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Rodney Serres and Brent McCown

The capability to uniformlyinduce flowering in cranberry (Vaccinium macrocarpon Ait. `Stevens') in < 1 year from microculture was investigated to accelerate cranberry breeding and to study woody plant reproductive biotechnology. Flower buds were induced on newly micropropagated cranberry plants during the first growing season. A treatment of 2.5 mg of paclobutrazol applied as a soil drench per 2- to 3-month-old potted plant in midsummer, when the plants were grown in coldframes under natural daylength and air temperatures, resulted in 70% of the plants flowering. Plants not treated with paclobutrazol did not flower. Reduced but significant flower bud set was observed on plants treated with paclobutrazol but grown in the greenhouse under natural daylength. Flowering was stimulated by cold treatment coupled with gibberellin sprays and/or repotting to nonpaclobutrazol-treated medium. Chemical name used: β -[(4-chlorophenyl)methyl]-ct-(1,1-dimethylethyl)-1H-1,2,4-triazole-1-ethanol (paclobutrazol).

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Rodney Serres and Brent McCown

The gene encoding β-glucuronidase, GUS, has been inserted into cranberry and is expressed in various tissues. Detectable expression of the GUS gene is enhanced up to 15x when the phenol-adsorbing compound, polyvinylpolypyrrolidone, is included in the extraction buffer of the fluorometric MUG assay, indicating that an endogenous, probably phenolic, compound is inactivating the foreign enzyme. Extracts from in vitro-grown cranberry leaves reduce the activity of purified β-glucuronidase in fluorometric assays. This is in contrast to extracts from other plants which have no affect on the enzyme. Detectable expression of the GUS gene for an individual transclone varies with the age of the tissue and the environment in which the plant is grown. The BT gene, which encodes for the Bacillus thuringiensis δ-endotoxin, was also inserted into cranberry with the purpose of incorporating lepidopteran insect resistance. Bioassays using an important insect pest on cranberry show generally inconsistent feeding patterns on transgenic plants. These results may be due to the interaction of the endogenous compounds and the B.t. δ-endotoxin.

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Dale T. Lindgren and Brent McCown

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Brent H. McCown and Eric L. Zeldin

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Eric L. Zeldin, Brent H. McCown, Elden Stang, and John Klueh

A project to determine the comparative growth response of micropropagated (MP) and field propagated (FP) cranberry plants was conducted in field plots at a commercial cranberry marsh. Microcuttings were derived from shoot culture and rooted in either plugs or peat pots filled with peat. Replicated 1 m2 plots of MP plants and 15 cm FP cuttings were planted in June. Survival of MP plants after one month was significantly greater than that of the FP plants. Significant growth differences were observed later in the season. The MP plants produced more branches and greater runner elongation, resulting in a much greater ground cover. Many of the FP plants flowered and produced fruit, while the MP plants produced neither. Far fewer new flower buds were set in the fall on the MP plants. Potential advantages of MP cranberries include the fast, uniform establishment of new marshes and consequently earlier achievement of full productivity, and the rapid introduction of new genotypes from breeding or genetic engineering.

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Tom J. Buechel, Edward R. Hasselkus, and Brent H. McCown

Root girdling and deformation are problems that occur in containerized woody ornamentals. Two small trees, the coarse-rooted Magnolia × loebneri `Leonard Messel' (grown in 5-gal containers), and the fine-rooted Crataegus × viridis `Winterking' (6' bare root), were transplanted into 10-gal containers, some of which were treated with “SpinOut” [a latex paint containing 100 g Cu(OH)2/liter]. The plants were grown in a pot-in-pot system for 3 months, after which new roots were analyzed for mat formation and branching pattern. No significant differences in shoot growth that could be attributed to the Cu(OH)2 treatment were observed. The treated containers prevented both root encirclement and reformation of matted roots, and resulted in a more-dense and fibrous root system than that observed in untreated containers. The differences were greatest with the coarse-rooted magnolia. The use of containers treated with copper compounds may be an effective means to reduce root problems commonly observed in modern containerized ornamental production.

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Rod Serres, David Russell, Dan Mahr, and Brent McCown

Genetically transformed Vaccinium macrocarpon `Stevens' and `Pilgrim' plants have been obtained using electric discharge particle acceleration. Three foreign genes, kan encoding a selectable marker, gus a reporter gene, and B.t.k. conferring lepidopteran resistance, were incorporated into the genome. Expression of kan was assayed by culturing shoots in vitro on media with several concentrations of kanamycin. Expression among transformed clones (transclones) varied from high resistance (normal growth at 300 mg/L kan) to no resistance. Histochemical analyses for gus expression revealed variability among transclones. Some transclones exhibited no gus expression, others had consistent area-specific expression while others displayed random expression. In preliminary feeding trials with blackheaded fireworm larvae, B.t.k. expression was found to be ineffective at controlling insect development. We have recovered plants transformed with a different promoter driving the B.t.k. gene in an effort to enhance expression.

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Eric L. Zeldin, David D. Ellis, and Brent H. McCown

Taxol, a promising anticancer drug, is limited by inadequate supply. The production of taxol and related compounds (taxanes) by Taxus tissue cultures has been reported, yet sustained production has not been demonstrated. One theory is that cell differentiation and/or tissue organization is required to sequester taxol and avoid autotoxicity. To investigate this, T. cuspidata shoot cultures were established and the taxane content of various culture stages compared to that of field needles. HPLC analysis identified two peaks which comigrated and had UV spectra identical to taxol and 10-deacetyl taxol. The levels of 10-deacetyl taxol were similar in all samples. Cultured shoots contained much less taxol than field needles, and the level of a third peak which migrates closely to taxol was inversely related to that of taxol. Taxol content was restored in the first flush out of culture. Shoot cultures of T. brevifolia, T. x media, and T. canadensis have also been analyzed. In addition to shoot cultures, nodule cultures, a biological unit that may be suitable for production of taxanes in plant bioreactors, have been initiated and characterized.

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William A. Hoch, Geunhwa Jung, and Brent H. McCown

A significant pest affecting commonly planted Betula spp. is the birch leafminer (Fenusa pusilla Lepeletier), an insect that can be present in large populations in the landscape and can greatly reduce the vigor and ornamental value of these trees. Twenty-two interspecific crosses were performed between leafminer resistant and susceptible Betula species in an attempt to create the novel combination of ornamental white bark and significant leafminer resistance. Of the nine successful crosses, two produced resistant offspring. Progeny of the diploid × hexaploid cross B. turkestanica Litvin (2x) × B. alleghaniensis Britt. (6x) displayed a broad range of resistance levels, likely the result of segregating alleles contributed by the hexaploid parent. All crosses involving highly resistant individuals of B. costata Trautv. (2x) yielded leafminer susceptible progeny. These results suggest that the larval antibiosis demonstrated by B. alleghaniensis and B. costata is inherited as a recessive trait, and exhibits a gene dosage effect as evidenced by the B. turkestanica × B. alleghaniensis offspring. While most progeny of the B. populifolia Marsh (2x) × B. maximowicziana Regal (2x) cross were susceptible, a single resistant offspring, which was found to be triploid (3x), displayed a mechanism of resistance similar to that of a hypersensitive response. No strong intersectional barriers to hybridization were observed and all interploidy crosses were successful. The chromosome numbers of B. costata (2n = 2x = 28) and B. turkestanica (2n = 2x = 28) are reported here for the first time. The results of this study indicate that the potential exists for the development of insect resistant, ornamental white-barked birch clones through the implementation of a planned, systematic breeding program.