CN110710566B - Application of yarrowia lipolytica in preventing and treating postharvest diseases of asparagus and storing and refreshing asparagus - Google Patents
Application of yarrowia lipolytica in preventing and treating postharvest diseases of asparagus and storing and refreshing asparagus Download PDFInfo
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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
- 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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- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
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- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention belongs to the field of postharvest disease biological control, and particularly relates to application of yarrowia lipolytica in postharvest disease control and storage and preservation of asparagus; the method comprises the following steps: activating yarrowia lipolytica, culturing, centrifuging to obtain thallus, washing with sterile normal saline, centrifuging, diluting with sterile normal saline to 1 × 10 concentration 7 Directly and uniformly spraying cell/mL bacterial suspension on the surface of the asparagus, and naturally drying; the disease control and storage and fresh keeping of the asparagus after picking can be realized; the method can effectively control the root rot and the natural rot of the picked asparagus, thereby reducing the loss caused by the disease of the picked asparagus; meanwhile, the method has no adverse effect on the main quality indexes of the picked asparagus, such as the content of soluble sugar, vitamin C and chlorophyll, is safe and environment-friendly, can replace a chemical bactericide to prevent and treat the picked asparagus diseases, and reduces the potential harm of the chemical bactericide to human bodies and the environment.
Description
Technical Field
The invention belongs to the field of biological prevention and control of postharvest diseases of vegetables, and particularly relates to application of Yarrowia lipolytica (Yarrowia lipolytica) in prevention and control of postharvest diseases of asparagus and storage and freshness preservation.
Background
Asparagus, the academic name of Asparagus (Asparagus officinalis), also known as Asparagus, is a perennial herb of the Asparagus genus of the liliaceae family. The green asparagus contains chlorophyll, amino acid, multivitamins, carbohydrate, calcium, phosphorus and other mineral substances and various active ingredients, has higher nutritional value, is considered as one of the medicine and food dual-purpose plants with the best nutritional balance in the world, and has curative effects on various diseases such as hypertension, hyperlipidemia, arteriosclerosis, heart disease, hepatitis, liver cirrhosis, nephritis, edema, cystitis and the like. The long-term eating of the health-care food can help digestion, enhance appetite, improve immunity of the human body, reduce toxicity of harmful substances, inhibit vitality of cancer cells, prevent generation of the cancer cells and have good anticancer effect.
At present, China has become the first world-wide Luzhou bamboo shoot production and export country, the sowing area and the export amount exceed 50% of the world total amount, and the annual output value reaches hundreds of billions of yuan. However, the harvested asparagus tissue is fragile and tender and is easily damaged mechanically and physiologically, and further infected by various pathogenic bacteria such as Fusarium (Fusarium), Alternaria (Alternaria) and Botrytis (Botrytis). After the picked asparagus is infected by pathogenic bacteria, the physiological and metabolic activities of the asparagus are deteriorated to different degrees, the nutritional value and the eating quality of the asparagus are greatly influenced, and huge economic loss is caused. Therefore, appropriate methods must be taken to control the postharvest disease.
At present, domestic and foreign control methods for postharvest diseases of fruits and vegetables are mainly divided into a physical control method, a chemical control method and a biological control method. The physical control method has the advantages of safety, harmlessness and no adverse effect on human bodies and environment. However, most physical control methods have high requirements on equipment, the cost is increased to a certain extent, improper condition control is extremely easy to have adverse effects on sensory quality of the picked asparagus, for example, the asparagus is frozen due to too low temperature, the hardness of the asparagus is increased due to too high temperature, the edible rate is reduced, the degradation of nutrient substances and the deterioration of flavor are accelerated, and the like. Since 1960, the use of chemical bactericides has become the most important means for controlling postharvest diseases of fruits and vegetables. The chemical bactericide has the advantages of high efficiency, low price, convenient use and the like, and is used in large quantities for a long time, but the method has the defects of causing pathogenic bacteria drug resistance, harming human health, polluting environment and the like. With the concern of people on environment, ecology and health, the policy for chemical sterilization is becoming stricter. Therefore, it is very important to actively find an efficient and safe method for preventing and treating postharvest diseases of fruits and vegetables, which can replace chemical bactericides.
The biological control method can effectively control the postharvest diseases of the fruits and the vegetables, has the advantages of safety, no toxicity, no pollution to the environment, no generation of drug resistance of pathogenic bacteria and the like, gradually becomes a research hotspot for controlling the postharvest diseases of the fruits and the vegetables, and is expected to replace chemical bactericides in the future. However, no biological control method for safely and effectively reducing the postharvest diseases of the asparagus exists at present, and no research report on the use of yarrowia lipolytica for preventing and treating the postharvest diseases of the asparagus is found.
Disclosure of Invention
The invention aims to provide a method for controlling postharvest diseases of asparagus by Yarrowia lipolytica (Yarrowia lipolytica), which can effectively control the occurrence of postharvest root rot and natural rot of asparagus and reduce the disease index of asparagus, thereby achieving the purposes of storing and refreshing asparagus and reducing the loss caused by postharvest diseases and having potential application value.
The technical scheme adopted by the invention
The Yarrowia lipolytica WSL-01 for preventing and treating postharvest diseases of asparagus provided by the invention is disclosed in the patent of application and using method of Yarrowia lipolytica in postharvest disease control of grapes (patent number: ZL 201410231417.9); is preserved in China general microbiological culture Collection center with the preservation number of CGMCC NO.8492, the preservation address is China general microbiological culture Collection center of China microbiological culture Collection management Committee of West Lu No. 1 Hospital, North West Chen, south China, Kyoho, Beijing, and the preservation time is 11 months and 21 days in 2013.
The yarrowia lipolytica WSL-01 is used for controlling the postharvest root rot and natural rot of the asparagus and is used for storing and refreshing the asparagus.
The yarrowia lipolytica is applied to the prevention and control of diseases after the asparagus is picked and the storage and the fresh-keeping of the asparagus, and the specific steps are as follows:
activating yarrowia lipolytica, culturing yarrowia lipolytica in NYDB culture medium, centrifuging to obtain thallus, washing with sterile physiological saline, centrifuging, repeating the washing and centrifuging steps for several times, and diluting the thallus with sterile physiological saline to 1 × 10 7 cell/mL of bacterial suspension; punching the surface of Germinatus Phragmitis with sterilized puncher, injecting appropriate volume of yarrowia lipolytica suspension into each hole, inoculating Fusarium spore suspension with the same volume as the yeast suspension into each hole after a period of time, wherein the concentration is 5 × 10 4 spores/mL; or directly and uniformly spraying the bacterial suspension on the surface of the asparagus; naturally drying; placing in plastic basket, sealing with plastic wrap, and storing at room temperature or in incubator.
Preferably, the culture conditions are: the temperature is 28-30 ℃, the rotating speed is 180-200 rpm, and the time is 24-28 h.
Preferably, the centrifugation conditions are: the rotating speed is 7000g, and the time is 10-15 min.
Preferably, the aperture of the punched hole is 3mm, and the depth is 2 mm; the period of time is 2 hours.
Preferably, the injection volume of the yarrowia lipolytica suspension and the injection volume of the fusarium sporophore suspension into the wells are both 20 μ L.
Preferably, the temperature of the incubator stored in the incubator is set to be 20-25 ℃, and the humidity is set to be 90%.
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.
The invention has the advantages that:
(1) the yarrowia lipolytica WSL-01 used in the invention can effectively control the postharvest root rot and natural rot of asparagus, thereby reducing the loss caused by postharvest diseases of asparagus.
(2) The yarrowia lipolytica WSL-01 used in the invention has no adverse effect on the content of soluble sugar, vitamin C and chlorophyll which are main quality indexes of the picked asparagus, and has the effect of slowing down the reduction of the quality indexes.
(3) The yarrowia lipolytica WSL-01 used in the invention has no report of application in asparagus postharvest disease control at present, and the yarrowia lipolytica WSL-01 used in the invention can replace a chemical bactericide to control asparagus postharvest diseases, reduce potential harm of the chemical bactericide to human bodies and environment, reduce economic and energy burden generated by physical sterilization and storage, and has remarkable economic and social benefits.
Drawings
FIG. 1 shows the control effect of yarrowia lipolytica WSL-01 with different concentrations on asparagus root rot; wherein: the disease index of asparagus tip part (I) and the disease index of asparagus basal part (II) are shown in the specification; the bar graphs corresponding to different days are CK, A, B, C and D from left to right in sequence;
CK is sterile normal saline treated group, namely a control group; A. b, B,C. D respectively represents different concentrations (unit: cells/mL) of the yarrowia lipolytica WSL-01 bacterial suspension, wherein A: 1X 10 6 ；B：1×10 7 ；C：1×10 8 ；D：1×10 9 (ii) a The different lower case letters represent significant differences (P)<0.05)。
FIG. 2 shows the control effect of yarrowia lipolytica WSL-01 on the natural decay of asparagus; wherein: CK is sterile normal saline treated group, namely a control group; y is 1X 10 7 cells/mL yeast suspension treatment group; the different lower case letters represent significant differences (P)<0.05)。
FIG. 3 is a graph showing the effect of yarrowia lipolytica WSL-01 on soluble sugar content in asparagus; wherein: the content of soluble sugar at the tip part of the asparagus (I) and the content of soluble sugar at the base part of the asparagus (II) are shown in the specification; CK is sterile normal saline treated group, namely a control group; y is 1X 10 7 cells/mL yeast suspension treatment group; the different lower case letters represent significant differences (P)<0.05)。
FIG. 4 is a graph showing the effect of yarrowia lipolytica WSL-01 on vitamin C content in asparagus; wherein: the vitamin C content of asparagus tips (I) and the vitamin C content of asparagus basal parts (II) are shown in the specification; CK is sterile normal saline treated group, namely a control group; y is 1X 10 7 cells/mL yeast suspension treatment group; the different lower case letters represent significant differences (P)<0.05)。
FIG. 5 is the effect of yarrowia lipolytica WSL-01 on the chlorophyll content of asparagus; wherein: the chlorophyll content of the asparagus tips (I) and the chlorophyll content of the asparagus basal parts (II) are shown in the specification; CK is sterile normal saline treated group, namely a control group; y is 1X 10 7 cells/mL yeast suspension treatment group; the different lower case letters represent significant differences (P)<0.05)。
Detailed Description
The invention is 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.
The culture program of yarrowia lipolytica WSL-01 is as follows: (1) solid activation: inoculating the yeast strain to NYDA culture medium, and culturing at 28 deg.C for 48 hr; (2) liquid culture: 50mL of NYDB seed culture medium is filled into a 250mL triangular flask, a loop is inoculated with a loop activated yeast, and the yeast is cultured for 24h at the temperature of 28 ℃ and the speed of 180 rpm; (3) centrifugal separation and resuspension: the yeast culture mixture was centrifuged for 15min at 7000g and washed 2 times with sterile physiological saline to remove the culture medium, and finally resuspended with sterile physiological saline and finally adjusted to the desired yeast cell concentration.
Pathogenic bacteria, Fusarium proliferatum (Fusarium proliferatum) used in the invention is obtained by self-screening in a laboratory, culturing for 7 days at 25 ℃ on a PDA culture medium, scraping hyphae into sterile physiological saline, filtering with sterile eight-layer gauze to obtain spore suspension, and finally adjusting to 5 × 10 4 spores/mL。
Example 1:
screening the concentration of yarrowia lipolytica WSL-01 for preventing and treating the postharvest diseases of asparagus;
firstly, a test scheme;
the asparagus to be tested has uniform thickness and consistent color, and has no mechanical damage and pest damage on the surface. Washing the soil on the surface of asparagus with clear water, then soaking the asparagus in 3% sodium hypochlorite solution for disinfection for 5 minutes, washing the asparagus with sterile water, and airing the asparagus at room temperature. Respectively punching a wound with the aperture of 3mm and the depth of 2mm on the tip part and the base part of the asparagus; the following treatment solutions were added to different wounds at 20 μ L: (1)1 x 10 6 cells/mL of yarrowia lipolytica WSL-01 suspension; (2)1 x 10 7 cells/mL of yarrowia lipolytica WSL-01 suspension; (3) 1X 10 8 cells/mL of yarrowia lipolytica WSL-01 suspension; (4) 1X 10 9 cells/mL of yarrowia lipolytica WSL-01 suspension; (5) sterile normal saline;
after standing for 2h, each wound was replated with 20. mu.L of 5X 10 4 spores/mL of a pathogen spore suspension; naturally drying, sealing the asparagus in a plastic basket by using a preservative film, and storing in a constant temperature and humidity incubator (the humidity is 90%) at 20 ℃; each treatment was repeated three times, 16 asparagus per time. The whole experiment was repeated 2 times.
The disease index is calculated as follows:
disease index ═ Σ (number of disease plants at each stage × relative stage number representative value)/(total number of plants × highest stage number representative value)
Secondly, testing results;
FIG. 1 shows the control effect of yarrowia lipolytica WSL-01 with different concentrations on asparagus root rot; wherein: the disease index of asparagus tip part (I) and the disease index of asparagus basal part (II) are shown in the specification; the bar graphs corresponding to different days are CK, A, B, C and D from left to right in sequence; CK is sterile normal saline treated group, namely a control group; A. b, C, D represent different concentrations of yarrowia lipolytica WSL-01 bacterial suspension (unit: cells/mL), wherein A: 1X 10 6 ；B：1×10 7 ；C：1×10 8 ；D：1×10 9 ；
As can be seen from FIG. 1 (I), when stored in storage spaces of 4d and 5d, the storage space is 1X 10 6 cells/mL、1×10 7 cells/mL and 1X 10 9 The disease index of asparagus tips treated by cells/mL yeast suspension is obviously lower than that of a control group and 1 × 10 8 cells/mL Yeast suspension treatment group (p)<0.05); and as can be seen from the diagram (II), 1X 10 7 The disease index of asparagus basal treated by yeast suspension of cells/mL is always significantly lower than that of other treatment groups (p)<0.05). Thus, 1X 10 7 The cells/mL yeast suspension has a remarkable control effect on the root rot of the asparagus after picking, so that the concentration can be used for subsequent experiments.
Example 2:
the yarrowia lipolytica WSL-01 has the control effect on the natural decay of the picked asparagus;
firstly, a test scheme;
the asparagus to be tested has uniform thickness and consistent color, and has no mechanical damage and pest damage on the surface. Uniformly spraying 1 × 10 spray on the surface of uncleaned and sterilized asparagus 7 cells/mL of yarrowia lipolytica WSL-01 suspension, and asparagus sprayed with sterile physiological saline as a control group. Naturally drying, sealing the asparagus in a plastic basket by using a preservative film, and storing in a constant temperature and humidity incubator (the humidity is 90%) at 20 ℃. Each treatment was repeated three times, 16 asparagus per time. The whole experiment was repeated 2 times.
The disease index is calculated as follows:
disease index ═ Σ (number of disease plants at each stage × relative stage number representative value)/(total number of plants × highest stage number representative value)
Secondly, testing results;
FIG. 2 shows the control effect of yarrowia lipolytica WSL-01 on the natural decay of asparagus; as shown in figure 2, the disease index of asparagus treated by yarrowia lipolytica WSL-01 is always remarkable (p is less than 0.05) and is lower than that of asparagus treated by sterile normal saline, thereby showing that the yarrowia lipolytica WSL-01 has a certain control effect on decay of the picked asparagus.
Example 3:
influence of yarrowia lipolytica WSL-01 on the content of soluble sugar, vitamin C and chlorophyll after the asparagus is picked;
firstly, a test scheme;
the asparagus to be tested has uniform thickness and consistent color, and has no mechanical damage and pest damage on the surface. Respectively and uniformly spraying the following treatment solutions on the surfaces of unwashed and disinfected asparagus: (1) 1X 10 7 cells/mL of yarrowia lipolytica yeast suspension; (2) the control group was sterile saline. Naturally drying, sealing the treated asparagus in a plastic basket by using a preservative film, and storing in a constant temperature and humidity incubator (humidity is 90%) at 20 ℃. Each treatment was repeated three times, 16 asparagus per time. The whole experiment was repeated 2 times.
The specific measurement method is as follows:
1. soluble sugar: weighing 2g of asparagus tissue by adopting an anthrone colorimetric method, shearing, placing in a 25mL graduated test tube, adding 10mL of distilled water, sealing by using a plastic film, extracting in a boiling water bath for 30min, and fixing the volume of an extracting solution to 100 mL. And (3) sucking 1mL of the supernatant into a test tube, adding 1mL of distilled water, 0.5mL of anthrone ethyl acetate and 5mL of concentrated sulfuric acid, fully shaking and uniformly mixing, immediately placing into a boiling water bath for heat preservation for 1min, taking out, naturally cooling to room temperature, taking a blank as a reference, and measuring the absorbance at the wavelength of 630 nm.
The soluble sugar content was calculated as follows:
where C is the sugar content (μ g) found from the standard curve, V is the extract volume (ml), a is the aspirated sample volume (ml), N is the dilution factor, and W is the sample weight (g).
2. Vitamin C: 2, 6-dichloro indophenol sodium method, taking 5g of green asparagus tissue sample, placing the green asparagus tissue sample in a mortar, adding a small amount of 2% oxalic acid solution, grinding the green asparagus tissue sample in ice bath and light-proof conditions, transferring the green asparagus tissue sample into a 50mL centrifuge tube after being ground into slurry, washing the mortar with the oxalic acid solution, pouring the green asparagus tissue sample into the centrifuge tube, centrifuging the green asparagus tissue sample for 15min at the temperature of 4 ℃ and 8000g, sucking supernatant into a 100mL volumetric flask, fixing the volume to the scale, shaking the supernatant evenly, and filtering the solution for later use (0.45 mu m filter membrane). 10mL of the filtrate was placed in a 100mL Erlenmeyer flask and the filtrate was titrated with a calibrated 2, 6-dichloroindophenol solution until a reddish color appeared and no discoloration occurred for 15 seconds. The amount was recorded while blanketing with 10mL of 2% oxalic acid solution, and titration was performed in the same manner and repeated three times. The content of ascorbic acid in green asparagus was calculated from the amount of 2, 6-dichloroindophenol, expressed as the mass of ascorbic acid contained per 100g of sample (fresh weight), i.e. mg/100 g.
The ascorbic acid content was calculated as follows:
wherein V is the total volume (ml) of the sample extract, V1 is the dye volume (ml) consumed by the titration of the sample, V2 is the dye volume (ml) consumed by the blank titration, V3 is the volume (ml) of the sample solution taken at the time of titration, P is the mass of 1ml of the dye solution corresponding to ascorbic acid in mg/ml, and M is the sample mass (g).
3. Chlorophyll: weighing 3g of cut fresh sample, adding three parts of the cut fresh sample into a mortar respectively, adding a small amount of quartz sand, calcium carbonate and 2-3ml of 95% ethanol, grinding into homogenate, adding 10ml of 95% ethanol, continuously grinding until the tissue turns white, and standing for 3-5 minutes; taking one piece of filter paper, putting the filter paper in a funnel, wetting the filter paper with ethanol, pouring the extracting solution into the funnel along a glass rod, filtering the extracting solution into a 25ml brown volumetric flask, washing a mortar with a small amount of 95% ethanol, pouring the mortar rod and residues for a plurality of times, and finally pouring the mortar rod and the residues into the funnel together; sucking ethanol by a dropper, washing all chloroplast pigments on the filter paper into a volumetric flask until no green color exists in the filter paper and residues, finally fixing the volume to 25ml by using ethanol, and shaking up; pouring the chloroplast pigment extract into a cuvette, and measuring the light absorption value at 665nm, 649nm and 470nm by using 95% ethanol as a blank.
The chlorophyll content was calculated as follows:
Ca＝13.95A 665 -6.88A 649
Cb＝24.96A 649 -7.32A 665
wherein C is pigment content (mg/L); v is volume of extractive solution (ml), N is dilution multiple, m is sample mass (g), and 1000 is 1L-1000 ml
II, testing results;
according to the test of the steps, the statistical test results are as follows:
FIG. 3 is the effect of yarrowia lipolytica WSL-01 on the soluble sugar content of asparagus; as can be seen from FIG. 3, the soluble sugar content in the tip and base of both treated asparagus was reduced after the harvested green asparagus was stored at 20 ℃ for 5 days. As shown in FIG. 3 (I), the soluble sugar content in the tip of asparagus treated with yarrowia lipolytica WSL-01 was significantly higher than that in the control group (p <0.05) after 5d storage; as can be seen from the graph (II), the soluble sugar content of the base of the yeast-treated asparagus is not significantly different from that of the control group; the result shows that the yarrowia lipolytica WSL-01 has no adverse effect on the content of the soluble sugar in the picked asparagus and can slow down the reduction of the content of the soluble sugar in the picked asparagus tips.
FIG. 4 is the effect of yarrowia lipolytica WSL-01 on the vitamin C content of asparagus; as can be seen from FIG. 4, after the harvested green asparagus were stored at 20 ℃ for 5 days, the vitamin C content in the tips and bases of both the treated asparagus was also reduced. As can be seen from FIG. 4 (I), the vitamin C content in the tip of asparagus treated with yarrowia lipolytica WSL-01 was not significantly different from that in the control group when stored for 5 d; as shown in the graph (II), the vitamin C content of the base of the yeast-treated asparagus was significantly higher than that of the control group (p < 0.05). This indicates that yarrowia lipolytica has no adverse effect on the vitamin C content in the tips of harvested asparagus and can slow down the decrease of the vitamin C content in the basal parts.
FIG. 5 is the effect of yarrowia lipolytica WSL-01 on the chlorophyll content of asparagus; wherein (I) is the influence of yarrowia lipolytica WSL-01 on the chlorophyll content of asparagus tips; (II) influence of yarrowia lipolytica WSL-01 on chlorophyll content of asparagus basal part; as can be seen in FIG. 5, after the harvested green asparagus were stored at 20 ℃ for 5 days, the chlorophyll content in the tips of both the treated asparagus was reduced, and the chlorophyll content in the bases of the asparagus of the control group was also reduced, but the chlorophyll content in the bases of the asparagus of the antagonistic yeast treated was not significantly changed. After 5 days of storage, the chlorophyll content of the tip and the basal part of the asparagus treated by the antagonistic yeast is obviously higher than that of the control group. Therefore, the antagonistic yeast treatment has no adverse effect on the chlorophyll content of the asparagus tips and basal parts, but can remarkably reduce the reduction of the chlorophyll content.
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 (6)
1. The application of yarrowia lipolytica in disease control and storage and preservation of asparagus after picking is characterized in that yarrowia lipolytica (S) (A)Yarrowia lipolytica) The WSL-01 is used for controlling the root rot and natural rot of the picked asparagus and is used for storing and preserving the asparagus.
2. The application of yarrowia lipolytica for postharvest disease control and storage and freshness preservation of asparagus according to claim 1 is characterized by comprising the following specific steps:
firstly, activating yarrowia lipolytica, and then culturing yarrowia lipolytica by using NYDB culture mediumCentrifuging to obtain thallus, washing with sterile physiological saline, centrifuging, repeating the washing and centrifuging steps for several times, and diluting the thallus with sterile physiological saline to 1 × 10 7 cell/mL of bacterial suspension; directly and uniformly spraying the bacterial suspension on the surface of the asparagus, and naturally drying; then the mixture is put into a plastic basket, sealed by a preservative film and stored under the condition of room temperature or in an incubator.
3. The application of yarrowia lipolytica in the postharvest disease control and storage and preservation of asparagus as claimed in claim 2, wherein the condition of yarrowia lipolytica cultured in NYDB medium is: the temperature is 28-30 ℃, the rotating speed is 180-200 rpm, and the time is 24-28 h.
4. The application of yarrowia lipolytica in preventing and treating postharvest diseases of asparagus as well as storing and refreshing the asparagus as claimed in claim 2, wherein the centrifugation conditions are as follows: the rotating speed is 7000g, and the time is 10-15 min.
5. The application of yarrowia lipolytica in asparagus postharvest disease control and storage fresh-keeping according to claim 2 is characterized in that the temperature of the incubator is set to be 20-25 ℃ and the humidity is set to be 90%.
6. The use of yarrowia lipolytica for postharvest disease control and storage and freshness preservation of asparagus as claimed in claim 2, wherein said NYDB 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.
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