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D.P. Miller, G.S. Howell and J.A. Flore

Chambers were constructed to measure gas exchange of entire potted grapevines (Vitis vinifera L.). The plant enclosures were constructed from Mylar film, which is nearly transparent to photosynthetically active radiation. Maintaining a slight, positive, internal pressure allowed the Mylar chambers to inflate like balloons and required no other means of support. The whole-plant, gas-exchange chamber design and construction were simple and inexpensive. They were assembled easily, equilibrated quickly, and did not require cooling. They allowed for the measurement of many plants in a relatively short period. This system would enable the researcher to make replicated comparisons of treatment influences on whole-plant CO2 assimilation throughout the growing season. While CO2 measurement was the focus of this project, it would be possible to measure whole-plant transpiration with this system.

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David P. Miller, G. Stanley Howell and James A. Flore

The measurement of whole-plant CO2 uptake integrates leaf-to-leaf variability, which arises from such sources as angle of incident radiation, source/sink relationships, age, and biotic or abiotic factors. Respiration of above-ground vegetative and reproductive sinks is also integrated into the final determination of whole-plant CO2 assimilation. While estimates of whole-plant CO2 uptake based on single-leaf determinations have been used, they do not accurately reflect actual whole-plant assimilation. Chambers were constructed to measure gas exchange of entire potted grapevines. The design and construction are simple, inexpensive, and easy to use, allowing for the measurement of many plants in a relatively short time. This enables the researcher to make replicated comparisons of the whole-plant CO2 assimilation of various treatments throughout the growing season. While CO2 measurement was the focus of this project, it is also possible to measure whole-plant transpiration with this system.

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G.C. Martin, A.N. Miller, L.A. Castle, J.W. Morris, R.O. Morris and A.M. Dandekar

The udiA gene encoding the enzyme β -glucuronidase (GUS) appears promising as a genetic marker for early confirmation of successful plant cell transformation. Two strains of Agrobacterium rhizogenes and eight strains of A. tumefaciens were selected as hosts to carry a binary plasmid (pBI121) containing the marker gene encoding the GUS marker that is controlled by the CaMV35S promoter. Presence of plasmid pBI121 in the bacteria was confirmed by resistance to kanamycin, plasmid re-isolation, and restriction enzyme analysis. When the GUS enzyme was expressed in transformed plant cells, reaction with the histochemical substrate 5-bromo-4-chloro-3-indolylglucuronide (X-gluc) lead to blue pigment development. Expression of GUS in viable bacteria that had not been eliminated from transformed tissue caused problems with the early transformation detection in radish, peach, and apple stem sections by also producing a positive X-gluc color reaction. Putative transformation of apple xylem parenchyma callus was accomplished, as judged by resistance to kanamycin, opine analysis, GUS marker gene expression, and presence of the APH(3')11 enzyme. In this system, elimination of bacterial contamination was accomplished during multiple culture transfers on selective media. To be more useful as a marker, the GUS gene should be coupled with a promoter that will not be expressed by Agrobacterium. Parenchyma callus may serve as a primary screen to provide an efficient way of determining the ideal strain for transformation of deciduous tree fruit genera. In our studies, strain A281 consistently proved to be a vector superior to others tested.

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Joshua H. Freeman, G.A. Miller, S.M. Olson and W.M. Stall

As triploid watermelons (Citrullus lanatus) increase in popularity, production has shifted away from seeded watermelons. To achieve successful fruit set in triploid watermelons, a diploid watermelon cultivar must be planted as a pollen source. Three diploid cultivars in 2005 and seven diploid cultivars in 2006 were evaluated at one and three locations, respectively, to determine their effectiveness as pollenizers. Each cultivar was planted within plots of the triploid watermelons ‘Tri-X 313’ (2005) and ‘Supercrisp’ (2006) with buffers on all sides of the plots to contain pollen flow within individual plots. Performance of pollenizers was based on triploid watermelon yield, soluble solids concentration, and incidence of hollowheart. In 2005, there were no significant differences in total weight, fruit per acre, average weight, or soluble solids concentration among pollenizers. In 2006, significant differences in yield were observed, and plots with ‘Sidekick’ as a pollenizer yielded the highest but were not significantly different from ‘Patron’, ‘SP-1’, ‘Jenny’, or ‘Mickylee’. In 2006, there were no significant differences in fruit per acre, soluble solids concentration, or incidence of hollowheart between pollenizers. The experimental design was successful in isolating pollenizers and there was minimal pollen flow outside of experimental plots as indicated by minimal fruit set in control plots.

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Aparna Gazula, Matthew D. Kleinhenz, John G. Streeter and A. Raymond Miller

Pigment concentrations in leaf tissue affect the visual and nutritional value-based indices of lettuce crop quality. To better discern the independent and interactive effects of temperature and cultivar on anthocyanin and chlorophyll b concentrations, three closely related Lolla Rosso lettuce cultivars (`Lotto', `Valeria', and `Impuls'), varying primarily in the number of genes controlling anthocyanin concentrations, were subjected to different air temperatures in controlled environments. Fifteen-day-old seedlings previously grown at 20 °C day/night (D/N) were transplanted into growth chambers maintained at 20 °C (D/N), 30/20 °C D/N and 30 °C D/N air temperatures. Twenty days later, leaf tissue was sampled for measures of pigment concentrations, calculated based on spectrophotometric absorbance readings taken at 530 nm (anthocyanin) and 660 nm (chlorophyll b) respectively. Although significant, the temperature × cultivar interaction resulted from differences in the magnitude (not direction) of the change in pigment concentrations among cultivars with changes in temperature. Regardless of cultivar, anthocyanin and chlorophyll b concentrations were highest, moderate and lowest after growth at 20 °C D/N, 30/20 °C D/N and 30 °C D/N respectively. Likewise, irrespective of temperature, anthocyanin and chlorophyll b concentrations followed the pattern `Impuls' (three genes) > `Valeria' (two genes) > `Lotto' (one gene). These data provide additional strong evidence that lettuce leaf pigment concentrations and growing temperatures are negatively related. The data also suggest that low temperatures during the dark phase may mitigate high temperature-driven reductions in lettuce leaf pigment levels.

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Theodore J.K. Radovich, Matthew D. Kleinhenz, John G. Streeter, A. Raymond Miller and Joseph C. Scheerens

Glucosinolates are secondary plant metabolites derived from amino acids and they influence human health, pest populations and crop flavor. Our primary objective was to determine the independent and interactive effects of planting date (PD) and cultivar (C) on total glucosinolate concentrations in cabbage, in part to help develop management systems that optimize them. A second objective was to explore the reported link between total glucosinolate concentrations and pungency in fresh cabbage. Six commercial fresh market cabbage cultivars were planted in May and June 2001 and 2002 at the Ohio Agricultural Research and Development Center (OARDC) Vegetable Crops Research Branch in Fremont, Ohio. Total glucosinolate concentrations in horticulturally mature heads were determined using a glucose evolution procedure. In 2001, 12 to 14 experienced panelists also scored sample pungency. Total glucosinolate concentrations were significantly affected by PD and C, but the PD × C interaction was not significant. Mean glucosinolate concentrations were greater in Maythan June-planted cabbage in both years. Cultivar ranking with regard to glucosinolate concentrations was similar between planting dates in both years. `Cheers' had the highest mean glucosinolate concentrations (23.1 and 29.5 mmol·kg-1 dry weight in 2001 and 2002, respectively) and `Solid Blue 790' the lowest (17.1 and 19.7 mmol·kg-1 dry weight in 2001 and 2002, respectively). In 2001, panelists generally scored cultivars highest in glucosinolates as more pungent than cultivars lowest in glucosinolates. These data suggest that planting date and cultivar effects on total glucosinolate concentrations in cabbage are largely independent. Climatic data suggest that higher air temperatures during head development of May-compared to June-planted cabbage induced plant stress and resulted in higher glucosinolate concentrations in May-planted cabbage.

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J.W. Scott, D.M. Francis, S.A. Miller, G.C. Somodi and J.B. Jones

Crosses were made between tomato (Lycopersicon esculentum Mill.) inbreds susceptible to races T2 and T3 of bacterial spot (Xanthomonas vesicatoria and Xanthomonas campestris pv. vesicatoria, respectively) and accession PI 114490 with resistance to races T1, T2, and T3. Resistance to race T2 was analyzed using the parents, F1, and F2 generations from one of the crosses. The F1 was intermediate between the parents for disease severity suggesting additive gene action. The segregation of F2 progeny fit a two-locus model (χ2 = 0.96, P = 0.9-0.5) where four resistance alleles are required for a high resistance level, two or three resistance alleles provide intermediate resistance, and zero or one resistance allele results in susceptibility. The narrow sense heritability of resistance to T2 strains was estimated to be 0.37 ± 0.1 based on F2 to F3 parent-offspring regression. A second cross was developed into an inbred backcross (IBC) population to facilitate multilocation replicated testing with multiple races. Segregation for T2 resistance in the inbred backcross population also suggested control was by two loci, lending support to the two-locus model hypothesized based on the F2 segregation. To determine if the same loci conferred resistance to the other races, selections for race T2 resistance were made in the F2 and F3 generations and for race T3 resistance in the F2 through F4 generations. Six T3 selections (F5), 13 T2 selections (F4's that diverged from seven F2 selections), and control lines were then evaluated for disease severity to races T1, T2, and T3 over two seasons. Linear correlations were used to estimate the efficiency of selecting for resistance to multiple races based on a disease nursery inoculated with a single race. Race T1 and race T2 disease severities were correlated (r ≥ 0.80, P< 0.001) within and between years while neither was correlated to race T3 either year. These results suggest that selecting for race T2 resistance in progeny derived from crosses to PI 114490 would be an effective strategy to obtain resistance to both race T1 and T2 in the populations tested. In contrast, selection for race T3 or T2 will be less likely to result in lines with resistance to the other race. PI 114490 had less resistance to T3 than to T2 or T1. Independent segregation of T2 and T3 resistance from the IBC population derived from PI 114490 suggests that T3 resistance is not controlled by the same genes as T2 resistance, supporting the linear correlation data.

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A.G. Gillaspie, O.L. Chambliss, R.L. Fery, A.E. Hall, J.C. Miller Jr. and T.E. Morelock

The Vigna Crop Germplasm Committee has established a core subset for the USDA cowpea germplasm collection. The subset consists of 9.3% (700 accessions) of the 7525 accessions currently contained in the collection. The subset was selected on the basis of country of origin, taxonomic characteristics, and known disease and pest resistance characteristics. Theoretically, the lines in the subset represent the genetic diversity present in the entire collection. A listing of the accessions in the subset is available from the Vigna germplasm curator (A.G. Gillaspie). The listing can also be accessed through the USDA's Germplasm Resources Information Network (GRIN).

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J.W. Scott, S.A. Miller, R.E. Stall, J.B. Jones, G.C. Somodi and V. Barbosa

Thirty-three tomato (Lycopersicon esculentum Mill.) or L. pimpinellifolium (L.) Mill. accessions were inoculated with race T2 of Xanthomonas campestris pv. vesicatoria (Xcv) in a field experiment at Wooster, Ohio, in Summer 1995. These included accessions selected for race T2 resistance in greenhouse tests in Florida, and accessions from Hawaii, Brazil, and Bulgaria. One L. esculentum (PI 114490-1-1) and three L. pimpinellifolium (PI 340905-S1, PI 128216-T2, and LA 442-1-BK) accessions had no Xcv symptoms. This is the first report of resistance to Xcv race T2. Partial resistance was found in PI 271385, PI 79532-S1, PI 155372-S1, PI 195002, and PI 126428. Most of the 33 genotypes were tested for race T1 resistance in Presidente Prudente, Sao Paulo, Brazil in summer 1993. Hawaii 7983, PI 155372-S1, PI 114490, PI 114490-S1, and PI 262173 had greater resistance to T1 than the susceptible control `Solar Set'. Comparisons with earlier experiments in which accessions were inoculated with race T1 or T3 indicated that the most consistent source of resistance to all three races was PI 114490 or selections from it.

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J.W. Scott, S.A. Miller, R.E. Stall, J.B. Jones, G.C. Somodi, V. Barbosa, D.L. Francis and F. Sahin

Thirty-two tomato (Lycopersicon esculentum Mill.) or L. pimpinellifolium (L.) Mill. accessions were inoculated with race T2 of Xanthomonas campestris pv. vesicatoria (Xcv) in a field experiment at Wooster, Ohio, in 1995. Plants from accessions which segregated for race T2 resistance in greenhouse tests were selected and these are designated by hyphenated extensions below. The eight most resistant accessions from 1995 and PI 262173 were retested in 1996. Lycopersicon esculentum accession PI 114490-1-1 had virtually no Xcv symptoms either year. Lycopersicon pimpinellifolium accessions LA 442-1-Bk and PI 128216-T2 expressed a high level of resistance in 1995, but only partial resistance in 1996. Accessions with partial resistance for both seasons were PI 79532-S1, PI 155372-S1, PI 126428, PI 271385, PI 195002, PI 262173, Hawaii 7998, and Hawaii 7983. PI 79532-S1 is a L. pimpinellifolium accession and the remaining seven are L. esculentum. Twenty accessions tested in 1995 for T2 plus 10 other accessions were also tested for race T1 resistance in Presidente Prudente, Sao Paulo, Brazil, in 1993. Hawaii 7983, PI 155372-S1, PI 114490, PI 114490-S1, and PI 262173 had greater resistance to T1 than the susceptible control, `Solar Set'. Comparisons with earlier experiments, in which accessions were inoculated with race T1 or T3, indicated that the most consistent source of resistance to all three races was PI 114490 or selections derived from it.