Broccoli is well recognized as a source of glucosinolates and their isothiocyanate breakdown products. Glucoraphanin is one of the most abundant glucosinolates present in broccoli and its cognate isothiocyanate is sulphoraphane, a potent inducer of mammalian detoxication (phase 2) enzyme activity and anti‐cancer agent. This study was designed to measure: glucosinolate levels in broccoli florets from an array of genotypes grown in several environments; the elevation of a key phase 2 enzyme, quinone reductase, in mammalian cells exposed to floret extracts; and total broccoli head content. There were significant environmental and genotype‐by‐environment effects on levels of glucoraphanin and quinone reductase induction potential of broccoli heads; however, the effect of genotype was greater than that of environmental factors. The relative rankings among genotypes for glucoraphanin and quinone reductase induction potential changed, when expressed on a per head basis, rather than on a concentration basis. Correlations of trait means in one environment vs. means from a second were stronger for glucoraphanin and quinone reductase induction potential on a per head basis than on a fresh weight concentration basis. Results of this study indicate that development of a broccoli phenotype with a dense head and a high concentration of glucoraphanin to deliver maximum chemoprotective potential (high enzyme induction potential/glucoraphanin content) is a feasible goal.

Publicly available genomic tools help researchers integrate information and make new discoveries. In this paper, we describe the development of immortal mapping populations of rapid cycling, self-compatible lines, molecular markers, and linkage maps for Brassica rapa and B. oleracea and make the data and germplasm available to the Brassica research community. The B. rapa population consists of 160 recombinant inbred (RI) lines derived from the cross of highly inbred lines of rapid cycling and yellow sarson B. rapa. The B. oleracea population consists of 155 double haploid (DH) lines derived from an F1 cross between two DH lines, rapid cycling and broccoli. A total of 120 RFLP probes, 146 SSR markers, and one phenotypic trait (flower color) were used to construct genetic linkage maps for both species. The B. rapa map consists of 224 molecular markers distributed along 10 linkage groups (A1-A10) with a total distance of 1125.3 cM and a marker density of 5.7 cM/marker. The B. oleracea genetic map consists of 279 molecular markers and one phenotypic marker distributed along nine linkage groups (C1-C9) with a total distance of 891.4 cM and a marker density of 3.2 cM/marker. A syntenic analysis with Arabidopsis thaliana identified collinear genomic blocks that are in agreement with previous studies, reinforcing the idea of conserved chromosomal regions across the Brassicaceae.

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In broccoli, sulforaphane forms when the glucosinolate glucoraphanin is hydrolyzed by the endogenous plant thiohydrolase myrosinase. A myrosinase cofactor directs hydrolysis away from the formation of bioactive sulforaphane and toward an inactive product, sulforaphane nitrile. The cofactor is more heat sensitive than myrosinase, presenting an opportunity to preferentially direct hydrolysis toward sulforaphane formation through regulation of thermal processing. Four broccoli cultivars were microwave heated, boiled, or steamed for various lengths of time. Production of nitrile during hydrolysis of unheated broccoli varied among cultivars from 91 to 52% of hydrolysis products (Pinnacle > Marathon > Patriot > Brigadier). Boiling and microwave heating caused an initial loss of nitrile, with a concomitant increase in sulforaphane, followed by loss of sulforaphane, all within 1 min. In contrast, steaming enhanced sulforaphane yield between 1.0 and 3.0 min in all but Brigadier. These data are proof of concept that steaming for 1.0-3.0 min provides less nitrile and more sulforaphane yield from a broccoli meal.

We constructed a 1,257-marker, high-density genetic map of Brassica oleracea spanning 703 cM in nine linkage groups, designated LG1-LG9. It was developed in an F2 segregating population of 143 individuals obtained by crossing double haploid plants of broccoli "Early-Big" and cauliflower "An-Nan Early". These markers are randomly distributed throughout the map, which includes a total of 1,062 genomic SRAP markers, 155 cDNA SRAP markers, 26 SSR markers, 3 broccoli BAC end sequences and 11 known Brassica genes: BoGSL-ALK, BoGSL-ELONG, BoGSL-PROa, BoGSL-PROb, BoCS-lyase, BoGS-OH, BoCYP79F1, BoS-GT (glucosinolate pathway), BoDM1 (resistance to downy mildew), BoCALa, BoAP1a (inflorescence architecture). BoDM1 and BoGSL-ELONG are linked on LG 2 at 0.8 cM, making it possible to use the glucosinolate gene as a marker for the disease resistance gene. By QTL analysis, we found three segments involved in curd formation in cauliflower. The map was aligned to the C genome linkage groups and chromosomes of B. oleracea and B. napus, and anchored to the physical map of A. thaliana. This map adds over 1,000 new markers to Brassica molecular tools.

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.

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).

A collection of collard (Brassica oleracea L., Acephala group) germplasm, including 13 cultivars or breeding lines and 5 landraces, was evaluated using randomly amplified polymorphic DNA (RAPD) markers and compared to representatives of kale (Acephala group), cabbage (Capitata group), broccoli (Italica group), Brussels sprouts (Gemmifera group), and cauliflower (Botrytis group). Objectives were to assess genetic variation and relationships among collard and other crop entries, evaluate intrapopulation variation of open-pollinated (OP) collard lines, and determine the potential of collard landraces to provide new B. oleracea genes. Two hundred nine RAPD bands were scored from 18 oligonucleotide decamer primers when collard and other B. oleracea entries were compared. Of these, 147 (70%) were polymorphic and 29 were specific to collard. Similarity indices between collard entries were computed from RAPD data and these ranged from 0.75 to 0.99 with an average of 0.83. Collard entries were most closely related to cabbage (similarity index = 0.83) and Brussels sprouts entries (index = 0.80). Analysis of individuals of an OP cultivar and landrace indicated that intrapopulation genetic variance accounts for as much variation as that observed between populations. RAPD analysis identified collard landraces as unique genotypes and showed them to be sources of unique DNA markers. The systematic collection of collard landraces should enhance diversity of the B. oleracea germplasm pool and provide genes for future crop improvement.