Storage Of Broccoli Research Articles

Modifying the Processing and Handling of Frozen Broccoli for Increased Sulforaphane Formation

Frozen broccoli can provide a cheaper product, with a longer shelf life and less preparation time than fresh broccoli. We previously showed that several commercially available frozen broccoli products do not retain the ability to generate the cancer-preventative agent sulforaphane. We hypothesized that this was because the necessary hydrolyzing enzyme myrosinase was destroyed during blanching, as part of the processing that frozen broccoli undergoes. This study was carried out to determine a way to overcome loss of hydrolyzing activity. Industrial blanching usually aims to inactivate peroxidase, although lipoxygenase plays a greater role in product degradation during frozen storage of broccoli. Blanching at 86 °C or higher inactivated peroxidase, lipoxygenase, and myrosinase. Blanching at 76 °C inactivated 92% of lipoxygenase activity, whereas there was only an 18% loss in myrosinase-dependent sulforaphane formation. We considered that thawing frozen broccoli might disrupt membrane integrity, allowing myrosinase and glucoraphanin to come into contact. Thawing frozen broccoli for 9 h did not support sulforaphane formation unless an exogenous source of myrosinase was added. Thermal stability studies showed that broccoli root, as a source of myrosinase, was not more heat stable than broccoli floret. Daikon radish root supported some sulforaphane formation even when heated at 125 °C for 10 min, a time and temperature comparable to or greater than microwave cooking. Daikon radish (0.25%) added to frozen broccoli that was then allowed to thaw supported sulforaphane formation without any visual alteration to that of untreated broccoli.

Time of day at harvest affects the expression of chlorophyll degrading genes during postharvest storage of broccoli

Broccoli is an important component of the human diet with a high nutritional value. This vegetable is harvested when its development is not complete, triggering a quick senescence that is accompanied by chlorophyll degradation and a shortening of shelf life. Recently it was shown that the time of the day at which the broccoli is harvested can influence in the rate of chlorophyll degradation. This report describes changes of expression of chlorophyll degrading related genes during postharvest storage of heads harvested at different moments of day. At harvest, the content of chlorophyll did not change among samples but the expression of many genes associated with chlorophyll degradation did. The level of chlorophyll diminished during storage, but samples harvested at 18:00h had a lower rate of degradation in comparison with heads harvested at 08:00h. Most of genes that were previously associated with chlorophyll degradation during senescence such as BoSGR, BoCLH2, BoPPH and BoPaO showed a lower expression or a delay in their mRNA level increments in samples harvested at 18:00h. Other genes related to chlorophyll degradation during senescence, like BoNYC or BoRCCR, showed an increased expression during senescence but there was no positive correlation between chlorophyll catabolism and expression of these genes. In conclusion, changes in the metabolism due to time of day at harvest not only influence the expression of genes during the day but also may cause different patterns of expression during postharvest.

Postharvest quality response of broccoli florets to combined application of 1-methylcyclopropene and modified atmosphere packaging

Broccoli is rich in health-promoting compounds such as ascorbic acid and glucosinolates. However, amount of such compounds inevitably decrease along with the storage. This study was conducted to investigate the effects of 1-methylcyclopropene (1-MCP) and Modified Atmosphere Packaging (MAP) treatments alone or in combination on postharvest life and quality of broccoli heads during cold storage. MAP and 1-MCP+MAP significantly reduced weight loss, delayed surface color changes and chlorophyll breakdown, maintained visual quality, ascorbic acid, total phenol, soluble solid contents and titratable acidity compared with control as well as 1-MCP treatment. In 1-MCP plus MAP treated florets, approximate 47% and more chlorophyll and 24% more total phenol contents were found compared to control florets. 1-MCP plus MAP application better performed in maintaining the quality of broccoli florets during 28-day-storage. Overall results revealed that combined treatment can be recommended for commercial broccoli storage at 0 °C.

Altered commercial controlled atmosphere storage conditions for ‘Parhenon’ broccoli plants (Brassica oleracea L. var. italica). Influence on the outer quality parameters and on the health-promoting compounds

Parameters such as bright and dark green compact heads, firmness to hand pressure and closed florets, and the evolution of main health-promoting compounds such as carotenoid and chlorophyll pigments, phenolic compounds and glucosinolates, were assessed to determine the effects of the atmosphere composition during the cold storage of broccoli ‘Parthenon’.The controlled atmosphere evaluated in this study was a gas mixture containing 10% of O2 and 5% of CO2, with 85–90% relative humidity. The storage temperature was established at 1 and 2 °C.We found that the loss of weight and firmness was greater in the reference samples compared to samples stored under controlled atmosphere conditions (20.60% and 84.88%, for loss of weight and firmness in reference samples respectively, and 4.80% and 16.48%, for loss of weight and firmness in controlled atmosphere samples respectively) after 9 days of storage, approximately. In addition, the proposed controlled atmosphere conditions helped to maintain the concentration of the main health-promoting compounds, such as ascorbic acid, carotenoid and chlorophylls pigments, total phenolic compounds and intact glucosinolates, and the values of the antioxidant activity in vitro were also maintained.

THE INFLUENCE OF CONTROLLED ATMOSPHERE STORAGE ON PHYTOSTEROLS IN BROCCOLI HEADS

The objective of this study was to analyze changes in sterols content during storage of broccoli. Broccoli heads of “Marathon” and “Cezar” cultivars were stored for 6 weeks in a cold store, under normal (0% CO2 + 21% O2) and controlled atmosphere conditions (CA, 5% CO2 + 2% O2) at the temperature of 0–1C. Sterols, in the forms of free sterols and glucosides, were determined by high-performance liquid chromatography. Color parameters of broccoli were determined spectrophotometrically using CIE L*a*b* system. Amount of free sterols in the freshly harvested heads reached 54.3 mg/100 g dw and was the highest in the heads of “Marathon.” The dominant free sterol was b-sitosterol. During the storage of broccoli, sterol content decreased but was higher in the heads stored in CA. The broccoli showed smaller changes of color when stored in CA than in normal atmosphere.

INVOLVEMENT OF CHLOROPHYLLASE ON CHLOROPHYLL DEGRADATION IN STORED BROCCOLI FLORETS AND ITS CONTROL BY UV TREATMENT

Yellowing is the most visible deterioration in broccoli (Brassica oleracea L. Italica Group) that usually occurs with the progress of chlorophyll (Chl) degradation. This study deals with the control of floret yellowing in stored broccoli by using UV-B irradiation at 0 (control) and 8.8 kJ.m -2 . The application of UV-B at 8.8 kJ.m -2 effectively delayed yellowing of broccoli florets. Chl a derivatives including chlorophyllide (Chlide) a, C13 2 -hydroxychlorophyll (C13 2 -OHChl) a and pheophytin (Phy) a levels in broccoli florets were decreased concomitantly with the enhancement of pheophorbide (Pheide) a and pyropheophorbide (Pyropheide) a levels especially in the control treatment. On the other hand, Chlide a, C13 2 -OHChl a and Phy a levels were increased in 8.8 kJ.m -2 UV-B treated broccoli during storage at 15°C. Two types of chlorophyllase (Chlase) were identified by molecular exclusion chromatography. Type I was suppressed in UV-B treated broccoli on day 4, and the Km value of Type I was lower than that of Type II. Thus, the Chlase activity, especially Type I, was effectively suppressed by UV-B treatment, whereas Type II Chlase may take in part of Chl degradation in stored broccoli florets.

PARTIALLY PURIFICATION OF MG-DECHELATASE IN RELATION TO CHLOROPHYLL DEGRADATION IN BROCCOLI (BRASSICA OLERACEA L. ITALICA GROUP) FLORETS

ISHS Southeast Asia Symposium on Quality and Safety of Fresh and Fresh-Cut Produce PARTIALLY PURIFICATION OF MG-DECHELATASE IN RELATION TO CHLOROPHYLL DEGRADATION IN BROCCOLI (BRASSICA OLERACEA L. ITALICA GROUP) FLORETS

COMPARING THE FUNCTION OF PERFORATED AND NON-PERFORATED FILM IN MODIFIED-ATMOSPHERE PACKAGING: IMPACT ON THE QUALITY OF BROCCOLI

A wide range of steady-state O 2 partial pressures (0.1 to 18 kPa) was obtained in perforated and non-perforated packages composed of low density polyethylene (LDPE) film at 0 and 10°C for the storage of broccoli (Brassica oleracea L. var. italica). Half of the broccoli was treated for 24 h at 10°C with 1 ppm I-MCP prior to storage. The combinations of steady-state O 2 and CO 2 partial pressures achievable depended on whether the film was perforated or not. The regression fitting the O 2 versus CO 2 headspace data for non-perforated packages was linear with a negative slope such that for every 4 kPa decrease in O 2 , there was a 1 kPa increase in CO 2 . The curve fitting the data for perforated packages was also linear and negative, but steeper, such that for every 4 kPa decrease in O 2 , there was a 4 kPa increase in CO 2 . The result was that for a given steady-state O 2 concentration, the CO 2 concentration was higher in the perforated package compared to the non-perforated packages. For florets stored at 10°C, the higher CO 2 in the perforated packages resulted in higher visual quality and better color retention was significantly better than in the non-perforated packages for packages with O 2 levels greater than approximately 2 kPa. There was no effect of the elevated CO 2 for the florets stored at 0°C. 1-MCP application only had a small positive impact on floret quality at 10°C and no effect at 0°C. The data suggest that combination of both perforated packages and 1-MCP application on broccoli florets may have the potential to provide some protection against rapid quality loss when the storage temperature exceeds the recommended 0°C.

Impact of UV-B irradiation on chlorophyll degradation and chlorophyll-degrading enzyme activities in stored broccoli (Brassica oleracea L. Italica Group) florets

UV-B irradiation was applied to broccoli florets to investigate its effect on chlorophyll degradation and chlorophyll-degrading enzyme activities in stored broccoli. Broccoli florets were irradiated with UV-B doses at 4.4, 8.8, and 13.1 kJ m−2 and then kept at 15 °C in darkness. We found that a UV-B dose of at least 8.8 kJ m−2 efficiently delayed the decrease of the hue angle value and the contents of chlorophylls a and b. Chlorophyllide a and 132-hydroxychlorohyll a gradually decreased with senescence. Pheophorbide a and pyropheophorbide a levels were significantly higher in broccoli without UV-B treatment. Chlorophyllase and chlorophyll-degrading peroxidase activities with UV-B treatment were suppressed, as well as the activity of Mg-dechelatase. Mg-dechelating substance activity was also suppressed with this treatment. We concluded that UV-B treatment effectively suppressed chlorophyll degradation in broccoli florets during storage, suggesting that the effect could be due to the suppression of chlorophyll-degrading enzyme activities.

Prediction of ascorbic acid content in broccoli using a model equation of respiration

The relationship between ascorbic acid (AA) and respiration of stored broccoli was investigated in an experiment involving various temperature conditions (5, 10, 20 and 30 °C), O 2 concentrations (21%, 10% and 5% O 2), cultivars (cv. ‘Pixcel’, ‘Morimidori’ and ‘Subaru’) and produce types (intact and cut). The relative AA (AA rel) in percent, which was normalized from the initial AA value, was plotted against the accumulated amount of CO 2 production ( A R C O 2 ). AA rel decreased with increasing A R C O 2 in each experiment. The relationship between AA rel and A R C O 2 of broccoli can be expressed by a single equation with a common parameter ( β), regardless of storage and produce variations, suggesting that decreasing AA content could be deduced from respiratory activity. Subsequently, the feasibility of applying the published respiration data to this equation to predict AA change during actual distribution processes was investigated. Cut broccoli was stored under fluctuating temperature conditions and modified atmosphere packaging (MAP). We calculated the change in AA values under these conditions using our proposed equation substituted in the Arrhenius and Michaelis–Menten models. Predicted values had good agreement with measured values. This finding suggests a new application of the respiration model for quality prediction. Our proposed model might also be able to predict quality for other commodities, by applying knowledge of respiration data that has been reported previously.

Effect of Heat Treatment on Catabolites Formation in Relation to Chlorophyll Degradation during Storage of Broccoli (Brassica olearacea L. Italica Group) Florets

The effects of heat treatment on the formation of chlorophyll (Chl) derivatives and Chl-degrading enzyme activities in stored broccoli (Brassica oleracea L. Italica Group ‘Ryokutei’) florets were determined. The Chl a level barely changed during heat treatment, but each Chl a derivative level changed. Chlorophyllide (Chlide) a, pheophorbide (Pheide) a, C132-hydroxychlorophyll (C132-OHChl) a, Chl a′, an isomer of Chl a, and pheophytin (Phy) a were detected as a Chl derivative during heat treatment and during storage after treatment. Chlide a, Pheide a, and C132-OHChl a levels decreased during 2-h heat treatment (50°C), whereas Chl a′ and Phy a levels increased. Chl-degrading enzyme activities, in particular, Mg-dechelation activity, were effectively suppressed after 2 h of heat treatment. The content of Chls a and b in control broccoli florets decreased greatly during storage at 15°C, while the content in heat-treated broccoli florets hardly changed at all. In Chl derivatives, the Pheide a level in broccoli florets treated with or without heat treatment, especially the former, increased appreciably during storage. The Chlide a level in control florets decreased markedly during storage. On the other hand, the Chlide a level in heat-treated broccoli florets did not change during storage. Mg-dechelation activity in control florets markedly increased after 4 days of storage at 15°C, but the enhancement of the activity was suppressed by heat treatment. These findings suggest that Chl derivatives, especially Pheide a, are accumulated as intermediates in heat-treated broccoli florets, and Mg-dechelating action, in conjunction with that of chlorophyllase and Chl-degrading peroxidase, could be involved in Chl degradation in stored broccoli florets.

KINETIC MODELINGS OF BROCCOLI COLOR CHANGES DURING CHILLED STORAGE

This work studied the kinetic models of broccoli color changes during storage. The Commission Internationale d'Eclairage ([CIE] International Commission on Illumination)-L*a*b* color system parameters a*, b*, H°, total color difference (TCD), chlorophyll content and yellowness degree were used to determine the color changes under the storage conditions at 0, 5 and 10C. The broccoli ball was packed with high-density polyethylene film. The experimental results showed that the package greatly promoted the activation energy of b* value in broccoli while delaying the respiration peak. The nonlinear regression analysis showed that the rate constants of color parameters b* and TCD were following a first-order Arrhenius-type reaction depending on temperature, and the polynomial model was suitable for the changes of a* and H° value. The observed results demonstrated that the variation of color parameters was in accordance with chlorophyll content and yellowness degree in stored broccoli, suggesting the possibility of a computer-vision system application on the color grading of broccoli.

Effects of heat treatment on an ascorbate–glutathione cycle in stored broccoli ( Brassica oleracea L.) florets

Effects of heat treatment on an ascorbate–glutathione cycle in stored broccoli ( Brassica oleracea L. cv Erude) florets were measured. Floret yellowing during storage at 15 °C was effectively retarded by hot air treatment at 50 °C for 2 h. Hydrogen peroxide contents in heat-treated broccoli florets were higher than in control florets. Ascorbic acid content with or without heat treatment decreased during storage, but the decrease in the control was faster than in heat-treated broccoli. Additionally, the reduced glutathione content in heat-treated florets showed almost no change during storage, whereas that in control florets decreased gradually. In heat-treated florets, both dehydroascorbate reductase and glutathione reductase activities were maintained for the first 2 days and then decreased slightly, while they decreased considerably in the control florets during storage. Cytosolic ascorbate peroxidase activity in heat-treated florets increased slightly during storage, whereas that in control florets showed almost no change. However, chloroplastic ascorbate peroxidase activity with or without heat treatment greatly decreased during 4 days of storage. These results indicate that hydrogen peroxide, which is generated by the heat treatment, might induce activation of enzymes related to the ascorbate–glutathione cycle, and the enhancement of the action of the cycle might relate to the suppression of senescence in heat-treated broccoli florets.

Control of isoperoxidases involved in chlorophyll degradation of stored broccoli ( Brassica oleracea) florets by heat treatment

Changes in isoperoxidases involved in chlorophyll (Chl) degradation of stored broccoli (Brassica oleracea L.) florets and their control by heat treatment (HT) were determined. Chl a and b contents in non-heat-treated broccoli florets decreased greatly after 2 days at 15 degrees C, whereas the contents in heat-treated florets (50 degrees C for 2 h) showed almost no change. Three isoperoxidases involved in Chl degradation were detected by means of molecular exclusion chromatography and the molecular weights of those isoperoxidases were about 95 (Type I), 67 (Type II) and 56 (Type III) kDa, respectively. Only Type I was detected in broccoli florets immediately after harvest, and its activity in non-heat-treated broccoli increased greatly during storage. Both Type II and Type III were present in non-heat-treated broccoli with floret senescence. HT suppressed the enhancement of all of the isoperoxidase activities. Cycloheximide treatment also effectively retarded the increase in Types I, II and III isoperoxidase activities concomitant with the suppression of floret yellowing. The K(m) values corresponding to Chl a of Type II and Type III were lower than Type I, and the V(max)/K(m) values corresponding to Chl a of Type II and Type III were higher than Type I. This suggests that both Types II and III could be closely associated with Chl degradation in broccoli florets and that HT might inhibit floret senescence by suppression of isoperoxidase activities.

Reversible and Irreversible Emission of Methanethiol and Dimethyl Disulfide from Anaerobically Stored Broccoli

The reversible and irreversible emission of methanethiol (MT) and dimethyl disulfide (DMDS) from broccoli florets was demonstrated during anaerobic storage at 20 degrees C for up to 24 h. Reversible emission of MT and DMDS was feasible only in broccoli stored for between 0 and 12 h under entirely anaerobic condition. Beyond that, the emission was completely irreversible. This irreversible process was demonstrated through significant reductions in the chlorophyll fluorescence values and rate of carbon dioxide production and significant increase in the membrane permeability of induced broccoli tissues after exposure to air and incubation. Irreversible emission was also demonstrated through significant change in color from the characteristic bright green to olive green as well as the conversion of chlorophyll a to pheophytin a and chlorophyll a' contents of the induced florets after hot-water treatment. These findings suggest that the irreversible emission of MT and DMDS is a function of permanent membrane damage and loss of intracellular compartmentation in the broccoli tissues as a result of the anaerobic induction. The off-odor formation can still be reversed if the affected tissue is only temporarily impaired by anaerobic condition, thereby maintaining the quality of stored broccoli.

Physiological changes in the antioxidant system of broccoli flower buds senescing during short-term storage, related to temperature and packaging

The superoxide dismutase (SOD) and the antiradical activity (RSA) together with the phenylopropanoids, flavonoids and chlorophyll levels in flower bud tissue were estimated during short-term storage of broccoli Lord cultivar. Catalase (CAT) and peroxidase (POD) activities were detected in broccoli flower branchlets. The activity of SOD increased gradually at both storage temperatures (20 and 5 °C), while the RSA increased only in higher temperature. Increase in catalase activity was observed after 1-day storage of non-packaged broccoli at 20 °C, followed by its decrease, and after 1- and 3-day storage of the packaged heads. In the case of low temperature the enzyme activity increased only after 10 days of storage. Storage of broccoli at room temperature caused the significant increase of peroxidase activity, while at low temperature (5 °C) activity of the enzyme rose after 10 days. During the storage, the accumulation of phenylopropanoids and flavonoids in broccoli flower buds was observed. The rapid decrease of chlorophyll was observed only in the case of broccoli heads stored at 20 °C. Applying of polyethylene foil to packaging of broccoli heads during storage in most cases decreased the SOD, CAT, POD antiradical activities and phenol content, and slowed down chlorophyll decomposition in flower buds. The obtained results emphasize the active role of antioxidant components of broccoli flower buds senescing during short-term storage, especially in intense stress conditions.

Involvement of peroxidase in chlorophyll degradation in stored broccoli (Brassica oleracea L.) and inhibition of the activity by heat treatment

Cell localization of chlorophyll (Chl)-degrading peroxidase and the effects of heat treatment on the activity were determined to elucidate the involvement of the enzyme with yellowing in stored broccoli. Subcellular fractions and intact chloroplasts were prepared by differential and Percoll gradient centrifugation, respectively. In fresh broccoli, only low levels of Chl-degrading peroxidase activity were located in the intact chloroplast, and the highest activity was present in the cytosolic fraction. The Chl content of broccoli without heat treatment decreased greatly after 4 days storage at 15 °C, whereas the content in broccoli treated at 50 °C for 2 h showed almost no change during storage. In microsomal and cytosolic fractions, the activity of C 2 ( R f =0.47) isoperoxidase, which is involved in Chl degradation, increased greatly with floret yellowing. The Chl-degrading peroxidase activities in microsomal and cytosolic fractions also increased strongly after 4 days of storage. In addition, the Chl-degrading peroxidase activity as well as C 2 isoperoxidase activity was suppressed by heat treatment. These results indicate that heat treatments may be effective in inhibiting senescence in part by suppressing the enhancement of Chl-degrading peroxidase activity in the microsomes and the cytosol.

The effect of post-harvest and packaging treatments on glucoraphanin concentration in broccoli (Brassica oleracea var. italica).

The effects of post-harvest and packaging treatments on glucoraphanin (4-methylsulfinylbutyl glucosinolate), the glucosinolate precursor of anticancer isothiocyanate sulforaphane [4-methylsulfinylbutyl isothiocyanate], were examined in broccoli (Brassica oleracea var. italica) during storage times. The results showed that at 20 degrees C, 55% loss of glucoraphanin concentration occurred in broccoli stored in open boxes during the first 3 days of the treatment and 56% loss was found in broccoli stored in plastic bags by day 7. Under both air and controlled atmosphere (CA) storage, glucoraphanin concentration appeared to fluctuate slightly during 25 days of storage and the concentrations under CA was significantly higher than those stored under air treatment. In modified atmosphere packaging (MAP) treatments, glucoraphanin concentration in air control packaging decreased significantly whereas there were no significant changes in glucoraphanin concentration in MAP with no holes at 4 degrees C and two microholes at 20 degrees C for up to 10 days. Decreases in glucoraphanin concentration occurred when the broccoli heads deteriorated. In the present study, the best method for preserving glucoraphanin concentration in broccoli heads after harvest was storage of broccoli in MAP and refrigeration at 4 degrees C. This condition maintained the glucoraphanin concentration for at least 10 days and also maintained the visual quality of the broccoli heads.

Effects of heat treatment on chlorophyll degrading enzymes in stored broccoli (Brassica oleracea L.)

Abstract Effects of heat treatment on chlorophyll (Chl) degrading enzyme activities in stored broccoli ( Brassica oleracea L.) were determined. Chl contents of broccoli treated at 50 °C for 1 h or without heat treatment (control) decreased after 4 days of storage at 15 °C, whereas the contents of broccoli treated at 50 °C for 2 h showed almost no change after 4 days of storage at 15 °C. Chlorophyllase activity of broccoli heat-treated for 1 or 2 h decreased during storage before the occurrence of yellowing, while the activity of the control showed a slight decrease on day 4. Chl oxidase activity of broccoli heat-treated for 2 h remained unchanged. Chl degrading peroxidase activity in the control markedly increased after 6 days of storage at 15 °C, but an increase in the activities in broccoli treated at 50 °C, especially for 2 h, was suppressed. Six anionic and two cationic isoperoxidases were detected in broccoli on day 0. One of the isoperoxidases, C 2 (Rf 0.3) cationic isoperoxidase, was involved in Chl degradation and the increase in C 2 level was greatly reduced by treatment at 50 °C for 2 h. These results indicate that heat treatment could reduce Chl degradation due to the suppression of Chl degrading enzyme activities.

Chlorophyll fluorescence, fermentation product accumulation, and quality of stored broccoli in modified atmosphere packages and subsequent air storage

Abstract Earlier work showed that chlorophyll fluorescence changes of broccoli (Brassica oleracea L., Italica group) were associated with the accumulation of CO2 in modified atmosphere packages (MAP) during storage. This work was initiated to determine whether the chlorophyll fluorescence changes in broccoli from high CO2 MAP are persistent after the packages are opened and the broccoli is allowed to aerate. PD-961EZ bags, which allow the CO2 to accumulate (∼11 kPa CO2), were used in this study. During 28 days in MAP at 1°C, the broccoli gradually developed slight to moderate alcoholic off-odors and accumulated ethanol, acetaldehyde, and ethyl acetate in the tissues. These levels dissipated slightly on opening of the bags and holding the broccoli in 1°C air storage for 4 days. Chlorophyll fluorescence measurements (Fv/Fm, T 1 2 , Fmd, and ΦPSII) declined for broccoli with the accumulation of these anaerobic products and the fluorescence measurements recovered slightly after the bags were opened and the broccoli was held in air at 1°C for 4 days. Chlorophyll fluorescence measurements were found to be highly correlated with the anaerobic volatile content in broccoli during MAP storage and after opening of the MAP. Chlorophyll fluorescence measurements were also highly correlated with the perceived off-odors that were noted for the broccoli after longer storage durations in MAP.