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Anna L. Hale and Mark W. Farnham

Over the last 3 decades, broccoli (Brassica oleracea L., Italica Group) hybrids made by crossing two inbred lines replaced open-pollinated populations to become the predominant type of cultivar. The change to hybrids evolved with little or no understanding of heterosis or hybrid vigor in this crop. Therefore, the purpose of the present study was to determine levels of heterosis expressed by a set of hybrids derived by crossing relatively elite, modern inbreds (n = 9). A total of 36 hybrids formed by crossing nine parents were evaluated for horticultural characters, including head weight, head stem diameter, plant height, plant width (in a row), and maturity (e.g., days from transplant to harvest) in four environments. When averaged across all four environments, roughly half of the hybrids exhibited high parent heterosis for head weight (1 to 30 g) and stem diameter (0.2 to 3.5 cm). Almost all hybrids showed high parent heterosis for plant height (1 to 10 cm) and width (2 to 13 cm). Unlike other traits, there was negative heterosis for maturity, indicating that heterosis for this character in hybrids is expressed as earliness. With modern broccoli inbreds, heterosis for head characteristics appears less important than for traits that measure plant vigor.

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Anna L. Hale, Mark W. Farnham and Monica A. Menz

Breeders of cole crops (Brassica oleracea L.) have an interest in utilizing current and emerging PCR-based marker systems to differentiate elite germplasm. However, until efficiency and cost-effectiveness are determined, most breeders are hesitant to change methods. In this study, our goal was to compare simple sequence repeat (SSR), amplified fragment-length polymorphism (AFLP), and sequence-related amplified polymorphism (SRAP) marker systems for their effectiveness in differentiating a diverse population of 24 elite broccoli (B. oleracea Italica Group) inbreds. Published SSR primer sequences for Brassica L. species were used along with AFLP and SRAP primer combinations. Several SSR primers failed to amplify DNA in the broccoli population, but all AFLP and SRAP primer combinations produced multiple bands. Twenty-nine percent of the SSR primers were monomorphic, while most of the remaining primers detected only one or two differences among inbreds. AFLP and SRAP methods produced multiple differences per primer in almost every case. Phenetic analysis revealed that the type of marker affected the classification of the genotypes. All three marker systems were able to successfully differentiate between the 24 elite inbreds, however, AFLPs and SRAPs were more efficient, making them better alternatives than SSRs over other established methods for fingerprinting B. oleracea inbreds.

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Lavanya Reddivari*, Anna L. Hale, Douglas C. Scheuring and J. Creighton Miller Jr.

In recent years, much emphasis has been placed on functional/antioxidant properties of various fruits and vegetables and their contribution to human health. Since average per capita consumption of potatoes in the United States is about 137 pounds, even moderate levels of antioxidants could be viewed as an important human health benefit. Variation in antioxidant activity has not been extensively investigated for colored potatoes (specialty selections). Therefore, the objective of this study was to evaluate antioxidant activity of specialty selections from the Texas Potato Variety Development Program and identify elite lines to use in breeding for improvement of this trait. Potato tubers were also assessed for their outer appearance, skin color, flesh color, spoilage and yield characteristics. Specialty potato selections (320 lines) were screened for total antioxidant activity using the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assay. After an initial screening, the top 10 % of selections were reevaluated in the following year. Significant (P ≤ 0.01) differences were found among selections and, for some selections, differences were found between seasons. Total antioxidant activity ranged from 27 μg/gfw to 832 μg/gfw. The specialty selection CO112F2-2 (purple flesh) had the highest antioxidant activity (832 μg/gfw) irrespective of season. In most cases, purple flesh selections produced the highest antioxidant activity, probably due to the presence of anthocyanins, followed by yellow selections.

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M. Ndambe Nzaramba, Anna L. Hale, Douglas C. Scheuring and J. Creighton Miller Jr.

The inheritance of antioxidant activity (AOA) and its association with seedcoat color was investigated in cowpea [Vigna unguiculata (L.) Walp.]. Four advanced cowpea lines, ARK95-356 (black seedcoat) and ARK98-348 (red seedcoat), which were high (H) in AOA, and ARK96-918 (cream seedcoat) and LA92-180 (cream seedcoat), which were low (L) in AOA, were selected from the 2002 Regional Southernpea Cooperative Trials. They were crossed in a complete diallel mating design, generating F1, F1′ (1st generation and 1st generation reciprocal cross, respectively), F2, F2′ (2nd generations from F1, F1′), BC1, and BC2 (backcrosses to parents 1 and 2, respectively) populations. Individual seeds were ground and samples were extracted in methanol and analyzed for AOA using the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. Combining ability tests using Griffing's Method I Model I indicated presence of highly significant general combining ability (GCA), specific combining ability (SCA), and reciprocal (RE) and maternal (MAT) effects, with pigmented lines exhibiting positive GCA and MAT, while nonpigmented lines exhibited negative GCA and MAT. AOA in the F1 was not significantly different from the maternal parent, with seedcoat color also resembling the maternal parent. Segregation for seedcoat color was observed in the F2 and F2′. Additive, dominance, and epistatic effects were significant. The broad sense heritability estimate was 0.87. Minimum number of genes responsible for AOA was estimated at five. Factors governing high AOA appeared to be the same as those responsible for seedcoat color, with apparent pleiotropic effects. In conclusion, breeding for high AOA in cowpea is possible using highly pigmented parental lines.

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Anna L. Hale, Douglas C. Scheuring, Thomas J. Gerik, Jeffrey D. Hart and J. Creighton Miller Jr.

Iron Deficiency Chlorosis (FeDC) is a problem in cowpea because it affects the ability of the plant to produce chlorophyll. Earlier studies indicated that FeDC was conditioned by a single gene. Pinkeye Purple Hull (PEPH), a susceptible variety, and Texas Pinkeye Purple Hull (TXPE), a resistant variety, were crossed and allowed to self for one generation. The F1s were backcrossed to the parents. SPAD readings were taken on each population. SPAD measures the transmission of light through the leaves at a wavelength where chlorophyll absorbs and a wavelength where it does not. The SPAD reading is calculated based on a ratio of these two numbers. Thus, the SPAD value is unitless and is an indication of the relative amount of chlorophyll present in the leaf. Chlorophyll was extracted from leaves, and regressed on the SPAD readings from the same leaves. An R 2 of .9102 was obtained as well as a regression equation of y = 12.8x + 54.5. Thus, a SPAD value of 1 corresponds with a chlorophyll content of ≈67.3 μg chlorophyll/gfw. The data was analyzed using a bootstrap method, and indicated that FeDC is not controlled by a single gene. A P-value of .0004 showed a highly significant difference between the expected and observed segregation ratios in the F2 plants. Narrow sense heritibility (Mather) was estimated at 0.3.

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Tyann Blessington*, Anna L. Hale, Douglas C. Scheuring and J. Creighton Miller Jr.

We have demonstrated that potatoes contain significant levels of antioxidants important to human health; however, since potatoes are not consumed raw, it is important to determine the effects of cooking/processing on these levels. Therefore, the changes in phenolic and carotenoid content and total antioxidant activity in potatoes were investigated using combinations of storage and cooking methods. Fresh and stored tubers (110 days at 4 °C) of 17 potato cultivars, both raw and cooked (microwaved, boiled, baked, fried), were analyzed for antioxidant activity using the DPPH method. In addition, carotenoid levels were determined for each treatment based on the absorbance of the methanol extraction (oxygenated phenolics and carotenoids) at 445 nm and the hexane extraction (non-oxygenated carotenoids) at 450 nm. Total antioxidant activity as well as carotenoid levels were significantly affected by both genotype and cooking method. Across extraction methods, the microwave and fry cooking treatments were generally highest in antioxidant activity, while boiling was the lowest. Oxygenated carotenoids were significantly affected by storage, while the non-oxygenated carotenoids were unaffected.

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Anna L. Hale, Mark W. Farnham, Michael A. Grusak and John W. Finley

Broccoli(Brassica oleracea L. Italica Group) can contain high levels of selenium (Se) in the form of selenium methyl selenocystine. This is a relatively unique Se compound that is found in certain plant species that accumulate this element. Several recent studies have shown that high Se broccoli can inhibit the development of certain cancers (e.g., colon and mammary) in rodents and this has led to increasing interest in broccoli as a vegetable that confers chemoprotective effects. The objective of this research was to determine the relative importance of genotype vs. environment in the expression of Se concentration in broccoli heads. A set of 15 broccoli inbreds and a set of 20 hybrids were evaluated in three different environments. Mature heads were harvested from plots, heads were dried and ground, and Se concentration was determined on a dry weight basis. Overall, Se levels measured in this study were low to moderate, typically ranging from about 20 to more than 100 ng/gdw of Se per head. For both inbreds and hybrids, the effect of environment on Se head concentration was highly significant and more than 10 times greater than the effect of genotype. When analyzed across all three environments, the genotypic effect on Se concentration was significant for hybrids only. However, when assessed for individual environments, the genotypic effect was significant in just one of three of the test environments with both inbreds and hybrids. Results indicate that genetic modification of broccoli to increase selenium concentration of heads will likely be difficult to achieve.

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Anna L. Hale, J. Creighton Miller Jr., K. Renganayaki, Alan K. Fritz, J.J. Coombs, L.M. Frank and D.S. Douches

The objective of this study was to differentiate six intraclonal variants of the potato (Solanum tuberosum L.) cultivar Russet Norkotah. One-hundred-twelve AFLP primer combinations producing 3755 bands and 79 microsatellite primers producing over 400 bands failed to identify any reproducible polymorphisms among the intraclonal variants and `Russet Norkotah'. The inability to detect differences between clones underscores the degree of genetic similarity between them, despite differences in phenotypic expression. This inability could be due to the tetraploid nature of the clones and/or to epigenetic differences not detected by the utilized procedures.