CN112342147A - Pichia guilliermondii strain capable of antagonizing various pathogenic bacteria and application thereof - Google Patents
Pichia guilliermondii strain capable of antagonizing various pathogenic bacteria and application thereof Download PDFInfo
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- CN112342147A CN112342147A CN202011264508.4A CN202011264508A CN112342147A CN 112342147 A CN112342147 A CN 112342147A CN 202011264508 A CN202011264508 A CN 202011264508A CN 112342147 A CN112342147 A CN 112342147A
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention provides a Pichia guilliermondii strain Meyerozyma guilliermondii lut-Y1 with the preservation number of CGMCC NO.20461, which can antagonize various pathogenic bacteria. The strain can antagonize Alternaria alternata (Alternaria alternata), Alternaria tenuissima (Alternaria tenuissima), Ascochyta leprospora and Deschltzilla (Drechslera spp.), and has wide application value in the aspects of preventing and treating mildew and rot of fruits and vegetables and plant diseases caused by the mildew.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a Pichia guilliermondii strain (Meyerozyma guilliermondii) lut-Y1 capable of antagonizing various pathogenic bacteria, and application thereof in inhibiting growth of plant pathogenic bacteria.
Background
Fruit and vegetable spoilage and plant diseases are two important factors causing yield reduction and threatening food safety, and both are caused by parasitism of pathogenic microorganisms such as mold, bacteria and the like. Pathogenic microorganisms parasitize on fruits and vegetables, which not only change the appearance, smell and color of the fruits and vegetables to cause putrefaction, but also secrete toxins to poison people eating the fruits and vegetables by mistake. The plant infection pathogenic microorganism can cause various plant diseases, such as plant dwarfing, generation of lesion leaves, yield reduction and the like. Therefore, the development of safe and highly effective biocontrol agents for inhibiting spoilage and plant disease using microbial antagonism is one of the most effective ways to solve the problem.
Pichia guilliermondii is a widely available yeast that can be isolated in the course of fermentation of coastal, deep sea, marine algae surfaces, high salt soy sauce and acidic juice beverages. The pichia mondii strains in different seasons have different characteristics, and some strains are used for brewing fruit wine to play a role in increasing the flavor; some can inhibit plant pathogenic bacteria or putrefying bacteria, can be made into biocontrol preparation for keeping loquat, strawberry, pear and other fruits fresh and inhibiting strawberry diseases, and some strains are used for producing xylitol by fermentation.
Alternaria alternata (Alternaria alternata) is one of Alternaria of the order of Trichosporon of the class of mitosporitic spore fungi, is also a main crop pathogenic bacterium, can cause tobacco brown spot, is generated in Shandong, Henan, Anhui, Jilin, Yunnan, Guizhou, Zhejiang and the like in China, and brings serious economic loss to tobacco production. Alternaria alternata is also a fungus causing decay of picked fruits such as pears, hami melons, jujubes and the like. The infection of fruit can cause the epidermis to appear brown and concave round spots, the flesh under the spots is in a black sponge shape, and the commodity is reduced. At present, chemical control is mainly used for preventing and treating alternaria fungal diseases, and the problems of drug resistance, environmental safety and the like caused by the chemical control are increasingly serious.
Alternaria tenuissima (Alternaria tenuissima) is also a main plant pathogenic bacterium of Alternaria, and can cause leaf spot of water wax (Ligustrum obtusifolia Sieb.) and black spot of Fritillaria thunbergii. Black spot is one of important diseases of fritillary, mainly harms leaves, the thunberg region usually begins to get ill in the late 3 th of the month, yellow brown water stain-like disease spots appear at the top of the leaves in the early stage of onset, the diseased part and the healthy part have obvious limits, the disease spots spread to the base part until the overground part of the plant dies early, the underground bulb is thin and small, the serious yield reduction is caused, and especially the disease is more serious in rainy years; meanwhile, the black spot easily overwinter on the damaged plants and the diseased leaves with hypha and conidia, and the damage is infected again in the next year. Therefore, the prevention and control of fritillaria black spot disease is the key point for guaranteeing the development of fritillaria industry.
Ascochyta leptospora (Ascochyta leptospora) is a fungus of Deuteromycotina, Ascochyta and Ascochyta, which can cause leaf spot of peanut and longan. The ascochyta microsporum can cause ryegrass wilt, and brings huge loss to production management of the lawn.
The genus Dermatopteria (Drechslera spp.) is a pathogenic microorganism that causes leaf spotting, leaf blight, root rot and stem rot of a variety of turf grasses. Under the appropriate environmental conditions, the disease condition is rapidly developed, and the lawn can be premature senility and bald spots, which seriously harm the lawn landscape.
Disclosure of Invention
The invention aims to provide a Pichia guilliermondii strain (Meyerozyma guilliermondii) lut-Y1 capable of inhibiting the growth of various pathogenic bacteria, which is preserved in China general microbiological culture Collection center (GCMCC) at 7-28 months in 2020 at the preservation address of: the invention also provides application of the compound in inhibiting various phytopathogens, wherein the number of the compound is CGMCC No. 1, and the number of the compound is CGMCC No. 3.
The invention provides a Pichia guilliermondii strain Meyerozyma guilliermondii lut-Y1 with the preservation number of CGMCCNO.20461, which can antagonize various pathogenic bacteria.
The invention also provides a preparation method of the pichia guilliermondii lut-Y1 fermentation product, which comprises the steps of inoculating the pichia guilliermondii lut-Y1 into a potato glucose liquid culture medium for culture to obtain a fermentation liquid, centrifuging the fermentation liquid, and taking a supernatant to obtain the pichia guilliermondii lut-Y1 fermentation product.
Preferably, the culture conditions are: at 25-30 ℃, 120--1Shaking and culturing for 18-35 h. When the resulting fermentation broth is centrifuged, the parameters conventional in the art may be selected so long as the majority of the fermentation product is present in the centrifuged supernatant, e.g., 4000 r.min-1And centrifuging for 10 min.
The invention also provides a biological control preparation, and the active ingredient of the biological control preparation is the fermentation product of the pichia guilliermondii lut-Y1 prepared by the method.
The invention also provides application of the thallus, the sprout or the biological control preparation of the pichia guilliermondii lut-Y1 strain in preparing products for inhibiting the mildew rot pathogen.
Preferably, the product comprises a fruit and vegetable fresh-keeping preservative, a plant disease control preparation and a medicament.
Preferably, the mould pathogenic fungus is alternaria, alternaria tenuis, ascochyta and/or a mould of the genus drechslera.
The Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 is separated from fruit mold rotten spots of Lycium barbarum and is determined to be the Pichia guilliermondii (Meyerozyma guilliermondii) by ITS region sequence sequencing analysis and morphological identification. The microbiological properties of the yeast strain are: on a yeast extract peptone glucose agar medium (YPDA), a bacterial colony is pink, round, neat in edge, large and thick, and bulged in the middle, is opaque, smooth and moist, and is easy to pick up; white, round, smooth and moist colonies were formed on potato dextrose agar medium (PDA) and wort agar Medium (MEA). The strain can tolerate NaCl with the mass percent of less than 3%, and when the concentration of NaCl is more than 3%, the growth of the yeast is obviously inhibited. The characteristics of the growth curve of the strain are as follows: 0-6h is latent period, 6-18h is exponential growth period, 18-35h is stable growth period, and the decline begins after 36 h.
The Pichia guilliermondii strain (Meyerozyma guilliermondii) lut-Y1 of the present invention is capable of antagonizing Alternaria (Alternaria alternata), Alternaria tenuissima (Alternaria tenuissima), Chaetomium microsporum (Ascocchyta leptospora) and a mold of the genus Derma (Drechslera spp.); the fermentation liquor can be used as a biological control preparation to obviously inhibit the growth of the 4 moulds, and has wide application value in the aspects of preventing and treating mildew and rot of fruits and vegetables and plant diseases caused by the moulds.
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 shows the colony morphology and cell morphology of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1.
FIG. 2 shows the comparison of rDNA-ITS sequence of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 with Gen Bank nucleic acid database.
FIG. 3 shows the colony morphology of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 on different media.
FIG. 4 shows the salt tolerance of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1.
FIG. 5 is a graph showing the growth curves of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 in YPDA medium.
FIG. 6 is a graph showing the antagonistic effect of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 against 4 pathogenic bacteria.
FIG. 7 is a graph showing the inhibitory effect of a Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 biocontrol agent on Alternaria alternata (Alternaria alternata).
FIG. 8 is a graph showing the inhibitory effect of a Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 biocontrol agent against a mold of the genus Dermatopteria (Drechslera spp.).
FIG. 9 shows the inhibitory effect of a Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 biocontrol agent against Chaetomium microsporus (Ascochyta leptospora) mold.
FIG. 10 is a graph showing the inhibitory effect of a Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 biocontrol agent on Alternaria tenuissima (Alternaria tenuissima).
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from conventional biochemicals, unless otherwise specified.
The preparation method of the culture medium comprises the following steps:
potato dextrose agar medium (PDA): weighing 200g, and cutting into 1cm3Boiling potato blocks with different sizes in boiling water for 20min, filtering with 4 layers of gauze, adding 20g of glucose and 15-20g of agar into the filtrate, adding water to 1L, adjusting pH to natural, and sterilizing to obtain PDA solid culture medium; the PDA liquid culture medium is prepared by the same method as the PDA solid culture medium except that agar powder is not added.
Yeast extract peptone glucose agar medium (YPDA): 20g of peptone, 20g of glucose, 10g of yeast extract powder and 15-20g of agar powder, adding water to 1L, naturally adjusting pH, and sterilizing. The YPDA liquid culture medium is prepared by the same method as YPDA solid culture medium except that no agar powder is added.
The formula of the wort agar culture Medium (MEA) is as follows: 130g of wort culture medium and 15-20g of agar powder, adding water to 1L, naturally adjusting the pH value, and sterilizing.
The strain source is as follows: alternaria alternata (Alternaria alternata) is separated from fresh fruit mildew rot spots of medlar produced by medlar production cooperative society of Yongden county, Lanzhou, Gansu province and stored in the laboratory, and the specific separation and identification process is shown in the literature (Yuanjun, Li Hujun, Jiahong Sha, and the like. separation and identification of mildew rot pathogenic fungi in the airing process of the fresh fruits of Yongden medlar. food industry science and technology 2016,37(21): 135. 138.).
Dermatopteria (Drechslera spp.) mold, Ascochyta leptospora (Ascochyta leptospora) and Alternaria tenuisiana (Alternaria tenuissima) were given by the plant pathology laboratory of the Soft. agri, Lanzhou university, and the specific isolation and identification process was described in the literature (Mashi, perennial ryegrass-endophytic fungi symbiont disease resistance and its research on the disease resistance mechanism of Helminthosporium rothii (Bipolaris sorokiniana) [ D ].2015 ]
Example 1
The Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 of the invention is isolated from the fresh fruit mold rot spot of Lycium barbarum produced by Gongdeng county Lycium production Cooperia, Lanzhou, Gansu. The separation process is as follows: the fermented fruit of Chinese wolfberry is collected from the co-production bed of Chinese wolfberry of Chuanzhen county of Yongdeng county of Lanzhou, Gansu province in 2014 8 months, is transported to a laboratory of the university for separation and purification on the same day, adopts a tissue separation method, selects and dries 3d diseased fruits typical of disease spots to be washed clean, cuts off a small tissue with the size of 5mm multiplied by 5mm at the boundary of disease keys, uses 2% sodium hypochlorite for surface disinfection for 8min, uses sterile water for 5 times, uses 75% alcohol for disinfection for 30s, uses sterile water for 8 times, moves to a sterilization culture dish, adds 1mL of sterile water for mashing, uses an inoculating loop to dip a tissue fluid containing bacteria on a PDA flat plate for scribing, cultures at 28 ℃ and picks up a single colony, and a pure culture after continuous purification for 3 times is stored on a slant PDA culture medium and is stored in a refrigerator at 4 ℃ for later use; representative strains were placed on different media, respectively. The yeast is cultured in malt extract agar medium at 28 deg.C for 3d, and the colony culture characteristics are observed and recorded. Selecting yeast, making into water-filled tablet, and observing shape under common optical microscope; the yeast is picked up, stuck on a sample table, placed in liquid nitrogen for quick freezing, and then observed under a scanning electron microscope for morphological characteristics such as cells and the like (figure 1). The test strains were inoculated to PDA medium and cultured at 25 ℃ for 3-5 days, and then sent to Dalibao bioengineering GmbH for strain genomic DNA extraction, PCR amplification and ITS region sequence sequencing (FIG. 2), and the obtained sequences were compared with GenBank nucleic acid database (www.ncbi.nlm.nih.gov/BLAST) (FIG. 3). Finally, the strain is determined to be Pichia guilliermondii (Meyerozyma guilliermondii) according to the sequence sequencing analysis and morphological identification of the ITS region. The strain is preserved in China general microbiological culture Collection center (GCMCC) at 7 months and 28 days in 2020, with the preservation addresses as follows: no. 3 of Xilu No. 1 of Beijing, Chaoyang, and the preservation number is CGMCC NO. 20461.
Example 2
The specific experimental process of the antibacterial performance of the Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 strain CGMCC NO.20461 comprises the following steps:
(1) activating strains: subpackaging the sterilized PDA solid culture medium in 50mL test tubes, pouring 15mL of the PDA solid culture medium in each test tube, obliquely placing the test tubes on a table top to prepare a slant culture medium, and cooling and solidifying the culture medium; pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 strain CGMCC NO.20461 was streaked on a slant medium with an inoculating needle and cultured at room temperature for 2-3 days. The preparation of the bacterial suspension comprises the following steps: 4mL of sterile water is added into the slant strains respectively, and the mixture is fully oscillated to prepare bacterial suspension.
(2) And (4) observing colony morphology: pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 were streaked onto 3 solid medium plates and observed for colony morphology after 2 days of inverted culture at 30 ℃. The 3 solid culture media are respectively: yeast extract peptone glucose agar medium (YPDA), malt extract agar Medium (MEA) and potato glucose agar medium (PDA).
(3) Determination of salt resistance: mu.l of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 bacterial suspension was inoculated into 50ml of YPDA-NaCl liquid medium containing 0%, 0.5%, 1%, 3%, 5% by mass, and subjected to shaking culture at 30 ℃ and 120r/min for 24 hours, and then the absorbance of the culture was measured at a wavelength of 660nm using a spectrophotometer. Each treatment was set to 5 replicates.
(4) And (3) measuring a growth curve: 50 μ l of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 bacterial suspension was added to 50ml of YPDA liquid medium, shaking-cultured at 30 ℃ for 24 hours at 160r/min in a shaker, and the absorbance of the culture solution was measured every 2 hours at a wavelength of 660nm by a spectrophotometer, and the growth curve was plotted with absorbance as the vertical axis and time as the horizontal axis. Each treatment was set to 5 replicates.
(5) Antagonism: adopting a flat plate opposing method. Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 and the test strains (Alternaria alternata, a mold of the genus Deschleria (Drechslera spp.), Chaetomium microsporus (Ascochyta leptospora) and Alternaria tenuissima) were inoculated on the same PDA plate, respectively, and the colony growth was observed.
(6) Preparation of the biological control preparation: inoculating 500 μ L of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 strain suspension into PDA liquid culture medium, and culturing at 30 deg.C for 160 r.min-1After shaking culture for 18h, the fermentation liquor is cultured at 4000 r.min-1Centrifuging for 10min to obtain supernatant as biological control preparation with OD value of 1.6-2.0.
(7) Preparing a fungus cake for inoculation: alternaria alternata (Alternaria alternate), Alternaria tenuissima (Alternaria tenuissima), Ascochyta leptospora, a mold of the genus Deschschltzilla (Drechslera spp.) were prepared as test bacteria. Respectively adding 4mL of sterile water into slant strains of the bacteria to be tested, fully oscillating, preparing bacterial suspension, coating 1mL of the bacterial suspension in a PDA solid culture medium flat plate, culturing at 24-28 ℃ for 5-7 days until the strains grow to fill the flat plate, and taking a bacteria cake to be tested with the diameter of 1cm from the center of the flat plate by using a sterile puncher.
(8) The biological control preparation has the following inhibition effect on bacteria to be tested: a biological control preparation with gradient concentration, namely PDA solid culture medium, is prepared according to the proportion of table 1, sterilized and then subpackaged in culture dishes with the diameter of 9cm, and each dish is 25 mL. Sticking the bacterial cake to be tested to the center of a biological control preparation-PDA solid culture medium flat plate, repeating each strain for 3 times, placing culture dishes inoculated with the bacterial and the fungal cake to a constant-temperature incubator at 30 ℃ for 5-7d respectively, observing the colony morphology, measuring the diameter of a growth ring of the bacteria to be tested, and performing single-factor variance analysis on the diameter change of the growth ring of the bacteria to be tested by using SPSS statics 21 statistical software.
TABLE 1 formulation ratio of PDA culture medium as biological control agent with gradient concentration
The experimental results are as follows:
(1) the Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 of the present invention was cultured on wort agar medium for 3d to form colonies with a diameter of 0.5-1 cm. The colony is pink, round, irregular in edge wrinkles, convex in surface, smooth and opaque (fig. 1). The cell shape is oval, and has the shape and size characteristics of yeast. The rDNA-ITS region primer is used for PCR amplification to obtain the rDNA-ITS sequence of 552bp of the strain, and BLAST comparison is carried out on the rDNA-ITS sequence and the Pichia guilliermondii (Meyerozyma guilliermondii) which is recorded in GenBank, and the sequence similarity reaches 100 percent (figure 2). Based on the above morphological and molecular biological characteristics, the strain was identified as Pichia guilliermondii (Meyerozyma guilliermondii).
The rDNA-ITS sequence of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 is as follows:
FIG. 1 shows the colony morphology and cell morphology of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1.
FIG. 2 shows the comparison of rDNA-ITS sequence of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 with Gen Bank nucleic acid database.
(2) The colony of the Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 on a yeast extract peptone glucose agar medium (YPDA) is pink, round, neat in edge, large and thick, and has a bulged middle part, is opaque, smooth and moist, and is easy to pick up; white, round, smooth and moist colonies were formed on potato dextrose agar medium (PDA) and on wort agar Medium (MEA) (fig. 3).
FIG. 3 shows the colony morphology of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 on different media.
(2) The Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 strain of the present invention was able to tolerate NaCl at a mass percentage of 3% or less, and significantly inhibited the growth of the yeast when the concentration of NaCl was greater than 3% by mass (FIG. 4).
FIG. 4 shows the salt tolerance of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1.
(3) The growth curve of the Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 strain is characterized in that: 0-6h is incubation period, 6-18h is exponential growth period, 18-35h is stationary growth period, and beginning to decline at 36h (fig. 5).
FIG. 5 is a graph showing the growth curves of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 in YPDA medium.
(4) The Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 of the present invention antagonized Alternaria alternata (Alternaria alternata), a mold of the genus Detrichella (Drechslera spp.), Septoria microsporus (Ascochyta leptospora) and Alternaria tenuissima (Alternaria tenuissima) (FIG. 6).
FIG. 6 is a graph showing the antagonistic effect of Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 against 4 pathogenic bacteria.
(5) The biological control preparation of the invention has obvious inhibition effect on the growth of alternaria alternate. Compared with the control, the culture medium is added with biological control preparations with the volume percentages of 20 percent, 30 percent and 40 percent, so that the diameters of Alternaria spores are respectively reduced by 26 percent, 75 percent and 91 percent (figure 7).
FIG. 7 is a graph showing the inhibitory effect of a Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 biocontrol agent on Alternaria alternata (Alternaria alternata).
(6) The biological control preparation of the invention has obvious inhibition effect on the growth of Deerhella (Drechslera spp.). Compared with the control, the culture medium is added with 30 percent and 40 percent of biological control agent by volume, so that the diameters of the mould colonies of the Dermatophagoides respectively are obviously reduced by 52 percent and 68 percent (figure 8).
FIG. 8 is a graph showing the inhibitory effect of a Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 biocontrol agent against a mold of the genus Dermatopteria (Drechslera spp.).
(7) The biological control preparation of the invention has obvious inhibition effect on the growth of Ascochyta leptospora. Addition of 10%, 20%, 30%, 40% by volume of biocontrol agent to the culture medium resulted in a significant reduction in colony diameter of Ascochyta leptospora (Ascochyta leptospora) of 51%, 90%, 97%, 99%, respectively, compared to the control (fig. 9).
FIG. 9 shows the inhibitory effect of a Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 biocontrol agent against Chaetomium microsporus (Ascochyta leptospora) mold.
(8) The biological control preparation of the invention has obvious inhibition effect on the growth of Alternaria tenuissima. Compared with a control, the culture medium is added with biological control preparations with the volume percentages of 10%, 20%, 30% and 40%, so that the diameters of Alternaria tenuissima colonies can be respectively and remarkably reduced by 90%, 96% and 100% (figure 10).
FIG. 10 is a graph showing the inhibitory effect of a Pichia guilliermondii (Meyerozyma guilliermondii) lut-Y1 biocontrol agent on Alternaria tenuissima (Alternaria tenuissima).
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Sequence listing
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Claims (7)
1. A Pichia guilliermondii strain Meyerozyma guilliermondii lut-Y1 with CGMCC NO.20461 is capable of antagonizing multiple pathogenic bacteria.
2. The method for preparing fermentation product of Pichia guilliermondii lut-Y1 of claim 1, wherein: the pichia guilliermondii lut-Y1 of claim 1 is inoculated in a potato dextrose liquid culture medium for culture to obtain a fermentation liquid, and the fermentation liquid is centrifuged to obtain a supernatant to obtain a pichia guilliermondii lut-Y1 fermentation product.
3. The method of claim 2, wherein: the culture conditions are as follows: at 25-30 ℃, 120--1Shaking and culturing for 18-35 h.
4. A biological control formulation characterized by: the active ingredient of the biological control preparation is a fermentation product of Pichia guilliermondii lut-Y1 prepared by the method of claim 2 or 3.
5. The use of a thallus, a sprout, or a biocontrol agent of claim 1 from the strain pichia guilliermondii lut-Y1 for the preparation of a product for inhibiting pathogenic fungi of mold rot.
6. Use according to claim 5, characterized in that: the product comprises a fruit and vegetable fresh-keeping preservative, a plant disease control preparation and a medicament.
7. Use according to claim 5 or 6, characterized in that: the mildew rot pathogen is Alternaria alternata, Alternaria tenuissima, Chaetomium microsporum and/or a mold of Deerhermella.
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