Broccoli Tissue Research Articles

Free Fatty Acid Determination in Broccoli Tissues Using Liquid Chromatography-High-Resolution Mass Spectrometry.

Broccoli (Brassica oleracea L. var. italica Plenck) is a widely consumed vegetable, very popular due to its various nutritional and bioactive components. Since studies on the lipid components of broccoli have been limited so far, the aim of the present work was the study of free fatty acids (FFAs) present in different broccoli parts, aerial and underground. The direct determination of twenty-four FFAs in broccoli tissues (roots, leaves, and florets) was carried out, using a liquid chromatography-high-resolution mass spectrometry (LC-HRMS) method in a 10 min single run. Linolenic acid was found to be the most abundant FFA in all different broccoli parts in quantities ranging from 0.76 to 1.46 mg/g, followed by palmitic acid (0.17-0.22 mg/g) and linoleic acid (0.06-0.08 mg/g). To extend our knowledge on broccoli's bioactive components, for the first time, the existence of bioactive oxidized fatty acids, namely hydroxy and oxo fatty acids, was explored in broccoli tissues adopting an HRMS-based lipidomics approach. 16- and 2-hydroxypalmitic acids were detected in all parts of broccoli studied, while ricinoleic acid was detected for the first time as a component of broccoli.

Determination of Phenethyl Isothiocyanate, Erucin, Iberverin, and Erucin Nitrile Concentrations in Healthy and Pieris rapae-Infected Broccoli Tissues Using Gas Chromatography-Mass Spectrometry

Cruciferous vegetables (Brassicaceae family) are a rich source of phytochemicals, in particular glucosinolates (GLS) and their hydrolysis products, isothiocyanates and nitriles. These phytochemicals may act as chemosensors, attracting insects, such as Pieris rapae, and stimulating oviposition. There is a lack of information on the concentrations of isothiocyanates and nitriles when an insect affects a Brassicaceae plant. In the current study, some GLS hydrolysis products were determined in healthy and Pieris rapae-infected organic cultivated broccoli plants, as well as the infesting insects’ larvae, using gas chromatography-mass spectrometry (GC-MS). This study investigated the following phytochemicals: phenethyl isothiocyanate (PEITC), erucin (ER), 3-(methylthio)propyl isothiocyanate (3MIC), and 1-cyano-4-(methylthio)butane (5MITN). All these components were quantified in the aerial and underground parts of the plants and were found in high concentrations in the roots. Among the phytochemicals studied, 5MITN presented the highest concentration in all the broccoli samples but was especially high in the stalks of the infected plants. Moreover, the analysis of a sample of Pieris rapae larvae, fed from the hosted broccoli, revealed the presence of PEITC and ER. These findings indicate that the infestation of broccoli with Pieris rapae may affect the distribution of PEITC, ER, 3MIC, and 5MITN throughout the plant. An extension of our study to conventional cultivated broccoli showed that the roots are indeed rich in GLS hydrolysis products.

Mechanical Digestion of Broccoli through Chewing and its Impact on Myrosinase Activity

Myrosinase activity on broccoli glucosinolates has been widely and extensively discussed. Purpose: Studies on cruciferous vegetables, especially broccoli, have gained significance in the fight against cancer. Glucosinolates in broccoli transformation into sulforaphane occur after its exposure through chewing. However, the relationship between individual chewing patterns and denture morphology has not been studied extensively. Research on human digestion has demonstrated how the mechanical breakdown of larger food particles into smaller ones is a crucial precursor to chemical food breakdown. This study is a comparative analysis of how chewing on broccoli tissues by different individuals enables the enzyme myrosinase to break down broccoli glucosinolates chemically. We investigate the individual chewing patterns linked to the surface anatomy of the pre-molars and molars and the myrosinase activity on broccoli glucosinolates. Methods: Three individuals chewed a 4-millimeter broccoli floret four times sequentially (2 grams), and we measured the floret length at each bite until the sample was ground. Then, we combined the chewed broccoli with distilled water, filtered it, and myrosinase activity was measured using photo spectrometric measurements and an agar diffusion test. Individual pre-molars and molars samples were measured in millimetres. Results: Data from the three human chewing mechanisms compared to the mechanical breakdown performed by an automatic mixer shows different and individually specific values. Conclusion: Individual chewing patterns link to the unique surface anatomy of the pre-molars and molars, subsequently impacting the myrosinase activity on broccoli glucosinolates’ breakdown individually

Evaluation of broccoli (Brassica oleracea var. italica) crop by-products as sources of bioactive compounds

This study was performed to evaluate different by-products of broccoli (Brassica oleracea var. italica) production (leaf, inflorescence and stem) as sources of valuable bioactive compounds, considering different cultivars and states of maturation. The total phenolic and chlorophyll contents as well as the antioxidant, ACE-inhibitory and antimicrobial activities were quantified from the ethanolic extracts of the different broccoli tissues at two different maturation stages from five cultivars (‘TSX 007′, ‘Monaco’, ‘BRO 2047′, ‘Parthenon’ and ‘Summer purple’). The major compounds in ethanolic extracts were identified by HPLC-UV-ESI-MS/MS, whereas chlorophylls were determined by UV–vis spectrometry. The leaf extracts showed the highest antioxidant activities and were the best sources of chlorophylls and phenolic compounds, constituting mainly kaempferol and quercetin glucosides. These compounds were more abundant in extracts from the ‘Summer Purple’ variety and commercial state. Glucosinolates were more abundant in the inflorescence extracts, principally in the flower bud state. The stem and inflorescence extracts, mainly from the ‘TSX 007′ variety, showed a strong inhibitory effect on the three bacteria studied (Bacillus cereus, Staphylococcus aureus and Listeria innocua), which was related to a higher concentration of fatty acid derivates. The findings suggest that broccoli by-products are useful and value-added products as sources of bioactive compounds, providing a sustainable alternative to reduce residues from this crop.

Exogenous Selenium Treatment Promotes Glucosinolate and Glucoraphanin Accumulation in Broccoli by Activating Their Biosynthesis and Transport Pathways

Supplementation using selenium (Se) on plants is an effective and widely used approach. It can not only be converted to more Se rich compounds but promote the accumulation of glucosinolates (GSLs) with anti-carcinogenic properties. However, the molecular mechanism of Se in regulating GSLs synthesis remains unclear. In the present study, we analyzed the effects of Se treatment (50 μM sodium selenite) on GSLs, glucoraphanin (4MSOB), and sulforaphane compounds in broccoli tissues. The transcript levels of genes involved in sulfur absorption and transport, GSLs biosynthesis, translocation, and degradation pathways were also evaluated. The study showed that Se treatment remarkably promoted the accumulation of total sulfur and total Se contents and increased Trp-derived GSLs levels in roots by 2 times. The 4MSOB concentration and sulforaphane content in fresh leaves was increased by 67% and 30% after Se treatment, respectively. For genes expressions, some genes involved in sulfate uptake and transporters, GSLs biosynthesis, and transporters were induced strongly upon Se exposure. Results revealed that exogenous Se treatment promotes the overaccumulation of GSLs and 4MSOB content in broccoli by activating the transcript levels of genes involved in sulfur absorption, GSLs biosynthesis, and translocation pathways.

Persistence of Fusarium oxysporum f. sp. fragariae in Soil Through Asymptomatic Colonization of Rotation Crops.

Asymptomatic plant colonization is hypothesized to enhance persistence of pathogenic forms of Fusarium oxysporum. However, a correlation between pathogen populations on living, asymptomatic plant tissues and soilborne populations after tillage has not been demonstrated. Living and dead tissues of broccoli, lettuce, spinach, wheat, cilantro, raspberry, and strawberry plants grown in soil infested with F. oxysporum f. sp. fragariae (the cause of Fusarium wilt of strawberry) were assayed to quantify the incidence of infection and extent of colonization by this pathogen. All crops could be infected by F. oxysporum f. sp. fragariae but the extent of colonization varied between plant species. Pathogen population densities on nonliving crown tissues incorporated into the soil matrix were typically greater than those observed on living tissues. Crop-dependent differences in the inoculum density of F. oxysporum f. sp. fragariae in soil were only observed after decomposition of crop residue. Forty-four weeks after plants were incorporated into the soil, F. oxysporum f. sp. fragariae soil population densities were positively correlated with population densities on plant tissue fragments recovered at the same time point. Results indicate that asymptomatic colonization can have a significant, long-term impact on soilborne populations of Fusarium wilt pathogens. Cultural practices such as crop rotation should be leveraged to favor pathogen population decline by planting hosts that do not support extensive population growth on living or decomposing tissues.

Agrobacterium-Mediated Insect Resistance Gene (cry1Aa) Transfer Studies Pertaining to Antibiotic Sensitivity on Cultured Tissues of Broccoli

ABSTRACTSuccessful plant transformation requires an efficient regeneration protocol and a suitable selection method. A genetically engineered, disarmed Agrobacterium tumefaciens strain containing a binary vector pBin-1Aa with cry1Aa (insect resistance) and npt-II (neomycin phosphotransferase-II) genes was used for genetic transformation studies. Effects of antibiotics kanamycin and cefotaxime were studied to determine the aptness of kanamycin resistance as a selectable marker and for cefotaxime in controlling excessive bacterial growth during genetic transformation studies using cultured hypocotyl, cotyledon, leaf, and petiole tissues of Broccoli (Brassica oleracea L var. italica). Leaf and petiole explants exhibited decreased fresh weight as kanamycin concentration increased, resulting in full or partial inhibition of shoot regeneration. A significant or nonsignificant negative correlation occurred between kanamycin concentration and explant fresh weight over the time. Cefotaxime had little effect on regeneration potential. Growth of Agrobacterium cells was controlled at a concentration of 400 mg·L−1 cefotaxime in explants, and maximum putative transgenic shoot regeneration in explants of hypocotyls (44.88%) and cotyledons (36.29%) was obtained on MS (Murashige and Skoog)-selective shoot regeneration medium. Polymerase chain reaction analysis of genomic DNA using specifically designed primers detected the presence of the cryIAa and npt-II genes in kanamycin-resistant broccoli plantlets. Of five randomly selected putative transgenic shoots, three were positive for presence/integration of cryIAa and npt-II genes during T-DNA transfer and integration into the plant genome. Kanamycin and cefotaxime act as effective selective agents during genetic transformation pertaining to antibiotic sensitivity on cultured tissues of broccoli.

Investigation on water status and distribution in broccoli and the effects of drying on water status using NMR and MRI methods

Many quality attributes of food products are influenced by the water status and the microstructure. Low-field nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI) methods are applied to non-destructively monitor the water status and structure of food. The aim of this study is to investigate the water status and distribution inside broccoli tissues and the effects of hot-air drying on the water status by using NMR and MRI methods. Transverse relaxation times (T2) provide the information of water status and water distribution. Results show that three water fractions with different T2 relaxation times were detected inside broccoli, which corresponded to different cell compartments. Proton weighted imaging could monitor the spatial distribution of water. Image analysis indicates that the water distribution inside broccoli was heterogeneous and the water content reduced from the stalk to the buds. During hot-air drying experiments, different drying kinetics were observed in the florets and stalks, which were related to their different structures. In addition, a detection limit of the moisture content was calculated for LF-NMR (about 11.35%). The results of this study show that the low-field NMR and MRI methods can precisely provide the quantitative information of water status inside food materials, and can be used to investigate the effects of food processing on product quality. The method provided in this study can be used to monitor changes of water status and distribution in a sample non-destructively during drying process.

Potential of different mechanical and thermal treatments to control off-flavour generation in broccoli puree

The aim of this study was scientifically investigate the impact of the sequence of different thermo-mechanical treatments on the volatile profile of differently processed broccoli puree, and to investigate if any relationship persists between detected off-flavour changes and microstructural changes as a function of selected process conditions. Comparison of the headspace GC–MS fingerprinting of the differently processed broccoli purees revealed that an adequate combination of processing steps allows to reduce the level of off-flavour volatiles. Moreover, applying mechanical processing before or after the thermal processing at 90°C determines the pattern of broccoli tissue disruption, resulting into different microstructures and various enzymatic reactions inducing volatile generation.These results may aid the identification of optimal process conditions generating a reduced level of off-flavour in processed broccoli. In this way, broccoli can be incorporated as a food ingredient into mixed food products with limited implications on sensorial consumer acceptance.

Seasonal Variation in Contents of Sugars in Different Parts of Broccoli

Seasonal variation in the contents of sugars (fructose, glucose, and sucrose) in the floret, leaf, and stem of broccoli were studied in ten commercial broccoli cultivars. Plants were grown in the spring and fall seasons in 2011. In both seasons, glucose was the major constituent, comprising about 50% of the total sugar content in the floret and leaf tissue of most cultivars, whereas the broccoli stem showed an unusual pattern of accumulation. Sucrose exhibited greater cultivar dependency as well as seasonal variation compared to fructose and glucose in floret and leaf tissues. The floret tissue had a higher total content of sugar in the spring compared to the fall due to an increase in glucose and fructose. However, most of the leaf and stem tissues of broccoli had a higher total sugar content in the fall compared to the spring. Furthermore, stem and leaf tissues possessed a relatively higher total sugar content compared to floret tissue in both seasons. ‘Grandeur’ broccoli contained a significantly greater amount of total sugar in both floret and leaf tissues in both seasons, whereas ‘YuDoRi No.1’ broccoli exhibited the highest total content of sugar in stem tissue. At overall, the results showed significant influences of genotype, plant part and growing season on sugar content in broccoli.

Sprouting Broccoli Accumulate Higher Concentrations of Nutritionally Important Metabolites under Narrow-band Light-emitting Diode Lighting

Previous research in our group demonstrated that short-duration exposure to narrow-band blue wavelengths of light can improve the nutritional quality of sprouting broccoli (Brassica oleacea var. italica) microgreens. The objective of this study was to measure the impact of different percentages of blue light on the concentrations of nutritional quality parameters of sprouting broccoli microgreens and to compare incandescent/fluorescent light with light-emitting diodes (LEDs). Microgreen seeds were cultured hydroponically on growing pads under light treatments of: 1) fluorescent/incandescent light; 2) 5% blue (442 to 452 nm)/95% red (622 to 632 nm); 3) 5% blue/85% red/10% green (525 to 535 nm); 4) 20% blue/80% red; and 5) 20% blue/70% red/10% green in controlled environments. Microgreens were grown at an air temperature of 24 °C and a 16-hour photoperiod using a light intensity of 250 μmol·m−2·s−1 for all light treatments. On emergence of the first true leaf, a nutrient solution of 42 mg·L−1 nitrogen (N) (20% Hoagland’s #2 solution) was used to submerge the growing pads. Microgreens were harvested after 20 days under the light treatments and shoot tissues were processed and measured for nutritionally important shoot pigments, glucosinolates, and mineral nutrients. Microgreens under the fluorescent/incandescent light treatment had significantly lower shoot fresh mass than plants under the 5% blue/95% red, 5% blue/85% red/10% green, and the 20% blue/80% red LED light treatments. The highest concentrations of shoot tissue chlorophyll, β-carotene, lutein, total carotenoids, calcium (Ca), magnesium (Mg), phosphorus (P), sulfur (S), boron (B), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo), zinc (Zn), glucoiberin, glucoraphanin, 4-methoxyglucobrassicin, and neoglucobrassicin were found in microgreens grown under the 20% blue/80% red light treatment. In general, the fluorescent/incandescent light treatment resulted in significantly lower concentrations of most metabolites measured in the sprouting broccoli tissue. Results from the current study clearly support data from many previous reports that describe stimulation of primary and secondary metabolite biosynthesis by exposure to blue light wavelengths from LEDs.

The effects of ultrasound-assisted freezing on the freezing time and quality of broccoli (Brassica oleracea L. var. botrytis L.) during immersion freezing

The application of ultrasound during immersion freezing of broccoli was studied and particular attention was given to the effects on freezing time, microstructure, firmness and drip loss of broccoli. Broccoli florets were immersion frozen in an ultrasound-assisted freezer at two frequencies and four different power levels. The results showed that the total freezing time and times required for pre-cooling, phase change and sub-cooling stages of broccoli were significantly reduced by the application of ultrasound-assisted freezing (UAF) at 150 (30 kHz) or 175 W (20 kHz) power level and with judicious combination of process parameters (exposure time, ultrasound irradiation temperature and pulse mode). The microstructure and the firmness of broccoli tissue were better preserved and the drip loss was significantly reduced by the application of optimized UAF compared to the normal immersion freezing. These findings indicate that there is a great potential of UAF in immersion freezing of food.

(+)-Pisatin biosynthesis: from (-) enantiomeric intermediates via an achiral 7,2'-dihydroxy-4',5'-methylenedioxyisoflav-3-ene.

(+)-Pisatin, produced by peas (Pisum sativum L.), is an isoflavonoid derivative belonging to the pterocarpan family. It was the first chemically identified phytoalexin, and subsequent research has demonstrated that most legumes produce pterocarpans with the opposite stereochemistry. Studies on the biosynthesis of (+)-pisatin have shown that (-) enantiomeric compounds are intermediates in (+)-pisatin synthesis. However, the steps from the (-)-7,2'-dihydroxy-4',5'-methylenedioxyisoflavanone [(-)-sophorol] intermediate to (+)-6a-hydroxymaackiain intermediate are undetermined. Chemical reduction of (-)-sophorol using sodium borohydride (NaBH4) produced two isomers of (-)-7,2'-dihydroxy-4',5'-methylenedioxyisoflavanol [(-)-DMDI] with optimal UV absorbance at 299.3 and 300.5 nm, respectively. In contrast, enzymatic reduction of (-)-sophorol by the pea enzyme sophorol reductase (SOR) produced only the 299.3 nm (-)-DMDI isomer. Proton nuclear magnetic resonance ((1)H NMR) analysis of the 299.3 nm (-)-DMDI isomer demonstrated that this isomer had the same NMR spectrum as previously reported for cis-isoflavanol isomers, indicating that cis-(-)-DMDI is an intermediate in (+)-pisatin biosynthesis. Enzyme assays using protein extracts from pea tissue treated with CuCl2 as an elicitor converted the cis-(-)-DMDI isomer into an achiral isoflavene, 7,2'-dihydroxy-4',5'-methylenedioxyisoflav-3-ene (DMDIF), and the trans-(-)-DMDI isomer was not metabolized by the same protein preparation. A comparison of the enzyme activities on cis-(-)-DMDI with protein preparations from elicited tissue versus non-elicited tissue showed a threefold increase in the amount of activity in the proteins from the elicited tissue. Proteins from the elicited tissues of alfalfa, bean, and chickpea converted cis-(-)-DMDI into either (-)-maackiain and/or (-)-sophorol, while proteins from the elicited tissues of broccoli and pepper produced no detectable product. These results are consistent with the involvement of cis-(-)-DMDI and the achiral DMDIF as intermediates in (+)-pisatin biosynthesis.

Influence of processing on the pectin structure–function relationship in broccoli purée

Abstract The structure–function relationship of pectin in broccoli puree as influenced by different types of (pre)processing was investigated. In particular, the effect of various broccoli pretreatments, including low-temperature and high-temperature blanching, two tissue disruption techniques, blending and high-pressure homogenisation, and a subsequent cooking step on the consistency and syneresis of broccoli puree was explored. The influence of these different puree preparation steps on pectin was examined via physicochemical analysis of fractionated cell walls and isolated polymers, and via anti-pectin antibodies. Puree prepared by blending non-pretreated or low-temperature blanched broccoli showed a rather high consistency and pronounced syneresis. High-temperature blanching, high-pressure homogenisation and cooking were all able to limit the degree of syneresis to a large extent. High-pressure homogenisation and cooking however resulted in an opposite effect on puree consistency: the former lowers the consistency of broccoli puree, presumably due to a reduction in broccoli tissue particle size, whereas the latter increases it. Cooking of broccoli puree led to a dramatic increase in pectin solubility which was attributed to the thermosolubilisation and β-eliminative depolymerisation of pectin at the high temperatures reached during this treatment. Moreover, it appeared that during the cooking step especially highly methyl-esterified homogalacturonan-rich components were leached out into the serum phase of the puree. Industrial relevance The deliberate application of particular thermal and mechanical processes on raw plant material makes it possible to design naturally structured/textured food products without the addition of texture-controlling agents such as starches, gums and stabilizers. In this study, research towards the role of in situ pectin on the consistency and degree of syneresis of differently (pre)processed broccoli purees is provided.

The Isolation and Determination of Sulforaphane from Broccoli Tissues by Reverse Phase‐High Performance Liquid Chromatography

AbstractSulforaphane is a cognate isothiocyanate of glucoraphanin, released by degradation of glucosinolate through myrosinase enzyme, when Brassica tissues are crushed or chewed. C8 analytical column and mixture of water/acetonitrile 65/35 (ν/ν) as mobile phase were used to separate sulforaphane in broccoli tissues (e.g., 7‐day‐seedling, seed, floret and leave). Confirmation of sulforaphane in crude extract of broccoli tissues was monitored using gas chromatography‐mass spectroscopy (GC‐MS). The highest amount of sulforaphane (1216 μg/g dry weight) found in 7‐day‐seedling. This method is suitable for routine screening of plant materials and proposes a low cost and robust technique for the analysis of sulforaphane.

Towards a better understanding of the pectin structure–function relationship in broccoli during processing: Part II — Analyses with anti-pectin antibodies

To investigate the structure–function relationship of pectin during (pre)processing, broccoli samples ( Brassica oleracea L. cultivar italica) were subjected to one of the following pretreatments: (i) low-temperature blanching (LTB), (ii) LTB in combination with Ca 2+ infusion, (iii) high-pressure pretreatment (HP), (iv) HP in combination with Ca 2+ infusion, or (v) no pretreatment (control sample), whether or not in combination with a thermal treatment of 15 min at 90 °C. Anti-homogalacturonan antibodies were used to perform in situ (microscopy) and ex situ (immuno-dot assays) analyses on broccoli pectin which resulted in information concerning the localisation of defined pectic domains in broccoli cell walls and pectin's structure. Water-soluble pectin appears to contain unbranched, high-esterified pectin and some pectic polymers with abundant side chains that are less esterified. Ionically cross-linked pectin, on the other hand, contains low-esterified pectin with either highly branched or unbranched domains. The in situ visualisation of pectin in broccoli suggested that de-esterification of pectin by PME during LTB as well as during HP mainly takes place in the tricellular junctions of adjacent cells in broccoli tissue. Ca 2+-cross-linked pectin could be found in cell walls lining intercellular spaces and was particularly abundant at the corners of intercellular spaces, indicating its important role in cell–cell adhesion. Both LTB and HP created pectin–Ca 2+-cross-links in parts of the cell wall where these cross-links were originally absent. The influence of thermal processing and the effect of pressurisation on the pectic components in the cell wall could also be visualised using the antibodies.

Anti-homogalacturonan antibodies: A way to explore the effect of processing on pectin in fruits and vegetables?

The potential of the anti-homogalacturonan(HG) antibodies JIM5, JIM7, LM7, LM18, LM19, LM20 and PAM1 for investigating the effect of processing on pectin in fruit and vegetable tissues was screened in this study. In this respect, the specificity of the antibodies towards pectin and methoxylated polygalacturonic acid with defined degrees and patterns of methylesterification was elaborated, leading to a substantial extension of the information already available in literature. Based on the obtained specificities, the distribution of pectin methylesterification in carrot and broccoli tissue was mapped. It was established that pectin with a low degree of esterification (DE) and a more blockwise distribution of the methylesters is, both in carrot and in broccoli, preferentially located at the tricellular junctions between adjacent cells. This cell wall region, however, is also likely to contain other structural pectic domains. The inner face of the cell wall adjacent to the plasma membrane seems to contain pectin with a medium DE. To evaluate the potential of the anti-HG antibodies to detect changes in degree and pattern of methylesterification caused by processing, carrot and broccoli were subjected to a thermal treatment aimed to stimulate the endogenous pectinmethylesterase (PME) activity. It was revealed that process-induced de-esterification by endogenous PME mainly tends to take place at discrete regions of the inner face of the cell wall adjacent to the plasma membrane for carrot and in tricellular junctions and the middle lamella for broccoli.

Chlorophyllase versus pheophytinase as candidates for chlorophyll dephytilation during senescence of broccoli

Degradation of chlorophylls during senescence is a highly regulated process which requires the concerted action of several enzymes. Traditionally, it has been stated that the dismantling process of the chlorophyll molecule begins with a dephytilation step, followed by Mg 2+ removal and other breakdown reactions. Recently, new evidence suggests the possibility of a rearrangement in the first two steps of this process, occurring Mg 2+ removal prior to the loss of the phytol side chain. With the purpose of approximating to the real sequential order of these reactions and to assess if dephytilation occurs on intact (catalyzed by chlorophyllase) or Mg-free (catalyzed by pheophytinase) chlorophyll, expression of both genes was analyzed in broccoli tissue during senescence. Samples of broccoli florets treated with plant hormones, such as cytokinin and ethylene were utilized, as to assess the effect of such compounds on the expression of these genes. Results showed that chlorophyllase expression did not correlate to typical expression patterns for genes related to senescence, since a decrease in expression during senescence was found for one of the two chlorophyllase genes analyzed, and the hormonal-treatment effects on gene expression did not match those observed on chlorophyll content for both chlorophyllase genes. Pheophytinase expression patterns, on the other hand, displayed an increase in the first 3 days of induced senescence, followed by lower expression values towards the end of the experiment. Samples subjected to postharvest treatments mostly showed an inhibition of pheophytinase expression, especially in samples in which degradation of chlorophylls had been delayed. These results suggest that pheophytinase expression correlates to the visual manifestation of postharvest treatments, supporting the possibility that this enzyme is responsible for the dephytilation step in chlorophyll breakdown.

Broccoli growth in response to increasing rates of pre-plant nitrogen. II. Dry matter and nitrogen accumulation

An understanding of plant nitrogen accumulation and soil nitrogen dynamics is needed to develop management practices that balance nitrogen requirements of vegetable crops with environmental protection. Field trials were conducted in 2001 and 2002 to determine the interaction of increasing rates of pre-plant nitrogen fertilizer with broccoli tissue nitrogen accumulation and soil nitrogen dynamics. Broccoli cultivars Decathlon and Captain were grown with seven rates of nitrogen (0, 50, 100, 150, 200, 300, 400 kg N ha-1) applied pre-plant as ammonium nitrate. Rate of nitrogen accumulation by the above-ground tissue biomass varied over time and among nitrogen treatments, ranging from 1 to 16 kg N ha-1 d-1. At harvest, tissue nitrogen was high, but nitrogen use efficiency was low when high rates of nitrogen were applied. Soil NO3--N content decreased from planting to harvest. At harvest, soil NO3--N increased with increasing rates of nitrogen, with the majority of NO3--N found in the top 0 to 30 cm of the soil. At 200 kg ha-1 applied nitrogen, plants recovered essentially all of the estimated available nitrogen and there appears to be little risk of nitrogen loss during the growing season. Approximately 130 kg N ha-1 was supplied by the soil during the cropping season. Soil and crop residues at harvest ranged from 96 to 330 kg N ha-1. This residual fertility poses a risk for nitrogen loss. Practical and cost-effective strategies are needed to manage residual nitrogen in the soil and crop residues to minimize loss and retain this nitrogen for subsequent crops. Key words: Brassica oleracea L. italica Plenck, nitrogen budget, nitrogen rates, nitrogen use efficiency, nutrient management

High-pressure treatments induce folate polyglutamate profile changes in intact broccoli ( Brassica oleraceae L. cv. Italica) tissue

In plant matrices, folates exist largely as folylpoly-γ-glutamates requiring deglutamylation to monoglutamates prior to absorption, which might impair dietary folate bioavailability. This study investigated folylpoly-γ-glutamate stability and conversions in broccoli tissue during thermal (25–90 °C, 30 min) and high-pressure treatments (0.1–600 MPa, 25–45 °C, 30 min) after vacuum packaging. Folates were analyzed based on poly-γ-glutamate side chain length by RP-HPLC. During thermal treatments, folates were stable up to 90 °C, whereas differences in folylpoly-γ-glutamate profiles towards higher conjugated folylpoly-γ-glutamates were observed at elevated temperatures (70–90 °C). High-pressure treatments resulted in significant folate losses (48–78%). Depending on the pressure–temperature combinations studied, folylpoly-γ-glutamates were converted to folylmono- and folyldi-γ-glutamates, which was shown to occur mainly during the initial stages of the high-pressure treatments, i.e. during pressure build-up and subsequent equilibration. Targeted application of high-pressure treatments can hence be applied to obtain broccoli with higher monoglutamate folate content. Implications towards folate bioavailability in relation to the observed folate degradation, however, requires further investigation.