CN115399366A - Blueberry fruit storage method - Google Patents
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- CN115399366A CN115399366A CN202211084345.0A CN202211084345A CN115399366A CN 115399366 A CN115399366 A CN 115399366A CN 202211084345 A CN202211084345 A CN 202211084345A CN 115399366 A CN115399366 A CN 115399366A
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- 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
- A23B7/153—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
- A23B7/154—Organic compounds; Microorganisms; Enzymes
- A23B7/155—Microorganisms; Enzymes; Antibiotics
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- 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
- A23B7/04—Freezing; Subsequent thawing; Cooling
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- 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
- A23B7/153—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
- A23B7/154—Organic compounds; Microorganisms; Enzymes
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Abstract
The invention discloses a blueberry fruit storage method which comprises the steps of pretreating picked blueberry fruits, treating the blueberry fruits by using a biocontrol bacterium liquid, and storing the blueberry fruits at a low temperature in the presence of an ethylene inhibitor. The method utilizes the ethylene inhibitor to inhibit the respiration of the blueberry fruits, and the biocontrol bacteria inhibit the infection of pathogenic bacteria after the fruits are picked, so that the ripening and aging of the fruits after the fruits are picked are delayed, the effect of improving the storage performance of the fruits is achieved, the method has the advantages of safety, high efficiency, environmental friendliness and the like, the storage and transportation cost of the picked blueberry fruits is reduced, the economic benefit is improved, and the method is a main trend of future development of the storage and preservation technology of the blueberry fruits.
Description
Technical Field
The invention relates to a storage method of crops, in particular to a storage method of blueberry fruits.
Background
Blueberry (Vaccinium spp), the scientific name of which is Vaccinium myrtillus, a plant of Vaccinium of Ericaceae, the fruit of which is berry, is dark blue, the surface of which is coated by white fruit powder, the pulp is fine and smooth, the product is sour, sweet and delicious, contains rich nutrient substances including anthocyanin, pectin, arbutin, tannic acid, folic acid, flavonoid and the like, and has the effects of resisting aging, preventing cancer, preventing heart and brain diseases, eliminating eye fatigue, improving eyesight and the like, so the blueberry is classified as one of five kinds of health food for human by food and agriculture organization of the United nations. At present, the rabbitia blueberry is a main variety group in the growing area of blueberries in Yangtze river basin in China, the maturation period is concentrated in the high-temperature rainy season in the middle and late 6 months, the fruits are soft and juicy, and are susceptible to fungus and rot, so that the storage and preservation technology of the blueberries is not beneficial to storage, and the research on the storage and preservation technology of the blueberries has important significance for the development of the whole blueberry industry.
At present, the blueberry preservation technology is mainly divided into a physical method, a chemical method and a biological method, wherein the physical preservation method mainly comprises a low-temperature refrigeration method, a radiation preservation method, a modified atmosphere preservation method, a low-pressure preservation method, an electrostatic preservation method, a film preservation method and the like. The chemical fresh-keeping method mainly uses fruit fresh-keeping agent to make film coating, sterilization and preservation, etc.. The physical fresh-keeping method has the defects of needing special mechanical equipment, having higher cost and more complex operation and being not suitable for being used in a large range. The existing chemical preservation method has short preservation time, so that the method for preserving the blueberries for a long time is needed by the current market.
Disclosure of Invention
The invention aims to provide a blueberry fruit storage method, which utilizes an ethylene inhibitor to inhibit the respiration of blueberry fruits and biocontrol bacteria to inhibit the infection of pathogenic bacteria after fruit picking, thereby delaying the ripening and aging of the picked fruits, achieving the effect of improving the storage performance of the fruits, having the advantages of safety, high efficiency, environmental protection and the like, reducing the storage and transportation cost of the picked blueberry fruits, improving the economic benefit and being the main trend of future development of the blueberry fruit storage and preservation technology.
In order to achieve the purpose, the invention provides a blueberry fruit storage method which comprises the steps of pretreating picked blueberry fruits, treating the blueberry fruits by using a biocontrol bacterium liquid, and storing the blueberry fruits at low temperature in the presence of an ethylene inhibitor.
Through the technical scheme, the invention has the following beneficial effects:
the method has the advantages of safety, high efficiency, environmental protection and the like, and is the main trend of future development of fruit storage and preservation technology; provides a new way for reducing the cost and improving the economic benefit of the storage and transportation of the picked blueberry fruits.
Additional features and advantages of the invention will be set forth in the detailed description which follows.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:
FIG. 1 is a graph showing the decay rate of "brilliant" blueberry fruits of experimental examples 1 to 3 and comparative example 1 under a storage condition of 4 ℃;
FIG. 2 shows the Malondialdehyde (MDA) content of "brilliant" blueberry fruits of examples 1-3 and comparative example 1 at 4 deg.C storage;
FIG. 3 shows Peroxidase (POD) contents of "brilliant" blueberry fruits of experimental examples 1 to 3 and comparative example 1 under storage conditions at 4 ℃;
FIG. 4 shows the superoxide dismutase (SOD) content of "brilliant" blueberry fruits of experimental examples 1-3 and comparative example 1 under the storage condition of 4 ℃;
FIG. 5 shows anthocyanin contents of "Brilliant" blueberry fruits of Experimental examples 1-3 and comparative example 1 under storage conditions at 4 ℃;
FIG. 6 is the vitamin C content of "Brilliant" blueberry fruit of Experimental examples 1-3 and comparative example 1 under storage conditions at 4 ℃;
FIG. 7 is a graph showing the acid stability ratio of "brilliant" blueberry fruit of experimental examples 1 to 3 and comparative example 1 under storage conditions of 4 ℃.
Detailed Description
The following describes in detail specific embodiments of the present invention. It should be understood that the detailed description and specific examples, while indicating the present invention, are given by way of illustration and explanation only, not limitation.
The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For numerical ranges, each range between its endpoints and individual point values, and each individual point value can be combined with each other to give one or more new numerical ranges, and such numerical ranges should be construed as specifically disclosed herein.
The invention provides a blueberry fruit storage method which comprises the steps of pretreating picked blueberry fruits, treating the blueberry fruits by using biocontrol bacterium liquid, and storing the blueberry fruits at low temperature in the presence of an ethylene inhibitor.
According to the invention, the blueberry fruits are soaked by using the biocontrol bacterium liquid, then the ethylene inhibitor is placed in the storage process of the blueberry fruits, under the combined action of the biocontrol bacterium liquid and the ethylene inhibitor, the blueberry fruits are stored at a low temperature, the respiration of the fruits is inhibited by using the ethylene inhibitor, and the biocontrol bacterium inhibits the infection of pathogenic bacteria after the fruits are picked, so that the ripening and the aging of the fruits after the fruits are picked are delayed, and the effect of improving the storage performance of the fruits is achieved.
In the above method, the species of the biocontrol bacterium can be selected from a wide range, but for better preservation, it is preferable that the biocontrol bacterium is selected from at least one of bacillus subtilis, bacillus cereus, bacillus alpina, bacillus atrophaeus and bacillus tropicalis.
In the above method, the species of the biocontrol bacterium can be selected within a wide range, but for better preservation, it is preferable that the biocontrol bacterium is bacillus alpina.
In the above method, the ethylene inhibitor may be selected within a wide range, but in order to secure the anticorrosive effect of the experiment, it is preferable that the ethylene inhibitor is selected from at least one of 1-methylcyclopropene, aminooxyethylvinylglycine, methylaminovinylglycine, and aminooxyacetic acid.
In the above process, the ethylene inhibitor may be selected within a wide range, but in order to secure the anticorrosive effect of the experiment, it is preferable that the ethylene inhibitor is 1-methylcyclopropene.
In the above method, the pretreatment conditions for the blueberry fruit may be selected within a wide range, but in order to secure the freshness of the blueberry fruit, it is preferable that the pretreatment conditions include: the blueberry fruit is pre-cooled after being harvested, wherein the pre-cooling temperature is 18-24 ℃, and the pre-cooling time is 1.5-3h.
In the above method, the conditions for the treatment with the biocontrol bacterium solution can be selected from a wide range, but in order to achieve a better preservative effect, it is preferable that the conditions for the treatment with the biocontrol bacterium solution include: and (3) soaking the blueberry fruits by adopting the biocontrol bacterium liquid for 2-4min.
In the above method, the preparation method of the biocontrol bacteria liquid can be selected in a wide range, but in order to ensure the activity and the antiseptic effect of the biocontrol bacteria liquid, it is preferable that the preparation method of the biocontrol bacteria liquid is: activating a biocontrol bacteria source, and then inoculating the activated biocontrol bacteria source into an LB liquid culture medium for culture to obtain biocontrol bacteria liquid;
the conditions of the culture include: the volume ratio of the bacterial source of the biocontrol bacteria to the LB liquid culture medium is 1X 10 3 -2×10 3 The temperature is 35-40 ℃, the rotating speed is 130-140r/min, and the culture time is 8-12h.
In the above method, the variety of blueberry fruit can be selected in a wide range, but in order to take the economic benefit of the experiment into consideration, the variety of blueberry fruit is preferably selected from the group consisting of "brilliant" and/or "aundel", more preferably "brilliant", and the blueberry fruit is selected from the group consisting of fruits having no stem, no mechanical damage, relatively uniform size and maturity, and uniform color.
In the above method, the conditions for cryopreservation may be selected within a wide range, but in order to achieve a better preservation effect, it is preferable that the conditions for cryopreservation include: the temperature is 2-6 ℃.
The present invention will be described in detail below by way of examples. In the following examples, the bacillus alpina was identified separately from rotten blueberries, pears, and rice, provided by the present group; the medicines and the medicaments are conventional commercial products.
Example 1
(1) Adding 500 μ L of activated Bacillus alpina into 500mL LB liquid medium (1% tryptone, 0.5% yeast extract, 1% inorganic salt), shaking and culturing at 37 deg.C and 135r/min for 10h to obtain bacterial liquid;
(2) Mature, bright and blue blueberries are harvested, placed in a pre-cooling chamber (at 21 ℃) for pre-cooling for 2 hours, then placed in a foam box with an ice bag inside and transported back to a laboratory. Selecting fruits without fruit stems, dry and comfortable surfaces, complete fruit powder and basically consistent size and maturity, and randomly putting the fruits into 120 pieces/boxes of low-density polyethylene plastic food packaging boxes with the specification of 109mm multiplied by 42mm (length multiplied by width multiplied by height) for standby;
(3) Soaking the blueberry fruits in bacillus alpina bacterial solution for 3min, fishing out, draining, putting into a packaging box, and then putting 2 paper type 1-MCPs with the specification of 10cm multiplied by 10cm into the packaging box;
(4) The fruits after the above treatment were stored under refrigeration at 4 ℃ and sampled every 5 days during storage, and the samples were continuously sampled 5 times and recorded as B1.
Example 2
(1) Adding 500 μ L of activated Bacillus alpine strain source into 750mL LB liquid medium (1% tryptone, 0.5% yeast extract, 1% inorganic salt), and performing shake culture at 35 deg.C and 130r/min for 8h to obtain a bacterial solution;
(2) After being harvested, the mature blueberry 'Aunei' fruits are placed in a pre-cooling chamber (at 18 ℃) for pre-cooling for 1.5 hours, and then placed in a foam box with an ice bag inside and transported back to a laboratory. Selecting fruits without fruit stems, dry and comfortable surfaces, complete fruit powder and basically consistent size and maturity, and randomly putting the fruits into 120 pieces/boxes of low-density polyethylene plastic food packaging boxes with the specification of 109mm multiplied by 42mm (length multiplied by width multiplied by height) for standby;
(3) Soaking the blueberry fruits in the bacillus alpina bacterial solution for 2min, taking out, draining, filling into a packaging box, and then putting 2 paper type 1-MCPs with the specification of 10cm multiplied by 10cm into the packaging box;
(4) The fruits after the above treatment were stored under refrigeration at 2 ℃ and sampled every 5 days during storage, and the samples were continuously sampled 5 times and recorded as B2.
Example 3
(1) Adding 500 μ L of activated Bacillus cereus into 1000mL LB liquid medium (1% tryptone, 0.5% yeast extract, 1% inorganic salt), and performing shake culture at 40 deg.C and 140r/min for 12h to obtain a bacterial solution;
(2) Mature, bright and blue blueberries are harvested, placed in a pre-cooling chamber (at 24 ℃) for pre-cooling for 3 hours, then placed in a foam box with an ice bag inside and transported back to a laboratory. Selecting fruits without fruit stems, dry and comfortable surfaces, complete fruit powder and basically consistent size and maturity, and randomly putting the fruits into 120 low-density polyethylene plastic food packaging boxes with the specifications of 109mm multiplied by 42mm (length multiplied by width multiplied by height) for standby;
(3) Soaking the blueberry fruits in bacillus cereus liquid for 4min, taking out, draining, filling into a packaging box, and then putting 2 paper sheets of 1-MCP with the specification of 10cm multiplied by 10cm into the packaging box;
(4) The fruits after the above treatment were stored under refrigeration at 6 ℃ and sampled every 5 days during storage, and the samples were continuously sampled 5 times and recorded as B3.
Comparative example 1
(1) The harvested ripe 'brilliant' fruits of the rabbit-eye blueberries are placed in a pre-cooling chamber (at 21 ℃) for pre-cooling for 2 hours, then placed in a foam box with an ice bag inside and transported back to a laboratory. Selecting fruits without fruit stems, dry and comfortable surfaces, complete fruit powder and basically consistent size and maturity, and randomly putting the fruits into 120 low-density polyethylene plastic food packaging boxes with the specifications of 109mm multiplied by 42mm (length multiplied by width multiplied by height) for standby;
(2) Soaking the blueberry fruits in ultrapure water for 3min, fishing out, draining, filling into a packaging box, and then putting 2 pieces of filter paper sheets with the specification of 10cm multiplied by 10cm into the packaging box;
(3) The fruits after the above treatment were stored under refrigeration at 4 ℃ and sampled every 5 days for analysis during storage, and the samples were continuously sampled 5 times and recorded as CK.
Comparative example 2
(1) The harvested ripe 'brilliant' fruits of the rabbit-eye blueberries are placed in a pre-cooling chamber (at 21 ℃) for pre-cooling for 2 hours, then placed in a foam box with an ice bag inside and transported back to a laboratory. Selecting fruits without fruit stems, dry and comfortable surfaces, complete fruit powder and basically consistent size and maturity, and randomly putting the fruits into 120 pieces/boxes of low-density polyethylene plastic food packaging boxes with the specification of 109mm multiplied by 42mm (length multiplied by width multiplied by height) for standby;
(2) Soaking the blueberry fruits in ultrapure water for 3min, taking out, draining, filling into a packaging box, and then putting 2 paper sheets of 1-MCP with the specification of 10cm × 10cm into the packaging box;
(3) The fruits after the above treatment were stored under refrigeration at 4 ℃ and sampled every 5 days during storage, and the samples were continuously sampled 5 times and recorded as D1.
Comparative example 3
(1) Mature, bright and blue blueberries are harvested, placed in a pre-cooling chamber (at 21 ℃) for pre-cooling for 2 hours, then placed in a foam box with an ice bag inside and transported back to a laboratory. Selecting fruits without fruit stems, dry and comfortable surfaces, complete fruit powder and basically consistent size and maturity, and randomly putting the fruits into 120 low-density polyethylene plastic food packaging boxes with the specifications of 109mm multiplied by 42mm (length multiplied by width multiplied by height) for standby;
(2) Soaking the blueberry fruits in the bacillus alpina bacterial solution for 3min, taking out, draining, filling into a packaging box, and then putting 2 pieces of filter paper sheets with the specification of 10cm multiplied by 10cm into the packaging box;
(3) The fruits after the above treatment were stored under refrigeration at 4 ℃ and sampled every 5 days during storage, and the samples were continuously sampled 5 times and recorded as D2.
Example of detection
The relevant indexes of example 1 and comparative examples 1 to 3 obtained as described above were measured, and the measurement items and standards were as follows:
the rot rate is based on the obvious juice or rot on the surface of the fruit, and the rot rate (%) = (number of rotted fruits/total number of fruits) × 100%;
the soluble solid is measured by a digital display glucometer;
titratable acid adopts an acid-base titration method;
the anthocyanin content is measured by a methanol extraction colorimetric method (Caojiankang, 2007);
vitamin C is measured by a 2, 4-dinitrophenylhydrazine colorimetric method (in terms of fresh mass) (GB/T, 2003);
malondialdehyde (MDA) adopts a thiobarbituric acid colorimetric method (MDA is an intermediate product of membrane lipid peroxidation during plant tissue aging, and is often used as an index for measuring the membrane lipid peroxidation degree to reflect the cell membrane permeability and the fruit aging degree);
peroxidase (POD) and superoxide dismutase (SOD) activity (Liheng, 2003),
the results obtained are shown in FIGS. 1 to 7.
As shown in fig. 1, example 1 significantly reduced the rotting rate of the blueberry fruits compared to comparative examples 1-3 under the storage condition of 4 ℃. At 25d, the rot rate of example 1 is much higher than that of comparative examples 1-3, so that it can be seen that example 1 has the best effect of reducing the rot rate of blueberries in the storage period, and the rot rate is only 8.33% when the blueberries are stored for 25d, while the rot rate of comparative example 1 is as high as 27.78%.
As shown in figure 2, under the storage condition of 4 ℃, the MDA content of the blueberry fruits shows a trend of increasing and then decreasing, and the peak of the MDA content appears at 15d. The MDA content of the blueberry fruits in the example 1 is changed more slowly than that in the comparative examples 1-3. At 25d, the MDA content of the blueberry fruits treated in the example 1 is remarkably lower than that of the blueberry fruits treated in the comparative examples 1-3.
As can be seen from fig. 3, the POD activity of the blueberry fruits shows a trend of increasing before decreasing under the storage condition of 4 ℃, and reaches a peak at 20 d. At 25d, the POD activity of the blueberry fruits of example 1 was significantly higher than that of comparative example 1.
As can be seen from FIG. 4, under the storage condition of 4 ℃, the SOD activity of the blueberry fruits is similar, the SOD activity of the blueberry fruits shows a trend of increasing first and then decreasing, the SOD peak value of the blueberry fruits all appears at the 20d, and the SOD activity peak value of the blueberry fruits treated in the example 1 is higher than that of the blueberry fruits treated in the comparative example 1. At 25d, the SOD activities of example 1 were all significantly higher than comparative example 1, with the SOD activity of example 1 being the highest.
As can be seen from FIG. 5, under the storage condition of 4 ℃, the anthocyanin content of the blueberry fruits is in a descending trend along with the prolongation of the storage period. The example 1 has obvious delaying effect on the reduction of the anthocyanin content of the blueberry fruits, and at the 25d, the anthocyanin content of the example 1 is obviously higher than that of the comparative example 1, wherein the content of the example 1 is the highest.
As can be seen from fig. 6, under the storage condition of 4 ℃, the Vc content of the blueberry fruits decreases with the increase of the storage time, the Vc content of the blueberry fruits is delayed in example 1, and when the storage time reaches 25d, compared with comparative example 1, the Vc content of the blueberry fruits is significantly delayed in example 1, wherein the effect of delaying the decrease in example 1 is the best.
As can be seen from FIG. 7, under the storage condition of 4 ℃, the solid acid ratio of the blueberry fruits is in an increasing trend along with the extension of the storage period, 0-5d at the early stage of the storage, and the solid acid ratio of the blueberry fruits is not significantly different from that of the blueberry fruits in the example 1 and the blueberry fruits in the comparative examples 1-3. At 25d, the acid fixing ratio of the blueberry fruit in the example 1 is obviously higher than that in the comparative examples 1-3.
The results of examples 2 to 3 were slightly inferior to those of example 1, and the tendency of each index was substantially the same as that of example 1 and was superior to those of comparative examples 1 to 3.
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.
In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.
Claims (10)
1. A blueberry fruit storage method is characterized by comprising the steps of pretreating picked blueberry fruits, treating the blueberry fruits by using a biocontrol bacterium liquid, and storing the blueberry fruits at a low temperature in the presence of an ethylene inhibitor.
2. The method of claim 1, wherein the biocontrol bacteria is selected from at least one of bacillus subtilis, bacillus cereus, bacillus alpina, bacillus atrophaeus, and bacillus tropicalis.
3. The method according to claim 1 or 2, wherein the biocontrol bacterium is bacillus alpina.
4. The method of any one of claims 1-3, wherein the ethylene inhibitor is selected from at least one of 1-methylcyclopropene, aminooxyethylvinylglycine, methylaminovinylglycine, and aminooxyacetic acid.
5. The process according to any one of claims 1 to 4, wherein the ethylene inhibitor in step (3) is 1-methylcyclopropene.
6. The method of any of claims 1-5, wherein the pre-treatment conditions comprise: the blueberry fruit is pre-cooled after being harvested, wherein the pre-cooling temperature is 18-24 ℃, and the pre-cooling time is 1.5-3h.
7. The method according to any one of claims 1 to 6, wherein the conditions for the treatment with the biocontrol bacterial liquid comprise: and (3) soaking the blueberry fruits by adopting the biocontrol bacterium liquid for 2-4min.
8. The method according to any one of claims 1 to 7, wherein the biocontrol bacterium liquid is prepared by a method comprising the following steps: and activating the biocontrol bacteria source, and inoculating the activated biocontrol bacteria source into an LB liquid culture medium for culture to obtain the biocontrol bacteria liquid.
9. Process according to any one of claims 1 to 8, wherein the variety of blueberry fruit is selected from "brilliant" and/or "aundel", preferably "brilliant".
10. The method of any one of claims 1-9, wherein the conditions of cryopreservation comprise: the temperature is 2-6 ℃.
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CN105784947A (en) * | 2016-03-01 | 2016-07-20 | 浙江农林大学 | Physiological and biochemical experiment method in blueberry preservation process |
CN110468075A (en) * | 2019-08-21 | 2019-11-19 | 华南农业大学 | One plant height mountain bacillus and its application in prevention and treatment lichee frost epidemic disease |
CN112608868A (en) * | 2020-12-23 | 2021-04-06 | 上海交通大学 | Bacillus altitudinis and application thereof |
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