, 2 lateral bracteoles, and 3-7 stamens. The initiation and time when stages of floral differentiation occur were determined for 1 protandrous and 2 protogynous cultivars. The time of early inflorescence development and the initiation of floral primordia and bracts were similar in both cultivar types, occurring about 12 months prior to staminate maturity. However, the initiation time of later floral development stages was divergent. The floral stages in the protandrous cultivar up to anther lobing occurred during the previous growing season. In the protogynous cultivars, initiation of bracteoles, the floral apex, stamen primordia and anther lobing took place in the spring of their anthesis.

Open Access