WO2023221378A1 - High-oxygen stress freshness preservation method suitable for fresh fruit storage, and application - Google Patents
High-oxygen stress freshness preservation method suitable for fresh fruit storage, and application Download PDFInfo
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- WO2023221378A1 WO2023221378A1 PCT/CN2022/124559 CN2022124559W WO2023221378A1 WO 2023221378 A1 WO2023221378 A1 WO 2023221378A1 CN 2022124559 W CN2022124559 W CN 2022124559W WO 2023221378 A1 WO2023221378 A1 WO 2023221378A1
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- fresh
- oxygen
- stress
- treatment
- fruits
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/14—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
- A23B7/144—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of gases, e.g. fumigation; Compositions or apparatus therefor
- A23B7/148—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of gases, e.g. fumigation; Compositions or apparatus therefor in a controlled atmosphere, e.g. partial vacuum, comprising only CO2, N2, O2 or H2O
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/85—Food storage or conservation, e.g. cooling or drying
Definitions
- the invention belongs to the technical field of food preservation, especially a high oxygen stress preservation method and application suitable for the storage of fresh fruits.
- this invention uses high-concentration gas short-term molecular stress stress response as a means to elucidate the systemic stress effect and response mechanism of ROS-Redox oxidation-reduction and enzymatic browning metabolism of fruits and vegetables in response to high oxygen/CO 2 stress after harvesting.
- innovative molecular stress preservation technology that couples gas stress to control color, taste and quality has clarified the gas stress concentration effects, time effects and suppression thresholds of different fruits and vegetables, breaking through the problem of controlling the balance point of MA preservation gases.
- controlled atmosphere preservation is considered to be the most convenient, safe, feasible, effective and environmentally friendly method for processing thin-skinned and juicy berries. Because compared with chemical treatment, soaking, and coating methods, gas stress preservation has fewer safety issues and is relatively easy to implement, which can reduce the damage caused to berries.
- a high-oxygen modified packaging gas composition for cooling beef storage and preservation (CN104621231A).
- the present invention appropriately reduces the O2 concentration in the mixed gas, appropriately increases CO2, and sets the gas volume ratio in the modified atmosphere packaging. It is 50% to 55% O2 + 35% to 40% CO2 + 10% N2, and the head space ratio is 2.5:1 to 3:1.
- the packaged beef is stored at 4 ⁇ 1°C.
- the gas composition of the present invention can not only maintain the same good color protection effect as that of 80% O2 high-oxygen packaging, but also extend the shelf life of cooled beef from 12 to 16 days under 80% O2 high-oxygen modified packaging to at least 20 days. , and can also reduce the loss of storage juice in the early stage of storage.
- a method of high-concentration oxygen modified atmosphere packaging combined with low-temperature storage and preservation of fresh-cut thorn sprouts (CN104957244A). The steps are as follows: (1) selection; (2) cleaning; (3) drying; (4) sorting; ( 5) Bagging; (6) Exhausting; (7) Inflating; (8) Sealing; (9) Storage.
- This method uses a high-barrier polyethylene film as a modified atmosphere packaging bag.
- the inflation ratio of the high-oxygen modified packaging is 75%-95% oxygen and 25%-5% carbon dioxide gas respectively.
- the modified atmosphere packaging is combined with low-temperature storage.
- the present invention is applied to the preservation of freshly cut thorn buds, which significantly reduces the toxic effects of anaerobic respiration, reduces respiration, prevents color change, reduces water evaporation and infection by diseases and insect pests, and maintains the original color of the thorn buds to the greatest extent.
- the unique flavor and nutritional content effectively extend the shelf life of fresh-cut thorn sprouts.
- This technical solution significantly inhibits the rot and browning of fresh-cut thorn shoots and the reduction of nutrients such as vitamin C and total phenols, and improves their hardness and antioxidant activity.
- the product has high sensory quality and hygienic quality, and the product shelf life is from 2 ⁇ 3 is extended to 10 ⁇ 12 days, and the price can be increased by 2 times.
- the purpose of the present invention is to fill the gap in the technical field of fresh wolfberry preservation and provide a high-oxygen stress preservation method and application suitable for fresh fruit storage.
- a high-oxygen stress preservation method suitable for the storage of fresh fruits uses 50%-100% high-concentration oxygen as the main gas and high-purity nitrogen as the auxiliary gas to stimulate the fresh fruits for 20-30 minutes for short-term preservation. .
- High oxygen stress treatment Put the selected fresh fruits into a sealed air-conditioned box, first use high-purity nitrogen to discharge the remaining gas in the sealed air-conditioned box, and then pass 50 %-100% O 2 (w/w) high-concentration oxygen stress treatment for 20-30 minutes, then open the sealed air-conditioning box and let it stand for 20-30 minutes to release the high-concentration oxygen remaining in the air-conditioning box and reduce the oxygen content. The concentration returns it to its natural state;
- the diameter of the hole of the PE fresh-keeping bag in step (1) is 0.013-0.020mm.
- step (4) the fresh fruits treated with high oxygen stress are stored under MAP spontaneous atmosphere control and 0 ⁇ 1°C conditions.
- the fresh fruits include fresh wolfberry, Lingwu jujube and Daqing grape.
- the conditions for high-oxygen stress treatment are: introducing high-concentration oxygen of 90% O 2 (w/w) into a sealed air-conditioning box for short-term stress treatment for 30 minutes. , then open the sealed air conditioning box and let it sit for 30 minutes;
- the conditions for high oxygen stress treatment are: introduce oxygen with a concentration of 50% O 2 (w/w) into the sealed air-conditioned box for short-term stress treatment for 30 minutes, and then open the seal The air-conditioning box is left for 30 minutes;
- the conditions for high-oxygen stress treatment are: introduce oxygen with a concentration of 80% O 2 (w/w) into the sealed air-conditioned box for short-term stress treatment for 30 minutes, and then open the sealed box. Atmospheric conditioning box, let stand for 30 minutes.
- the present invention puts the perforated PE fresh-keeping bag on the outside of the fruit and vegetable box for pre-cooling, which can not only achieve a rapid cooling effect, but also maintain good air permeability. , effectively alleviate the serious water loss problem caused by the external environment, maintain the moisture content of fresh wolfberry, and achieve better preservation effect.
- the present invention performs 50%-100% high-concentration oxygen stress treatment on fresh wolfberry at low temperature, effectively inhibiting the growth of microorganisms on the surface of fresh fruits such as fresh wolfberry, reducing the decay of fresh fruits during storage, and having excellent preservation effects. Based on the blank control group without any treatment, it is a new preservation technology that is low-cost, easy to operate, safe, effective and environmentally friendly.
- the present invention treats fresh wolfberry with high-concentration oxygen for a short period of 20-30 minutes, effectively inducing the gene expression level related to antioxidant enzymes in fresh fruits such as fresh wolfberry, and activating the antioxidant enzyme activity of fresh fruits such as fresh wolfberry with ascorbic acid and glutathione.
- the cyclic regeneration of glycopeptide antioxidant components inhibits the accumulation of reactive oxygen species and malondialdehyde, reduces membrane damage and oxidative damage to tissue cells, and maintains the color, hardness, taste and nutritional content of fresh wolfberry.
- the high-oxygen stress fresh fruit preservation method of the present invention is easy and simple to operate and has obvious preservation effects.
- This technical method utilizes high-oxygen abiotic stress (50%-100% O 2 ) and short-term treatment (20-20%) under low temperature conditions. 30min), the fresh fruit can develop resistance after rapid oxidative stress, acting as a "plant vaccine” to achieve the effect of "suppressing oxygen and gaining freshness", effectively solving the problem of mildew, rot, softening, discoloration and loss of taste during the storage process of fresh fruit. It extends the storage period of fresh fruits to 25-30 days, and the good fruit rate is over 90%. The effect is obviously better than that of fresh fruits such as fresh wolfberries that are only refrigerated at low temperatures without high oxygen stress.
- the high oxygen stress fresh fruit preservation method of the present invention is not limited to the preservation of fresh wolfberry, but is also applicable to other fruits and vegetables.
- the optimal high oxygen stress concentration, short treatment time and frequency are different for different fruits and vegetables.
- the method of the present invention can well maintain the color, hardness, taste, nutrients and good fruit rate of the fruit, and by activating the antioxidant system, enhance the anti-browning ability and reduce the tissue damage caused by the accumulation of active oxygen during the storage process of the fruit. Oxidative damage can effectively delay the ripening and aging of fruits.
- the method of the present invention is convenient and safe to operate, and has low cost. It uses high-concentration oxygen to stimulate the fresh fruit for a short time, causing the body to produce a stress response, changing metabolic pathways, increasing the body's resistance, effectively inhibiting the growth of microorganisms on the surface of the fresh fruit during storage, and reducing It reduces the decay rate of fresh fruits, activates the body's antioxidant enzyme activity and antioxidant components, inhibits the accumulation of reactive oxygen species and malondialdehyde content, reduces oxidative damage to tissue cells, and delays the ripening and aging of fresh fruits.
- the fresh fruits are packed in fruit and vegetable boxes after being harvested.
- a perforated PE fresh-keeping bag is placed on the outside of the fruit and vegetable box, and then the whole is blasted at low temperature in the cold storage
- fresh fruits, fruit and vegetable boxes and PE fresh-keeping bags are placed in a cold storage for pre-cooling.
- the cold storage is equipped with air blast to enhance the pre-cooling effect and avoid water loss caused by air blast.
- a perforated PE fresh-keeping bag is placed on the outside of the fruit and vegetable box, which will not prevent the cool air from the cold storage from entering the fruit and vegetable box, and will keep the moisture of the fresh wolfberry fruits as much as possible.
- the applicant has tested during the preliminary experiment that if the fruit and vegetable boxes are not covered with PE fresh-keeping bags and are directly placed in the cold storage for pre-cooling, water loss will be very serious. The effect of using perforated fresh-keeping bags is much improved.
- Figure 1 is a diagram showing the effects of different temperature-mediated hyperoxic stress on changes in the decay rate of fresh wolfberry in Example 1 of the present invention
- Figure 2 is a diagram showing the influence of different temperature-mediated hyperoxic stress on the weight loss rate of fresh wolfberry in Example 1 of the present invention
- Figure 3 is a diagram showing the influence of different temperature-mediated hyperoxic stress on changes in hardness of fresh wolfberry in Example 1 of the present invention
- Figure 4 is a diagram showing the influence of different temperature-mediated hyperoxic stress on the color L* value of fresh wolfberry in Example 1 of the present invention
- Figure 5 is a diagram showing the effect of different durations of hyperoxic stimulation on the decay rate of fresh wolfberry in Example 1 of the present invention
- Figure 6 is a graph showing the effect of different durations of hyperoxic stimulation on the weight loss rate of fresh wolfberry in Example 1 of the present invention.
- Figure 7 is a graph showing the effect of different durations of hyperoxic stimulation on changes in hardness of fresh wolfberry in Example 1 of the present invention.
- Figure 8 is a graph showing the influence of different durations of hyperoxic stimulation on the color L* value of fresh wolfberry in Example 1 of the present invention.
- Figure 9 is a diagram showing the preservation effect and total number of bacterial colonies of fresh wolfberry treated with 90% O 2 short-term stress in Example 1 of the present invention; wherein, A1 is the 30th day of storage, fresh wolfberry without high oxygen stress and single low temperature Storage blank treatment effect diagram, A 2 is the total number of bacterial colonies (10 ⁇ -3) in the culture medium of single low-temperature storage blank treatment of fresh wolfberry on the 30th day of storage without high oxygen stress, B 1 is the fresh wolfberry on the 30th day of storage The effect of 90% O 2 short-term stress treatment of wolfberry for 30 minutes. B 2 is the total number of colonies (10 ⁇ -3) in the medium of fresh wolfberry treated with 90% O 2 short-term stress for 30 minutes on the 30th day of storage. C is the picture of bacterial colonies. Total change chart;
- Figure 10 is a graph showing changes in superoxide anion release, hydrogen peroxide and malondialdehyde content of fresh wolfberry treated with 90% O 2 short-term stress in Example 1 of the present invention; where A is the superoxide anion release Content change chart, B is hydrogen peroxide (H 2 O 2 ) content change chart, C is malondialdehyde (MDA) content change chart;
- A is the superoxide anion release Content change chart
- B hydrogen peroxide (H 2 O 2 ) content change chart
- C malondialdehyde (MDA) content change chart
- Figure 11 is a graph showing changes in antioxidant components of fresh wolfberry treated with 90% O 2 short-term stress in Example 1 of the present invention
- A is a graph showing changes in ascorbic acid (ASA) content
- B is a graph showing changes in glutathione (GSH) content picture
- ASA ascorbic acid
- GSH glutathione
- Figure 12 is a diagram showing changes in antioxidant enzymes of fresh wolfberry treated with 90% O 2 short-term stress in Example 1 of the present invention; wherein A is superoxide dismutase (SOD), B is catalase (CAT), C is ascorbate peroxidase (APX), D is dehydroascorbate reductase (DHAR), E is monodehydroascorbate reductase (MDHAR), and F is glutathione reductase (GR);
- A superoxide dismutase
- B catalase
- C ascorbate peroxidase
- DHAR dehydroascorbate reductase
- DHAR dehydroascorbate reductase
- MDHAR monodehydroascorbate reductase
- F glutathione reductase
- Figure 13 is a diagram of the appearance and internal structure microstructure of Lingwu Changzao treated with short-term stress treatment with oxygen at different concentrations in Example 2 of the present invention
- A is the appearance quality picture of Lingwu Changzao
- B is the internal structure of Lingwu Changzao microstructure diagram
- Figure 14 is a graph showing changes in hardness of Lingwu jujube treated with short-term stress treatment with oxygen at different concentrations in Example 2 of the present invention.
- Figure 15 is a graph showing changes in electrical conductivity of Lingwu jujube treated with short-term stress treatment with oxygen at different concentrations in Example 2 of the present invention.
- Figure 16 is a graph showing the changes in enzyme activity related to the quality of Lingwu jujube treated with short-term stress treatment with oxygen at different concentrations in Example 2 of the present invention.
- Figure 17 is a graph showing changes in hardness of Lingwu jujube treated with different high-oxygen methods in Example 3 of the present invention.
- Figure 18 is a graph showing changes in conductivity of Lingwu Changzao treated with different high-oxygen methods in Example 3 of the present invention.
- Figure 19 is a graph showing changes in enzyme activity related to the quality of Lingwu jujube treated with different high-oxygen methods in Example 3 of the present invention.
- Figure 20 is a diagram showing the preservation effect, color a* value and hardness change of Daqing grapes treated with 80% O 2 short-term stress in Example 4 of the present invention; where A is the 80% O 2 short-term stress treatment on the 35th day of storage. Picture of the preservation effect of green grapes after 30 minutes of stimulation treatment. B is the picture of the preservation effect of the blank control group on the 35th day of storage. C is the change of color a* value. D is the change of hardness.
- Figure 21 is a diagram showing changes in soluble solids in green grapes treated with 80% O 2 short-term stress in Example 4 of the present invention.
- Figure 22 is a graph showing changes in electrical conductivity, antioxidant capacity, superoxide dismutase (SOD) activity and lipoxygenase (LOX) activity of green grapes treated with 80% O 2 short-term stress in Example 4 of the present invention; wherein , A is the change of conductivity, B is the change of free radical scavenging rate (DPPH), C is the change of SOD enzyme activity, D is the change of LOX enzyme activity;
- Figure 23 is a graph showing changes in total phenols and browning-related enzyme activities of green grapes treated with 80% O 2 short-term stress in Example 4 of the present invention
- A is a graph showing changes in total phenols
- B is phenylalanine ammonia lyase.
- C is the polyphenol oxidase (PPO) enzyme activity change chart
- D is the peroxidase (POD) enzyme activity change chart.
- the raw materials used in the present invention are all conventional commercially available products; the methods used in the present invention, unless otherwise specified, are conventional methods in the field.
- a high-oxygen stress preservation method suitable for the storage of fresh fruits uses 50%-100% high-concentration oxygen as the main gas and high-purity nitrogen as the auxiliary gas to stimulate the fresh fruits for 20-30 minutes for short-term preservation. .
- the present invention mainly optimizes the optimal high oxygen stress concentration, time and other process parameters of fresh wolfberry, and can realize standardized enterprise demonstration applications, improve the post-harvest storage quality of fresh wolfberry, reduce the rot rate of fresh fruit, etc.
- Post-harvest pre-cooling treatment of fresh fruits Fresh fruits are packed in fruit and vegetable boxes after being harvested. In order to prevent excessive water loss, a perforated PE fresh-keeping bag is placed on the outside of the fruit and vegetable box, and then the whole is blasted at low temperature in the cold storage to remove field heat. The temperature is 0 ⁇ 1°C and the time is 18-24 hours;
- High oxygen stress treatment Put the selected fresh fruits into a sealed air-conditioned box, first use high-purity nitrogen to discharge the remaining gas in the sealed air-conditioned box, and then pass 50 %-100% O 2 (w/w) high-concentration oxygen stress treatment for 20-30 minutes, then open the sealed air-conditioning box and let it stand for 20-30 minutes to release the high-concentration oxygen remaining in the air-conditioning box and reduce the oxygen content. The concentration returns it to its natural state;
- the diameter of the hole of the PE fresh-keeping bag in step (1) is 0.013-0.020mm.
- step (4) the fresh fruits treated with high oxygen stress are stored under MAP spontaneous atmosphere control and 0 ⁇ 1°C.
- the fresh fruits are fresh wolfberry, Lingwu jujube and Daqing grape.
- the conditions for high-oxygen stress treatment are: introducing high-concentration oxygen of 90% O 2 (w/w) into a sealed air-conditioned box for short-term stress treatment 30min, then open the sealed air conditioning box and let it sit for 30min;
- the conditions for high oxygen stress treatment are: introduce oxygen with a concentration of 50% O 2 (w/w) into the sealed air-conditioned box for short-term stress treatment for 30 minutes, and then open the seal The air-conditioning box is left for 30 minutes;
- the conditions for high-oxygen stress treatment are: introduce oxygen with a concentration of 80% O 2 (w/w) into the sealed air-conditioned box for short-term stress treatment for 30 minutes, and then open the sealed box. Atmospheric conditioning box, let stand for 30 minutes.
- a high-oxygen stress preservation method suitable for storing fresh wolfberry the specific steps are as follows:
- Post-harvest pre-cooling treatment of fresh wolfberry After the fresh fruits are harvested, they are first blasted at low temperature in the cold storage to remove field heat for 18-24 hours. To prevent excessive water loss, put a perforated PE fresh-keeping bag (0.014mm) on the outside of the fruit and vegetable box. .
- High oxygen stress treatment Put the selected fresh wolfberry into a sealed air-conditioned box, first use high-purity nitrogen to discharge the remaining gas from the sealed air-conditioned box, and then pass it into the sealed air-conditioned box. 90% O 2 (w/w) high-concentration oxygen was subjected to short-term stress treatment for 30 minutes and long-term treatment for 60 minutes respectively, and then the sealed air-conditioning box was opened and left to stand for 30 minutes to release the high-concentration oxygen remaining in the air-conditioning box. The oxygen concentration was lowered to restore it to its natural state, and fresh wolfberry fruits without any treatment were used as the control group.
- Static storage fresh wolfberry that has been pre-cooled and subjected to short-term hyperoxic stress treatment at low temperature for 30 minutes and long-term hyperoxic stress treatment for 60 minutes, fresh wolfberry without any treatment and short-term hyperoxic stress treatment without pre-cooling Fresh wolfberry treated with time stress for 30 minutes was stored in PE fresh-keeping bags at 0 ⁇ 1°C.
- Quality evaluation The quality indicators of fresh wolfberry are measured regularly.
- the 90% O 2 short-term stress treatment after pre-cooling of the present invention effectively inhibited the increase in weight loss rate and was significantly (P ⁇ 0.05) lower than that without pre-cooling.
- Cold direct 90% O 2 short-term stress treatment and blank control low-temperature storage treatment groups In addition, the weight loss rate of fresh wolfberry fruits treated with 90% O 2 short-term stress without pre-cooling was higher than that of the blank control low-temperature storage treatment group.
- the above results show that there is a synergistic effect between the high-oxygen short-term stress treatment of the present invention under low-temperature conditions, that is, 90% O 2 +30 min, pre-cooling treatment, and storage under 0 ⁇ 1°C.
- the method of the present invention simultaneously uses high-oxygen short-term stress treatment. Temporal stress treatment, pre-cooling treatment and storage at 0 ⁇ 1°C can better maintain fruit moisture.
- the hardness value of fresh wolfberry fruits treated with 90% O 2 for short-term emergency treatment without pre-cooling was significantly (P ⁇ 0.05) lower than that of fresh wolfberry fruits treated with low-temperature storage as the blank control.
- the above results show that there is a synergistic effect between the high-oxygen short-term stress treatment of the present invention under low-temperature conditions, that is, 90% O 2 +30 min, pre-cooling treatment, and storage under 0 ⁇ 1°C.
- the method of the present invention simultaneously uses high-oxygen short-term stress treatment. Temporal stress treatment, pre-cooling treatment and storage at 0 ⁇ 1°C can better maintain fruit hardness.
- the L* values of fresh wolfberry fruits treated with 90% O 2 short-term stress after pre-cooling of the present invention were significantly (P ⁇ 0.05) higher than those in other treatment groups, while those without pre-cooling
- the L* value of fresh wolfberry fruits treated directly with 90% O 2 for short-term emergency treatment was significantly (P ⁇ 0.05) lower than that of fresh wolfberry fruits treated with low-temperature storage in the blank control.
- the above results show that there is a synergistic effect between the high-oxygen short-term stress treatment of the present invention under low-temperature conditions, that is, 90% O 2 +30 min, pre-cooling treatment, and storage under 0 ⁇ 1°C.
- the method of the present invention simultaneously uses high-oxygen short-term stress treatment. Temporal stress treatment, pre-cooling treatment and storage at 0 ⁇ 1°C can better maintain the bright color of the fruit and maintain a high level of antioxidant components such as carotenoids in the fruit.
- the 90% hyperoxic stress treatment of the present invention decreased by approximately 66.7% on the 10th day (P ⁇ 0.05) Release amount.
- Figure 10B shows that with The release trend is similar.
- the H 2 O 2 content of the fruits treated with 90% high oxygen stress of the present invention is lower than that of the blank control group.
- Figure 10C shows that the MDA content showed a trend of first increasing and then decreasing in both groups. With the extension of storage time, the MDA content of the 90% hyperoxic stress treated group of the present invention was also significantly lower than that of the control group (P ⁇ 0.05).
- the 90% hyperoxic stress treated group of the present invention was significantly lower than the control group.
- the high oxygen stress treatment of the present invention reduces the accumulation of ROS and MDA in the fruit, helps to alleviate membrane damage of wolfberry fruits, and maintains post-harvest quality.
- Ascorbic acid (ASA) and reduced glutathione (GSH) are potent antioxidants in plants and are important components of the ASA-GSH pathway, which protect against oxidative damage by scavenging ROS.
- the ASA content in the blank control group showed a downward trend throughout the storage period.
- the 90% hyperoxic stress treatment of the present invention reduced the ASA content due to the body's reaction in the early stages of stress, it rapidly changed the body's pathways and delayed the ASA content. The content showed a downward trend and remained at a higher level compared with the control group.
- the 90% hyperoxic stress treatment of the present invention quickly activated the accumulation of GSH content to resist the damage caused by reactive oxygen species, and showed an upward trend in the later stage of storage, with the level basically higher than that of the blank control group .
- the above results show that the high oxygen stress treatment of the present invention promotes the production of antioxidant components in fresh wolfberry and improves the ability to remove harmful reactive oxygen species.
- the antioxidant system plays an important role in the ripening and aging process of fruits and vegetables.
- SOD enzyme can serve as an efficient scavenger of intracellular reactive oxygen species, catalytically decomposing it into H 2 O 2 , which is then converted into oxygen and non-toxic water by CAT and APX enzymes. It can be seen from Figure 12A-C that the SOD, CAT, and APX enzyme activities of fresh wolfberry show an upward trend in the early stages to resist the H 2 O 2 content and It increases rapidly to control changes in fruit quality and plays a key role in the defense response of fresh wolfberry berries against oxidative damage, which can better remove reactive oxygen species and maintain storage quality.
- the 90% hyperoxic stress treatment of the present invention improved the antioxidant capacity of the fruit, not only increased the activities of SOD, CAT and APX, but also inhibited the H 2 O 2 content and Release amount.
- non-enzymatic endogenous antioxidants such as ASA and GSH, are also crucial for controlling the accumulation of ROS in plants, and they are regulated by APX, DHAR, MDHAR and GR activities. It is worth noting that, as shown in Figure 12C-F, the APX, DHAR, MDHAR and GR enzyme activities of the 90% hyperoxic stress treatment group of the present invention are all higher than those of the blank control group, which promotes the ASA-GSH cycle.
- the high oxygen stress treatment of the present invention can effectively improve the activity of reactive oxygen scavenging enzymes in fresh wolfberry fruits, promote the cyclic regeneration of ASA and GSH, thereby enhancing the scavenging ability of reactive oxygen species in fresh wolfberry fruits and reducing the accumulation of reactive oxygen species. Improve the cold storage quality of fresh wolfberry fruit.
- a high-oxygen stress preservation method suitable for Lingwu jujube storage The specific steps are as follows:
- Post-harvest pre-cooling treatment of Lingwu Changzao After harvesting, the fresh fruits are first blasted at low temperature in the cold storage to remove field heat for 18-24 hours. In order to prevent excessive water loss, a perforated PE fresh-keeping bag (0.014mm) is placed in the fruit and vegetable box. outside.
- Oxygen stress treatment Put the selected Lingwu Changzao into a sealed air-conditioned box, first use high-purity nitrogen to discharge the remaining gas from the sealed air-conditioned box, and then pass it into the sealed air-conditioned box. Short-term stress treatment with oxygen at concentrations of 0% (w/w), 50% O 2 (w/w), and 90% O 2 (w/w) for 30 minutes, then open the sealed air conditioning box and let it stand for 30 minutes. The high-concentration oxygen remaining in the controlled atmosphere box is released, and the oxygen concentration is reduced to return to its natural state. The Lingwu jujube fruits without any treatment are used as the control group.
- Quality evaluation The quality indicators of Lingwu Changzao are measured regularly.
- FIG. 13A The effects of short-term stress treatment with different concentrations of oxygen on the appearance quality of Lingwu Changzao jujube are shown in Figure 13A.
- the fresh fruit color of Lingwu Changzao on the 56th day was relatively plump and bright regardless of whether it was treated with 50% high oxygen stress or 90% high oxygen stress.
- the control group and the 0% hypoxia treatment group began to experience peel shrinkage from the 28th and 35th days respectively. Scanning electron microscopy was used to observe the microstructure of Lingwu jujube pulp. The results are shown in Figure 13B. In the early stage of storage, fresh jujube pulp cells showed crystalline arrangement, small intercellular gaps, and complete cell structure.
- the cell structure showed an unfavorable structure in the early stage due to the effect of high oxygen stress, the damage to the tissue structure was slowed down in the later stage.
- the 50% hyperoxic stress treatment group not only delayed the red-turning period of fresh dates compared to other treatment groups, but also delayed internal tissue damage.
- Hardness is one of the important indicators to measure storage quality, which can reveal the commercial value and maturity of fruits during storage to a certain extent.
- the hardness of Lingwu Changzao gradually decreased during storage.
- the fruits in the 50% high oxygen stress treatment group could maintain a significantly higher hardness level during the entire storage period than the other treatment groups.
- the 90% high oxygen stress treatment can maintain a high hardness value in the first 35 days of storage, but the hardness drops seriously in the later stage of storage. It may be because the oxygen concentration is too high, which causes certain damage to the cell membrane in the later stage of storage.
- Electrical conductivity is an important indicator that reflects the integrity of fruit and vegetable cell membranes. Higher electrical conductivity usually indicates lower integrity of the cell membrane and accelerated loss of nutrients in jujube fruits. As can be seen from Figure 15, an overall observation of the changes in electrical conductivity of Lingwu Changzao fruits shows that the electrical conductivity of the fruits generally shows an upward trend with the increase of storage time, indicating that the integrity of the cell membrane gradually increases with the increase of storage time. reduce.
- POD's main task is to help fruits and vegetables remove free radicals and inhibit the oxidation of fruits and vegetables. It can be seen from Figure 16A that as the storage time increases, the POD enzyme activity of Lingwu Changzao shows a trend of first increasing and then decreasing. On the 28th day, the fruits treated with 0% hypoxia and 90% high oxygen short shock showed a peak of enzyme activity. On the 35th day, the fruits of the blank control group Lingwu Changzao showed a peak of enzyme activity. On the 42nd day , the peak of enzyme activity appeared in the 50% hyperoxic short-shock treatment group.
- the antioxidant enzyme activity of Lingwu Changzao in the 50% hyperoxic short-shock treatment group lasts longer and the antioxidant capacity is stronger; after the enzyme peak appears, the activity decreases, indicating that as the fruit matures, the number of free radicals increases. increase, destroying the resistance of antioxidants and causing fruit senescence.
- the appropriate concentration of high oxygen stress treatment that is, 50% O 2 +30min, can slow down the oxidation rate of Lingwu Changzao fruit and play a role in preservation.
- the main task of PPO is to transmit information and oxidize phenolic substances into quinones.
- the quinone compounds then form brown polymers, which are associated with tissue browning during post-harvest storage of fruits and vegetables. close.
- the PPO enzyme activity was low 28 days before storage, even tending to 0. Starting from the 28th day, the PPO enzyme activity of the fruits in the 0% hypoxia treatment group began to show an upward trend, while starting from the 35th day, the PPO enzyme activity of the fruits in the blank control group and the 90% high oxygen short stimulation treatment group began to increase.
- the PPO enzyme activity of Lingwu Changzao in the 50% high oxygen short-shock treatment group showed a significant upward trend only after the 49th day, indicating that this treatment delayed the oxidation process of the fruit; while the Lingwu Changzao in the 0% low oxygen treatment group Jujube has the fastest oxidation rate, followed by 90% high oxygen short stimulation treatment. Therefore, the appropriate concentration of high oxygen stress treatment, that is, 50% O 2 +30min, can slow down the oxidation rate of Lingwu Changzao fruit, reduce browning, and play a certain role in preservation.
- the main working process of ADH is to use ethanol as a substrate to decompose and convert ethanol, which can avoid the damage of ethanol to the fruit and vegetable body to a certain extent.
- Figure 16C It can be seen from Figure 16C that as the storage time increases, the changes in ADH enzyme activity first increase and then decrease. At the 21st day, the fruits treated with 0% hypoxia and 90% high oxygen short shock showed a peak of enzyme activity. On the 35th day, the blank control group showed a peak of enzyme activity, while the fruit treated with 50% high oxygen short shock showed a peak of enzyme activity. The fruits of the group only showed the peak of enzyme activity on the 35th day.
- a high-oxygen stress preservation method suitable for Lingwu jujube storage The specific steps are as follows:
- Post-harvest pre-cooling treatment of Lingwu Changzao After harvesting, the fresh fruits are first blasted at low temperature in the cold storage to remove field heat for 18-24 hours. In order to prevent excessive water loss, a perforated PE fresh-keeping bag (0.014mm) is placed in the fruit and vegetable box. outside.
- Oxygen treatment Put the selected Lingwu Changzao into a sealed air-conditioned box, first use high-purity nitrogen to discharge the rest of the gas in the sealed air-conditioned box, and then introduce 50% of the gas into the sealed air-conditioned box. Perform a short-term stress treatment with O 2 (w/w) oxygen concentration for 30 minutes, then open the sealed air conditioning box and let it stand for 30 minutes to release the high concentration of oxygen remaining in the air conditioning box and reduce the oxygen concentration to return to its natural state. ; The other group was continuously treated with oxygen at a concentration of 50% O 2 (w/w) to keep it in a high oxygen atmosphere; the Lingwu Changzao fruit without any treatment was used as a blank control group.
- Static storage Put the Lingwu Changzao treated with 50% high-concentration oxygen for short-term stress for 30 minutes, the continuous treatment with 50% high-concentration oxygen and the blank control group without any treatment in PE fresh-keeping bags at 0 ⁇ 1 Store at °C.
- Quality evaluation The quality indicators of Lingwu Changzao are measured regularly.
- Hardness is one of the important indicators to measure storage quality, which can reveal the commercial value and maturity of fruits during storage to a certain extent. As shown in Figure 17, the hardness of Lingwu Changzao gradually decreased during storage. Among them, the fruits in the 50% hyperoxic stress treatment group of the present invention can maintain a significantly higher hardness level during the entire storage period than other treatment groups, and there is not much difference in hardness during the entire storage period between the 50% hyperoxic continuous treatment and the blank control group. . The results show that under low-temperature storage conditions, the high-oxygen stress treatment of the present invention can better maintain the fresh and crisp taste of Lingwu Changzao fruits than the continuous high-oxygen treatment and the blank control group. There is a synergistic effect between the high oxygen stress treatment of the present invention, that is, oxygen with a concentration of 50% O 2 (w/w) and short-term stress treatment for 30 minutes.
- Electrical conductivity is an important indicator that reflects the integrity of fruit and vegetable cell membranes. Higher electrical conductivity usually indicates lower integrity of the cell membrane and accelerated loss of nutrients in jujube fruits. As can be seen from Figure 18, an overall observation of the changes in electrical conductivity of Lingwu Changzao fruits shows that the electrical conductivity of the fruits generally shows an upward trend with the increase of storage time, indicating that the integrity of the cell membrane gradually increases with the increase of storage time. reduce.
- the electrical conductivity of the fruits in the 50% high oxygen stress treatment group of the present invention has been at a relatively low level compared with other treatment groups, and the 50% high oxygen continuous treatment although in storage
- the conductivity was suppressed in the early stage, but the conductivity increased rapidly in the middle and late stages of storage, and the level was higher than that of the blank control group.
- the integrity of the cell membrane of Lingwu Changzao was obviously protected within a certain period of time, the rate of loss of nutrients inside the jujube fruit was slowed down, and the quality of Lingwu Changzao was ensured.
- the high-oxygen stress treatment of the present invention can better ensure the integrity of the cell membrane of Lingwu Changzao fruit than the continuous high-oxygen treatment and the blank control group.
- POD's main task is to help fruits and vegetables remove free radicals and inhibit the oxidation of fruits and vegetables. It can be seen from Figure 19A that as the storage time increases, the POD enzyme activity of Lingwu Changzao shows a trend of first increasing and then decreasing. On the 35th day, the Lingwu Changzao fruits in the blank control group showed a peak of enzyme activity. On the 42nd day, the 50% high oxygen short-shock treatment group and the 50% high oxygen continuous treatment group of the present invention showed a peak in enzyme activity. Peak, and the POD level of the 50% hyperoxic short-shock treatment group of the present invention is higher than that of the 50% hyperoxic continuous treatment group.
- the antioxidant enzyme activity of Lingwu Changzao in the 50% hyperoxic short-shock treatment group of the present invention is maintained The longer the time, the stronger the antioxidant capacity; the activity decreases after the enzyme peak appears, indicating that as the fruit matures, the number of free radicals increases greatly, destroying the antioxidant capacity and causing the aging of the fruit.
- the hyperoxic stress treatment of the present invention can enhance the antioxidant capacity of Lingwu Changzao fruit and play a role in preservation than the continuous hyperoxic treatment and the blank control group.
- the main task of PPO is to transmit information and oxidize phenolic substances into quinones.
- the quinone compounds then form brown polymers, which are associated with tissue browning during post-harvest storage of fruits and vegetables. close.
- the PPO enzyme activity was low 28 days before storage, even tending to 0. Starting from the 28th day, the PPO enzyme activity of the fruits in the 50% high oxygen continuous treatment group began to show an upward trend, and starting from the 35th day, the PPO enzyme activity of the fruits in the blank control group began to increase.
- the 50% high oxygen short-term treatment group of the present invention The PPO enzyme activity of Lingwu Changzao in the stimulation treatment group showed a significant upward trend only after the 49th day, indicating that the treatment delayed the oxidation process of the fruit; while the oxidation rate of Lingwu Changzao in the 50% high oxygen continuous treatment group was the fastest. Fast, followed by blank control processing. Therefore, the hyperoxic stress treatment of the present invention can slow down the oxidation rate of Lingwu Changzao fruit, reduce browning, and play a certain role in preserving freshness than the continuous hyperoxic treatment and the blank control group.
- the main working process of ADH is to use ethanol as a substrate to decompose and convert ethanol, which can avoid the damage of ethanol to the fruit and vegetable body to a certain extent.
- Figure 19C it can be seen from Figure 19C that as the storage time increases, the changes in ADH enzyme activity first increase and then decrease.
- the fruits treated with 50% high oxygen continuously showed a peak of enzyme activity.
- the blank control group showed a peak of enzyme activity, while the fruits of the 50% high oxygen short-shock treatment group of the present invention The peak of enzyme activity appeared on the 35th day.
- a high oxygen stress preservation method suitable for the storage of green grapes The specific steps are as follows:
- Post-harvest pre-cooling treatment of Daqing grapes After harvest, the fresh fruits are first blasted at low temperature in the cold storage to remove field heat for 18-24 hours. To prevent excessive water loss, put a perforated PE fresh-keeping bag (0.014mm) on the outside of the fruit and vegetable box. .
- High oxygen stress treatment Put the selected green grapes into a sealed air-conditioned box, first use high-purity nitrogen to discharge the remaining gas from the sealed air-conditioned box, and then pass it into the sealed air-conditioned box. Short-term stress treatment with oxygen at a concentration of 80% O 2 (w/w) for 30 minutes, then open the sealed air conditioning box and let it stand for 30 minutes to release the high concentration of oxygen remaining in the air conditioning box and reduce the oxygen concentration to return to Fresh wolfberry fruits in their natural state without any treatment were used as the control group.
- Static storage The green grapes treated with high-concentration oxygen for short-term stress for 30 minutes and the blank control group without any treatment were placed in PE fresh-keeping bags and stored at 0 ⁇ 1°C.
- Quality evaluation The quality indicators of Daqing grapes are measured regularly.
- Appearance quality is an important indicator of the commercial value of fruits and vegetables. It can be seen from Figure 20A-B that on the 35th day of storage of Daqing grapes, compared with the blank control group without hyperoxic stress treatment, after 80% hyperoxic stress treatment of the present invention, The fruits treated with stimulation significantly inhibited the rot of the fruits and maintained the commercial value of the fruits. From the perspective of color, the lower the color difference a* value, the greener the fruit, and the higher the fruit, the redder. It can be observed from Figure 17C that the color difference a* value shows an overall upward trend, indicating that as the storage time increases, Daqing grapes will Gradually fade away.
- the hardness value of the fruit treated with 80% high oxygen stress of the present invention was significantly higher than that of the blank group from the 15th day onwards, indicating that the high oxygen stress treatment of the present invention can slow down the softening of the fruit and maintain the hardness of the fruit. .
- the soluble solid content (SSC) of fruit is an important indicator of fruit maturity and quality, and directly determines the taste of fruits and vegetables. It can be observed from Figure 21 that the SSC of the blank group did not change significantly during storage, but the SSC of the Daqing grapes after the 80% hyperoxic stress treatment of the present invention increased compared with the blank group, indicating that this treatment effectively increased the SSC of the Daqing grape. The sweetness of green grapes improves the quality of the fruit.
- LOX specifically catalyzes the conversion of membrane lipid unsaturated fatty acids into saturated fatty acids, accelerates the accumulation of peroxidized metabolites, destroys cell membrane structure, and induces membrane lipid peroxidation. In addition, LOX also promotes the loss of cell membrane function, thereby reducing postharvest storage capacity. It can be seen from Figure 20D that the overall LOX content of Daqing grape first increased and then decreased. The peak value of the 80% hyperoxic stress treatment group of the present invention appeared 7 days later than that of the blank group, indicating that the indirect hyperoxic treatment delayed the LOX peak value. appearance, and maintain a low level of LOX activity. Therefore, the high oxygen stress treatment of the present invention can maintain cell membrane function and is an effective treatment technology to improve the postharvest storage capacity of fruits.
- the antioxidant capacity of Daqing grape is expressed by DPPH free radical scavenging rate.
- DPPH free radical scavenging rate As can be seen from Figure 20B, during the 35-day storage period, compared with the blank group, the 80% hyperoxic stress treatment of the present invention The fruit showed the highest level of DPPH, and its ability to scavenge free radicals was significantly higher than that of the blank group in the last week, indicating that the 80% high oxygen stress treatment had the highest ability to scavenge free radicals.
- ROS reactive oxygen species
- Phenolic substances are the main nutrients of fresh fruits and vegetables. They can give fruits and vegetables the function of promoting human health.
- PAL is a key enzyme in the phenylpropanoid metabolism pathway in plants. Its activity controls a variety of phenolic compounds in plants.
- both phenolics and PAL play important roles in plants themselves as secondary metabolites produced in response to biotic and abiotic stresses. It can be observed from Figure 21A that in the first 14 days, after the 80% hyperoxic stress treatment of the present invention, the changes in fruit phenolic substances fluctuated greatly, which shows that the fluctuation trend of phenolic substances is caused by the plant's defense response and high oxygen stimulation. caused together.
- PAL participates in the browning substrate ( Synthesis of phenolic compounds), therefore, while the fruit treated with 80% high oxygen stress of the present invention maintains a lower total phenolic content, the PAL level remains lower than that of the blank group. It shows that the high oxygen stress treatment of the present invention inhibits the synthesis of browning substrates in green grape fruits.
- PPO Polyphenol oxidase
- the high oxygen stress treatment of the present invention effectively inhibits the browning of postharvest Daqing grape fruits because it acts as one of the three modes of anti-browning inhibitors and reduces the risk of fruit damage.
- the content of total phenols increases the activities of PPO and POD.
- the other two modes are: first, increasing the content of total phenols and reducing the activities of PAL, PPO and POD; second, increasing the content of total phenols and the activities of PAL, PPO and POD. .
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Abstract
A high-oxygen stress freshness preservation method suitable for fresh fruit storage: picking fruits, then performing precooling treatment, selecting a sample, placing the sample in a sealed box, introducing high-purity nitrogen to discharge other gases, introducing 50%-100% (w/w) of high-concentration oxygen for stress treatment for 20-30 min, opening the sealed box, standing for 20-30 min, and then performing static storage.
Description
本发明属于食品保鲜技术领域，尤其是一种适用于鲜果贮藏的高氧应激保鲜方法和应用。The invention belongs to the technical field of food preservation, especially a high oxygen stress preservation method and application suitable for the storage of fresh fruits.
我国果蔬年产量约10.2亿吨，但损失高达20％-30％，是欧美国家的2-3倍，严重困扰农民增收。疫情后，我国社区生鲜团购电商净增新用户约2542万，生鲜食品的贮运已成为保证产品品质的关键因素之一。然而，果蔬贮运销过程“预冷-诱抗-贮藏-运输-销售”环节中“诱抗”被忽视。本发明首次以高浓度气体短时分子胁迫应激响应为手段，在阐明果蔬采后响应高氧/CO
2应激的ROS-Redox氧化还原和酶促褐变代谢的系统应激效应与应答机制基础上，创新气体应激耦控色味质的分子应激保鲜技术，明确不同果蔬气体应激浓度效应、时间效应和抑控阈值，突破了MA保鲜气体平衡点掌控难题。
my country's annual fruit and vegetable output is about 1.02 billion tons, but the loss is as high as 20%-30%, which is 2-3 times that of European and American countries, seriously hampering farmers' income growth. After the epidemic, my country's community fresh food group buying e-commerce has a net increase of approximately 25.42 million new users. The storage and transportation of fresh food has become one of the key factors to ensure product quality. However, "induced resistance" is ignored in the "pre-cooling-induced resistance-storage-transportation-sales" process of fruit and vegetable storage, transportation and marketing. For the first time, this invention uses high-concentration gas short-term molecular stress stress response as a means to elucidate the systemic stress effect and response mechanism of ROS-Redox oxidation-reduction and enzymatic browning metabolism of fruits and vegetables in response to high oxygen/CO 2 stress after harvesting. On this basis, innovative molecular stress preservation technology that couples gas stress to control color, taste and quality has clarified the gas stress concentration effects, time effects and suppression thresholds of different fruits and vegetables, breaking through the problem of controlling the balance point of MA preservation gases.
近年来，国内外有许多用于果蔬保鲜的物理和化学技术，如气调、紫外辐射联合壳聚糖涂层、壳聚糖纳米二氧化硅薄膜、氯化钙浸渍法、香精油(百里香-迷迭香、百里香-肉桂等)、1-甲基环丙烯(1-MCP)、抗氧化剂(褪黑素、抗坏血酸、氨基酸、马齿苋提取物)等。其中，气调保鲜被认为是处理皮薄多汁的浆果果实最方便、安全、可行、有效、环保的方法。因为与化学处理及浸泡、涂膜方式相比，气体应激保鲜有较少的安全问题且相对容易实施，可减轻处理对浆果带来的伤害。In recent years, there have been many physical and chemical technologies used for fruit and vegetable preservation at home and abroad, such as controlled atmosphere, ultraviolet radiation combined with chitosan coating, chitosan nano-silica film, calcium chloride impregnation method, essential oil (thyme- Rosemary, thyme-cinnamon, etc.), 1-methylcyclopropene (1-MCP), antioxidants (melatonin, ascorbic acid, amino acids, purslane extract), etc. Among them, controlled atmosphere preservation is considered to be the most convenient, safe, feasible, effective and environmentally friendly method for processing thin-skinned and juicy berries. Because compared with chemical treatment, soaking, and coating methods, gas stress preservation has fewer safety issues and is relatively easy to implement, which can reduce the damage caused to berries.
通过检索，发现如下几篇与本发明专利申请相关的专利公开文献：Through the search, the following patent publications related to the patent application of the present invention were found:
1、一种用于冷却牛肉贮藏保鲜的高氧气调包装气体组成(CN104621231A)，本发明适当降低了混合气体中的O2浓度，并适当提高CO2，将气调包装中的气体体积配比设定为50％～55％O2+35％～40％CO2+10％N2，顶隙空间比为2.5:1～3:1，包装后的牛肉贮藏在4±1℃的条件下。本发明的气体组成既能保持与80％O2高氧包装等同的良好护色效果，又能将冷却牛肉的货架期由80％O2高氧气调包装下的12～16天延长到至少20天以上，还能减少贮藏前期的贮藏汁液损失。1. A high-oxygen modified packaging gas composition for cooling beef storage and preservation (CN104621231A). The present invention appropriately reduces the O2 concentration in the mixed gas, appropriately increases CO2, and sets the gas volume ratio in the modified atmosphere packaging. It is 50% to 55% O2 + 35% to 40% CO2 + 10% N2, and the head space ratio is 2.5:1 to 3:1. The packaged beef is stored at 4±1°C. The gas composition of the present invention can not only maintain the same good color protection effect as that of 80% O2 high-oxygen packaging, but also extend the shelf life of cooled beef from 12 to 16 days under 80% O2 high-oxygen modified packaging to at least 20 days. , and can also reduce the loss of storage juice in the early stage of storage.
2、一种鲜切刺嫩芽高浓度氧气气调包装结合低温贮藏保鲜方法(CN104957244A)，其步骤如下：(1)挑选；(2)清洗；(3)晾干；(4)整理；(5)装袋；(6)抽气；(7)充气；(8)密封；(9)贮藏。该方法采用高阻隔性聚乙烯薄膜作为气调包装袋，高氧气调包装充气比例分别为75％-95％氧气和25％-5％二氧化碳气体，气调包装后再结合低温贮藏。本发明应用于鲜切刺嫩芽的保鲜中，显著了降低无氧呼吸产生毒害影响，同时降低呼吸作用，防止颜色改变，减少水分蒸发和病虫害的侵染，最大程度保持刺嫩芽原有的独特风味和营养成分，有效延长鲜切刺嫩芽货架期。该技术方案显著抑制鲜切刺嫩芽腐烂、褐变以及维生素C和总酚等营养成分降低，提高其硬度和抗氧化活性，其产品具有较高感观品质和卫生质量，产品保鲜期从2～3延长至10～12天，价格可以提高2倍。2. A method of high-concentration oxygen modified atmosphere packaging combined with low-temperature storage and preservation of fresh-cut thorn sprouts (CN104957244A). The steps are as follows: (1) selection; (2) cleaning; (3) drying; (4) sorting; ( 5) Bagging; (6) Exhausting; (7) Inflating; (8) Sealing; (9) Storage. This method uses a high-barrier polyethylene film as a modified atmosphere packaging bag. The inflation ratio of the high-oxygen modified packaging is 75%-95% oxygen and 25%-5% carbon dioxide gas respectively. The modified atmosphere packaging is combined with low-temperature storage. The present invention is applied to the preservation of freshly cut thorn buds, which significantly reduces the toxic effects of anaerobic respiration, reduces respiration, prevents color change, reduces water evaporation and infection by diseases and insect pests, and maintains the original color of the thorn buds to the greatest extent. The unique flavor and nutritional content effectively extend the shelf life of fresh-cut thorn sprouts. This technical solution significantly inhibits the rot and browning of fresh-cut thorn shoots and the reduction of nutrients such as vitamin C and total phenols, and improves their hardness and antioxidant activity. The product has high sensory quality and hygienic quality, and the product shelf life is from 2 ~3 is extended to 10~12 days, and the price can be increased by 2 times.
通过对比，本发明专利申请与上述专利公开文献存在本质的不同。By comparison, the patent application of the present invention is essentially different from the above-mentioned patent publications.
发明内容Contents of the invention
本发明目的在于填补鲜食枸杞保鲜技术领域的空缺，提供一种适用于鲜果贮藏的高氧应激保鲜方法和应用。The purpose of the present invention is to fill the gap in the technical field of fresh wolfberry preservation and provide a high-oxygen stress preservation method and application suitable for fresh fruit storage.
本发明解决其技术问题所采用的技术方案是：The technical solutions adopted by the present invention to solve the technical problems are:
一种适用于鲜果贮藏的高氧应激保鲜方法，所述方法是以50％-100％高浓度氧气为主要气体，以高纯氮气为辅助气体对鲜果进行20-30min短时刺激进行保鲜的。A high-oxygen stress preservation method suitable for the storage of fresh fruits. The method uses 50%-100% high-concentration oxygen as the main gas and high-purity nitrogen as the auxiliary gas to stimulate the fresh fruits for 20-30 minutes for short-term preservation. .
进一步地，具体步骤如下：Further, the specific steps are as follows:
(1)挑拣样品：预冷结束后，挑选无病害，无磕碰，大小、颜色均匀的鲜果；(1) Pick samples: After pre-cooling, select fresh fruits that are disease-free, bump-free, and uniform in size and color;
(2)鲜果采后预冷处理：新鲜果实采后盛装在果蔬箱内，为防止失水过多，将打孔的PE保鲜袋套在果蔬箱外侧，然后整体在冷库低温鼓风去除田间热，温度为0±1℃，时间为18-24小时；(2) Post-harvest pre-cooling treatment of fresh fruits: Fresh fruits are packed in fruit and vegetable boxes after being harvested. In order to prevent excessive water loss, a perforated PE fresh-keeping bag is placed on the outside of the fruit and vegetable box, and then the whole is blasted at low temperature in the cold storage to remove field heat. The temperature is 0±1℃ and the time is 18-24 hours;
(3)高氧应激处理：将挑拣好的鲜果放入到密封的气调箱内，先用高纯氮气将密封的气调箱其余气体排出，再向密封的气调箱内通入50％-100％O
2(w/w)的高浓度氧气应激处理20-30min，然后打开密封的气调箱，静置20-30min，使气调箱内残留的高浓度氧释放，降低氧浓度使其恢复到自然状态；
(3) High oxygen stress treatment: Put the selected fresh fruits into a sealed air-conditioned box, first use high-purity nitrogen to discharge the remaining gas in the sealed air-conditioned box, and then pass 50 %-100% O 2 (w/w) high-concentration oxygen stress treatment for 20-30 minutes, then open the sealed air-conditioning box and let it stand for 20-30 minutes to release the high-concentration oxygen remaining in the air-conditioning box and reduce the oxygen content. The concentration returns it to its natural state;
(4)静态贮藏：将高氧应激处理好的鲜果放在PE保鲜袋内于0±1℃条件下贮藏。(4) Static storage: Store the fresh fruits treated with high oxygen stress in PE fresh-keeping bags at 0±1°C.
进一步地，所述步骤(1)中PE保鲜袋的孔的直径为0.013-0.020mm。Further, the diameter of the hole of the PE fresh-keeping bag in step (1) is 0.013-0.020mm.
进一步地，所述步骤(4)中将高氧应激处理好的鲜果放在MAP自发气调及0±1℃条件下贮藏。Further, in step (4), the fresh fruits treated with high oxygen stress are stored under MAP spontaneous atmosphere control and 0±1°C conditions.
进一步地，所述鲜果为鲜食枸杞、灵武长枣、大青葡萄。Further, the fresh fruits include fresh wolfberry, Lingwu jujube and Daqing grape.
进一步地，当鲜果为鲜食枸杞时，高氧应激处理时的条件为：向密封的气调箱内通入90％O
2(w/w)的高浓度氧气分别短时应激处理30min，然后打开密封的气调箱，静置30min；
Further, when the fresh fruit is fresh wolfberry, the conditions for high-oxygen stress treatment are: introducing high-concentration oxygen of 90% O 2 (w/w) into a sealed air-conditioning box for short-term stress treatment for 30 minutes. , then open the sealed air conditioning box and let it sit for 30 minutes;
当鲜果为灵武长枣时，高氧应激处理时的条件为：向密封的气调箱内通入50％O
2(w/w)浓度的氧气短时应激处理30min，然后打开密封的气调箱，静置30min；
When the fresh fruit is Lingwu jujube, the conditions for high oxygen stress treatment are: introduce oxygen with a concentration of 50% O 2 (w/w) into the sealed air-conditioned box for short-term stress treatment for 30 minutes, and then open the seal The air-conditioning box is left for 30 minutes;
当鲜果为大青葡萄时，高氧应激处理时的条件为：向密封的气调箱内通入80％O
2(w/w)浓度的氧气短时应激处理30min，然后打开密封的气调箱，静置30min。
When the fresh fruit is green grapes, the conditions for high-oxygen stress treatment are: introduce oxygen with a concentration of 80% O 2 (w/w) into the sealed air-conditioned box for short-term stress treatment for 30 minutes, and then open the sealed box. Atmospheric conditioning box, let stand for 30 minutes.
如上所述的保鲜方法在鲜果贮藏方面中的应用。The application of the above-mentioned preservation method in the storage of fresh fruits.
本发明取得的优点和积极效果为：The advantages and positive effects achieved by the present invention are:
1、本发明在鲜食枸杞采后低温差压预冷处理过程中，将打了孔的PE保鲜袋套在果蔬箱外侧进行预冷处理，既能达到快速降温的效果，又能保持良好的透气性，有效缓解外界环境对其引起的严重失水问题，保持鲜食枸杞的水分含量，达到较好保鲜效果。1. In the process of post-harvest low-temperature differential pressure pre-cooling of fresh wolfberry, the present invention puts the perforated PE fresh-keeping bag on the outside of the fruit and vegetable box for pre-cooling, which can not only achieve a rapid cooling effect, but also maintain good air permeability. , effectively alleviate the serious water loss problem caused by the external environment, maintain the moisture content of fresh wolfberry, and achieve better preservation effect.
2、本发明在低温状态下对鲜食枸杞进行50％-100％高浓度氧气应激处理，有效抑制鲜果如鲜食枸杞表面微生物的生长，减少了鲜果在贮藏过程中的腐烂，保鲜效果优于未做任何处理的空白对照组，是一种低成本、操作方便安全、有效环保的新型保鲜技术。2. The present invention performs 50%-100% high-concentration oxygen stress treatment on fresh wolfberry at low temperature, effectively inhibiting the growth of microorganisms on the surface of fresh fruits such as fresh wolfberry, reducing the decay of fresh fruits during storage, and having excellent preservation effects. Based on the blank control group without any treatment, it is a new preservation technology that is low-cost, easy to operate, safe, effective and environmentally friendly.
3、本发明对鲜食枸杞进行高浓度氧气20-30min短时处理，有效诱导鲜果如枸杞鲜果体内抗氧化酶相关的基因表达水平，激活鲜果如鲜食枸杞抗氧化酶活性与抗坏血酸和谷胱甘肽抗氧化成分的循环再生，抑制活性氧和丙二醛的积累，减轻对组织细胞的膜伤害及氧化损伤，保持了鲜食枸杞的色泽，硬度，口感及营养成分。3. The present invention treats fresh wolfberry with high-concentration oxygen for a short period of 20-30 minutes, effectively inducing the gene expression level related to antioxidant enzymes in fresh fruits such as fresh wolfberry, and activating the antioxidant enzyme activity of fresh fruits such as fresh wolfberry with ascorbic acid and glutathione. The cyclic regeneration of glycopeptide antioxidant components inhibits the accumulation of reactive oxygen species and malondialdehyde, reduces membrane damage and oxidative damage to tissue cells, and maintains the color, hardness, taste and nutritional content of fresh wolfberry.
4、本发明的高氧应激鲜果保鲜方法，操作方便简单且保鲜效果明显，该技术方法在低温条件下利用高氧非生物胁迫(50％-100％O
2)与短时处理(20-30min)结合，使鲜果快速氧化应激后产生抗性，起到了“植物疫苗”的作用，达到“抑氧夺鲜”的效果，有效解决了鲜果贮藏过程中发霉腐烂、软化、失色、失味等问题，延长了鲜果的贮藏期，保鲜时长为25-30天，好果率90％以上，效果明显优于对鲜果如鲜食枸杞未经高氧应激而只在低温状态下进行单一冷藏的处理及低温条件下高氧非生物胁迫(50％-100％O
2)和长时处理(＞60min)结合的效果。在第30天时，未经高氧应激只在低温状态下进行单一冷藏处理与高氧长时处理鲜果腐烂率约为25-30％，而高氧短时应激处理组腐烂率仅约为10％。
4. The high-oxygen stress fresh fruit preservation method of the present invention is easy and simple to operate and has obvious preservation effects. This technical method utilizes high-oxygen abiotic stress (50%-100% O 2 ) and short-term treatment (20-20%) under low temperature conditions. 30min), the fresh fruit can develop resistance after rapid oxidative stress, acting as a "plant vaccine" to achieve the effect of "suppressing oxygen and gaining freshness", effectively solving the problem of mildew, rot, softening, discoloration and loss of taste during the storage process of fresh fruit. It extends the storage period of fresh fruits to 25-30 days, and the good fruit rate is over 90%. The effect is obviously better than that of fresh fruits such as fresh wolfberries that are only refrigerated at low temperatures without high oxygen stress. The treatment and the effect of combining high oxygen abiotic stress (50%-100% O 2 ) and long-term treatment (>60min) under low temperature conditions. On the 30th day, the rot rate of fresh fruits in the single refrigeration treatment and high oxygen long-term treatment at low temperature without hyperoxic stress was about 25-30%, while the rot rate of the high-oxygen short-term stress treatment group was only about 10%.
5、本发明的高氧应激鲜果保鲜方法，不限于对鲜食枸杞的保鲜，对于其他果蔬同样适用，不同果蔬最佳高氧应激浓度、短处理时长及频率不同。5. The high oxygen stress fresh fruit preservation method of the present invention is not limited to the preservation of fresh wolfberry, but is also applicable to other fruits and vegetables. The optimal high oxygen stress concentration, short treatment time and frequency are different for different fruits and vegetables.
6、本发明方法能够很好地保持果实色泽，硬度，口感，营养成分及好果率，并通过激活抗氧化系统，增强抗褐变能力，减轻果实在贮藏过程中由于活性氧积累造成的组织氧化损伤，有效延缓果实的成熟衰老。6. The method of the present invention can well maintain the color, hardness, taste, nutrients and good fruit rate of the fruit, and by activating the antioxidant system, enhance the anti-browning ability and reduce the tissue damage caused by the accumulation of active oxygen during the storage process of the fruit. Oxidative damage can effectively delay the ripening and aging of fruits.
7、本发明方法操作便捷安全，成本低，利用高浓度氧气短时刺激鲜果，使机体产生应激反应，改变代谢通路，机体抗性增加，高效抑制了贮藏过程中鲜果表面微生物的生长，减少了鲜果的腐烂率，激活了机体抗氧化酶活性与抗氧化成分，抑制了活性氧与丙二醛含量的积累，减轻了组织细胞氧化损伤，延缓了鲜果的成熟与衰老。7. The method of the present invention is convenient and safe to operate, and has low cost. It uses high-concentration oxygen to stimulate the fresh fruit for a short time, causing the body to produce a stress response, changing metabolic pathways, increasing the body's resistance, effectively inhibiting the growth of microorganisms on the surface of the fresh fruit during storage, and reducing It reduces the decay rate of fresh fruits, activates the body's antioxidant enzyme activity and antioxidant components, inhibits the accumulation of reactive oxygen species and malondialdehyde content, reduces oxidative damage to tissue cells, and delays the ripening and aging of fresh fruits.
8、本发明方法在鲜果采后预冷处理时，新鲜果实采后盛装在果蔬箱内，为防止失水过多，将打孔的PE保鲜袋套在果蔬箱外侧，然后整体在冷库低温鼓风去除田间热，该步骤中将新鲜果实、果蔬箱和PE保鲜袋整体放在冷库里预冷，预冷的同时冷库带有鼓风，增强预冷效果，同时为避免鼓风引起的失水问题在果蔬箱外侧套了打孔的PE保鲜袋，不会阻止冷库的凉气进入到果蔬箱内，且尽可能保住鲜食枸杞果的水分。本申请人在预实验的时候试验过，如果果蔬箱外不套PE保鲜袋并直接将其放在冷库预冷的话，失水非常严重，套了打孔的保鲜袋效果提高了很多。8. During the post-harvest pre-cooling treatment of fresh fruits according to the method of the present invention, the fresh fruits are packed in fruit and vegetable boxes after being harvested. In order to prevent excessive water loss, a perforated PE fresh-keeping bag is placed on the outside of the fruit and vegetable box, and then the whole is blasted at low temperature in the cold storage To remove field heat, in this step, fresh fruits, fruit and vegetable boxes and PE fresh-keeping bags are placed in a cold storage for pre-cooling. During pre-cooling, the cold storage is equipped with air blast to enhance the pre-cooling effect and avoid water loss caused by air blast. A perforated PE fresh-keeping bag is placed on the outside of the fruit and vegetable box, which will not prevent the cool air from the cold storage from entering the fruit and vegetable box, and will keep the moisture of the fresh wolfberry fruits as much as possible. The applicant has tested during the preliminary experiment that if the fruit and vegetable boxes are not covered with PE fresh-keeping bags and are directly placed in the cold storage for pre-cooling, water loss will be very serious. The effect of using perforated fresh-keeping bags is much improved.
图1为本发明实施例1中不同温度介导高氧应激对鲜食枸杞腐烂率变化影响图；Figure 1 is a diagram showing the effects of different temperature-mediated hyperoxic stress on changes in the decay rate of fresh wolfberry in Example 1 of the present invention;
图2为本发明实施例1中不同温度介导高氧应激对鲜食枸杞失重率变化影响图；Figure 2 is a diagram showing the influence of different temperature-mediated hyperoxic stress on the weight loss rate of fresh wolfberry in Example 1 of the present invention;
图3为本发明实施例1中不同温度介导高氧应激对鲜食枸杞硬度变化影响图；Figure 3 is a diagram showing the influence of different temperature-mediated hyperoxic stress on changes in hardness of fresh wolfberry in Example 1 of the present invention;
图4为本发明实施例1中不同温度介导高氧应激对鲜食枸杞色泽L*值变化影响图；Figure 4 is a diagram showing the influence of different temperature-mediated hyperoxic stress on the color L* value of fresh wolfberry in Example 1 of the present invention;
图5为本发明实施例1中不同时长高氧刺激对鲜食枸杞腐烂率变化影响图；Figure 5 is a diagram showing the effect of different durations of hyperoxic stimulation on the decay rate of fresh wolfberry in Example 1 of the present invention;
图6为本发明实施例1中不同时长高氧刺激对鲜食枸杞失重率变化影响图；Figure 6 is a graph showing the effect of different durations of hyperoxic stimulation on the weight loss rate of fresh wolfberry in Example 1 of the present invention;
图7为本发明实施例1中不同时长高氧刺激对鲜食枸杞硬度变化影响图；Figure 7 is a graph showing the effect of different durations of hyperoxic stimulation on changes in hardness of fresh wolfberry in Example 1 of the present invention;
图8为本发明实施例1中不同时长高氧刺激对鲜食枸杞色泽L*值变化影响图；Figure 8 is a graph showing the influence of different durations of hyperoxic stimulation on the color L* value of fresh wolfberry in Example 1 of the present invention;
图9为本发明实施例1中90％O
2短时应激处理鲜食枸杞保鲜效果及菌落总数变化图；其中，A
1为贮藏第30天时，鲜食枸杞未经高氧应激单一低温贮藏空白处理效果图，A
2为贮藏第30天时，鲜食枸杞未经高氧应激单一低温贮藏空白处理培养基菌落总数(10^-3)图，B
1为贮藏第30天时，鲜食枸杞90％O
2短时应激处理30min效果图，B
2为贮藏第30天时，鲜食枸杞90％O
2短时应激处理30min培养基菌落总数(10^-3)图；C为菌落总数变化图；
Figure 9 is a diagram showing the preservation effect and total number of bacterial colonies of fresh wolfberry treated with 90% O 2 short-term stress in Example 1 of the present invention; wherein, A1 is the 30th day of storage, fresh wolfberry without high oxygen stress and single low temperature Storage blank treatment effect diagram, A 2 is the total number of bacterial colonies (10^-3) in the culture medium of single low-temperature storage blank treatment of fresh wolfberry on the 30th day of storage without high oxygen stress, B 1 is the fresh wolfberry on the 30th day of storage The effect of 90% O 2 short-term stress treatment of wolfberry for 30 minutes. B 2 is the total number of colonies (10^-3) in the medium of fresh wolfberry treated with 90% O 2 short-term stress for 30 minutes on the 30th day of storage. C is the picture of bacterial colonies. Total change chart;
图10为本发明实施例1中90％O
2短时应激处理鲜食枸杞超氧阴离子释放量、过氧化氢及丙二醛含量变化图；其中，A为超氧阴离子释放量
含量变化图，B为过氧化氢(H
2O
2)含量变化图，C为丙二醛(MDA)含量变化图；
Figure 10 is a graph showing changes in superoxide anion release, hydrogen peroxide and malondialdehyde content of fresh wolfberry treated with 90% O 2 short-term stress in Example 1 of the present invention; where A is the superoxide anion release Content change chart, B is hydrogen peroxide (H 2 O 2 ) content change chart, C is malondialdehyde (MDA) content change chart;
图11为本发明实施例1中90％O
2短时应激处理鲜食枸杞抗氧化成分变化图；其中，A为抗坏血酸(ASA)含量变化图，B为谷胱甘肽(GSH)含量变化图；
Figure 11 is a graph showing changes in antioxidant components of fresh wolfberry treated with 90% O 2 short-term stress in Example 1 of the present invention; A is a graph showing changes in ascorbic acid (ASA) content, and B is a graph showing changes in glutathione (GSH) content picture;
图12为本发明实施例1中90％O
2短时应激处理鲜食枸杞抗氧化酶变化图；其中，A为超氧化物歧化酶(SOD)，B为过氧化氢酶(CAT)，C为抗坏血酸过氧化物酶(APX)，D为脱氢抗坏血酸还原酶(DHAR)，E为单脱氢抗坏血酸还原酶(MDHAR)，F为谷胱甘肽还原酶(GR)；
Figure 12 is a diagram showing changes in antioxidant enzymes of fresh wolfberry treated with 90% O 2 short-term stress in Example 1 of the present invention; wherein A is superoxide dismutase (SOD), B is catalase (CAT), C is ascorbate peroxidase (APX), D is dehydroascorbate reductase (DHAR), E is monodehydroascorbate reductase (MDHAR), and F is glutathione reductase (GR);
图13为本发明实施例2中经不同浓度氧气短时应激处理的灵武长枣外观及内部组织微观结构图；其中A为灵武长枣外观品质图，B为灵武长枣内部组织微观结构图；Figure 13 is a diagram of the appearance and internal structure microstructure of Lingwu Changzao treated with short-term stress treatment with oxygen at different concentrations in Example 2 of the present invention; A is the appearance quality picture of Lingwu Changzao, and B is the internal structure of Lingwu Changzao microstructure diagram;
图14为本发明实施例2中经不同浓度氧气短时应激处理的灵武长枣硬度变化图；Figure 14 is a graph showing changes in hardness of Lingwu jujube treated with short-term stress treatment with oxygen at different concentrations in Example 2 of the present invention;
图15为本发明实施例2中经不同浓度氧气短时应激处理的灵武长枣电导率变化图；Figure 15 is a graph showing changes in electrical conductivity of Lingwu jujube treated with short-term stress treatment with oxygen at different concentrations in Example 2 of the present invention;
图16为本发明实施例2中经不同浓度氧气短时应激处理的灵武长枣品质相关酶活变化图；Figure 16 is a graph showing the changes in enzyme activity related to the quality of Lingwu jujube treated with short-term stress treatment with oxygen at different concentrations in Example 2 of the present invention;
图17为本发明实施例3中经不同高氧方式处理的灵武长枣硬度变化图；Figure 17 is a graph showing changes in hardness of Lingwu jujube treated with different high-oxygen methods in Example 3 of the present invention;
图18为本发明实施例3中经不同高氧方式处理的灵武长枣电导率变化图；Figure 18 is a graph showing changes in conductivity of Lingwu Changzao treated with different high-oxygen methods in Example 3 of the present invention;
图19为本发明实施例3中经不同高氧方式处理的灵武长枣品质相关酶活变化图；Figure 19 is a graph showing changes in enzyme activity related to the quality of Lingwu jujube treated with different high-oxygen methods in Example 3 of the present invention;
图20为本发明实施例4中80％O
2短时应激处理大青葡萄保鲜效果、色泽a*值及硬度变化图；其中，A为贮藏第35天时，80％％O
2短时应激处理30min大青葡萄保鲜效果图，B为贮藏第35天时，空白对照组保鲜效果图，C为色泽a*值变化图，D为硬度变化图；
Figure 20 is a diagram showing the preservation effect, color a* value and hardness change of Daqing grapes treated with 80% O 2 short-term stress in Example 4 of the present invention; where A is the 80% O 2 short-term stress treatment on the 35th day of storage. Picture of the preservation effect of green grapes after 30 minutes of stimulation treatment. B is the picture of the preservation effect of the blank control group on the 35th day of storage. C is the change of color a* value. D is the change of hardness.
图21为本发明实施例4中80％O
2短时应激处理大青葡萄可溶性固形物变化图；
Figure 21 is a diagram showing changes in soluble solids in green grapes treated with 80% O 2 short-term stress in Example 4 of the present invention;
图22为本发明实施例4中80％O
2短时应激处理大青葡萄电导率、抗氧化能力、超氧化物歧化酶(SOD)活性及脂氧合酶(LOX)活性变化图；其中，A为电导率变化图，B为自有基清除率(DPPH)变化图，C为SOD酶活变化图，D为LOX酶活变化图；
Figure 22 is a graph showing changes in electrical conductivity, antioxidant capacity, superoxide dismutase (SOD) activity and lipoxygenase (LOX) activity of green grapes treated with 80% O 2 short-term stress in Example 4 of the present invention; wherein , A is the change of conductivity, B is the change of free radical scavenging rate (DPPH), C is the change of SOD enzyme activity, D is the change of LOX enzyme activity;
图23为本发明实施例4中80％O
2短时应激处理大青葡萄总酚及褐变相关酶活变化图；其中，A为总酚变化图，B为苯丙氨酸解氨酶(PAL)酶活变化图，C为多酚氧化酶(PPO)酶活变化图，D为过氧化物酶(POD)酶活变化图。
Figure 23 is a graph showing changes in total phenols and browning-related enzyme activities of green grapes treated with 80% O 2 short-term stress in Example 4 of the present invention; A is a graph showing changes in total phenols, and B is phenylalanine ammonia lyase. (PAL) enzyme activity change chart, C is the polyphenol oxidase (PPO) enzyme activity change chart, D is the peroxidase (POD) enzyme activity change chart.
下面详细叙述本发明的实施例，需要说明的是，本实施例是叙述性的，不是限定性的，不能以此限定本发明的保护范围。The embodiments of the present invention are described in detail below. It should be noted that this embodiment is illustrative, not restrictive, and cannot be used to limit the scope of the present invention.
本发明中所使用的原料，如无特殊说明，均为常规的市售产品；本发明中所使用的方法，如无特殊说明，均为本领域的常规方法。The raw materials used in the present invention, unless otherwise specified, are all conventional commercially available products; the methods used in the present invention, unless otherwise specified, are conventional methods in the field.
一种适用于鲜果贮藏的高氧应激保鲜方法，所述方法是以50％-100％高浓度氧气为主要气体，以高纯氮气为辅助气体对鲜果进行20-30min短时刺激进行保鲜的。本发明主要优化了鲜食枸杞的最佳高氧应激浓度、时间等工艺参数，可实现标准化企业示范应用，提高鲜食枸杞采后贮藏品质，降低鲜果腐烂率等。A high-oxygen stress preservation method suitable for the storage of fresh fruits. The method uses 50%-100% high-concentration oxygen as the main gas and high-purity nitrogen as the auxiliary gas to stimulate the fresh fruits for 20-30 minutes for short-term preservation. . The present invention mainly optimizes the optimal high oxygen stress concentration, time and other process parameters of fresh wolfberry, and can realize standardized enterprise demonstration applications, improve the post-harvest storage quality of fresh wolfberry, reduce the rot rate of fresh fruit, etc.
较优地，具体步骤如下：Preferably, the specific steps are as follows:
(1)鲜果采后预冷处理：新鲜果实采后盛装在果蔬箱内，为防止失水过多，将打孔的PE保鲜袋套在果蔬箱外侧，然后整体在冷库低温鼓风去除田间热，温度为0±1℃，时间为18-24小时；(1) Post-harvest pre-cooling treatment of fresh fruits: Fresh fruits are packed in fruit and vegetable boxes after being harvested. In order to prevent excessive water loss, a perforated PE fresh-keeping bag is placed on the outside of the fruit and vegetable box, and then the whole is blasted at low temperature in the cold storage to remove field heat. The temperature is 0±1℃ and the time is 18-24 hours;
(2)挑拣样品：预冷结束后，挑选无病害，无磕碰，大小、颜色均匀的鲜果；(2) Pick samples: After pre-cooling, select fresh fruits that are disease-free, bump-free, and uniform in size and color;
(3)高氧应激处理：将挑拣好的鲜果放入到密封的气调箱内，先用高纯氮气将密封的气调箱其余气体排出，再向密封的气调箱内通入50％-100％O
2(w/w)的高浓度氧气应激处理20-30min，然后打开密封的气调箱，静置20-30min，使气调箱内残留的高浓度氧释放，降低氧浓度使其恢复到自然状态；
(3) High oxygen stress treatment: Put the selected fresh fruits into a sealed air-conditioned box, first use high-purity nitrogen to discharge the remaining gas in the sealed air-conditioned box, and then pass 50 %-100% O 2 (w/w) high-concentration oxygen stress treatment for 20-30 minutes, then open the sealed air-conditioning box and let it stand for 20-30 minutes to release the high-concentration oxygen remaining in the air-conditioning box and reduce the oxygen content. The concentration returns it to its natural state;
(4)静态贮藏：将高氧应激处理好的鲜果放在PE保鲜袋内于0±1℃条件下贮藏。(4) Static storage: Store the fresh fruits treated with high oxygen stress in PE fresh-keeping bags at 0±1°C.
较优地，所述步骤(1)中PE保鲜袋的孔的直径为0.013-0.020mm。Preferably, the diameter of the hole of the PE fresh-keeping bag in step (1) is 0.013-0.020mm.
较优地，所述步骤(4)中将高氧应激处理好的鲜果放在MAP自发气调及0±1℃条件下贮藏。Preferably, in step (4), the fresh fruits treated with high oxygen stress are stored under MAP spontaneous atmosphere control and 0±1°C.
较优地，所述鲜果为鲜食枸杞、灵武长枣、大青葡萄。Preferably, the fresh fruits are fresh wolfberry, Lingwu jujube and Daqing grape.
较优地，当鲜果为鲜食枸杞时，高氧应激处理时的条件为：向密封的气调箱内通入90％O
2(w/w)的高浓度氧气分别短时应激处理30min，然后打开密封的气调箱，静置30min；
Preferably, when the fresh fruit is fresh wolfberry, the conditions for high-oxygen stress treatment are: introducing high-concentration oxygen of 90% O 2 (w/w) into a sealed air-conditioned box for short-term stress treatment 30min, then open the sealed air conditioning box and let it sit for 30min;
当鲜果为灵武长枣时，高氧应激处理时的条件为：向密封的气调箱内通入50％O
2(w/w)浓度的氧气短时应激处理30min，然后打开密封的气调箱，静置30min；
When the fresh fruit is Lingwu jujube, the conditions for high oxygen stress treatment are: introduce oxygen with a concentration of 50% O 2 (w/w) into the sealed air-conditioned box for short-term stress treatment for 30 minutes, and then open the seal The air-conditioning box is left for 30 minutes;
当鲜果为大青葡萄时，高氧应激处理时的条件为：向密封的气调箱内通入80％O
2(w/w)浓度的氧气短时应激处理30min，然后打开密封的气调箱，静置30min。
When the fresh fruit is green grapes, the conditions for high-oxygen stress treatment are: introduce oxygen with a concentration of 80% O 2 (w/w) into the sealed air-conditioned box for short-term stress treatment for 30 minutes, and then open the sealed box. Atmospheric conditioning box, let stand for 30 minutes.
如上所述的保鲜方法在鲜果贮藏方面中的应用。The application of the above-mentioned preservation method in the storage of fresh fruits.
具体地，相关制备及检测实施例如下：Specifically, relevant preparation and detection examples are as follows:
实施例1Example 1
一种适于鲜食枸杞贮藏的高氧应激保鲜方法，具体步骤如下：A high-oxygen stress preservation method suitable for storing fresh wolfberry, the specific steps are as follows:
1、鲜食枸杞采后预冷处理：新鲜果实采后先在冷库低温鼓风去除田间热18-24h，为防止失水过多，将打孔的PE保鲜袋(0.014mm)套在果蔬箱外侧。1. Post-harvest pre-cooling treatment of fresh wolfberry: After the fresh fruits are harvested, they are first blasted at low temperature in the cold storage to remove field heat for 18-24 hours. To prevent excessive water loss, put a perforated PE fresh-keeping bag (0.014mm) on the outside of the fruit and vegetable box. .
2、挑拣样品：预冷结束后，挑选无病害，无磕碰，大小、颜色均匀的鲜果。2. Pick samples: After pre-cooling, select fresh fruits that are disease-free, bump-free, and uniform in size and color.
3、高氧应激处理：将挑拣好的鲜食枸杞放入到密封的气调箱内，先用高纯氮气将密封的气调箱其余气体排出，在向密封的气调箱内通入90％O
2(w/w)的高浓度氧气分别短时应激处理30min、长时处理60min，然后打开密封的气调箱，静置30min，使气调箱内残留的高浓度氧释放，降低氧浓度使其恢复到自然状态，不做任何处理的鲜食枸杞果实做为对照组。
3. High oxygen stress treatment: Put the selected fresh wolfberry into a sealed air-conditioned box, first use high-purity nitrogen to discharge the remaining gas from the sealed air-conditioned box, and then pass it into the sealed air-conditioned box. 90% O 2 (w/w) high-concentration oxygen was subjected to short-term stress treatment for 30 minutes and long-term treatment for 60 minutes respectively, and then the sealed air-conditioning box was opened and left to stand for 30 minutes to release the high-concentration oxygen remaining in the air-conditioning box. The oxygen concentration was lowered to restore it to its natural state, and fresh wolfberry fruits without any treatment were used as the control group.
4、静态贮藏：将经预冷处理后低温条件下高氧短时应激30min处理与高氧长时处理60min的鲜食枸杞，未做任何处理的鲜食枸杞及未经预冷直接高氧短时应激30min处理的鲜食枸杞放在PE保鲜袋内于0±1℃条件下贮藏。4. Static storage: fresh wolfberry that has been pre-cooled and subjected to short-term hyperoxic stress treatment at low temperature for 30 minutes and long-term hyperoxic stress treatment for 60 minutes, fresh wolfberry without any treatment and short-term hyperoxic stress treatment without pre-cooling Fresh wolfberry treated with time stress for 30 minutes was stored in PE fresh-keeping bags at 0±1°C.
品质评价：定期对鲜食枸杞的品质指标进行测定。Quality evaluation: The quality indicators of fresh wolfberry are measured regularly.
相关检测如下：The relevant tests are as follows:
1、不同温度介导高氧应激对鲜食枸杞腐烂率变化影响1. Effects of different temperatures mediated by hyperoxic stress on changes in the decay rate of fresh wolfberry
由图1可以看出，贮藏第5d时，在常温条件下的枸杞鲜果的腐烂率已高达33.46％，而在低温贮藏条件下的各处理组均未出现腐烂现象，由此可见常温条件下其内部生理代谢活性和果实表面微生物的繁殖极快导致枸杞鲜果在常温条件保鲜期短于5天。此外，在低温贮藏环境下的各处理组，未经预冷处理直接经90％O
2短时应激处理的枸杞鲜果在贮藏第10天时出现了腐烂现象，早于本发明经预冷处理后90％O
2短时应激处理组与低温贮藏空白对照组，且腐烂率保持较高水平。本发明预冷后经90％O
2短时应激处理的枸杞鲜果在贮藏第20天时才出现了腐烂现象，且在第30天时，其腐烂率显著(P≤0.05)低于其他处理组，可见在低温条件下进行本发明的高氧短时应激处理即90％O
2+30min、预冷处理与0±1℃条件下贮藏之间具有协同作用，本发明方法同时使用高氧短时应激处理、预冷处理与0±1℃条件下贮藏能够更好地抑制贮藏环境中微生物的生长繁殖。
It can be seen from Figure 1 that on the 5th day of storage, the rot rate of fresh wolfberry fruits under normal temperature conditions has reached as high as 33.46%, while no rot phenomenon occurred in each treatment group under low temperature storage conditions. This shows that under normal temperature conditions, The internal physiological metabolic activity and the rapid reproduction of microorganisms on the fruit surface cause the fresh wolfberry fruit to be kept fresh for less than 5 days at room temperature. In addition, in each treatment group under a low-temperature storage environment, the fresh wolfberry fruits that were directly treated with 90% O 2 short-term stress treatment without pre-cooling treatment showed rottenness on the 10th day of storage, which was earlier than 90% of the pre-cooling treatment in the present invention. O 2 short-term stress treatment group and low-temperature storage blank control group, and the decay rate remained at a high level. The fresh wolfberry fruit treated with 90% O 2 short-term stress after pre-cooling of the present invention did not rot until the 20th day of storage, and on the 30th day, its rot rate was significantly (P≤0.05) lower than that of other treatment groups. It can be seen that there is a synergistic effect between the high-oxygen short-term stress treatment of the present invention under low-temperature conditions, that is, 90% O 2 +30min, pre-cooling treatment, and storage under 0±1°C. The method of the present invention simultaneously uses high-oxygen short-term stress treatment. Shock treatment, pre-cooling treatment and storage at 0±1°C can better inhibit the growth and reproduction of microorganisms in the storage environment.
2、不同温度介导高氧应激对鲜食枸杞失重率变化影响2. Effects of different temperatures mediated by hyperoxic stress on changes in weight loss rate of fresh wolfberry
由图2可以看出，贮藏于常温条件下的枸杞鲜果在贮藏第0-5d时，其失重率显著(P≤0.05)高于其他低温贮藏处理组，这是由于在贮藏过程中，果实采后的呼吸作用和水分蒸腾作用仍在进行，加之常温贮藏并且未散去“田间热”进而促进呼吸作用和水分蒸腾作用，同时无法适应高氧应激反应产生抗性，品质骤然劣变。低温贮藏条件下的枸杞鲜果的失重率均呈上升趋势，其中本发明预冷后经过90％O
2短时应激处理有效抑制了失重率的上升并且显著(P≤0.05)低于未经预冷直接90％O
2短时应激处理和空白对照低温贮藏处理组；此外，未经预冷直接90％O
2短时应激处理的枸杞鲜果其失重率高于空白对照低温贮藏处理组。以上结果表明，低温条件下进行本发明的高氧短时应激处理即90％O
2+30min、预冷处理与0±1℃条件下贮藏之间具有协同作用，本发明方法同时使用高氧短时应激处理、预冷处理与0±1℃条件下贮藏能够更好地保持果实水分。
It can be seen from Figure 2 that the weight loss rate of fresh wolfberry fruits stored under normal temperature conditions on the 0th to 5th day of storage was significantly (P≤0.05) higher than that of other low-temperature storage treatment groups. This is due to the fact that during the storage process, the fruit is harvested. The subsequent respiration and water transpiration are still going on. In addition, storage at room temperature and the "field heat" has not dissipated, thereby promoting respiration and water transpiration. At the same time, it cannot adapt to the high oxygen stress reaction and develop resistance, and the quality suddenly deteriorates. The weight loss rate of fresh wolfberry fruits under low-temperature storage conditions all showed an upward trend. Among them, the 90% O 2 short-term stress treatment after pre-cooling of the present invention effectively inhibited the increase in weight loss rate and was significantly (P≤0.05) lower than that without pre-cooling. Cold direct 90% O 2 short-term stress treatment and blank control low-temperature storage treatment groups; In addition, the weight loss rate of fresh wolfberry fruits treated with 90% O 2 short-term stress without pre-cooling was higher than that of the blank control low-temperature storage treatment group. The above results show that there is a synergistic effect between the high-oxygen short-term stress treatment of the present invention under low-temperature conditions, that is, 90% O 2 +30 min, pre-cooling treatment, and storage under 0±1°C. The method of the present invention simultaneously uses high-oxygen short-term stress treatment. Temporal stress treatment, pre-cooling treatment and storage at 0±1°C can better maintain fruit moisture.
3、不同温度介导高氧应激对鲜食枸杞硬度变化影响3. Effects of different temperatures mediated by hyperoxic stress on changes in hardness of fresh wolfberry
由图3可以看出，贮藏于常温条件下的枸杞鲜果在贮藏第0-5d时，由于常温条件下细胞壁代谢相关酶活性均处于极其活跃的状态，导致硬度急剧下降并失去了鲜食价值。值得注意的是，在整个贮藏过程中，本发明预冷后经90％O
2短时应激处理并且贮藏于低温条件下贮藏的枸杞鲜果硬度值均显著(P≤0.05)高于其他处理组，可能抑制了细胞壁代谢相关酶活性，有效保持果实原果胶等营养成分，进而延缓了枸杞鲜果硬度的下降。而未经预冷直接90％O
2短时应急处理的枸杞鲜果其硬度值明显(P≤0.05)低于空白对照低温贮藏处理的枸杞鲜果。以上结果表明，低温条件下进行本发明的高氧短时应激处理即90％O
2+30min、预冷处理与0±1℃条件下贮藏之间具有协同作用，本发明方法同时使用高氧短时应激处理、预冷处理与0±1℃条件下贮藏能够更好地保持果实硬度。
It can be seen from Figure 3 that the hardness of fresh wolfberry fruits stored under normal temperature conditions dropped sharply and the fresh food value was lost on the 0th to 5th day of storage due to the extremely active enzyme activities related to cell wall metabolism under normal temperature conditions. It is worth noting that during the entire storage process, the hardness values of the fresh wolfberry fruits treated with 90% O 2 short-term stress after pre-cooling and stored under low temperature conditions were significantly (P≤0.05) higher than those of other treatment groups. , may inhibit the activity of enzymes related to cell wall metabolism, effectively maintain the fruit's protopectin and other nutrients, thereby delaying the decline in the hardness of fresh wolfberry fruits. The hardness value of fresh wolfberry fruits treated with 90% O 2 for short-term emergency treatment without pre-cooling was significantly (P≤0.05) lower than that of fresh wolfberry fruits treated with low-temperature storage as the blank control. The above results show that there is a synergistic effect between the high-oxygen short-term stress treatment of the present invention under low-temperature conditions, that is, 90% O 2 +30 min, pre-cooling treatment, and storage under 0±1°C. The method of the present invention simultaneously uses high-oxygen short-term stress treatment. Temporal stress treatment, pre-cooling treatment and storage at 0±1℃ can better maintain fruit hardness.
4、不同温度介导高氧应激对鲜食枸杞色泽L*值变化影响4. Effects of different temperatures mediated by hyperoxic stress on the color L* value of fresh wolfberry
由图4可知，随着贮藏时间的增加，枸杞鲜果的色泽由鲜亮逐渐变暗。其中，在贮藏第5d时，常温贮藏条件下的枸杞鲜果的L*值降低了33％左右，其下降速度均显著(P≤0.05)大于其他低温贮藏处理组，这是由于在常温条件下更加促进了细胞壁代谢酶活性以及类胡萝卜素的降解，无法适应高氧应激带来的胁迫。而在低温贮藏条件下的处理组中，本发明预冷后经90％O
2短时应激处理的枸杞鲜果L*值均显著(P≤0.05)高于其他处理组，而未经预冷直接90％O
2短时应急处理的枸杞鲜果其L*值明显(P≤0.05)低于空白对照低温贮藏处理的枸杞鲜果。以上结果表明，低温条件下进行本发明的高氧短时应激处理即90％O
2+30min、预冷处理与0±1℃条件下贮藏之间具有协同作用，本发明方法同时使用高氧短时应激处理、预冷处理与0±1℃条件下贮藏能更好的保持果实鲜亮的色泽，使果实内类胡萝卜素等抗氧化成分保持较高水平。
It can be seen from Figure 4 that as the storage time increases, the color of fresh wolfberry fruits gradually changes from bright to dark. Among them, on the 5th day of storage, the L* value of fresh wolfberry fruits under normal temperature storage conditions decreased by about 33%, and their decreasing speeds were significantly (P≤0.05) greater than those of other low-temperature storage treatment groups. It promotes the activity of cell wall metabolic enzymes and the degradation of carotenoids, and cannot adapt to the stress caused by hyperoxic stress. In the treatment group under low-temperature storage conditions, the L* values of fresh wolfberry fruits treated with 90% O 2 short-term stress after pre-cooling of the present invention were significantly (P≤0.05) higher than those in other treatment groups, while those without pre-cooling The L* value of fresh wolfberry fruits treated directly with 90% O 2 for short-term emergency treatment was significantly (P≤0.05) lower than that of fresh wolfberry fruits treated with low-temperature storage in the blank control. The above results show that there is a synergistic effect between the high-oxygen short-term stress treatment of the present invention under low-temperature conditions, that is, 90% O 2 +30 min, pre-cooling treatment, and storage under 0±1°C. The method of the present invention simultaneously uses high-oxygen short-term stress treatment. Temporal stress treatment, pre-cooling treatment and storage at 0±1°C can better maintain the bright color of the fruit and maintain a high level of antioxidant components such as carotenoids in the fruit.
5、不同时长高氧刺激对鲜食枸杞腐烂率变化影响5. Effects of different durations of hyperoxic stimulation on changes in the decay rate of fresh wolfberry
由图5可以看出，高氧刺激30min与60min的处理组与空白对照组腐烂率在贮藏期间均呈上升趋势，在贮藏15天之后，与对照组相比，明显看出，高氧长时60min刺激的鲜食枸杞腐烂率在中后期保持较低水平，但贮藏末期坏果率快速上升，而本发明高氧短时30min刺激的鲜食枸杞腐烂率在整个贮藏期间一直保持较低的腐烂率水平，因此本发明的高氧在短时间内刺激果蔬可以更好地起到保鲜效果。It can be seen from Figure 5 that the decay rates of the treatment groups stimulated by high oxygen for 30 minutes and 60 minutes and the blank control group showed an upward trend during the storage period. After 15 days of storage, compared with the control group, it was obvious that the long-term high oxygen The decay rate of fresh wolfberry stimulated for 60 minutes remained at a low level in the middle and late stages, but the bad fruit rate increased rapidly at the end of storage. However, the decay rate of fresh wolfberry stimulated by high oxygen for 30 minutes in the present invention remained low during the entire storage period. Therefore, the high oxygen of the present invention can stimulate fruits and vegetables in a short time to achieve a better preservation effect.
6、不同时长高氧刺激对鲜食枸杞失重率变化影响6. Effects of different durations of hyperoxic stimulation on changes in weight loss rate of fresh wolfberry
由图6可以看出，高氧刺激30min与60min的处理组与空白对照组失重率在贮藏期间均呈上升趋势，值得注意的是，高氧长时60min刺激的鲜食枸杞失重率水平不仅高于高氧短时30min刺激的鲜食枸杞果实，还高于空白对照组。而本发明高氧短时30min刺激的鲜食枸杞失重率在整个贮藏期间一直保持最低的水平，因此本发明的高氧在短时间内刺激果蔬可以更好地起到保鲜效果，保持果蔬水分。It can be seen from Figure 6 that the weight loss rates of the treatment groups stimulated by hyperoxia for 30 minutes and 60 minutes and the blank control group showed an upward trend during storage. It is worth noting that the weight loss rate of fresh wolfberry stimulated by hyperoxia for 60 minutes was not only high. The results of fresh wolfberry fruits stimulated by high oxygen for 30 minutes were higher than those of the blank control group. The weight loss rate of fresh wolfberry stimulated by high oxygen for 30 minutes in the present invention has remained at the lowest level throughout the storage period. Therefore, the high oxygen stimulation of fruits and vegetables in a short time of the present invention can better preserve the freshness of fruits and vegetables and maintain the moisture of fruits and vegetables.
7、不同时长高氧刺激对鲜食枸杞硬度变化影响7. Effects of different durations of hyperoxic stimulation on changes in hardness of fresh wolfberry
由图7可以看出，高氧刺激30min与60min的处理组与空白对照组硬度在贮藏期间整体均呈下降趋势，值得注意的是，高氧长时60min刺激的鲜食枸杞硬度水平在贮藏前20天一直高于空白对照组，但贮藏后期明显下降趋势变快，表明品质劣变加快。与高氧长时60min刺激的鲜食枸杞和空白对照组相比，本发明高氧短时30min刺激的鲜食枸杞硬度在整个贮藏期间一直保持最高的水平，因此本发明的高氧在短时间内刺激果蔬可以更好的起到保鲜效果，保持果蔬硬度。It can be seen from Figure 7 that the hardness of the treatment groups and the blank control group stimulated by high oxygen for 30 minutes and 60 minutes showed an overall downward trend during the storage period. It is worth noting that the hardness level of fresh wolfberry stimulated by high oxygen for 60 minutes was before storage. It has been higher than the blank control group for 20 days, but the obvious downward trend became faster in the later period of storage, indicating that the quality deterioration accelerated. Compared with the fresh wolfberry stimulated by hyperoxia for 60 minutes and the blank control group, the hardness of the fresh wolfberry stimulated by hyperoxia for 30 minutes remained at the highest level throughout the storage period. Internal stimulation of fruits and vegetables can better preserve the freshness of fruits and vegetables and maintain the hardness of fruits and vegetables.
8、不同时长高氧刺激对鲜食枸杞色泽L*值变化影响8. Effects of different durations of hyperoxic stimulation on the color L* value of fresh wolfberry
由图8可以看出，高氧刺激30min与60min的处理组与空白对照组色泽L*值在贮藏期间整体呈相同的变化趋势，值得注意的是，与空白对照组相比，在贮藏早期高氧刺激降低了L*值，而贮藏中后期经高氧刺激的鲜食枸杞果实L*值高于空白对照组。其中，经高氧长时60min刺激的处理组L*值下降较快，且水平低于本发明高氧短时30min刺激的鲜食枸杞L*值。总体来说，本发明的高氧在短时间内刺激果蔬可以更好地起到保鲜效果，保持果蔬鲜亮的色泽。It can be seen from Figure 8 that the color L* values of the treatment groups stimulated for 30 minutes and 60 minutes by hyperoxia and the blank control group showed the same trend during storage. It is worth noting that compared with the blank control group, the color L* value was higher in the early storage period. Oxygen stimulation reduced the L* value, and the L* value of fresh wolfberry fruits stimulated by high oxygen in the middle and late stages of storage was higher than that of the blank control group. Among them, the L* value of the treatment group stimulated by hyperoxia for 60 minutes dropped rapidly, and the level was lower than the L* value of fresh wolfberry stimulated by hyperoxia for 30 minutes. Generally speaking, the high oxygen in the present invention can stimulate fruits and vegetables in a short period of time to achieve a better preservation effect and maintain the bright color of fruits and vegetables.
9、高氧应激处理对鲜食枸杞保鲜效果及菌落总数变化影响9. Effect of high oxygen stress treatment on the preservation effect of fresh wolfberry and changes in the total number of bacterial colonies
由图9可以看出，贮藏第30天时，本发明90％高氧应激处理组鲜食枸杞(B
2)保鲜效果明显优于空白处理组(A
1)，经90％高氧应激处理的鲜果颜色更鲜亮，腐烂程度更低，与图5、图8表现结果一致。与空白对照组相比，菌落总数的测定(图A
2,B
2和C)也证明了本发明高氧应激处理组菌落总数水平在整个贮藏期间均低于空白对照组，说明本发明的高氧应激处理能够显著抑制果实表面微生物的生长，抑制真菌对果实造成的侵染，提高枸杞果实在冷藏过程中的好果率，维持枸杞果实的品质。
It can be seen from Figure 9 that on the 30th day of storage, the preservation effect of fresh wolfberry (B 2 ) in the 90% hyperoxic stress treatment group of the present invention was significantly better than that in the blank treatment group (A 1 ). After 90% hyperoxic stress treatment, The fresh fruits are brighter in color and less rotten, which is consistent with the results shown in Figures 5 and 8. Compared with the blank control group, the measurement of the total number of bacterial colonies (Figures A 2 , B 2 and C) also proved that the level of the total number of bacterial colonies in the hyperoxic stress treatment group of the present invention was lower than that of the blank control group throughout the storage period, indicating that the present invention High oxygen stress treatment can significantly inhibit the growth of microorganisms on the fruit surface, inhibit fungal infection of the fruit, improve the good fruit rate of wolfberry fruits during the cold storage process, and maintain the quality of wolfberry fruits.
10、高氧应激处理对鲜食枸杞超氧阴离子释放量
过氧化氢(H
2O
2)及丙 二醛(MDA)含量变化影响
10. Effect of high oxygen stress treatment on superoxide anion release of fresh wolfberry Effects of changes in hydrogen peroxide (H 2 O 2 ) and malondialdehyde (MDA) content
果实衰老和品质下降可能归因于ROS(如，
和H
2O
2)引起的细胞内蛋白质、DNA和脂质氧化损伤而细胞膜脂质的氧化损伤主要通过MDA来评价。由图10A可知，果实经本发明90％高氧应激处理后
释放量显著低于对照组(P≤0.05)。且对照组在10d时出现了一个峰值，而本发明的高氧应激处理组抑制了峰值的出现。贮藏10d时，本发明的高氧应激处理和对照果实的最大产率分别为31.7nmol min
-1g
-1和95.0nmol min
-1g
-1。即与空白对照组相比，本发明的90％高氧应激处理在第10天降低了约66.7％(P≤0.05)
释放量。同时，由图10B可知，与
释放量趋势相似，除第20天外，本发明的90％高氧应激处理的果实H
2O
2含量均低于空白对照组。图10C显示，MDA含量在两组中均呈现先升高后降低的趋势。随着贮藏时间的延长，本发明的90％高氧应激处理组MDA含量也明显低于对照组(P≤0.05)，在第30天时，本发明的90％高氧应激处理组比对照组下降36.7％。由以上结果可知，与对照组相比，本发明的高氧应激处理降低了果实ROS和MDA的积累，有助于缓解枸杞果实的膜损伤，维持采后品质。
Fruit senescence and quality decline may be attributed to ROS (e.g., and H 2 O 2 ) caused oxidative damage to intracellular proteins, DNA, and lipids, while oxidative damage to cell membrane lipids was mainly evaluated by MDA. As can be seen from Figure 10A, after the fruit was treated with 90% hyperoxic stress of the present invention, The release amount was significantly lower than that of the control group (P≤0.05). And the control group had a peak at 10 days, while the hyperoxic stress treatment group of the present invention inhibited the peak. When stored for 10 days, the maximum yields of the high oxygen stress treated and control fruits of the present invention were 31.7 nmol min -1 g -1 and 95.0 nmol min -1 g -1 respectively. That is, compared with the blank control group, the 90% hyperoxic stress treatment of the present invention decreased by approximately 66.7% on the 10th day (P≤0.05) Release amount. At the same time, it can be seen from Figure 10B that with The release trend is similar. Except for the 20th day, the H 2 O 2 content of the fruits treated with 90% high oxygen stress of the present invention is lower than that of the blank control group. Figure 10C shows that the MDA content showed a trend of first increasing and then decreasing in both groups. With the extension of storage time, the MDA content of the 90% hyperoxic stress treated group of the present invention was also significantly lower than that of the control group (P≤0.05). On the 30th day, the 90% hyperoxic stress treated group of the present invention was significantly lower than the control group. group fell by 36.7%. It can be seen from the above results that compared with the control group, the high oxygen stress treatment of the present invention reduces the accumulation of ROS and MDA in the fruit, helps to alleviate membrane damage of wolfberry fruits, and maintains post-harvest quality.
11、高氧应激处理对鲜食枸杞抗坏血酸(ASA)和谷胱甘肽(GSH)含量变化影响11. Effect of hyperoxic stress treatment on changes in ascorbic acid (ASA) and glutathione (GSH) contents of fresh wolfberry
抗坏血酸(ASA)和还原型谷胱甘肽(GSH)是植物中有效的抗氧化剂，是ASA-GSH途径的重要组成部分，通过清除ROS来防御氧化损伤。由图11A可知，空白对照组ASA含量整个贮藏期间呈下降趋势，而本发明的90％高氧应激处理虽然在应激初期由于机体反应降低了ASA含量，但快速改变机体通路，延缓了ASA含量的下降的趋势，且与对照组相比保持在较高水平。由图11B可知，贮藏初期，本发明的90％高氧应激处理快速激活了GSH含量的积累，以抵抗活性氧带来的伤害，并且在贮藏后期呈上升趋势，水平基本高于空白对照组。以上结果说明，本发明的高氧应激处理促进了鲜食枸杞体内抗氧化剂成分的生成，提高了清除有害活性氧的能力。Ascorbic acid (ASA) and reduced glutathione (GSH) are potent antioxidants in plants and are important components of the ASA-GSH pathway, which protect against oxidative damage by scavenging ROS. As can be seen from Figure 11A, the ASA content in the blank control group showed a downward trend throughout the storage period. However, although the 90% hyperoxic stress treatment of the present invention reduced the ASA content due to the body's reaction in the early stages of stress, it rapidly changed the body's pathways and delayed the ASA content. The content showed a downward trend and remained at a higher level compared with the control group. As can be seen from Figure 11B, in the early stage of storage, the 90% hyperoxic stress treatment of the present invention quickly activated the accumulation of GSH content to resist the damage caused by reactive oxygen species, and showed an upward trend in the later stage of storage, with the level basically higher than that of the blank control group . The above results show that the high oxygen stress treatment of the present invention promotes the production of antioxidant components in fresh wolfberry and improves the ability to remove harmful reactive oxygen species.
12、高氧应激处理对鲜食枸杞抗氧化酶活变化影响12. Effect of hyperoxic stress treatment on changes in antioxidant enzyme activity of fresh wolfberry
抗氧化系统在果蔬的成熟和衰老过程中起着重要作用。SOD酶作为抗氧化损伤的第一道防线，可以作为细胞内活性氧的高效清除剂，催化分解为H
2O
2，再被CAT和APX酶转化为氧和无毒的水。由图12A-C可知，鲜食枸杞SOD,CAT,APX酶活在早期阶段呈上升趋势,用以抵抗H
2O
2含量和
迅速增加来控制水果品质变化，在枸杞鲜莓抗氧化损伤的防御反应中发挥关键作用，能够更好地清除活性氧，维持贮藏质量。且在整个贮藏期间，与对照组相比,本发明的90％高氧应激处理改善果实的抗氧化能力,不仅提高了SOD、CAT和APX的活性，还抑制了H
2O
2含量和
释放量。此外，非酶性内源抗氧化剂，如ASA和GSH，对控制植物体内ROS的积累也至关重要，他们受APX、DHAR、MDHAR和GR活性的调控。值得注意的是,由图12C-F可知，本发明的90％高氧应激处理组的APX、DHAR，MDHAR和GR酶活均高于空白对照组，促进了ASA-GSH循环。以上结果表明，本发明的高氧应激处理可有效提高枸杞鲜果中活性氧清除酶的活性，促进ASA和GSH的循环再生，从而增强枸杞鲜果中活性氧的清除能力，减少活性氧的积累，提高枸杞鲜果冷藏品质。
The antioxidant system plays an important role in the ripening and aging process of fruits and vegetables. As the first line of defense against oxidative damage, SOD enzyme can serve as an efficient scavenger of intracellular reactive oxygen species, catalytically decomposing it into H 2 O 2 , which is then converted into oxygen and non-toxic water by CAT and APX enzymes. It can be seen from Figure 12A-C that the SOD, CAT, and APX enzyme activities of fresh wolfberry show an upward trend in the early stages to resist the H 2 O 2 content and It increases rapidly to control changes in fruit quality and plays a key role in the defense response of fresh wolfberry berries against oxidative damage, which can better remove reactive oxygen species and maintain storage quality. And during the entire storage period, compared with the control group, the 90% hyperoxic stress treatment of the present invention improved the antioxidant capacity of the fruit, not only increased the activities of SOD, CAT and APX, but also inhibited the H 2 O 2 content and Release amount. In addition, non-enzymatic endogenous antioxidants, such as ASA and GSH, are also crucial for controlling the accumulation of ROS in plants, and they are regulated by APX, DHAR, MDHAR and GR activities. It is worth noting that, as shown in Figure 12C-F, the APX, DHAR, MDHAR and GR enzyme activities of the 90% hyperoxic stress treatment group of the present invention are all higher than those of the blank control group, which promotes the ASA-GSH cycle. The above results show that the high oxygen stress treatment of the present invention can effectively improve the activity of reactive oxygen scavenging enzymes in fresh wolfberry fruits, promote the cyclic regeneration of ASA and GSH, thereby enhancing the scavenging ability of reactive oxygen species in fresh wolfberry fruits and reducing the accumulation of reactive oxygen species. Improve the cold storage quality of fresh wolfberry fruit.
实施例2Example 2
一种适于灵武长枣贮藏的高氧应激保鲜方法，具体步骤如下：A high-oxygen stress preservation method suitable for Lingwu jujube storage. The specific steps are as follows:
1、灵武长枣采后预冷处理：新鲜果实采后先在冷库低温鼓风去除田间热18-24h，为防止失水过多，将打孔的PE保鲜袋(0.014mm)套在果蔬箱外侧。1. Post-harvest pre-cooling treatment of Lingwu Changzao: After harvesting, the fresh fruits are first blasted at low temperature in the cold storage to remove field heat for 18-24 hours. In order to prevent excessive water loss, a perforated PE fresh-keeping bag (0.014mm) is placed in the fruit and vegetable box. outside.
2、挑拣样品：预冷结束后，挑选无病害，无磕碰，大小、颜色均匀的鲜果。2. Pick samples: After pre-cooling, select fresh fruits that are disease-free, bump-free, and uniform in size and color.
3、氧气应激处理：将挑拣好的灵武长枣放入到密封的气调箱内，先用高纯氮气将密封的气调箱其余气体排出，在向密封的气调箱内通入0％(w/w)、50％O
2(w/w)、90％O
2(w/w)浓度的氧气短时应激处理30min，然后打开密封的气调箱，静置30min，使气调箱内残留的高浓度氧释放，降低氧浓度使其恢复到自然状态，不做任何处理的灵武长枣果实做为对照组。
3. Oxygen stress treatment: Put the selected Lingwu Changzao into a sealed air-conditioned box, first use high-purity nitrogen to discharge the remaining gas from the sealed air-conditioned box, and then pass it into the sealed air-conditioned box. Short-term stress treatment with oxygen at concentrations of 0% (w/w), 50% O 2 (w/w), and 90% O 2 (w/w) for 30 minutes, then open the sealed air conditioning box and let it stand for 30 minutes. The high-concentration oxygen remaining in the controlled atmosphere box is released, and the oxygen concentration is reduced to return to its natural state. The Lingwu jujube fruits without any treatment are used as the control group.
4、静态贮藏：将0％(w/w)与50％O
2(w/w)、90％O
2(w/w)低浓度氧与高浓度氧短时应激30min处理的灵武长枣与未做任何处理的空白对照组放在PE保鲜袋内于0±1℃条件下贮藏。
4. Static storage: Lingwu Chang was treated with 0% (w/w), 50% O 2 (w/w), 90% O 2 (w/w) low concentration oxygen and high concentration oxygen for 30 minutes. Jujube and the blank control group without any treatment were placed in PE fresh-keeping bags and stored at 0±1°C.
品质评价：定期对灵武长枣的品质指标进行测定。Quality evaluation: The quality indicators of Lingwu Changzao are measured regularly.
相关检测如下：The relevant tests are as follows:
1、不同浓度氧气短时应激处理对灵武长枣外观品质及内部组织微观结构的变化影响1. Effects of short-term stress treatment with different concentrations of oxygen on the appearance quality and internal tissue microstructure of Lingwu jujube.
不同浓度氧气短时应激处理对灵武长枣枣外观品质的影响如图13A所示。从图13A可以看出，灵武长枣在第56天时，无论是50％高氧应激处理，还是90％高氧应激处理，鲜果颜色都比较饱满鲜艳。而对照组和0％低氧处理组分别从第28天和第35天开始出现果皮皱缩现象。利用扫描电镜观察灵武长枣果肉的微观结构，结果如图13B所示，贮藏初期，鲜枣果肉细胞呈现晶态排列、细胞间隙小、细胞结构完整等特点。但随着贮存时间的延长，细胞间隙逐渐变大，甚至出现明显的细胞破裂。值得注意的是，虽然在高氧应激处理的样品中，由于高氧胁迫的作用，细胞结构在早期表现出不利的结构，但后期组织结构的损伤却减缓了。尤其是50％高氧应激处理组，相比其他处理组不仅延缓了鲜枣的转红期，还延缓了组织内部的损伤。The effects of short-term stress treatment with different concentrations of oxygen on the appearance quality of Lingwu Changzao jujube are shown in Figure 13A. As can be seen from Figure 13A, the fresh fruit color of Lingwu Changzao on the 56th day was relatively plump and bright regardless of whether it was treated with 50% high oxygen stress or 90% high oxygen stress. The control group and the 0% hypoxia treatment group began to experience peel shrinkage from the 28th and 35th days respectively. Scanning electron microscopy was used to observe the microstructure of Lingwu jujube pulp. The results are shown in Figure 13B. In the early stage of storage, fresh jujube pulp cells showed crystalline arrangement, small intercellular gaps, and complete cell structure. However, as the storage time prolongs, the intercellular spaces gradually become larger, and even obvious cell rupture occurs. It is worth noting that although in the samples treated with high oxygen stress, the cell structure showed an unfavorable structure in the early stage due to the effect of high oxygen stress, the damage to the tissue structure was slowed down in the later stage. In particular, the 50% hyperoxic stress treatment group not only delayed the red-turning period of fresh dates compared to other treatment groups, but also delayed internal tissue damage.
2、不同浓度氧气短时应激处理对灵武长枣硬度的变化影响2. Effects of short-term stress treatment with different concentrations of oxygen on changes in hardness of Lingwu jujube
硬度是衡量贮藏质量的重要指标之一，可以在一定程度上揭示贮藏期间果实的商业价值和成熟度。如图14可知，灵武长枣的硬度在贮藏过程中逐渐降低。其中，50％高氧应激处理组的果实相比其他处理组在整个贮藏期间能显著保持较高的硬度水平。90％高氧应激处理在贮藏前35天能够保持较高的硬度值，但是贮藏后期硬度下降严重，可能因为氧气浓度过高，在贮藏后期对其细胞膜造成一定伤害，0％低氧处理整个贮藏期间与空白对照硬度差别不大。因此，在低温贮藏条件下，适宜浓度的高氧应激处理即50％O
2+30min能保持灵武长枣果实的鲜脆口感。
Hardness is one of the important indicators to measure storage quality, which can reveal the commercial value and maturity of fruits during storage to a certain extent. As shown in Figure 14, the hardness of Lingwu Changzao gradually decreased during storage. Among them, the fruits in the 50% high oxygen stress treatment group could maintain a significantly higher hardness level during the entire storage period than the other treatment groups. The 90% high oxygen stress treatment can maintain a high hardness value in the first 35 days of storage, but the hardness drops seriously in the later stage of storage. It may be because the oxygen concentration is too high, which causes certain damage to the cell membrane in the later stage of storage. The 0% low oxygen treatment throughout During storage, there was little difference in hardness from the blank control. Therefore, under low-temperature storage conditions, a suitable concentration of high oxygen stress treatment, that is, 50% O 2 +30min, can maintain the fresh and crisp taste of Lingwu Changzao fruits.
3、不同浓度氧气短时应激处理对灵武长枣电导率的变化影响3. Effects of short-term stress treatment with different concentrations of oxygen on changes in conductivity of Lingwu Changzao
电导率是反应果蔬细胞膜完整性的一个重要指标，较高的电导率通常表示细胞膜的完整性较低，枣果实营养物质流失速度加快。从图15可以看出，从整体上观察灵武长枣果实电导率的变化表明，随着贮藏时间的增加果实的电导率总体呈上升趋势，说明随着贮藏时间的增加细胞膜的完整性逐渐在降低。与图13B电镜扫描组织结构结果一致，值得注意的是，在整个贮藏过程中，50％高氧应激处理组果实的电导率与其他处理组相比一直处于相对较低的水平，而90％高氧应激处理组和0％低氧处理组的果实电导率在贮藏前28天低于空白对照组，28天之后电导率增加较快，电导率值高于空白对照组。可见，在50％高氧短激处理下，在一定时间内明显保护了灵武长枣的细胞膜的完整程度，减缓了枣果实内部营养物质流失的速度，保证了灵武长枣的品质质量；而在0％低氧和90％高氧短激处理下，后期会对鲜枣果实产生一定的损伤，加快了灵武长枣的氧化速度，破坏了果实细胞膜的完整程度，从而造成了枣果果肉组织内营养物质的流失；因此，在一定的时间内，适宜浓度的高氧应激处理即50％O
2+30min能保证灵武长枣果实细胞膜的完整程度。
Electrical conductivity is an important indicator that reflects the integrity of fruit and vegetable cell membranes. Higher electrical conductivity usually indicates lower integrity of the cell membrane and accelerated loss of nutrients in jujube fruits. As can be seen from Figure 15, an overall observation of the changes in electrical conductivity of Lingwu Changzao fruits shows that the electrical conductivity of the fruits generally shows an upward trend with the increase of storage time, indicating that the integrity of the cell membrane gradually increases with the increase of storage time. reduce. Consistent with the results of electron microscopy scanning tissue structure in Figure 13B, it is worth noting that during the entire storage process, the conductivity of the fruits in the 50% high oxygen stress treatment group has always been at a relatively low level compared with other treatment groups, while the 90% The conductivity of fruits in the high oxygen stress treatment group and the 0% hypoxia treatment group was lower than that of the blank control group 28 days before storage. After 28 days, the conductivity increased rapidly and the conductivity value was higher than that of the blank control group. It can be seen that under the 50% high oxygen short stimulation treatment, the integrity of the cell membrane of Lingwu Changzao was obviously protected within a certain period of time, slowing down the loss of nutrients inside the jujube fruit, and ensuring the quality of Lingwu Changzao; Under 0% hypoxia and 90% high oxygen short-term stimulation treatment, the fresh jujube fruit will be damaged to a certain extent in the later stage, accelerating the oxidation rate of Lingwu Changzao, destroying the integrity of the fruit cell membrane, thus causing the jujube fruit to be damaged. The loss of nutrients in the pulp tissue; therefore, within a certain period of time, high oxygen stress treatment with an appropriate concentration, that is, 50% O 2 +30min, can ensure the integrity of the cell membrane of Lingwu Changzao fruit.
4、不同浓度氧气短时应激处理对灵武长枣PPO、POD、ADH酶活的变化影响4. Effects of short-term stress treatment with different concentrations of oxygen on changes in PPO, POD and ADH enzyme activities of Lingwu Changzao
作为果蔬组织内重要的抗氧化酶之一，POD的主要任务是帮助果蔬清除自由基，抑制果蔬的氧化现象。从图16A可知，随着贮藏时间的增加，灵武长枣POD酶活呈先增加后减小的趋势。在第28天时，0％低氧和90％高氧短激处理的果实出现了酶活的高峰，在第35天，空白对照组的灵武长枣果实出现酶活的高峰，在第42天时，50％高氧短激处理组才出现了酶 活的高峰。可见，50％高氧短激处理组灵武长枣的抗氧化酶活性维持时间较长，抗氧化能力较强；酶峰出现过后活性又降低，说明了随着果实的成熟，自由基数量大增，破坏了抗氧化剂的抵御能力，造成了果实的衰老。由此可见，适宜浓度的高氧应激处理能即50％O
2+30min减缓灵武长枣果实氧化的速度，起到保鲜作用。
As one of the important antioxidant enzymes in fruit and vegetable tissues, POD's main task is to help fruits and vegetables remove free radicals and inhibit the oxidation of fruits and vegetables. It can be seen from Figure 16A that as the storage time increases, the POD enzyme activity of Lingwu Changzao shows a trend of first increasing and then decreasing. On the 28th day, the fruits treated with 0% hypoxia and 90% high oxygen short shock showed a peak of enzyme activity. On the 35th day, the fruits of the blank control group Lingwu Changzao showed a peak of enzyme activity. On the 42nd day , the peak of enzyme activity appeared in the 50% hyperoxic short-shock treatment group. It can be seen that the antioxidant enzyme activity of Lingwu Changzao in the 50% hyperoxic short-shock treatment group lasts longer and the antioxidant capacity is stronger; after the enzyme peak appears, the activity decreases, indicating that as the fruit matures, the number of free radicals increases. increase, destroying the resistance of antioxidants and causing fruit senescence. It can be seen that the appropriate concentration of high oxygen stress treatment, that is, 50% O 2 +30min, can slow down the oxidation rate of Lingwu Changzao fruit and play a role in preservation.
作为果蔬呼吸作用中重要的氧化酶之一，PPO的主要任务是传递信息，氧化酚类物质变成醌，醌类化合物接下来形成褐色聚合物，在果蔬采后贮藏过程中与组织褐变联系紧密。由图16B可知，在贮藏前28天PPO酶活性较低，甚至趋于0。从第28天开始，0％低氧处理组的果实PPO酶活活性开始处于上升趋势，而从第35天开始，空白对照组和90％高氧短激处理组的果实PPO酶活开始上升，50％高氧短激处理组灵武长枣的PPO酶活性在第49天之后才有了明显上升的趋势，说明该处理延缓了果实的氧化进程；而0％低氧处理组的灵武长枣氧化速度最快，其次是90％高氧短激处理。所以，适宜浓度的高氧应激处理即50％O
2+30min能减缓灵武长枣果实氧化的速度，减轻褐变，起到一定的保鲜作用。
As one of the important oxidases in the respiration of fruits and vegetables, the main task of PPO is to transmit information and oxidize phenolic substances into quinones. The quinone compounds then form brown polymers, which are associated with tissue browning during post-harvest storage of fruits and vegetables. close. It can be seen from Figure 16B that the PPO enzyme activity was low 28 days before storage, even tending to 0. Starting from the 28th day, the PPO enzyme activity of the fruits in the 0% hypoxia treatment group began to show an upward trend, while starting from the 35th day, the PPO enzyme activity of the fruits in the blank control group and the 90% high oxygen short stimulation treatment group began to increase. The PPO enzyme activity of Lingwu Changzao in the 50% high oxygen short-shock treatment group showed a significant upward trend only after the 49th day, indicating that this treatment delayed the oxidation process of the fruit; while the Lingwu Changzao in the 0% low oxygen treatment group Jujube has the fastest oxidation rate, followed by 90% high oxygen short stimulation treatment. Therefore, the appropriate concentration of high oxygen stress treatment, that is, 50% O 2 +30min, can slow down the oxidation rate of Lingwu Changzao fruit, reduce browning, and play a certain role in preservation.
作为鲜枣果实组织内重要的乙醇催化酶，ADH的主要工作流程是以乙醇作为底物，分解转化乙醇，从某种程度上可以避免乙醇对果蔬机体的伤害。由图16C可知，随着贮藏时间的增加，ADH酶活的变化呈先增加后减小的趋势。在第21时，0％低氧和90％高氧短激处理过的果实出现了酶活的高峰，在第35天时，空白对照组出现了酶活的高峰，而50％高氧短激处理组的果实在第35天才出现了酶活的高峰。可以看出，0％低氧和90％高氧短激处理过的灵武长枣果实组织内部乙醇的累积迅速，从而使乙醇催化酶的酶活性很快增强，但也很快丧失活性；相反，50％高氧短激处理组果实的ADH酶活性维持时间较长，从而减少乙醇的累积，因此灵武长枣酒化速度低于其他三组；酶峰出现过后活性又降低，说明了随着果实的成熟，乙醇含量大增，破坏了乙醇催化酶的抵御能力，造成了鲜枣果实的酒化现象。由此可见，适宜浓度的高氧应激处理即50％O
2+30min更能减缓灵武长枣果实酒化的速度，起到较好的保鲜作用。
As an important ethanol catalyzing enzyme in fresh jujube fruit tissue, the main working process of ADH is to use ethanol as a substrate to decompose and convert ethanol, which can avoid the damage of ethanol to the fruit and vegetable body to a certain extent. It can be seen from Figure 16C that as the storage time increases, the changes in ADH enzyme activity first increase and then decrease. At the 21st day, the fruits treated with 0% hypoxia and 90% high oxygen short shock showed a peak of enzyme activity. On the 35th day, the blank control group showed a peak of enzyme activity, while the fruit treated with 50% high oxygen short shock showed a peak of enzyme activity. The fruits of the group only showed the peak of enzyme activity on the 35th day. It can be seen that ethanol accumulates rapidly in the Lingwu jujube fruit tissue treated with 0% hypoxia and 90% hyperoxia short shock, so that the enzyme activity of the ethanol catalytic enzyme is quickly enhanced, but the activity is also quickly lost; on the contrary , the ADH enzyme activity of the fruits in the 50% hyperoxic short-shock treatment group was maintained for a longer time, thereby reducing the accumulation of ethanol, so the alcoholization speed of Lingwu Changzao was lower than that of the other three groups; the activity decreased after the enzyme peak appeared, indicating that the As the fruit matures, the ethanol content increases significantly, destroying the resistance of ethanol catalytic enzymes and causing the alcoholization of fresh jujube fruits. It can be seen that the appropriate concentration of high oxygen stress treatment, that is, 50% O 2 +30min, can slow down the alcoholization speed of Lingwu Changzao fruit and play a better preservation role.
实施例3Example 3
一种适于灵武长枣贮藏的高氧应激保鲜方法，具体步骤如下：A high-oxygen stress preservation method suitable for Lingwu jujube storage. The specific steps are as follows:
1、灵武长枣采后预冷处理：新鲜果实采后先在冷库低温鼓风去除田间热18-24h，为防止失水过多，将打孔的PE保鲜袋(0.014mm)套在果蔬箱外侧。1. Post-harvest pre-cooling treatment of Lingwu Changzao: After harvesting, the fresh fruits are first blasted at low temperature in the cold storage to remove field heat for 18-24 hours. In order to prevent excessive water loss, a perforated PE fresh-keeping bag (0.014mm) is placed in the fruit and vegetable box. outside.
2、挑拣样品：预冷结束后，挑选无病害，无磕碰，大小、颜色均匀的鲜果。2. Pick samples: After pre-cooling, select fresh fruits that are disease-free, bump-free, and uniform in size and color.
3、氧气处理：将挑拣好的灵武长枣放入到密封的气调箱内，先用高纯氮气将密封的气调箱其余气体排出，在向密封的气调箱内通入50％O
2(w/w)浓度的氧气短时应激处理30min，然后打开密封的气调箱，静置30min，使气调箱内残留的高浓度氧释放，降低氧浓度使其恢复到自然状态；另一组为50％O
2(w/w)浓度的氧气持续处理，使其一直处于高氧氛围；不做任何处理的灵武长枣果实做为空白对照组。
3. Oxygen treatment: Put the selected Lingwu Changzao into a sealed air-conditioned box, first use high-purity nitrogen to discharge the rest of the gas in the sealed air-conditioned box, and then introduce 50% of the gas into the sealed air-conditioned box. Perform a short-term stress treatment with O 2 (w/w) oxygen concentration for 30 minutes, then open the sealed air conditioning box and let it stand for 30 minutes to release the high concentration of oxygen remaining in the air conditioning box and reduce the oxygen concentration to return to its natural state. ; The other group was continuously treated with oxygen at a concentration of 50% O 2 (w/w) to keep it in a high oxygen atmosphere; the Lingwu Changzao fruit without any treatment was used as a blank control group.
4、静态贮藏：将50％高浓度氧短时应激30min处理的灵武长枣，50％高浓度的氧气持续处理与未做任何处理的空白对照组放在PE保鲜袋内于0±1℃条件下贮藏。4. Static storage: Put the Lingwu Changzao treated with 50% high-concentration oxygen for short-term stress for 30 minutes, the continuous treatment with 50% high-concentration oxygen and the blank control group without any treatment in PE fresh-keeping bags at 0±1 Store at ℃.
品质评价：定期对灵武长枣的品质指标进行测定。Quality evaluation: The quality indicators of Lingwu Changzao are measured regularly.
相关检测如下：The relevant tests are as follows:
1、不同高氧处理方式对灵武长枣硬度的变化影响1. Effects of different high oxygen treatment methods on the hardness of Lingwu jujube
硬度是衡量贮藏质量的重要指标之一，可以在一定程度上揭示贮藏期间果实的商业价值和成熟度。如图17可知，灵武长枣的硬度在贮藏过程中逐渐降低。其中，本发明的50％高氧应激处理组的果实相比其他处理组在整个贮藏期间能显著保持较高的硬度水平，50％高氧持续处理与空白对照组整个贮藏期间硬度差别不大。结果表明，在低温贮藏条件下，本发明的高氧应激处理比高氧持续处理与空白对照组能更好地保持灵武长枣果实的鲜脆口感。本发明的高氧应激处理即50％O
2(w/w)浓度的氧气、短时应激处理30min之间具有协同作用。
Hardness is one of the important indicators to measure storage quality, which can reveal the commercial value and maturity of fruits during storage to a certain extent. As shown in Figure 17, the hardness of Lingwu Changzao gradually decreased during storage. Among them, the fruits in the 50% hyperoxic stress treatment group of the present invention can maintain a significantly higher hardness level during the entire storage period than other treatment groups, and there is not much difference in hardness during the entire storage period between the 50% hyperoxic continuous treatment and the blank control group. . The results show that under low-temperature storage conditions, the high-oxygen stress treatment of the present invention can better maintain the fresh and crisp taste of Lingwu Changzao fruits than the continuous high-oxygen treatment and the blank control group. There is a synergistic effect between the high oxygen stress treatment of the present invention, that is, oxygen with a concentration of 50% O 2 (w/w) and short-term stress treatment for 30 minutes.
2、不同高氧处理方式对灵武长枣电导率的变化影响2. Effects of different high oxygen treatment methods on the conductivity of Lingwu Changzao
电导率是反应果蔬细胞膜完整性的一个重要指标，较高的电导率通常表示细胞膜的完整性较低，枣果实营养物质流失速度加快。从图18可以看出，从整体上观察灵武长枣果实电导率的变化表明，随着贮藏时间的增加果实的电导率总体呈上升趋势，说明随着贮藏时间的增加细胞膜的完整性逐渐在降低。值得注意的是，在整个贮藏过程中，本发明的50％高氧应激处理组果实的电导率与其他处理组相比一直处于相对较低的水平，而50％高氧持续处理虽然在贮藏早期抑制了电导率，但贮藏中后期电导率上升较快，且水平高于空白对照组。可见，在本发明的50％高氧短激处理下，在一定时间内明显保护了灵武长枣的细胞膜的完整程度，减缓了枣果实内部营养物质流失的速度，保证了灵武长枣的品质质量；而50％高氧持续处理，后期会对鲜枣果实产生一定的损伤，加快了灵武长枣的氧化速度，破坏了果实细胞膜的完整程度，从而造成了枣果果肉组织内营养物质的流失；因此，在长期贮藏下，本发明的高氧应激处理比高氧持续处理和空白对照组更能保证灵武长枣果实细胞膜的完整程度。本发明的高氧应激处理即50％O
2(w/w)浓度的氧气、短时应激处理30min之间具有协同作用。
Electrical conductivity is an important indicator that reflects the integrity of fruit and vegetable cell membranes. Higher electrical conductivity usually indicates lower integrity of the cell membrane and accelerated loss of nutrients in jujube fruits. As can be seen from Figure 18, an overall observation of the changes in electrical conductivity of Lingwu Changzao fruits shows that the electrical conductivity of the fruits generally shows an upward trend with the increase of storage time, indicating that the integrity of the cell membrane gradually increases with the increase of storage time. reduce. It is worth noting that during the entire storage process, the electrical conductivity of the fruits in the 50% high oxygen stress treatment group of the present invention has been at a relatively low level compared with other treatment groups, and the 50% high oxygen continuous treatment although in storage The conductivity was suppressed in the early stage, but the conductivity increased rapidly in the middle and late stages of storage, and the level was higher than that of the blank control group. It can be seen that under the 50% high oxygen short stimulation treatment of the present invention, the integrity of the cell membrane of Lingwu Changzao was obviously protected within a certain period of time, the rate of loss of nutrients inside the jujube fruit was slowed down, and the quality of Lingwu Changzao was ensured. Quality and quality; continuous treatment with 50% high oxygen will cause certain damage to fresh jujube fruits in the later stage, speeding up the oxidation rate of Lingwu Changzao, destroying the integrity of the fruit cell membrane, thus causing the loss of nutrients in the jujube pulp tissue. loss; therefore, under long-term storage, the high-oxygen stress treatment of the present invention can better ensure the integrity of the cell membrane of Lingwu Changzao fruit than the continuous high-oxygen treatment and the blank control group. There is a synergistic effect between the high oxygen stress treatment of the present invention, that is, oxygen with a concentration of 50% O 2 (w/w) and short-term stress treatment for 30 minutes.
3、不同高氧处理方式对灵武长枣PPO、POD、ADH酶活的变化影响3. Effects of different high oxygen treatment methods on changes in PPO, POD and ADH enzyme activities of Lingwu Changzao
作为果蔬组织内重要的抗氧化酶之一，POD的主要任务是帮助果蔬清除自由基，抑制果蔬的氧化现象。从图19A可知，随着贮藏时间的增加，灵武长枣POD酶活呈先增加后减小的趋势。在第35天时，空白对照组的灵武长枣果实出现酶活的高峰，在第42天时，本发明的50％高氧短激处理组与50％高氧持续处理组才出现了酶活的高峰，且本发明的50％高氧短激处理组比50％高氧持续处理组POD水平更高，可见，本发明的50％高氧短激处理组灵武长枣的抗氧化酶活性维持时间较长，抗氧化能力较强；酶峰出现过后活性又降低，说明了随着果实的成熟，自由基数量大增，破坏了抗氧化剂的抵御能力，造成了果实的衰老。由此可见，本发明的高氧应激处理比高氧持续处理和空白对照组更能增强灵武长枣果实抗氧化能力，起到保鲜作用。本发明的高氧应激处理即50％O
2(w/w)浓度的氧气、短时应激处理30min之间具有协同作用。
As one of the important antioxidant enzymes in fruit and vegetable tissues, POD's main task is to help fruits and vegetables remove free radicals and inhibit the oxidation of fruits and vegetables. It can be seen from Figure 19A that as the storage time increases, the POD enzyme activity of Lingwu Changzao shows a trend of first increasing and then decreasing. On the 35th day, the Lingwu Changzao fruits in the blank control group showed a peak of enzyme activity. On the 42nd day, the 50% high oxygen short-shock treatment group and the 50% high oxygen continuous treatment group of the present invention showed a peak in enzyme activity. Peak, and the POD level of the 50% hyperoxic short-shock treatment group of the present invention is higher than that of the 50% hyperoxic continuous treatment group. It can be seen that the antioxidant enzyme activity of Lingwu Changzao in the 50% hyperoxic short-shock treatment group of the present invention is maintained The longer the time, the stronger the antioxidant capacity; the activity decreases after the enzyme peak appears, indicating that as the fruit matures, the number of free radicals increases greatly, destroying the antioxidant capacity and causing the aging of the fruit. It can be seen that the hyperoxic stress treatment of the present invention can enhance the antioxidant capacity of Lingwu Changzao fruit and play a role in preservation than the continuous hyperoxic treatment and the blank control group. There is a synergistic effect between the high oxygen stress treatment of the present invention, that is, oxygen with a concentration of 50% O 2 (w/w) and short-term stress treatment for 30 minutes.
作为果蔬呼吸作用中重要的氧化酶之一，PPO的主要任务是传递信息，氧化酚类物质变成醌，醌类化合物接下来形成褐色聚合物，在果蔬采后贮藏过程中与组织褐变联系紧密。由图19B可知，在贮藏前28天PPO酶活性较低，甚至趋于0。从第28天开始，50％高氧持续处理组的果实PPO酶活活性开始处于上升趋势，而从第35天开始，空白对照组的果实PPO酶活开始上升，本发明的50％高氧短激处理组灵武长枣的PPO酶活性在第49天之后才有了明显上升的趋势，说明该处理延缓了果实的氧化进程；而50％高氧持续处理组的灵武长枣氧化速度最快，其次是空白对照处理。所以，本发明的高氧应激处理比高氧持续处理和空白对照组更能减缓灵武长枣果实氧化的速度，减轻褐变，起到一定的保鲜作用。As one of the important oxidases in the respiration of fruits and vegetables, the main task of PPO is to transmit information and oxidize phenolic substances into quinones. The quinone compounds then form brown polymers, which are associated with tissue browning during post-harvest storage of fruits and vegetables. close. It can be seen from Figure 19B that the PPO enzyme activity was low 28 days before storage, even tending to 0. Starting from the 28th day, the PPO enzyme activity of the fruits in the 50% high oxygen continuous treatment group began to show an upward trend, and starting from the 35th day, the PPO enzyme activity of the fruits in the blank control group began to increase. The 50% high oxygen short-term treatment group of the present invention The PPO enzyme activity of Lingwu Changzao in the stimulation treatment group showed a significant upward trend only after the 49th day, indicating that the treatment delayed the oxidation process of the fruit; while the oxidation rate of Lingwu Changzao in the 50% high oxygen continuous treatment group was the fastest. Fast, followed by blank control processing. Therefore, the hyperoxic stress treatment of the present invention can slow down the oxidation rate of Lingwu Changzao fruit, reduce browning, and play a certain role in preserving freshness than the continuous hyperoxic treatment and the blank control group.
作为鲜枣果实组织内重要的乙醇催化酶，ADH的主要工作流程是以乙醇作为底物，分解转化乙醇，从某种程度上可以避免乙醇对果蔬机体的伤害。由图19C可知，随着贮藏时间的增加，ADH酶活的变化呈先增加后减小的趋势。在第21时，50％高氧持续处理过的果实出现了酶活的高峰，在第35天时，空白对照组出现了酶活的高峰，而本发明的50％高氧短激处理组的果实在第35天才出现了酶活的高峰。可以看出，50％高氧持续处理过的灵武长枣果实组织内部乙醇的累积迅速，从而使乙醇催化酶的酶活性很快增强，但也很快丧失活性；相反，本发明的50％高氧短激处理组果实的ADH酶活性维持时间较长，从而减少乙醇的累积，因此灵武长枣酒化速度低于其他两组；酶峰出现过后活性又降低，说明了随着果实的成熟，乙醇含量大增，破坏了乙醇催化酶的抵御能力，造成了鲜枣果实的酒化现象。由此可见，本发明的高氧应激处理比高氧持续处理和空白对照组更能减缓灵武长枣果实酒化的速度，起到较好的保鲜作用。As an important ethanol catalyzing enzyme in fresh jujube fruit tissue, the main working process of ADH is to use ethanol as a substrate to decompose and convert ethanol, which can avoid the damage of ethanol to the fruit and vegetable body to a certain extent. It can be seen from Figure 19C that as the storage time increases, the changes in ADH enzyme activity first increase and then decrease. On the 21st day, the fruits treated with 50% high oxygen continuously showed a peak of enzyme activity. On the 35th day, the blank control group showed a peak of enzyme activity, while the fruits of the 50% high oxygen short-shock treatment group of the present invention The peak of enzyme activity appeared on the 35th day. It can be seen that ethanol accumulates rapidly in the Lingwu jujube fruit tissue treated with 50% high oxygen continuously, so that the enzyme activity of the ethanol catalytic enzyme is quickly enhanced, but the activity is also quickly lost; on the contrary, the 50% of the present invention The ADH enzyme activity of the fruits in the high-oxygen short-shock treatment group was maintained for a longer time, thereby reducing the accumulation of ethanol. Therefore, the alcoholization speed of Lingwu Changzao was lower than that of the other two groups; the activity decreased after the appearance of the enzyme peak, indicating that as the fruit ages When ripe, the ethanol content increases significantly, destroying the resistance of the ethanol catalytic enzyme, causing the alcoholization of fresh jujube fruits. It can be seen that the hyperoxic stress treatment of the present invention can slow down the alcoholization speed of Lingwu Changzao fruit and play a better preservation effect than the continuous hyperoxic treatment and the blank control group.
实施例4Example 4
一种适于大青葡萄贮藏的高氧应激保鲜方法，具体步骤如下：A high oxygen stress preservation method suitable for the storage of green grapes. The specific steps are as follows:
1、大青葡萄采后预冷处理：新鲜果实采后先在冷库低温鼓风去除田间热18-24h，为防止失水过多，将打孔的PE保鲜袋(0.014mm)套在果蔬箱外侧。1. Post-harvest pre-cooling treatment of Daqing grapes: After harvest, the fresh fruits are first blasted at low temperature in the cold storage to remove field heat for 18-24 hours. To prevent excessive water loss, put a perforated PE fresh-keeping bag (0.014mm) on the outside of the fruit and vegetable box. .
2、挑拣样品：预冷结束后，挑选无病害，无磕碰，大小、颜色均匀的鲜果。2. Pick samples: After pre-cooling, select fresh fruits that are disease-free, bump-free, and uniform in size and color.
3、高氧应激处理：将挑拣好的大青葡萄放入到密封的气调箱内，先用高纯氮气将密封的气调箱其余气体排出，在向密封的气调箱内通入80％O
2(w/w)浓度的氧气短时应激处理30min，然后打开密封的气调箱，静置30min，使气调箱内残留的高浓度氧释放，降低氧浓度使其恢复到自然状态，不做任何处理的鲜食枸杞果实做为对照组。
3. High oxygen stress treatment: Put the selected green grapes into a sealed air-conditioned box, first use high-purity nitrogen to discharge the remaining gas from the sealed air-conditioned box, and then pass it into the sealed air-conditioned box. Short-term stress treatment with oxygen at a concentration of 80% O 2 (w/w) for 30 minutes, then open the sealed air conditioning box and let it stand for 30 minutes to release the high concentration of oxygen remaining in the air conditioning box and reduce the oxygen concentration to return to Fresh wolfberry fruits in their natural state without any treatment were used as the control group.
4、静态贮藏：将高浓度氧短时应激30min处理的大青葡萄与未做任何处理的空白对照组放在PE保鲜袋内于0±1℃条件下贮藏。4. Static storage: The green grapes treated with high-concentration oxygen for short-term stress for 30 minutes and the blank control group without any treatment were placed in PE fresh-keeping bags and stored at 0±1°C.
品质评价：定期对大青葡萄的品质指标进行测定。Quality evaluation: The quality indicators of Daqing grapes are measured regularly.
相关检测如下：The relevant tests are as follows:
1、高氧应激处理对大青葡萄保鲜效果、色泽及硬度的变化影响1. Effects of high oxygen stress treatment on the preservation effect, color and hardness of Daqing grapes
外观品质是果蔬商品价值的重要指标，由图20A-B可知，在大青葡萄贮藏第35天时，与未经高氧应激处理的空白对照组相比，经本发明的80％高氧应激处理的果实显著抑制了果实的腐烂，保持了果实的商品价值。从色泽上看，色差a*值越低代表果实偏绿，越高代表果实偏红，从图17C可以观察到色差a*值整体呈现上升趋势，说明大青葡萄随着贮藏时间的增加，会逐渐失色。但从图中很明显能看出，整个贮藏期间经本发明的80％高氧应激处理的果实色差a*值明显低于空白组，说明本发明的80％高氧应激处理更能保持大青葡萄的绿色。果蔬的硬度(Firmness)往往可以反映其新鲜程度，从图20D中可以观察到硬度值整体呈现下降趋势，说明大青葡萄随着贮藏时间的增加，大青葡萄果肉变软。值得注意的是，经本发明的80％高氧应激处理的果实硬度值从第15天以后均显著高于空白组，说明本发明的高氧应激处理能够减缓果实变软，保持果实硬度。Appearance quality is an important indicator of the commercial value of fruits and vegetables. It can be seen from Figure 20A-B that on the 35th day of storage of Daqing grapes, compared with the blank control group without hyperoxic stress treatment, after 80% hyperoxic stress treatment of the present invention, The fruits treated with stimulation significantly inhibited the rot of the fruits and maintained the commercial value of the fruits. From the perspective of color, the lower the color difference a* value, the greener the fruit, and the higher the fruit, the redder. It can be observed from Figure 17C that the color difference a* value shows an overall upward trend, indicating that as the storage time increases, Daqing grapes will Gradually fade away. However, it can be clearly seen from the figure that the color difference a* value of the fruit treated with 80% high oxygen stress of the present invention during the entire storage period is significantly lower than that of the blank group, indicating that the 80% high oxygen stress treatment of the present invention can better maintain the Big green grapes. The firmness of fruits and vegetables can often reflect their freshness. It can be observed from Figure 20D that the overall hardness value shows a downward trend, indicating that the Daqing grape pulp becomes softer as the storage time increases. It is worth noting that the hardness value of the fruit treated with 80% high oxygen stress of the present invention was significantly higher than that of the blank group from the 15th day onwards, indicating that the high oxygen stress treatment of the present invention can slow down the softening of the fruit and maintain the hardness of the fruit. .
2、高氧应激处理对大青葡萄可溶性固形物的变化影响2. Effect of high oxygen stress treatment on changes in soluble solids of Daqing grape
果实的可溶性固形物(Soluble Solid Content,SSC)是衡量果实成熟度和品质的重要指标，且直接决定果蔬的味道。从图21可以观察到空白组的SSC在贮藏期间没有显著性变化，而经过本发明的80％高氧应激处理后的大青葡萄比空白组SSC增大了，说明该处理有效增加了大青葡萄的甜度，改善了果实的品质。The soluble solid content (SSC) of fruit is an important indicator of fruit maturity and quality, and directly determines the taste of fruits and vegetables. It can be observed from Figure 21 that the SSC of the blank group did not change significantly during storage, but the SSC of the Daqing grapes after the 80% hyperoxic stress treatment of the present invention increased compared with the blank group, indicating that this treatment effectively increased the SSC of the Daqing grape. The sweetness of green grapes improves the quality of the fruit.
3、高氧应激处理对大青葡萄细胞膜透性、抗氧化能力的变化影响3. Effects of hyperoxic stress treatment on cell membrane permeability and antioxidant capacity of green grapes
果实膜透性的变化由相对电导率来表达，膜透性越大，膜的损伤程度越大。从图22A中可以看出，电导率在整个贮藏期间呈先下降后上升的趋势。随着果实的成熟，机体内糖含量增加，酸含量减少，导致氢离子减少，贮藏后期因为果实组织细胞膜受到损伤，电导率呈上升趋势。而经本发明的80％高氧应激处理后的大青葡萄远低于空白组电导率水平，这说明本发明的80％高氧应激处理能够延缓果实成熟，减少大青葡萄果实膜的损伤。LOX特异性催化膜脂不饱和脂肪酸向饱和脂肪酸转化，加速过氧化代谢物的积累，破坏细胞膜结构，诱导膜脂过氧化。此外，LOX还会促进细胞膜功能的丧失，从而降低采后贮藏能力。从图20D中可以看出大青葡萄整体LOX成先上升在下降的趋势，本发明的80％高氧应激处理组比空白组峰值晚出现7天，说明间接性高氧处理延缓了LOX峰值的出现，并且保持了较低水平的LOX活性。所以本发明的高氧应激处理能够维持细胞膜功能，是提高果实采后贮藏能力的有效处理技术。Changes in fruit membrane permeability are expressed by relative conductivity. The greater the membrane permeability, the greater the degree of membrane damage. It can be seen from Figure 22A that the conductivity first decreased and then increased during the entire storage period. As the fruit matures, the sugar content in the body increases and the acid content decreases, resulting in a decrease in hydrogen ions. In the later stages of storage, due to damage to the cell membrane of the fruit tissue, the conductivity shows an upward trend. The electrical conductivity of Daqing grapes treated with 80% high oxygen stress of the present invention is much lower than that of the blank group. This shows that the 80% high oxygen stress treatment of the present invention can delay fruit ripening and reduce the damage of Daqing grape fruit membranes. damage. LOX specifically catalyzes the conversion of membrane lipid unsaturated fatty acids into saturated fatty acids, accelerates the accumulation of peroxidized metabolites, destroys cell membrane structure, and induces membrane lipid peroxidation. In addition, LOX also promotes the loss of cell membrane function, thereby reducing postharvest storage capacity. It can be seen from Figure 20D that the overall LOX content of Daqing grape first increased and then decreased. The peak value of the 80% hyperoxic stress treatment group of the present invention appeared 7 days later than that of the blank group, indicating that the indirect hyperoxic treatment delayed the LOX peak value. appearance, and maintain a low level of LOX activity. Therefore, the high oxygen stress treatment of the present invention can maintain cell membrane function and is an effective treatment technology to improve the postharvest storage capacity of fruits.
实施例中大青葡萄的抗氧化能力通过DPPH清除自由基速率来表达，从图20B中可以看出，在35天贮藏期内，与空白组相比，本发明的80％高氧应激处理的果实DPPH表现出最高水平，且在最后一周其清除自由基的能力显著高于空白组，说明80％高氧应激处理具有最高清除自由基的能力。众所周知，植物衰老的主要原因是活性氧(ROS)的产生，活性氧对植物细胞的蛋白质和脂质产生氧化损伤，导致果实软化和衰老，因此，控制活性氧的产生是 果蔬采后贮藏延缓衰老的重要手段。而活性氧对机体的损坏可以通过活性氧保护酶来调节。SOD是植物体内一种重要的活性氧清除酶，具有抗氧化、抗衰老的作用。多项研究表明，高SOD活性可抑制活性氧对果实的伤害。因此，本文研究了抗氧化酶SOD对大青葡萄衰老品质的影响，如图20C所示，在整个贮藏期间本发明的80％高氧应激处理的SOD水平一开始由于果实自身的应激反应低于空白组，但在第10天左右之后都高于空白组，有效地提高了SOD活性，增强了果实的抗氧化性。In the embodiment, the antioxidant capacity of Daqing grape is expressed by DPPH free radical scavenging rate. As can be seen from Figure 20B, during the 35-day storage period, compared with the blank group, the 80% hyperoxic stress treatment of the present invention The fruit showed the highest level of DPPH, and its ability to scavenge free radicals was significantly higher than that of the blank group in the last week, indicating that the 80% high oxygen stress treatment had the highest ability to scavenge free radicals. As we all know, the main cause of plant senescence is the production of reactive oxygen species (ROS). Reactive oxygen species cause oxidative damage to the proteins and lipids of plant cells, leading to fruit softening and senescence. Therefore, controlling the production of reactive oxygen species is an important factor in delaying senescence during post-harvest storage of fruits and vegetables. important means. The damage caused by reactive oxygen species to the body can be regulated by reactive oxygen species protective enzymes. SOD is an important active oxygen scavenging enzyme in plants and has antioxidant and anti-aging effects. Multiple studies have shown that high SOD activity can inhibit the damage of reactive oxygen species to fruits. Therefore, this article studied the effect of antioxidant enzyme SOD on the aging quality of Daqing grape. As shown in Figure 20C, the SOD level of the 80% hyperoxic stress treatment of the present invention during the entire storage period was initially due to the stress response of the fruit itself. Lower than the blank group, but higher than the blank group after about the 10th day, which effectively increased SOD activity and enhanced the antioxidant properties of the fruit.
4、高氧应激处理对大青葡萄总酚及抗褐变相关酶活的变化影响4. Effects of hyperoxic stress treatment on changes in total phenolics and anti-browning-related enzyme activities of green grapes
酚类物质是新鲜果蔬的主要营养物质，它可以赋予果蔬促进人体健康的功能，而PAL是植物体内苯丙烷代谢途径的关键酶，其活性高低控制植物体内多种生酚类化合物。此外，酚类物质与PAL均在植物本身作为应对生物和非生物胁迫产生的次生代谢物中发挥着重要的作用。从图21A中可以观察出前14天，经过本发明的80％高氧应激处理后引起果实酚类物质变化波动较大，这说明酚类物质的波动趋势是由植物的防御反应和高氧刺激一起造成的。在14天之后，两个处理组总酚含量表现为相同的变化趋势，先增加后减小，并且可以很明显的看出，经本发明的80％高氧应激处理的果实比空白组总酚含量低，这说明高氧应激处理可以减缓酚类物质的增加。从图21B中可以观察到不同组处理PAL值基本呈现相同的变化趋势。有趣的是，与图21A相比较，空白组及本发明的80％高氧应激处理果实对应的总酚含量与PAL值的表现出相同的水平，这是因为PAL参与了褐变底物(酚类化合物)的合成，因此，经过本发明的80％高氧应激处理的果实保持较低总酚含量的同时，PAL水平较空白组与保持了较低水平。说明本发明的高氧应激处理抑制了大青葡萄果实褐变底物的合成。Phenolic substances are the main nutrients of fresh fruits and vegetables. They can give fruits and vegetables the function of promoting human health. PAL is a key enzyme in the phenylpropanoid metabolism pathway in plants. Its activity controls a variety of phenolic compounds in plants. In addition, both phenolics and PAL play important roles in plants themselves as secondary metabolites produced in response to biotic and abiotic stresses. It can be observed from Figure 21A that in the first 14 days, after the 80% hyperoxic stress treatment of the present invention, the changes in fruit phenolic substances fluctuated greatly, which shows that the fluctuation trend of phenolic substances is caused by the plant's defense response and high oxygen stimulation. caused together. After 14 days, the total phenolic content of the two treatment groups showed the same change trend, increasing first and then decreasing. It can be clearly seen that the fruits treated with 80% hyperoxic stress of the present invention had a higher total phenolic content than the blank group. Phenol content was low, suggesting that hyperoxic stress treatment can slow down the increase in phenolics. It can be observed from Figure 21B that the PAL values of different groups of treatments basically show the same changing trend. Interestingly, compared with Figure 21A, the total phenolic content and PAL value of the blank group and the 80% hyperoxic stress-treated fruits of the present invention showed the same level. This is because PAL participates in the browning substrate ( Synthesis of phenolic compounds), therefore, while the fruit treated with 80% high oxygen stress of the present invention maintains a lower total phenolic content, the PAL level remains lower than that of the blank group. It shows that the high oxygen stress treatment of the present invention inhibits the synthesis of browning substrates in green grape fruits.
多酚氧化酶(PPO)与果实组织的衰老和褐变密切相关，其活性已成为衡量果实衰老的重要指标，当氧气存在时，PPO催化酚类化合物转化为醌类，然后次生的非酶反映积累黑色素导致褐变。从图23C中可以看出，两个处理组的PPO表现为相同的变化趋势，且本发明的80％高氧应激处理比空白组PPO表现出较高水平。POD活性随贮藏时间的延长而减小，如图20D所示，本发明的80％高氧应激处理的果实比空白组POD表现出较高水平。从图23中可以看出，本发明的高氧应激处理有效的抑制了大青葡萄采后果实的褐变，因为它起到了抗褐变抑制剂作用的三种模式之一，降低了果实总酚的含量，增加了PPO和POD的活性，另外两种模式分别为：一，增加总酚的含量，降低PAL、PPO及POD活性；二，增加总酚的含量及PAL、PPO和POD活性。Polyphenol oxidase (PPO) is closely related to the senescence and browning of fruit tissues. Its activity has become an important indicator of fruit senescence. When oxygen is present, PPO catalyzes the conversion of phenolic compounds into quinones, and then secondary non-enzymatic Reflects the accumulation of melanin leading to browning. As can be seen from Figure 23C, the PPO of the two treatment groups showed the same change trend, and the 80% hyperoxic stress treatment of the present invention showed a higher level of PPO than the blank group. POD activity decreases with the prolongation of storage time. As shown in Figure 20D, the 80% hyperoxic stress-treated fruits of the present invention showed a higher level of POD than the blank group. As can be seen from Figure 23, the high oxygen stress treatment of the present invention effectively inhibits the browning of postharvest Daqing grape fruits because it acts as one of the three modes of anti-browning inhibitors and reduces the risk of fruit damage. The content of total phenols increases the activities of PPO and POD. The other two modes are: first, increasing the content of total phenols and reducing the activities of PAL, PPO and POD; second, increasing the content of total phenols and the activities of PAL, PPO and POD. .
尽管为说明目的公开了本发明的实施例，但是本领域的技术人员可以理解：在不脱离本发明及所附权利要求的精神和范围内，各种替换、变化和修改都是可能的，因此，本发明的范围不局限于实施例所公开的内容。Although the embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various substitutions, changes and modifications are possible without departing from the spirit and scope of the present invention and the appended claims. , the scope of the present invention is not limited to the contents disclosed in the embodiments.
Claims (7)
- 一种适用于鲜果贮藏的高氧应激保鲜方法，其特征在于：所述方法是以50％-100％高浓度氧气为主要气体，以高纯氮气为辅助气体对鲜果进行20-30min短时刺激诱抗。A high-oxygen stress preservation method suitable for fresh fruit storage, which is characterized in that: the method uses 50%-100% high-concentration oxygen as the main gas and high-purity nitrogen as the auxiliary gas to perform a short-term 20-30min period on the fresh fruit. Stimulation induces resistance.
- 根据权利要求1所述的适用于鲜果贮藏的高氧应激保鲜方法，其特征在于：具体步骤如下：The high oxygen stress preservation method suitable for fresh fruit storage according to claim 1, characterized in that: the specific steps are as follows:(1)挑拣样品：预冷结束后，挑选无病害，无磕碰，大小、颜色均匀的鲜果；(1) Pick samples: After pre-cooling, select fresh fruits that are disease-free, bump-free, and uniform in size and color;(2)鲜果采后预冷处理：新鲜果实采后挑选后的果蔬盛装在应激保鲜箱内，为防止失水过多，将打孔的PE保鲜袋套在果蔬箱外侧，然后箱整体开盖在冷库低温鼓风去除田间热，温度为0±1℃，时间为18-24小时；(2) Post-harvest pre-cooling treatment of fresh fruits: The fruits and vegetables selected after the fresh fruits are harvested are packed in stress preservation boxes. In order to prevent excessive water loss, a perforated PE preservation bag is placed on the outside of the fruit and vegetable box, and then the box is opened as a whole. Use low-temperature blast in the cold storage to remove field heat, the temperature is 0±1°C, and the time is 18-24 hours;(3)高氧应激处理：将挑拣好的鲜果放入到密封的气调箱内，先用高纯氮气将密封的气调箱其余气体排出，再向密封的应激箱内通入50％-100％O 2(w/w)的高浓度氧气应激处理20-30min，然后打开密封的应激箱，静置20-30min，使气调箱内残留的高浓度氧释放，降低氧浓度使其恢复到自然状态； (3) High oxygen stress treatment: Put the selected fresh fruits into a sealed air-conditioned box, first use high-purity nitrogen to discharge the remaining gas in the sealed air-conditioned box, and then pass 50 into the sealed stress box. %-100% O 2 (w/w) high-concentration oxygen stress treatment for 20-30 minutes, then open the sealed stress box and let it stand for 20-30 minutes to release the high-concentration oxygen remaining in the air-conditioning box and reduce the oxygen content. The concentration returns it to its natural state;(4)静态贮藏：将高氧应激处理好的鲜果放在PE保鲜袋内于0±1℃条件下贮藏。(4) Static storage: Store the fresh fruits treated with high oxygen stress in PE fresh-keeping bags at 0±1°C.
- 根据权利要求1所述的适用于鲜果贮藏的高氧应激保鲜方法，其特征在于：所述步骤(1)中PE保鲜袋的孔的直径为0.013-0.020mm。The high oxygen stress preservation method suitable for fresh fruit storage according to claim 1, characterized in that: in step (1), the diameter of the hole of the PE preservation bag is 0.013-0.020 mm.
- 根据权利要求1所述的适用于鲜果贮藏的高氧应激保鲜方法，其特征在于：所述步骤(4)中将高氧应激处理好的鲜果放在MAP自发气调及0±1℃条件下贮藏。The high-oxygen stress preservation method suitable for fresh fruit storage according to claim 1, characterized in that: in the step (4), the fresh fruits treated with high oxygen stress are placed in MAP spontaneous atmosphere control at 0±1°C stored under conditions.
- 根据权利要求1至4任一项所述的适用于鲜果贮藏的高氧应激保鲜方法，其特征在于：所述鲜果为鲜食枸杞、鲜枣、葡萄、枇杷等果蔬。The high-oxygen stress preservation method suitable for fresh fruit storage according to any one of claims 1 to 4, characterized in that: the fresh fruit is fresh wolfberry, fresh jujube, grape, loquat and other fruits and vegetables.
- 根据权利要求5所述的适用于鲜果贮藏的高氧应激保鲜方法，其特征在于：当鲜果为鲜食枸杞时，高氧应激处理时的条件为：向密封的气调箱内通入90％O 2(w/w)的高浓度氧气分别短时应激处理30min，然后打开密封的气调箱，静置30min； The high-oxygen stress preservation method suitable for fresh fruit storage according to claim 5, characterized in that: when the fresh fruit is fresh wolfberry, the conditions during the high-oxygen stress treatment are: passing into the sealed atmosphere-controlled box 90% O 2 (w/w) high-concentration oxygen was subjected to short-term stress treatment for 30 minutes, and then the sealed air conditioning box was opened and left to stand for 30 minutes;当鲜果为灵武长枣时，高氧应激处理时的条件为：向密封的气调箱内通入50％O 2(w/w)浓度的氧气短时应激处理30min，然后打开密封的气调箱，静置30min； When the fresh fruit is Lingwu jujube, the conditions for high oxygen stress treatment are: introduce oxygen with a concentration of 50% O 2 (w/w) into the sealed air-conditioned box for short-term stress treatment for 30 minutes, and then open the seal The air-conditioning box is left for 30 minutes;当鲜果为大青葡萄时，高氧应激处理时的条件为：向密封的气调箱内通入80％O 2(w/w)浓度的氧气短时应激处理30min，然后打开密封的气调箱，静置30min。 When the fresh fruit is green grapes, the conditions for high-oxygen stress treatment are: introduce oxygen with a concentration of 80% O 2 (w/w) into the sealed air-conditioned box for short-term stress treatment for 30 minutes, and then open the sealed box. Atmospheric conditioning box, let stand for 30 minutes.
- 如权利要求1至6任一项所述的保鲜方法在鲜果贮藏方面中的应用。Application of the preservation method according to any one of claims 1 to 6 in the storage of fresh fruits.
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