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  • Author or Editor: Mark W. Farnham x
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Broccoli (Brassica oleracea L. Italica Group) is a rich source of the aliphatic glucosinolate glucoraphanin. The glucoraphanin breakdown product, sulforaphane, has been shown to induce Phase II detoxication enzymes (e.g., Quinone Reductase) and has attracted attention as a potential chemoprotector against cancer. The objectives of this research were to evaluate the concentration of glucoraphanin in an array of diverse broccoli inbreds (doubled-haploids) largely derived from commercial germplasm and to determine if expression of glucoraphanin level in this initial evaluation is correlated with expression in a subsequent environment. In 1996, individual florets from single broccoli heads were sampled from 75 inbred lines grown in the field at Charleston, S.C., and glucoraphanin concentration was assayed. In this test, concentrations ranged from 0.04 to 2.94 μmol glucoraphanin per g fresh weight of florets and the mean concentration was 0.86. In 1997, a subset of 22 inbreds analyzed the first year were grown again in a replicated field trial. This inbred subset was made up of lines with diverse pedigrees and with high, low, or intermediate glucoraphanin concentrations. In this second year, glucoraphanin concentration had a range from 0.24 to 2.99 μmol per g fresh weight of florets and a mean of 1.37. Correlation of entry mean glucoraphanin concentration in 1997 with that in 1996 was positive (r = 0.79) and highly significant (P < 0.001) indicating that floret glucoraphanin concentration was relatively consistent between years. These observations provide evidence that floret glucoraphanin concentration has a significant genetic component.

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Broccoli (Brassica oleracea L., Italica Group) seed and resulting sprouts can contain high levels of glucoraphanin, a glucosinolate, which can be converted to sulforaphane, a compound with cancer protective and antioxidant properties. This observation has stimulated interest in broccoli seed production. In this study, inbred lines, which produce relatively high yields of homogeneous, selfed-seed across different environments in the absence of insect pollinators, were used to evaluate the relative importance of genotype versus environment as a determinant of glucoraphanin concentration in broccoli seed. Glucoraphanin and glucoiberin were measured in broccoli seed lots generated from ten broccoli inbred lines grown in two greenhouse and two screen cage environments. Typically, seed glucoraphanin level ranged from 5 to 100 μmol·g-1 seed and glucoiberin ranged from 0 to about 40 μmol·g-1 seed, regardless of the environment in which seed was produced. Analysis of variance indicated that genotype was the most significant factor influencing levels of the two glucosinolates. Although significant environmental and genotype × environment effects were observed for glucoraphanin and a significant genotype × environment effect was observed for glucoiberin, these effects were small compared to the genotype effects. Results indicate that it is possible to identify broccoli inbreds that consistently produce relatively high yields of seed with a high glucoraphanin content across different environments.

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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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Broccoli (Brassica oleracea L., Italica Group) is a good vegetable source of Ca and Mg, two critical minerals in human nutrition. Studies have shown that bioavailability of Ca from broccoli is comparable to that from milk. Thus, broccoli is an important alternative source of Ca in segments of the population that consume limited amounts of dairy products. Essentially nothing is known about the genetic influence on variation in Ca or Mg concentration of broccoli heads. Thus, the goal of this research was to examine variation in Ca, and also Mg concentrations, in a collection of USDA inbreds and commercial F1 hybrids. In 1996 and 1997 field studies, significant differences among inbred entries and among hybrid entries were observed for Ca and Mg concentrations of broccoli heads. With hybrids and inbreds, mean head Ca concentrations were ≈3.0 mg·g-1 dry weight (DW), and entries with lowest and highest Ca concentrations differed >2-fold. Mean Mg concentrations of hybrid heads was 2.3 mg·g-1 DW (range 1.8 to 2.6) and 2.8 mg·g-1 DW (range 2.2 to 3.7) in 1996 and 1997, respectively. Inbred lines had mean head Mg concentrations of 2.0 and 2.6 mg·g-1 DW in the two respective years and ranges in concentration were similar as for hybrids. Analysis of variance indicated significant environment and entry by environment effects for Ca and Mg concentrations of hybrids. With inbreds, a significant entry by environment effect for Ca concentration and environment effect for Mg concentration was also observed. Significant environment and entry by environment effects indicate that the environmental influence on phenotypic expression of Ca and Mg concentrations may complicate genetic improvement of head mineral concentration.

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Downy mildew, incited by the biotrophic fungal parasite, Peronospora parasitica (Pers. Fr.) Fr., is one of the most destructive diseases of broccoli (Brassica oleracea L., Italica Group) and other related crop species throughout the world. Cultivation of resistant cultivars is the most desirable control method because it provides a practical, long-term, and environmentally benign means of limiting damage from this disease. The commercial hybrid cultivar, Everest, has been shown previously to contain a high level of downy mildew resistance. Doubled-haploid (DH) lines developed from that hybrid were also shown to exhibit a similar, high level of resistance at the three- to four-leaf stage. To determine the mode of inheritance of this true leaf resistance, the resistant DH line was crossed to a susceptible line (derived from `Marathon') to produce an F1 hybrid. Subsequently, F2 and backcross (BC) populations were developed from the hybrid. In addition, a DH population of ≈100 lines was developed from the same F1 used to create the F2 and BC. All populations were evaluated for response to artificial inoculation with P. parasitica at the three- to four-leaf stage. F1 plants were resistant like the resistant parent and F2 populations segregated approximately nine resistant to seven susceptible. Using the resistant parent as recurrent parent, BC populations contained all resistant plants, while the BC to the susceptible parent fit a 1 resistant: 3 susceptible segregation ratio. These results can be explained by a model with two complementary dominant genes. This model was confirmed by the DH population that segregated ≈1:3, resistant to susceptible. Due to the dominant nature of this resistance, controlling genes should be easily incorporated into F1 hybrids and used commercially to prevent downy mildew.

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Broccoli (Brassica oleracea L. Italica group) breeders routinely use anther or microspore culture to produce dihaploid (diploid), homozygous lines. During the culture process, polyploidization occurs and diploid regenerants can result. However, polyploidization may not occur at all, or it may involve a tripling or quadrupling of the chromosome complement. Thus, regenerated populations must be screened to identify the diploids that are the regenerants most likely to set seed and serve as inbred lines. DNA flow cytometry has proven a useful procedure for determining ploidy of anther derived regenerants. This study was undertaken to evaluate the effect of leaf age and sampling procedures on ploidy determination via flow cytometry. Anther-derived plants were analyzed at a four- to five-leaf stage (transplant stage) and at time of heading (mature plant stage). In addition, leaves were sampled on a given date and stability of the flow cytometry preparations was evaluated over 7 days. Lastly, the stability of ploidy readings of leaves stored at 4°C was examined over a 7-day period. In only one case out of 123 comparative assays did leaf age affect ploidy determination. For that exception, a haploid at transplant stage was a diploid at the mature plant stage. Flow cytometry preparations and also leaves stored at 4°C gave consistent ploidy determinations up to four days after preparations were made or tissue was refrigerated, respectively. These results indicate that broccoli breeders can make flow cytometry preparations on site and send them offsite for flow cytometry analysis. Alternatively, leaves could be refrigerated, sent offsite, and then prepared and analyzed at another location.

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Downy mildew, caused by Peronospora parasitica (Pers. Fr.) Fr., is one of the most economically important diseases of broccoli (Brassica oleracea L. Italica group). Previous studies have shown that resistance to downy mildew in broccoli is dependent on plant age with seedling resistance being independent of mature-plant resistance. The objectives of this study were to: 1) determine if valid evaluations for downy mildew resistance can be conducted at both the cotyledon and the three to four true-leaf stages on the same plants of a given broccoli entry; 2) determine if doubled-haploid (DH) lines derived from the resistant hybrid `Everest' also exhibit resistance to downy mildew and if so, characterize the resistance phenotype(s) in these lines; and 3) determine if identified resistant DH lines exhibit resistance to isolates of P. parasitica acquired from different geographic regions of the United States. Twenty-three DH broccoli inbreds and two commercial hybrids were evaluated for reaction at different developmental stages to infection by P. parasitica in a controlled environment. Results showed that broccoli plants can be evaluated for downy mildew resistance in a two-stage process. Inoculation at the cotyledon stage did not offer any cross-protection or otherwise influence the expression of reaction phenotype (RP) when the same plants were subsequently inoculated at the three to four true-leaf stage. Three different RPs to infection by P. parasitica were identified in DH inbreds. These were: 1) susceptibility at both the cotyledon stage and the true-leaf stage; 2) resistance at both the cotyledon and true-leaf stage; and 3) susceptibility at the cotyledon stage but resistance at the true-leaf stage. There was no effect of two pathogenic isolates from different geographic regions on RP of DH broccoli inbreds. Selection of plant resistance to downy mildew at the cotyledon stage will effectively identify plants with high levels of resistance at subsequent developmental stages.

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Selection of superior broccoli hybrids involves multiple considerations, including optimization of head quality traits. Quality assessment of broccoli heads is often confounded by relatively subjective human preferences for optimal appearance of heads. To assist the selection process, we assessed five candidate head quality indices that make use of a set of individual and distinct ratings for traits such as head color, head smoothness, bead size, bead uniformity, and others. The head quality indices were tested for both a) the ability to reduce interobserver rating variability and b) the ability to emphasize specific attributes that display the greatest associations with overall horticultural quality of heads. Index development was based on datasets generated from quality evaluations by three independent raters of two replicated variety trials in Spring 2014. Relative-importance analysis was used to identify specific traits most associated with overall quality. Developed models were subsequently tested and compared using data collected by three raters evaluating two similar trials in Spring 2015. Head smoothness, bead uniformity, head color, and holding ability were found to account for 78% of the model variation in overall head quality. Intraclass correlation coefficients (ICCs), which measure the degree of concordance among raters, were increased from 0.71 to 0.88 (P < 0.05) in one 2015 trial and from 0.67 to 0.80 (P < 0.05) in the second when comparing the simple overall quality assessment to the use of the index weighted by the most important individual head attributes. Thus, results showed that a quality index taking into account the relative importance of individual traits should enhance the identification of the best hybrids adapted to target conditions. This method can be used to improve concordance for subjective ratings in general.

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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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Brassica leafy greens (Brassica juncea L. and Brassica rapa L.) represent one of the most economically important vegetable crop groups in the southeastern United States. In the last 10 years, numerous occurrences of a leaf-spot disease on these leafy vegetables have been reported in several states. This disease, known as peppery leaf spot, is now causing serious crop losses and has been attributed to the bacterial phytopathogen Pseudomonas syringae pv. maculicola (Psm). To date, it appears that all cultivars of the Brassica leafy greens are susceptible, and available pesticides for control of this disease appear unable to reduce the disease to acceptable levels. Thus, we undertook a search for potential resistance to this disease among accessions of B. juncea and B. rapa included in the U.S. Plant Introduction (PI) collection. In greenhouse trials, we screened commercial cultivars and 672 U.S. PIs (226 B. juncea and 446 B. rapa) for resistance to Psm with artificial inoculation. Although severity of disease symptoms was significantly different among inoculated accessions, no acceptable levels of resistance were found in any of the more than 400 B. rapa accessions tested. Only two B. juncea accessions (PI 195553 and G 30988) of 226 tested had acceptable levels of resistance that might prove economically useful.

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