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Ryan R.P. Noble, C.S. Charron and C.E. Sams

The development of alternative methods for control of soilborne pathogens is imperative since the U.S. Clean Air Act bans the use of methyl bromide after 2005. One possibility is to exploit the pesticidal properties of compounds released by macerated Brassica tissues. In this study, masked chaffer beetle larvae were placed in sealed 473-mL jars with 335 g of soil amended with 1%, 2%, 4%, or 8% (g·g–1) Brassica tissue. The most prevalent volatile toxic compound of Brassica juncea (PI 458934) is allyl isothiocyanate (AITC). AITC production was measured in the jars at 0.25, 4, 8, 24, and 48 h using a solid-phase microextraction device (SPME) and gas chromatography. After 7 days, larvae mortality was determined. Control treatments included untreated soil, soil amended with 8% tomato plant tissue, soil amended with pure AITC, and untreated soil with an atmosphere of ≈20% O2 and 0% CO2 changing over 48 h to 2% O2 and 20% CO2. AITC levels were positively correlated to larvae mortality. The estimated lethal concentration for 50% kill (LC50) was 3.6 μg AITC/L soil atmosphere. AITC levels may be influenced by Brassica mass added, soil bulk density, and environmental factors including temperature and moisture. B. juncea has a high tissue AITC concentration. However, the mass of Brassica tissue required for insecticidal application against Cyclocephala sp. is also high, between 4% and 8% of soil mass. Development and selection of Brassica species that produce higher concentrations of isothiocyanate would increase the effectiveness of Brassica biofumigation as an alternative to methyl bromide for controlling soilborne insects.

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Usha Palaniswamy, Richard McAvoy and Bernard Bible

Watercress plants were grown in growth chambers at 15°C or 25°C and either an 8- or 12-hour photoperiod (PP). The photosynthetic photon flux (PPF) was 265 μmol·m–2·s–1 in all chambers, but beginning 1 week before harvest, half of the plants in each chamber were subjected to a higher PPF (434 μmol·m–2·s–1). At harvest, watercress leaves and stems were analyzed for phenethyl isothiocyanate (PEITC) content. Watercress grown at 25°C, the 12-hour PP, and 1 week of high PPF produced the highest PEITC concentration in leaves and stems, and plants grown at 15°C, the 8-hour PP, and the low PPF until harvest produced the lowest PEITC concentration. Plants grown at the 8-hour PP, then exposed to 1 week of high PPF, produced 57.3% and 45.9% greater PEITC at 25 and 15°C, respectively, then plants exposed to the low PPF until harvest. However, plants grown at the 12-hour PP and subjected to 1 week of high PPF produced PEITC levels similar to plants grown under the low PPF at 25 and 15°C. At 25°C, plants grown under the low PPF and the 12-hour PP produced 62% greater dry mass than plants exposed to 1 week of high PPF and the 8-hour PP, but did not differ in PEITC content. Thus, the effect of 1 week of high PPF on PEITC concentration depended on photoperiod.

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Veronica L. Justen, Jerry D. Cohen, Gary Gardner and Vincent A. Fritz

glucohydrolase, electrical conductivity (EC)], which hydrolyzes the thioester bond, releasing glucose and creating unstable byproducts such as isothiocyanates, thiocyanates, and nitriles ( Bones and Rossiter, 1996 ). Identifying vegetable production

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Veronica L. Justen and Vincent A. Fritz

correlated with cancer and disease prevention ( Hecht, 2000 ; Talalay and Fahey, 2001 ) Glucosinolates and their hydrolyzed derivatives, mainly isothiocyanates, have been shown to serve as chemopreventive agents by stimulating both protection from and

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Mark W. Farnham, Katherine K. Stephenson and Jed W. Fahey

Broccoli (Brassica oleracea L., Italica Group) has been recognized as a source of glucosinolates and their isothiocyanate metabolites that may be chemoprotective against human cancer. A predominant glucosinolate of broccoli is glucoraphanin and its cognate isothiocyanate is sulforaphane. Sulforaphane has been shown to be a potent inducer of mammalian detoxication (Phase 2) enzyme activity and to inhibit chemical-induced tumorigenesis in animal models. Little is known about phenotypic variation in broccoli germplasm for Phase 2 enzyme (e.g., quinone reductase) induction potential. Thus, this study was undertaken to evaluate: 1) quinone reductase induction potential (QRIP) diversity among a population of broccoli inbreds; 2) QRIP levels in selected lines; 3) correlation of QRIP with other horticultural characteristics; and 4) QRIP expression in a sample of synthesized hybrids. In 1996, 71 inbreds and five hybrid checks (all field-grown), ranged from a QRIP of nearly zero to 150,000 units/g fresh weight (FW) (mean of 34,020 units/g FW). These values were highly correlated with methylsulphinylalkyl glucosinolate (MSAG; primarily glucoraphanin) concentrations that ranged from 0.04 to 2.94 μmol.g-1 FW. A select subset of lines evaluated in 1996 were reevaluated in 1997. QRIP and MSAG values in this second year were similar to and correlated with those observed in 1996 (r = 0.73, P < 0.0001 and r = 0.79, P < 0.0001, respectively). In addition, both QRIP and MSAG concentration were highly correlated with days from transplant to harvest. Average F1 hybrid values for QRIP and MSAG in 1997 fell typically between their parental means, but were often closer to the mean of the low parent. Results of this study indicate that divergent QRIP expression can effectively be used to select enhanced inbred lines to use in development of value-added hybrids. Evidence is also provided that there is a significant genetic component to both QRIP and MSAG concentration, and that selection for either one may provide an effective means for developing broccoli hybrids with enhanced chemoprotective attributes. Chemical names used: 4-methylsulphinylbutyl glucosinolate (glucoraphanin) and 4-methylsulphinylbutyl isothiocyanate (sulforaphane).

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Usha Rani Palaniswamy, Richard McAvoy, Bernard Bible, Suman Singha and Dennis Hill

A study was initiated to identify cultural conditions that optimize the production of important chemopreventive agents in watercress. Chemopreventatives are chemical compounds that reduce or prevent diseases such as cancer. Watercress (Nasturtium officinale) contains phenylethyl glucosinolate that, on hydrolysis, yields PEITC, and PEITC is one of the most-important anti-carcinogens among the cruciferous chemopreventatives tested. Watercress was grown in closed hydroponic systems containing 200 ppm nitrogen and either 64, 128, and 192 ppm sulfur to yield N:S ratios of 1:0.3, 1:0.6, and 1:0.9. The experiment was laid out as RCBD in the greenhouse with six replications. PEITC levels in leaf and stem tissue was assayed using gas chromatograph. After 36 days in the treatment solutions, watercress grown at a N:S ratio of 1:0.6 produced 90.1% and 65.3% (in repeated experiments) more PEITC than plants grown at a N:S ratio of 1:0.3. Plants grown in nutrient solution with a N:S ratio of 1:0.9 produced 57.4% and 24.2% greater PEITC than those grown with a N:S ratio of 1:0.3. Plants grown in a nutrient solution with a N:S ratio of 1:0.9 produced 17.2% to 24.2% less PEITC than those grown with a N:S ratio of 1:0.6. Leaves contained 54% to 70% more PEITC per unit dry mass than stems, suggesting that the leaf is the major site of synthesis and storage of PEITC.

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G. Li, A. Riaz, S. Goyal, S. Abel and C.F. Quiros

Inheritance of three major genes involved in synthesis of aliphatic glucosinolates (GSL) was followed in segregating populations of Brassica oleracea L. generated from three crosses: broccoli × cauliflower, collard × broccoli, and collard × cauliflower. Two of these genes, GSL-PRO and GSL-ELONG, regulate sidechain length. The action of the former results in three-carbon GSL, whereas action of the latter produces four-carbon GSL. We determined that these two genes act and segregate independently from each other in B. oleracea. The double recessive genotype produces only trace amounts of aliphatic GSL. The third gene, GSL-ALK controls sidechain desaturation and, as it has been observed in Arabidopsis thaliana (L.) Heynh., we found that this gene cosegregates with a fourth gene, GSL-OH, that is responsible for sidechain hydroxylation. Elucidation of the inheritance of major genes controlling biosynthesis of GSL will allow for manipulation of these genes and facilitate development of lines with specific GSL profiles. This capability will be important for improvement of Brassica breeding lines with high content of desirable GSL, like glucoraphanin, a demonstrated precursor of anticarcinogenic compounds. Additionally, this work is the first step towards cloning the major genes of the aliphatic GSL pathway, and to use these clones in transformation strategies for further crop enhancement.

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Theodore J.K. Radovich, Matthew D. Kleinhenz and John G. Streeter

To better understand the influence of environmental factors on components of crop productivity and nutritional and sensory quality parameters, the fresh-market cabbage (Brassica oleracea L. Capitata Group) `Bravo' was irrigated at different periods relative to head development in 2002 and 2003 at the Ohio Agricultural Research and Development Center in Wooster. Irrigation was provided to plots either: 1) from planting to maturity, 2) during frame development only, or 3) during head development only. Control plants received no irrigation after plant establishment. Irrigation timing relative to crop stage significantly affected all head characteristics with the greatest differences between cabbage receiving irrigation during head development and cabbage not irrigated during head development. On average, heads from cabbage irrigated during head development were heavier, larger, less pointed, and had less volume occupied by the core than heads from cabbage not irrigated during head development. A positive, linear relationship (r 2 = 0.89) was found between head volume and head weight. Across years, combined head fructose and glucose concentrations were significantly greater and sucrose concentrations significantly lower in cabbage receiving irrigation during head development than in cabbage not irrigated during head development. Total and individual glucosinolate levels were greater in cabbage not irrigated during head development relative to cabbage receiving irrigation during head development. Head weight, fructose and glucose were positively related to the proportion of estimated crop evapotranspiration replaced by irrigation during head development, while the opposite response was observed in head sucrose and total and indole glucosinolate concentrations.

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Brian A. Kahn, Niels O. Maness, Donna R. Chrz and Lynda K. Carrier

identified by Friis and Kjaer (1966) as a glucoside undergoing rapid enzymatic hydrolysis to 4-methylthio-3- trans -butenyl isothiocyanate, abbreviated MTBITC. There is some evidence that levels of glucosinolates in Brassicaceae vegetables can be influenced

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James P. Gilreath, Bielinski M. Santos and Timothy N. Motis

allyl isothiocyanate (AITC) break down in the soil, producing isothiocyanate, which is a potent biocide ( Ajwa et al., 2003b ; Rodriguez-Kabana (2005) ). Commercially, dazomet is mostly sold as solid granules and powder, which must be incorporated into