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Clifford W. Beninger, George L. Hosfield, Mark J. Bassett, and Shirley Owens

Three common bean (Phaseolus vulgaris L.) seedcoat color (or glossiness) genotypes, differing from each other by a single substitution at a seedcoat locus, were analyzed for presence and concentration of three anthocyanins: delphinidin 3-O-glucoside, petunidin 3-O-glucoside, and malvidin 3-O-glucoside. The three anthocyanins were present in Florida common bean breeding line 5-593 (P C J G B V Asp), matte black (P C J G B V asp), and dark brown violet (P C J G b V Asp), but the amounts varied greatly depending on the genotype. Dark brown violet had 19% of the total anthocyanin content when compared to 5-593, whereas matte black had amounts intermediate between the two other genotypes. The B gene acts to regulate the production of precursors of anthocyanins in the seedcoat color pathway above the level of dihydrokaempferol formation, perhaps at the chalcone synthase or chalcone isomerase steps in the biosynthetic pathway. We hypothesize that B regulates simultaneously the flavonoid (color) and isoflavonoid (resistance) pathways. The I gene for resistance to bean common mosaic virus (BCMV) is known to be linked closely to B. It is therefore hypothesized that the I gene function may be to respond to BCMV infection by dramatically increasing (over a low constituitive level) production in the 5-dehydroxy isoflavonoid pathway, which leads to synthesis of the major phytoalexin, phaseollin, for resistance to BCMV. Alternatively, the B and I genes may be allelic. The Asp gene affects seedcoat glossiness by means of a structural change to the seedcoat. We demonstrate that Asp in the recessive condition (asp/asp) changes the size and shape of the palisade cells of the seedcoat epidermis, making them significantly smaller than either 5-593 or dark brown violet. Asp, therefore, limits the amounts of anthocyanins in the seedcoat by reducing the size of palisade cells.

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P. Gercheva, R.H. Zimmerman, L.D. Owens, C. Berry, and F.A. Hammerschlag

Shoot regeneration from apple (Malus domestica Borkh.) leaf explants following particle bombardment at various acceleration pressures was studied. Basal leaf segments of micropropagated `Royal Gala' apple were bombarded with 1 μm gold particles, accelerated using helium pressures of 4.5, 6.2, 7.6, 9.3, or 13.8 MPa (650–2000 psi), and cultured on shoot regeneration medium consisting of N6 salts supplemented with 10 μM TDZ for 5, 10, or 20 days in darkness. Bombarded and control explants exhibited 63% to 100% shoot regeneration. With a 5-day dark period, average shoot production per explant ranged from 6.1 to 14; bombardments of 4.5 and 6.2 MPa significantly increased shoot production over the controls. With a 10-day dark period, average shoot production per explant ranged from 9.1 to 22 following bombardment at 9.3 and 6.2 MPa, respectively. Following bombardment at 6.2 MPa, 75% of the explants produced more than 20 regenerants per explant. With a 20-day dark period, average shoot production per explant ranged from 8.9 to 19 following bombardment at 13.8 MPa and following no bombardment, respectively. Shoot production per explant was significantly less than the controls following bombardments ranging from 6.2 to 13.8 MPa. Shoot production was highest per explant with particle bombardment at 6.2 MPa followed by incubation in darkness for 10 days. Chemical name used: thidiazuron (TDZ).

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Ted E. Bilderback, Stuart L. Warren, James S. Owen Jr., and Joseph P. Albano

Many research studies have evaluated potential organic and mineral container substrate components for use in commercial potting substrates. Most studies report results of plant growth over a single production season and only a few include physical properties of the substrates tested. Furthermore, substrates containing predominantly organic components decompose during crop production cycles producing changes in air and water ratios. In the commercial nursery industry, crops frequently remain in containers for longer periods than one growing season (18 to 24 months). Changes in air and water retention characteristics over extended periods can have significant effect on the health and vigor of crops held in containers for 1 year or more. Decomposition of organic components can create an overabundance of small particles that hold excessive amounts of water, thus creating limited air porosity. Mineral aggregates such as perlite, pumice, coarse sand, and calcined clays do not decompose, or breakdown slowly, when used in potting substrates. Blending aggregates with organic components can decrease changes in physical properties over time by dilution of organic components and preserving large pore spaces, thus helping to maintain structural integrity. Research is needed to evaluate changes in container substrates from initial physical properties to changes in air and water characteristics after a production cycle.

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Christopher L. Owens, Michael F. Thomashow, James F. Hancock, and Amy F. Iezzoni

Orthologs of CBF1, a cold-induced transcription factor important in the cold acclimation response in Arabidopsis thaliana were cloned from strawberry (Fragaria × ananassa Duchesne) and sour cherry (Prunus cerasus L.) with degenerate PCR primers. The putative orthologs [Fragaria ×ananassa CBF1 (FaCBF1) and Prunus cerasus CBF1 (PcCBF1)] have 48% amino acid identity to CBF1 and mRNA levels were up-regulated in leaves of both crops following exposure to 4 °C from 15 minutes to 24 hours. However, mRNA of FaCBF1 and PcCBF1 was not detected in pistils of strawberry and sour cherry following 4 °C exposure. Agrobacterium-mediated transformation of a CaMV35S-CBF1 construct was conducted on Fragaria ×ananassa `Honeoye' crown discs. Two transgenic lines were regenerated that expressed the transgene at low levels in both leaves and receptacles. Receptacles of the transgenic lines showed no significant change in freezing tolerance when compared to wild type plants, although the temperature at which 50 % electrolyte leakage occurred in detached leaf discs from the two transgenic lines was -8.2 °C and -10.3 °C, respectively. These freezing tolerance values were significantly greater than the value for the wild-type `Honeoye' leaf discs of -6.4 °C.

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Q. Liu, S. Salih, J. Ingersoll, R. Meng, L. Owens, and F. Hammerschlag

Transgenic `Royal Gala' apple (Malus × domestica Borkh.) shoots were obtained by Agrobacterium-mediated gene transfer using the plasmid binary vector pGV-osm-AC with a T-DNA encoding a chimeric gene consisting of a secretory sequence from barley-amylase joined to the modified cecropin MB39 coding sequence. Shoots were placed under the control of a wound-inducible, osmotin promoter from tobacco. The integration of the cecropin MB39 gene into apple was confirmed by Southern blot analysis. The transformation efficiency was 1.5% when internodes from etiolated shoots were used as explants and 2% when leaf explants were used. Both non- and transgenic tetraploid plants were produced by treatment of leaf explants with colchicine at 25 mg·L-1, and polyploidy was confirmed by flow cytometry. Of the diploid transgenics, three of seven were significantly more resistant to Erwinia amylovora than the non-transgenic `Royal Gala' control. Also, in one instance, a tetraploid transgenic was significantly more resistant than the diploid shoot from which it was derived.

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Jeb S. Fields, William C. Fonteno, Brian E. Jackson, Joshua L. Heitman, and James S. Owen Jr.

Pine tree substrates (PTSs) may provide growers with sustainable substrate component options. Improved processing of PTS components has provided new materials with little scientific evaluation or understanding of their hydrophysical behavior and properties. Moisture retention characteristics were developed for two PTSs and four traditional greenhouse components: sphagnum peat, coconut coir, perlite, pine bark, shredded-pine-wood (SPW), and pine-wood-chips (PWC). Mixtures of peat containing 10%, 20%, 30%, 40%, and 50% of perlite, SPW, or PWC were also characterized. Hydrophysical properties were measured, allowing for comparison of the PTS components to the more traditional substrate components (peat, coir, perlite, and pine bark). The SPW was constructed to retain water similarly to peat and pine bark, whereas the PWC was made to increase drainage like perlite. Shredded pine wood had higher total porosity and more easily available water than did PWC components. Total porosities of SPW and PWC were similar to pine bark and coir; air space and drainage were higher than peat and coir because of the lower percentage of fine particles in the PTS components. The two PTS components had a greater influence on water drainage and retention dynamics than did perlite when amended with peat as an aggregate. Water release patterns of SPW or PWC components at low tensions were lower than peat and greater than pine bark; drainage was similar to perlite at higher tensions. Equilibrium capacity variable models predicted similar physical properties (and trends) across multiple container sizes for peat mixes amended with perlite, SPW, or PWC. The impact of PWC on drainage and aeration was similar to perlite in all containers, but these effects were greater in smaller containers.

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James S. Owen Jr, Stuart L. Warren, Ted E. Bilderback, and Joseph P. Albano

Production of containerized nursery crops requires high inputs of water and mineral nutrients to maximize plant growth to produce a salable plant quickly. However, input efficiencies remain below 50% resulting in major quantities of water and nutrients leached. This study was conducted to determine if production factors could be altered to increase water and phosphorus uptake efficiency (PUE) without sacrificing plant growth. The effects of a pine bark substrate amendment (clay or sand) and a 50% reduction in both P application rate (1.0 g or 0.5 g) and leaching fraction (LF = effluent ÷ influent) (0.1 or 0.2) were investigated. Containerized Skogholm cotoneaster (Cotoneaster dammeri Schnied. ‘Skogholm’) was grown on gravel floor effluent collection plots that allowed for calculation of water and nutrient budgets. Pine bark amended with 11% (by vol.) Georgiana 0.25 to 0.85 mm calcined palygorksite-bentonite mineral aggregate (clay) increased available water 4% when compared with pine bark amended with 11% (by volume) coarse sand. Decreasing LF from 0.2 to 0.1 reduced cumulative container influent 25% and effluent volume 64%, whereas total plant dry weight was unaffected by LF. Reduction of target LF from 0.2 to 0.1 reduced dissolved reactive P concentration and content by 8% and 64%, respectively. In a sand-amended substrate, total plant dry weight decreased 16% when 1.0× P rate was reduced to 0.5× P, whereas total plant dry weight was unaffected by rate of P when pine bark was amended with clay. Plant content of all macronutrients, with the exception of N, increased when pine bark was amended with clay versus sand. Reducing P rate from 1.0× to 0.5× increased PUE 54% or 11% in a clay or sand-amended substrate, respectively. Amending pine bark with 11% (by volume) 0.25 to 0.85 mm calcined palygorksite-bentonite mineral aggregate produced an equivalent plant with half the P inputs and a 0.1 LF, which reduced water use 25% and P effluent losses 42% when compared with an industry representative substrate [8 pine bark : 1 sand (11% by volume)].

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Chad E. Finn and John R. Clark