CN113349250A - Application of brown algae oligosaccharide and pichia guilliermondii combined in prevention and control of penicilliosis and storage and preservation of pear fruit after picking - Google Patents

Application of brown algae oligosaccharide and pichia guilliermondii combined in prevention and control of penicilliosis and storage and preservation of pear fruit after picking Download PDF

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CN113349250A
CN113349250A CN202110501833.6A CN202110501833A CN113349250A CN 113349250 A CN113349250 A CN 113349250A CN 202110501833 A CN202110501833 A CN 202110501833A CN 113349250 A CN113349250 A CN 113349250A
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pichia guilliermondii
pear
penicilliosis
oligosaccharide
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张红印
韩俊佳
赵利娜
王远见
兰晨
周游
张晓云
王凯利
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Jiangsu University
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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
    • 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
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

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Abstract

The invention belongs to the technical field of biological control, and particularly relates to application of brown algae oligosaccharide and pichia guilliermondii combined in control of penicilliosis and storage and preservation of pear fruits after picking; the method comprises the following steps: activating and culturing Pichia guilliermondii, centrifuging to obtain thallus, preparing yeast suspension and alginate oligosaccharide solution with sterile water, and mixing the two solutions to obtain a mixed solution; after punching holes on the surface of the pear, injecting a mixed solution, and injecting a penicillium expansum spore suspension with the same volume after 2 hours to realize effective control on penicillium disease; or directly spraying the mixed solution on the surface of the pome, and naturally drying; the purpose of preventing and treating the penicilliosis of the picked pear and storing and refreshing the pear can be realized; the invention can effectively control penicilliosis and natural rot of the picked pear, thereby reducing the loss caused by diseases of the picked pear; meanwhile, the method has no adverse effect on the main quality index of the picked pome, is convenient to operate, safe and environment-friendly, and has remarkable economic and social benefits.

Description

Application of brown algae oligosaccharide and pichia guilliermondii combined in prevention and control of penicilliosis and storage and preservation of pear fruit after picking
Technical Field
The invention belongs to the technical field of biological prevention and control of postharvest diseases of fruits, and particularly relates to application of brown algae oligosaccharide and pichia guilliermondii in prevention and control of penicilliosis and storage and preservation of postharvest diseases of pears.
Background
The pome is one of five fruits in China, is widely loved by consumers and has great medicinal value. The picked pome has quick physiological decay under natural conditions, and the tissue is easy to be damaged and infected by pathogenic microorganisms. According to investigation, in China, due to the fact that the storage and preservation technical level is low, equipment conditions are limited, and loss of pears from packaging, storage and transportation to marketing due to physiological diseases is as high as about 30%. The diseases in the storage period of the pear mainly include physiological diseases such as black heart disease and pulp browning caused by water loss or low-temperature cold damage of the pear fruit, and infectious diseases such as Penicillium expansum (Penicillium expansum) caused by fungi. Penicillium expansum not only causes the decay of pear fruits from outside to inside, but also secretes Patulin (PAT) in the course of infection. Patulin has various toxicities, and PAT remained in polluted fruits and derivative products thereof can cause diseases such as acute diseases, chronic diseases, pathological changes at cell level and the like of organisms, threaten human health, and easily cause serious food safety problems.
At present, the control method for fruit postharvest diseases mainly comprises a physical method, a chemical method and a biological control method. The physical method can inhibit the activity and toxin accumulation of pathogenic bacteria to different degrees, but has great limitation in practical application due to high equipment cost, great industrialization difficulty, unfavorable changes of physicochemical properties and sensory quality of fruits caused by improper treatment and other problems. Although the chemical bactericide has remarkable effect, the long-term use of the chemical bactericide can cause pathogenic bacteria to generate drug resistance, cause environmental pollution and have potential safety hazard. The biological control method is to control the postharvest diseases of fruits by using antagonistic microorganisms (bacteria, mold and yeast), and compared with physical methods and chemical methods, the biological control method is safe and harmless, has no side effect and has good control effect.
Compared with other antagonistic microorganisms, the antagonistic yeast has the advantages of rapid proliferation, no generation of antibiotics and mycotoxins, genetic stability, high safety and the like. The partially antagonistic yeast has good prevention and treatment effects on postharvest diseases of fruits, can degrade toxins generated by pathogenic bacteria, and has great significance for large-scale practical application of biological prevention and treatment. The relevant literature shows that: the use of Cryptococcus albidus (Cryptococcus albicans) or Rhodosporidium rubrum (Rhodosporidium paludigenum) can inhibit penicilliosis of pear fruit after harvest; but the effect is not ideal when the yeast is used alone for prevention and control; in order to further improve the control effect of the antagonistic yeast, other methods can be used in combination with the antagonistic yeast, and chemical additives, other antagonistic bacteria, chemical bactericides and the like are commonly used in combination with the antagonistic yeast. If the method is researched in the literature; the control effects of the Pichia membranaeaciens and the low-dose carbendazim or thiophanate-methyl are better than the control effects of P.
The brown Algae Oligosaccharide (AOS) can be dissolved in water, has strong stability, has biological functions of reducing blood sugar, resisting inflammation, regulating immunity, regulating growth and the like, is safe and environment-friendly, and is widely applied to the aspects of medicines, foods, daily chemicals, agriculture and the like. However, in the aspect of biological control of penicillium piriformis, no related research report for the combined application of brown algae oligosaccharide and antagonistic yeast is found at present.
Disclosure of Invention
The invention aims to apply brown algae oligosaccharide and pichia guilliermondii (Meyerozyma guilliermondii) to biological control of pear fruit postharvest, can effectively control occurrence of penicilliosis of pear fruit postharvest, reduces loss caused by postharvest diseases, and has potential application value.
In order to achieve the above object, the present invention provides the following solutions:
the pichia guilliermondii is screened by the laboratory, has stronger antagonistic effect and is safe and harmless to the human body. The brown algae oligosaccharide is a functional oligosaccharide, has wide application and is non-toxic and harmless. The research proves that the brown algae oligosaccharide and the pichia guilliermondii are combined, the occurrence of penicilliosis after the pear fruit is picked can be obviously reduced, the natural rotting rate of the pear fruit after being picked is reduced, the quality of the pear fruit is not affected, the purpose of storing and preserving the fruit can be achieved, and the unexpected outstanding effect is achieved.
The preservation information of the pichia guilliermondii is as follows: the preservation name is: pichia guilliermondii (Meyerozyma guilliermondii); the preservation number is CCTCC NO: M2017270; the preservation unit: china Center for Type Culture Collection (CCTCC); and (4) storage address: eight Lopa in Wuchang region of Wuhan city, Hubei province; the preservation date is as follows: year 2017, month 5 and day 17.
The brown algae oligosaccharide and pichia guilliermondii combined application to control, store and keep freshness of penicilliosis after pear picking comprises the following specific steps:
firstly, activating the pichia guilliermondii, then culturing the pichia guilliermondii by using an NYDB culture medium, centrifuging to obtain thalli, and then washing and centrifuging by using sterile water; repeating the washing and centrifuging steps for several times, and diluting the obtained thalli into yeast suspension with a certain concentration by using sterile water; preparing a brown algae oligosaccharide solution by using sterile water, and then mixing the yeast suspension and the brown algae oligosaccharide solution to obtain a mixed solution; punching the surface of pear by using a sterilized puncher, then respectively injecting a proper volume of mixed liquor into each hole, and respectively inoculating penicillium expansum spore suspension with the same volume as the mixed liquor into each hole after a period of time, wherein the concentration of the penicillium expansum spore suspension is 5 multiplied by 104spores/mL, and realizes effective prevention and treatment of penicillium expansum spore caused penicillium disease; or the pears are not treated, the natural condition of the pears is kept and the pears are put into a plastic basket, the mixed liquid is uniformly sprayed on the surfaces of the pears, the pears are put into a freshness protection bag after being naturally dried, then the pears are put into the plastic basket, and the pears are sealed by a freshness protection film and then stored at room temperature, so that the purposes of controlling, storing and keeping freshness of the penicilliosis after picking the pears can be realized.
Preferably, the culture conditions in the NYDB culture medium are as follows: the temperature is 28 ℃, the rotating speed is 180-200 rpm, and the time is 20-22 h.
Preferably, the centrifugation conditions are: the centrifugal force is 8000g, and the time is 10-15 min.
Preferably, the aperture of the punch is 4mm, and the depth is 5 mm; the period of time is 2 hours.
Preferably, the volumes of the mixture and the penicillium expansum spore suspension injected into the wells are both 30. mu.L.
Preferably, the NYDB culture medium (calculated by 1L) is: 5g of yeast extract, 10g of glucose, 8g of beef extract and distilled water are added to a constant volume of 1000mL, the pH is natural, and the mixture is sterilized at 115 ℃ for 20 min.
Preferably, the concentration of yeast in the mixed solution is 1X 108cells/mL, the final concentration of the brown algae oligosaccharide is 5 g/L; the preparation method comprises the following steps: the cells obtained by centrifugation and repeated washing were diluted with sterile water to 2X 108cells/mL bacterial suspension; and preparing the brown algae oligosaccharide with the concentration of 10g/L by using sterile water, and mixing the yeast suspension and the brown algae oligosaccharide solution according to the volume ratio of 1:1 to obtain a mixed solution.
The invention has the advantages that:
1. the pichia guilliermondii is screened by the laboratory, has stronger antagonistic effect and is safe and harmless to human bodies. The brown algae oligosaccharide used in the invention is a functional oligosaccharide, has wide application, and is non-toxic and harmless. At present, the related research of applying pichia guilliermondii combined brown alginate oligosaccharides to penicilliosis after pear fruit harvest does not exist.
2. According to the method, the brown algae oligosaccharide is combined with the pichia guilliermondii to treat the pear, compared with the single treatment of the pichia guilliermondii, the combined treatment can obviously inhibit the occurrence of penicilliosis after the pear is picked, the natural rot rate of the pear after the pear is picked is reduced, and the method has a prominent effect; the concentration of the brown algae oligosaccharide is important, the more the brown algae oligosaccharide is, the better the brown algae oligosaccharide is, the more the brown algae oligosaccharide is, the too high or too low brown algae oligosaccharide cannot achieve remarkable effect, and the most remarkable effect is achieved when the final concentration is 5 g/L.
3. Compared with a control group, the brown alga oligosaccharide combined with the Mongolian pichia pastoris has no adverse effect on main quality indexes of the harvested pome, such as weight loss rate, hardness, soluble solid and titratable acid, and the like, and has the effect of slowing down the reduction of the quality indexes, such as the weight loss rate of the pome treated by the brown alga oligosaccharide combined with the Mongolian pichia pastoris is obviously reduced.
4. The brown alga oligosaccharide and the pichia guilliermondii are used for replacing a chemical bactericide to prevent and treat the fruit postharvest diseases, the chemical bactericide can be replaced to prevent and treat the pear fruit postharvest diseases, the potential harm of the chemical bactericide to human bodies and the environment is reduced, the economic and energy burden caused by physical sterilization and storage is reduced, and the method has obvious economic benefit and social benefit.
Drawings
FIG. 1 shows the effect of brown algae oligosaccharide and Pichia guilliermondii on the decay rate of penicilliosis after pear harvest. Note: CK represents sterile water treatment, and is a control group; A. b, C Brown algae oligosaccharide 1g/L, 5g/L, 10g/L, respectively, and D represents Pichia Quinquisensis single treatment (concentration 1 × 10)8cells/mL), E, F, G is the treatment of the mixed solution of alginate oligosaccharide and Pichia guilliermondii, wherein the concentration of Pichia guilliermondii in the mixed solution is 1 × 108cells/mL, the final concentration of the brown alginate oligosaccharide in the E mixed solution is 1G/L, the final concentration of the brown alginate oligosaccharide in the F mixed solution is 5G/L, and the final concentration of the brown alginate oligosaccharide in the G mixed solution is 10G/L; different lower case letters represent significant differences between treatments (P)<0.05)。
FIG. 2 shows the effect of brown algae oligosaccharide and Pichia guilliermondii on the decay diameter of penicilliosis after pear harvest. Note: CK represents sterile water treatment, and is a control group; A. b, C brown algae oligosaccharide 1g/L, 5g/L, and 10g/L, respectively, and D represents Pichia Quinwensis (concentration 1 × 10)8cells/mL) and E, F, G is the mixed solution treatment composed of alginate oligosaccharide and pichia guilliermondii, wherein the concentration of the pichia guilliermondii in the mixed solution is 1 multiplied by 108cells/mL, the final concentration of the brown alginate oligosaccharide in the E mixed solution is 1G/L, the final concentration of the brown alginate oligosaccharide in the F mixed solution is 5G/L, and the final concentration of the brown alginate oligosaccharide in the G mixed solution is 10G/L; different lower case letters represent significant differences between treatments (P)<0.05). Different lower case letters represent significant differences between treatments (P)<0.05)。
The specific implementation mode is as follows:
the invention will be explained in more detail by means of the following examples. The following examples are illustrative only, and the present invention is not limited by these examples.
Example 1:
pichia guilliermondii (Meyerozyma guilliermondii) is a safe and nontoxic antagonistic yeast screened by the laboratory, and the strain is applied for patent with the patent application numbers: 201710513315. X.
The culture procedure is as follows: (1) solid activation: inoculating Pichia guilliermondii into NYDA culture medium, and culturing at 28 deg.C for 48 hr; (2) liquid culture: inoculating a ring of activated Pichia guilliermondii into a 250mL triangular flask containing 50mL NYDB culture medium by using an inoculating ring, and culturing for 20h (180rpm, 28 ℃); (3) preparing a bacterial suspension: centrifuging (8000 Xg, 10min), collecting thallus, repeatedly washing with sterile distilled water to remove culture medium, adding appropriate amount of sterile distilled water, counting with blood count plate, and adjusting bacterial suspension concentration to 1X 108cells/mL、2×108cells/mL are ready for use.
The NYDA culture medium is: on the basis of the NYDB medium, 2% agar (by mass) was added.
Example 2:
control effect of brown algae oligosaccharide and pichia guilliermondii on pear penicilliosis
One) test protocol
The test variety is crystal pear, and pear fruits which are free from mechanical damage, uniform in size and at normal commercial maturity are selected. The pome is soaked in 0.2% sodium hypochlorite solution for 2min, washed by tap water and dried for later use.
The mould activation steps before use are as follows: (1) 1mL of the mold spore suspension was inoculated into sterilized PDB medium and cultured for 24 hours on a constant temperature shaker (120rpm, 25 ℃). (2) Taking a proper amount of mould to scribe on a PDA culture medium, culturing in a constant temperature and humidity box for 7d, taking mould spores on a flat plate, washing twice in sterile physiological saline, and adjusting to the required concentration by adopting a binocular microscope.
Three are prepared at each pear equator using a sterile puncherUniformly distributed wound (diameter 4 mm. times.depth 5mm), 30 μ L alginate-derived oligosaccharide solution (1g/L, 5g/L, 10g/L), and yeast suspension (1X 10)8cells/mL), mixed solution of yeast suspension and alginate-derived oligosaccharide (final concentration of alginate-derived oligosaccharide is 1g/L, 5g/L, 10g/L, respectively, and final concentration of Pichia guilliermondii is 1 × 108cells/mL); sterile water. After standing at room temperature for 2h, 30. mu.L of Penicillium expansum spore suspension (5X 10) was injected into the wound4spores/mL). Airing at room temperature, storing in a constant-temperature constant-humidity incubator (20 ℃, RH 95%), observing the rotting condition of the pomes after 4 days, and recording the rotting rate and the rotting diameter. Each treatment was performed in 3 replicates, each replicate 12 pomes, and the experiment was repeated twice.
The formula for the incidence of disease is as follows:
incidence (%) of disease ═ total number of fruits/fruit affected × 100%
II) test results
According to the test steps, the results of counting the rotting rate and the lesion diameter of the fruits are as follows:
as shown in figure 1, after storage for 4d, when the rotting rate of the control group and the brown alginate oligosaccharide single-treatment group reaches 100%, the rotting rate of the pear fruit treated by the pichia guilliermondii single-treatment group is 36.67%; the rot rate of the pear fruits can be further reduced by using the pichia guilliermondii and the alginate oligosaccharide in a combined mode, wherein the rot rate of the pear fruits is the lowest by using the pichia guilliermondii and 5g/L of alginate oligosaccharide in a combined mode, and the rot rate is only 8.33%.
As shown in FIG. 2, compared to the control group (14.33mm), the diameter of rot of the treated pear fruit was 8.37mm for Pichia guilliermondii alone and 6.59mm for Pichia guilliermondii in combination with 5g/L alginate oligosaccharide.
Example 3:
influence of combination of alginate oligosaccharides and pichia guilliermondii on natural decay and storage quality of pear fruits after picking
One) test protocol
The pear fruits which are not mechanically damaged, have uniform sizes and are in normal commercial maturity are selected for testing, do not need to be treated, and are directly sprayed on the surface of the pear fruitsPichia pastoris (1X 10) in season (1)8cells/mL), (2) alginate-derived oligosaccharide (5g/L), (3) mixed solution of Pichia guilliermondii and alginate-derived oligosaccharide (final concentration of alginate-derived oligosaccharide is 5g/L, and final concentration of Pichia guilliermondii is 1 × 10)8cells/mL)), (4) sterile water.
After the pears are dried at room temperature, the pears are independently packaged by a freshness protection package, then are placed in a plastic basket, are sealed by a freshness protection film, are placed in a constant temperature and humidity incubator (20 ℃, RH 95 percent) for storage, and are observed after 40 days, and the natural rot condition of the pears and the quality indexes such as weight loss rate, titratable acid and the like are measured. Each treatment was performed in 3 replicates, each replicate 12 pomes, and the experiment was repeated twice.
The inhibitory effect of pichia guilliermondii in combination with fucoidan on the natural decay of pear fruit is expressed in terms of incidence.
(1) The formula for the incidence of disease is as follows:
incidence (%) of disease ═ total number of fruits/fruit affected × 100%
(2) The storage quality was measured as follows:
1. weight loss rate: weight loss rate (mass before treatment-mass after storage)/mass before treatment × 100%
2. Soluble solid matter: the measurement was carried out by a hand-held sugar meter, and the measurement result was expressed as g/100g of sample.
3. Titratable acid: homogenizing 20g pulp tissue with 20mL distilled water, leaching in 80 deg.C water bath for 30min, diluting to 100mL, and filtering to obtain filtrate. 20mL of the filtrate was taken, phenolphthalein indicator was added dropwise, and titration was performed with a standard solution (0.1M NaOH), and the results were expressed as percentage of sorbic acid.
4. Brown degree: 10g of pome sample is taken and homogenized together with 20mL of distilled water, 10mL of supernatant is taken after centrifugation and mixed with 15mL of 95% ethanol, centrifugation is carried out again, and the absorbance value of the supernatant at 420nm is measured to indicate the browning degree.
5. Relative conductivity: taking 1g of pear tissue, placing in an erlenmeyer flask containing 20mL of deionized water, shaking for 20min (25 ℃, 75rpm), and measuring the conductivity of the leachate (C0); after boiling water bath for 10min, the temperature is cooled to room temperature, and the conductivity of the leachate (C1) is measured. The relative conductivity is C0/C1 × 100%.
6. Ascorbic acid (Vc): a20 g fruit sample is taken to be homogenized with 20mL of 1% hydrochloric acid, and the homogenate is transferred to a 50mL volumetric flask to be constant volume to a scale mark. Centrifuging at 10000rpm and 4 deg.C for 10min, and collecting supernatant for determination. 0.4mL of the supernatant and 0.8mL of 10% HCl were added to a 10mL graduated tube, and water was added to the graduated tube and shaken well. After blank correction, the absorbance of the solution was measured at 243 nm. And adding 4mL of distilled water and 1.6mL of 1mol/L NaOH into another 0.4mL of supernatant to a 10mL graduated test tube, shaking uniformly, and standing for 15 min. Then 1.6mL of 10% HCl is added, mixed evenly and added with distilled water to a constant volume to be calibrated. The blank was corrected with distilled water, and the absorbance at 243nm was measured. Preparing Vc standard curve, calculating twice to obtain A243And calculating the difference value of the values, and calculating the content of Vc in the sample. Each treatment was repeated 3 times.
7. Hardness: the maximum resistance (in N) of the probe when the probe is inserted into the fruit is defined as the hardness of the fruit by measuring the probe diameter by a TA-XT2Plus texture analyzer to be 5mm and the measuring depth to be 10 mm.
II) test results
TABLE 1 influence of Pichia guilliermondii in combination with fucoidan oligosaccharide on the natural decay and storage quality of pear
Figure BDA0003056710630000061
Note: CK is a control group treated with sterile water, AOS represents a single treatment of alginate-derived oligosaccharide solution with a final concentration of 5g/L, and Y represents a Pichia guilliermondii suspension (1X 10)8cells/mL) alone, and Y + AOS represents the mixture of alginate oligosaccharide solution and Pichia guilliermondii suspension (final concentration of alginate oligosaccharide is 5g/L, concentration of Pichia guilliermondii is 1 × 10)8cells/mL). Different lower case letters represent significance of difference (P)<0.05)。
As can be seen from Table 1, after storing for 40 days at 20 ℃, compared with the control group (31.82%), the natural rotting rate of the pear fruit is reduced after the brown alga oligosaccharide is singly treated (25.73%), the natural rotting rate of the pear fruit is 15.91% after the pear fruit is treated by the pichia guilliermondii, the natural rotting rate of the pear fruit is obviously reduced after the pear fruit is treated by the combination of the pichia guilliermondii and the brown alga oligosaccharide (9.09%), and a remarkable effect is achieved. Soluble solids, relative conductivity and hardness of the pome are not changed basically in different treatment modes, and compared with a control group, the content of titratable acid and ascorbic acid is increased when the pome is treated by the alginate oligosaccharide alone; compared with the two previous treatment modes, the pear fruit treated by the pichia guilliermondii has the advantages that the weight loss rate and the browning degree are reduced, and the content of ascorbic acid is increased; compared with the other three treatment modes, the pear fruit treated by combining the pichia guilliermondii and the brown alginate oligosaccharide has the advantages of reduced weight loss rate and browning degree and increased ascorbic acid content. In general, the natural rotting rate of the pear fruits is obviously reduced and the storage quality is not changed badly by the combined treatment of the pichia guilliermondii and the brown algae oligosaccharide.
Example 4:
preservation of pichia guilliermondii:
optionally, NYDA culture medium can be stored at 4 deg.C in refrigerator, or yeast can be stored at-80 deg.C in 25% glycerol tube for a long period.
Description of the drawings: the above embodiments are only used to illustrate the present invention and do not limit the technical solutions described in the present invention; thus, although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted; all such modifications and variations that do not depart from the spirit and scope of the invention are intended to be included within the scope of the appended claims.

Claims (7)

1. The application of brown algae oligosaccharide and pichia guilliermondii combined with season to prevention, storage and preservation of penicilliosis after pear picking is characterized by comprising the following specific steps:
firstly, activating the pichia guilliermondii, then culturing the pichia guilliermondii by using an NYDB culture medium, centrifuging to obtain thalli, and then washing and centrifuging by using sterile water; repeating the washing and centrifuging steps for several times, and diluting the obtained thalli into yeast suspension with a certain concentration by using sterile water; preparing alginate-derived oligosaccharide solution with sterile water, and mixing the yeast suspension and the alginate-derived oligosaccharide solution to obtain the final productMixing the solution; punching the surface of pear by using a sterilized puncher, then respectively injecting a proper volume of mixed liquor into each hole, and respectively inoculating penicillium expansum spore suspension with the same volume as the mixed liquor into each hole after a period of time, wherein the concentration of the penicillium expansum spore suspension is 5 multiplied by 104spores/mL, and realizes effective prevention and treatment of penicillium expansum spore caused penicillium disease;
or the pears are not treated, the natural condition of the pears is kept and the pears are put into a plastic basket, the mixed liquid is uniformly sprayed on the surfaces of the pears, the pears are put into a freshness protection bag after being naturally dried, then the pears are put into the plastic basket, and the pears are sealed by a freshness protection film and then stored at room temperature, so that the purposes of controlling, storing and keeping freshness of the penicilliosis after picking the pears can be realized; the pichia guilliermondii is particularly pichia guilliermondii (Meyerozyma guilliermondii), is preserved in the Chinese type culture collection with the preservation number of: CCTCC M2017270.
2. The application of the brown alginate oligosaccharide and the pichia guilliermondii combined with the pichia guilliermondii in the prevention and the storage and the preservation of the penicilliosis after pear fruit harvest is characterized in that the conditions for culturing the pichia guilliermondii in the NYDB culture medium are as follows: the temperature is 28 ℃, the rotating speed is 180-200 rpm, and the time is 20-22 h.
3. The application of the brown algae oligosaccharide and pichia guilliermondii combined with the pichia guilliermondii in the prevention and treatment of penicilliosis and storage and preservation after pear harvest according to claim 1 is characterized in that the centrifugation conditions are as follows: the centrifugal force is 8000g, and the time is 10-15 min.
4. The application of the brown algae oligosaccharide and pichia guilliermondii combined with the pichia guilliermondii in the prevention, storage and preservation of penicilliosis after pear fruit harvest is characterized in that the aperture of the punched hole is 4mm and the depth is 5 mm; the period of time is 2 hours.
5. The use of the brown algae oligosaccharide and pichia guilliermondii combined with the pichia guilliermondii in the prevention and the preservation of penicilliosis after pear harvest and the preservation and the freshness preservation of the pears is characterized in that the volumes of the mixed liquid and the penicillium expansum spore suspension injected into the holes are both 30 mu L.
6. The application of the brown alginate oligosaccharide and the pichia guilliermondii combined with the pichia guilliermondii in the prevention and the preservation of penicilliosis after pear fruit harvest and the preservation of the penicilliosis, which are disclosed by claim 1, is characterized in that the NYDB culture medium is: 5g of yeast extract, 10g of glucose, 8g of beef extract and distilled water are added to a constant volume of 1000mL, the pH is natural, and the mixture is sterilized at 115 ℃ for 20 min.
7. The use of the brown algae oligosaccharide and pichia guilliermondii combined with the pichia guilliermondii as claimed in claim 1 for preventing and treating penicilliosis and storing and refreshing pear fruit after picking, wherein the concentration of yeast in the mixed solution is 1 x 108cells/mL, the final concentration of the brown algae oligosaccharide is 5 g/L; the preparation method comprises the following steps: the cells obtained by centrifugation and repeated washing were diluted with sterile water to 2X 108cells/mL bacterial suspension; and preparing the brown algae oligosaccharide with the concentration of 10g/L by using sterile water, and mixing the yeast suspension and the brown algae oligosaccharide solution according to the volume ratio of 1:1 to obtain a mixed solution.
CN202110501833.6A 2021-05-08 2021-05-08 Application of brown algae oligosaccharide and pichia guilliermondii combined in prevention and control of penicilliosis and storage and preservation of pear fruit after picking Pending CN113349250A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114717122A (en) * 2022-05-23 2022-07-08 江苏大学 Method for preventing and treating penicilliosis of picked kiwi fruits and storing and preserving kiwi fruits by using Kalibik Meiyer yeast Q11
CN115918720A (en) * 2023-01-06 2023-04-07 福州大学 Application of brown algae oligosaccharide in improving prevention and treatment effect of yeast Lg 3 on postharvest diseases of fruits and vegetables

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107460134A (en) * 2017-06-29 2017-12-12 江苏大学 The Pichia guilliermondii of one plant of control the operatic circle postharvest disease
CN107494699A (en) * 2017-07-05 2017-12-22 福州大学 A kind of bio-preservative based on brown alga oligose and its application on prawn is fresh-keeping
CN111705005A (en) * 2020-05-20 2020-09-25 江苏大学 Yeast for controlling pear patulin and application thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107460134A (en) * 2017-06-29 2017-12-12 江苏大学 The Pichia guilliermondii of one plant of control the operatic circle postharvest disease
CN107494699A (en) * 2017-07-05 2017-12-22 福州大学 A kind of bio-preservative based on brown alga oligose and its application on prawn is fresh-keeping
CN111705005A (en) * 2020-05-20 2020-09-25 江苏大学 Yeast for controlling pear patulin and application thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ZHENSHUO WANG，等: "Management of blue mold (Penicillium italicum) on mandarin fruit with a combination of the yeast, Meyerozyma guilliermondiiand an alginate oligosaccharide", 《BIOCONTROL》 *
刘同梅，等: "海藻酸钠寡糖采前处理对猕猴桃贮藏品质的影响", 《辽宁农业科学》 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114717122A (en) * 2022-05-23 2022-07-08 江苏大学 Method for preventing and treating penicilliosis of picked kiwi fruits and storing and preserving kiwi fruits by using Kalibik Meiyer yeast Q11
CN114717122B (en) * 2022-05-23 2023-09-26 江苏大学 Method for preventing and treating penicilliosis of kiwi fruits after picking by using yarrowia callicarpa Q11 and storing and preserving method
CN115918720A (en) * 2023-01-06 2023-04-07 福州大学 Application of brown algae oligosaccharide in improving prevention and treatment effect of yeast Lg 3 on postharvest diseases of fruits and vegetables
CN115918720B (en) * 2023-01-06 2024-07-09 福州大学 Application of brown alginate oligosaccharides in improving control effect of saccharomycetes Lg 3 on fruit and vegetable postharvest diseases

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Application publication date: 20210907