CN107937290A - One plant of Bayer Zygosaccharomyces and its preparation and application method for being used for postharvest disease of fruits and vegetables prevention - Google Patents
One plant of Bayer Zygosaccharomyces and its preparation and application method for being used for postharvest disease of fruits and vegetables prevention Download PDFInfo
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Abstract
The invention discloses one plant of antimicrobial spectrum for being used for postharvest disease of fruits and vegetables prevention is wide, the Bayer Zygosaccharomyces Zygosaccharomyces bailii bacterial strains BY39 of effect stability and its application method and purposes.The bacterial strain is CGMCC No.14912 in the preservation strain numbering of China Committee for Culture Collection of Microorganisms's common micro-organisms center.The application method of the Wine-making yeast：Bacterial strain is activated, with YPD fermented and cultureds, centrifugation, thalline is made into 1 × 10 with sterile water8The bacteria suspension of cell/mL；The fruits and vegetables such as apple, grape, strawberry, citrus or cherry tomato are put into bacteria suspension, immersion is taken out after 30 seconds, is air-dried；It is put into crisper, room temperature storage.The Wine-making yeast strain can control melanose or canker of apple, grape grey mould, aspergillosis and fusarium mildew, grey mould fruit rot of strawberry and aspergillosis at the same time, Penicillium italicum, and cherry tomato gray mold and aspergillosis, loss caused by reducing postharvest disease, has good application prospect.
Description
Technical field
The present invention relates to diseases biological control of postharvest fruits and vegetables field, more particularly to one plant to be used for postharvest disease of fruits and vegetables biology anti-
The Bayer Zygosaccharomyces (Zygosaccharomyces bailii) controlled, the bacterial strain is to apple, grape, strawberry, citrus, cherry tomato
Main postharvest disease be respectively provided with significant prevention effect.
Background technology
Although fresh fruit of vegetables quality deterioration is influenced by factors, disease is most important reason.Wherein, fungoid
It is the factor of most serious in being lost after fruit is adopted to rot caused by disease.Although can by cultural control, physical control,
Many approach such as chemical prevention and biological control carry out prevention postharvest disease of fruits and vegetables, but current major measure is chemical prevention
(Eckert＆Ogawa, 1985,1988).However, long-time service chemical pesticide not only causes pathogen to produce drug resistance and reduce
Bactericidal effect (Prusky et al., 1985；As et al., 1991；Holmes＆Eckert, 1999), and continually
The persticide residue on fruit, serious threat people's health are also increased using the chemical agent of high concentration, and causes environment
Pollute (Gullion＆Kuijipers, 1994).Therefore, the postharvest diseases of fruit control of safe efficient, nontoxic, low resistance is developed
New technology become our times various countries research emphasis (Falik et al., 1995；Tian et al., 2001；
Kulakiotuet al., 2004), wherein being to be proved to safely and effectively new method at present using antagonist prevention
(Wilson＆Wisniewski, 1989；Janisiewicz＆Koersten, 2002).
So far, filtered out both at home and abroad many thin with obvious fungistatic effect to fruit postharvest pathogenic fungi
Bacterium, saccharomycete and small-sized filamentous fungi, wherein being the peace that currently has been demonstrated using antagonistic yeast prevention postharvest diseases of fruit
Entirely, efficient new technology, this mainly due to most of pathogens by wound invade fruit, and saccharomycete mainly by with cause of disease
Bacterium carries out nutrition and competition for space carrys out controlling disease, and antagonistic yeast can adapt to the fruits such as low temperature, hypoxemia, high carbon dioxide
Real postharvest storage condition (Wang Yousheng, 2012).
However, although the antagonistic yeast reported both at home and abroad has nearly hundred kinds at present, the biological and ecological methods to prevent plant disease, pests, and erosion of most of antagonistic yeasts
Effect is only verified on a small number of fruits.And since the biocontrol effect between the different strains of same saccharomycete has very big difference
(Filonowet al., 1996), therefore, most of antagonistic yeasts lack that antimicrobial spectrum is wide, the bacterial strain of effect stability.
The content of the invention
For above-mentioned technical barrier, it is an object of the invention to provide the suppression that one plant is used for diseases biological control of postharvest fruits and vegetables
Bacterium composes wide, the Bayer Zygosaccharomyces Zygosaccharomyces bailii of effect stability and its preparation and application.This kind
It is micro- in China for the Bayer Zygosaccharomyces Zygosaccharomyces bailii BY39 of diseases biological control of postharvest fruits and vegetables
The preservation strain numbering of biological inoculum preservation administration committee common micro-organisms center is CGMCC No.14912.
The Bayer Zygosaccharomyces bacterial strain carry out postharvest disease of fruits and vegetables prevention and preservation and freshness method, according to following steps into
OK：Bayer Zygosaccharomyces is activated, with YPD solution culture fermentation cultures, centrifugation obtains thalline, thalline is prepared with sterile water
It is 1 × 10 into concentration8The bacteria suspension of cell/mL；The fruits and vegetables such as apple, grape, strawberry, citrus and cherry tomato are put into bacteria suspension
In, immersion is taken out after 30 seconds, is air-dried；It is put into crisper, after sealing, is put into room temperature storage.
The preparation method of Bayer Zygosaccharomyces BY39 is to take out bacterial strain from -80 DEG C of refrigerators, is lived with YPDA culture mediums
Change, picking single bacterium is dropped down onto in YPD fluid nutrient mediums, and 24h is cultivated under the conditions of 26 DEG C, 200r/min, and 4000rpm centrifugations 5min is received
Collect thalline, washed 3 times with sterile.The YPDA culture mediums are：Yeast extract 10g, peptone 20g, glucose 20g, agar
18g, deionized water 1000ml, natural pH, 121 DEG C of sterilizing 30min.
The Bayer Zygosaccharomyces bacterial strain can be used to control melanose or canker of apple, grape grey mould, aspergillosis and sickle at the same time
Spore mildew, grey mould fruit rot of strawberry and aspergillosis, Penicillium italicum, and cherry tomato gray mold and aspergillosis.
Bacterial strain provided by the present invention be from the vinifera mash of spontaneous fermentation screening be separated to apple, Portugal
Grape, strawberry, citrus, cherry tomato postharvest disease have the Bayer Zygosaccharomyces BY39 of notable prevention effect, can be widely used for fruits and vegetables
The prevention of postharvest disease, loss caused by reducing postharvest disease, has good application prospect.
Advantages of the present invention：(1) Bayer Zygosaccharomyces BY39 provided by the present invention is this laboratory from vinous fermentation
Screening obtains in mash, its is harmless, safe.(2) Bayer Zygosaccharomyces BY39 antimicrobial spectrums used in the present invention
Extensively, melanose or canker of apple, grape grey mould, aspergillosis and fusarium mildew, grey mould fruit rot of strawberry and aspergillosis, citrus can be controlled at the same time
Penicilliosis, and cherry tomato gray mold and aspergillosis.(3) Bayer Zygosaccharomyces BY39 provided by the present invention is in YPD culture mediums
Middle well-grown, is easy to cultivate, and character is stablized, and a certain concentration bacteria suspension is used alone just can effectively prevent a variety of fruits and vegetables and adopt
Disease afterwards, use cost is low, wide market.(4) Bayer Zygosaccharomyces BY39 provided by the present invention can replace chemistry
Bactericidal agent for preventing and treating fruit postharvest diseases, avoid using harm of the chemical bactericide to people, and free from environmental pollution, society and Ecological Effect
It is beneficial notable.
Pass through the more detailed explanation present invention of following embodiment.Following embodiment is merely illustrative, the present invention
And from the limitation of these embodiments.
Brief description of the drawings
Fig. 1 is the 26S rDNA D1/D2 of Bayer Zygosaccharomyces Zygosaccharomyces bailii BY39 of the present invention
Area's Evolution of Nucleic Acid Sequences graph of a relation.
Fig. 2 is inhibitions of the Bayer Zygosaccharomyces BY39 to melanose or canker of apple.Note：Control：Sterile water, that is, compare
Group；Z.b：1×108The Bayer Zygosaccharomyces BY39 bacteria suspensions of cell/mL.Different letters represent significant difference (P ＜ 0.05).
Fig. 3 is inhibitions of the Bayer Zygosaccharomyces BY39 to grape grey mould, aspergillosis and fusarium mildew.Note：
Control：Sterile water, i.e. control group；Z.b：1×108The Bayer Zygosaccharomyces BY39 bacteria suspensions of cell/mL.Different letter generations
Table significant difference (P ＜ 0.05).
Fig. 4 is inhibitions of the Bayer Zygosaccharomyces BY39 to grey mould fruit rot of strawberry and aspergillosis.Note：Control：It is sterile
Water, i.e. control group；Z.b：1×108The Bayer Zygosaccharomyces bacteria suspension of cell/mL.Different letters represent significant difference (P ＜
0.05)。
Fig. 5 is inhibitions of the Bayer Zygosaccharomyces BY39 to Penicillium italicum.Note：Control：Sterile water, that is, compare
Group；Z.b：1×108The Bayer Zygosaccharomyces bacteria suspension of cell/mL.Different letters represent significant difference (P ＜ 0.05).
Fig. 6 is inhibitions of the Bayer Zygosaccharomyces BY39 to cherry tomato gray mold and aspergillosis.Note：Control：It is sterile
Water, i.e. control group；Z.b：1×108The Bayer Zygosaccharomyces bacteria suspension of cell/mL.Different letters represent significant difference (P ＜
0.05)。
Embodiment
Embodiment 1：The biological characteristics of Bayer Zygosaccharomyces Zygosaccharomyces bailii bacterial strains BY39
1. morphological feature
(1) YPDA culture mediums (yeast extract 1%, peptone 2%, glucose 2%, agar 1.8%, 121 DEG C of sterilizings
20min) upper 26 DEG C of cultures 48h, bacterium colony is rounded, white, and colony edge is smooth mellow and full.Cellular morphology is in spheroid shape.
(2) after cultivating 24h in YPDA fluid nutrient mediums, mould is not formed, bacterium solution is muddy, there is precipitation, microscopy yeast cells
It is oval, budding.
2. molecular biology identification
With general forward primer NL-1 (5 '-GCATATCAATAAGCGGAGGAAAAG-3 ') and reverse primer NL-4 (5 '-
GGTCCGTGTTTCAAGACGG-3 ') PCR amplification yeast 26S rDNA D1/D2 areas nucleotide sequence is carried out, by the survey of PCR product
Sequence result input website www.NCBI.nlm.nih.gov carries out BLAST, downloads homologous sequence from GenBank databases, leads to
MEGA6 software buildings chadogram such as Fig. 1 is crossed, the bacterial strain for determining to screen is Bayer Zygosaccharomyces (Zygosaccharomyces
bailii)。
The Bayer Zygosaccharomyces BY39 of the present invention has been preserved in city of BeiJing, China Chaoyang District North Star West Road 1 institute 3
The China Microbiological bacterial strain preservation committee common micro-organisms center of institute of microbiology of the academy of sciences of state, preservation time are 2017
November 15, deposit number are CGMCC No.14912, it is proposed that Classification And Nomenclature be Bayer Zygosaccharomyces
Zygosaccharomyces bailii。
Inhibitions of the 2 Bayer Zygosaccharomyces BY39 of embodiment to melanose or canker of apple
1. experimental program
Bayer Zygosaccharomyces BY39 is taken out from -80 DEG C of refrigerators, with YPDA culture mediums (yeast extract 10g, peptone 20g,
Glucose 20g, agar 18g, deionized water 1000ml, natural pH, 121 DEG C of sterilizing 30min) activation, picking single bacterium drops down onto YPD liquid
In body culture medium, 24h is cultivated under the conditions of 26 DEG C, 200r/min, 4000rpm centrifugation 5min, abandon supernatant, the thalline nothing of collection
Bacterium water cleans 3 times repeatedly, and blood counting chamber, which counts to be made into, is configured to concentration as 1 × 108The Bayer Zygosaccharomyces BY39 of cell/mL
Bacteria suspension.
Alternaric bacteria (Alternaria tenuissima) is activated on PDA culture medium tablet, at 26 DEG C cultivate 7~
14d, scrapes appropriate spore, and concentration is configured to as 5 × 10 with sterile water4The alternaric bacteria spore suspension of cell/mL.
By health 2% hypochlorite disinfectant 5min of lossless Apple, with deionized water rinsing, dry, use is sterile
Card punch makes a call to 5 holes at fruit equator, and superficial cut is 2mm (diameter) × 2mm (depth).Each wound equivalent adds 20 μ L
Following treatment fluid：(1)1×108The Bayer Zygosaccharomyces BY39 bacteria suspensions of cell/mL；(2) sterile distilled water.20 μ are inoculated with after 4h
L alternaric bacteria spore suspensions.After drying, fruit is put into plastic box, relative humidity 95% is kept, is placed in room temperature (25 DEG C) 4
Fruit incidence is recorded after it, the fungistatic effect of Bayer Zygosaccharomyces BY39 is evaluated with this.The calculation formula of incidence is：Morbidity
The fruit of rate (%)=morbidity/fruit sum × 100%.
2. result of the test
Tested according to above-mentioned steps, counting the incidence of apple, the results are shown in Figure 2, control group melanose or canker of apple incidence
For 100%, the melanose or canker of apple incidence handled by Bayer Zygosaccharomyces BY39 is 80%, therefore Bayer Zygosaccharomyces BY39
Melanose or canker of apple disease can effectively be controlled.
Inhibitions of the 3 Bayer Zygosaccharomyces BY39 of embodiment to grape postharvest disease
1. experimental program
Bayer Zygosaccharomyces BY39 is taken out from -80 DEG C of refrigerators, with YPDA culture mediums (yeast extract 10g, peptone 20g,
Glucose 20g, agar 18g, deionized water 1000ml, natural pH, 121 DEG C of sterilizing 30min) activation, picking single bacterium drops down onto YPD liquid
In body culture medium, 24h is cultivated under the conditions of 26 DEG C, 200r/min, 4000rpm centrifugation 5min, abandon supernatant, the thalline nothing of collection
Bacterium water cleans 3 times repeatedly, and blood counting chamber, which counts to be made into, is configured to concentration as 1 × 108The Bayer Zygosaccharomyces BY39 of cell/mL
Bacteria suspension.
By botrytis cinerea (Botrytis porri), aspergillus echinulatus (Aspergillus aculeatus) or fragrant Fusariumsp
(Fusarium redolens) is activated on PDA culture medium tablet, and 7~14d is cultivated at 26 DEG C, scrapes appropriate spore, use is sterile
Water is configured to concentration as 5 × 104Botrytis cinerea, aspergillus echinulatus or the fragrant Fusariumsp spore suspension of cell/mL.
By health 2% hypochlorite disinfectant 5min of lossless pear fruit, with deionized water rinsing, dry, beaten with sterile
Hole device makes a call to 1 hole at fruit equator, and superficial cut is 2mm (diameter) × 2mm (depth).Each wound equivalent add 20 μ L with
Lower treatment fluid：(1)1×108The Bayer Zygosaccharomyces BY39 bacteria suspensions of cell/mL；(2) sterile distilled water.20 μ L are inoculated with after 4h
Botrytis cinerea, aspergillus echinulatus or fragrant Fusariumsp spore suspension.After drying, fruit is put into plastic box, keeps relative humidity
95%, it is placed in room temperature (25 DEG C) and fruit incidence is recorded after 4 days, the fungistatic effect of Bayer Zygosaccharomyces BY39 is evaluated with this.Hair
The calculation formula of sick rate is：The fruit of incidence (%)=morbidity/fruit sum × 100%.
2. result of the test
Tested according to above-mentioned steps, the incidence result for counting grape fruit is as follows：
(1) inhibitions of the Bayer Zygosaccharomyces BY39 to grape fruit gray mold
As shown in figure 3, control group grape fruit gray mold incidence is 100%, by Bayer Zygosaccharomyces BY39 processing
Grape fruit gray mold incidence be 0, therefore Bayer Zygosaccharomyces BY39 can effectively control grape fruit gray mold disease.
(2) inhibitions of the Bayer Zygosaccharomyces BY39 to grape fruit aspergillosis
As shown in figure 3, the incidence of control group grape fruit aspergillosis is 100%, at Bayer Zygosaccharomyces BY39
The grape fruit aspergillosis incidence of reason is 46%, therefore Bayer Zygosaccharomyces BY39 can effectively control the grape fruit as caused by
Real aspergillosis disease.
(3) inhibitions of the Bayer Zygosaccharomyces BY39 to grape fruit fusarium mildew
As shown in figure 3, the incidence of control group grape fusarium mildew is 100%, by Bayer Zygosaccharomyces BY39 processing
Grape fusarium mildew incidence be 80%, therefore Bayer Zygosaccharomyces BY39 can effectively control grape fusarium mildew disease.
Inhibitions of the 4 Bayer Zygosaccharomyces BY39 of embodiment to strawberry fruit gray mold and aspergillosis
1. experimental program
Bayer Zygosaccharomyces BY39 is taken out from -80 DEG C of refrigerators, with YPDA culture mediums (yeast extract 10g, peptone 20g,
Glucose 20g, agar 18g, deionized water 1000ml, natural pH, 121 DEG C of sterilizing 30min) activation, picking single bacterium drops down onto YPD liquid
In body culture medium, 24h is cultivated under the conditions of 26 DEG C, 200r/min, 4000rpm centrifugation 5min, abandon supernatant, the thalline nothing of collection
Bacterium water cleans 3 times repeatedly, and blood counting chamber, which counts to be made into, is configured to concentration as 1 × 108The Bayer Zygosaccharomyces BY39 of cell/mL
Bacteria suspension.
Ash arrhizus bacteria (Botrytis porri) or aspergillus echinulatus (Aspergillus aculeatus) are cultivated in PDA
Activated on base tablet, 7~14d is cultivated at 26 DEG C, scrapes appropriate spore, concentration is configured to as 5 × 10 with sterile water4Cell/mL
Ash arrhizus bacteria or aspergillus echinulatus spore suspension.By health 2% hypochlorite disinfectant 5min of lossless strawberry fruit, spend
Ionized water rinses, and dries, makes a call to 1 hole at fruit equator with aseptic card punch, superficial cut is 2mm (diameter) × 2mm (depth).
Each wound equivalent adds the 20 following treatment fluids of μ L：(1)1×108The Bayer Zygosaccharomyces BY39 bacteria suspensions of cell/mL；(2)
Sterile distilled water.20 μ L ash arrhizus bacterias or aspergillus echinulatus spore suspension are inoculated with after 4h.After drying, fruit is put into plastic box,
Relative humidity 95% is kept, room temperature (25 DEG C) is placed in and fruit incidence is recorded after 4 days, evaluate Bayer Zygosaccharomyces BY39's with this
Fungistatic effect.The calculation formula of incidence is：The fruit of incidence (%)=morbidity/fruit sum × 100%.
2. result of the test
Tested according to above-mentioned steps, the incidence result for counting strawberry fruit is as follows
(1) inhibitions of the Bayer Zygosaccharomyces BY39 to strawberry fruit gray mold
Shown in Fig. 4, control group strawberry fruit gray mold incidence is 80%, the grass handled by Bayer Zygosaccharomyces BY39
Certain kind of berries fruit gray mold incidence is 30, therefore Bayer Zygosaccharomyces can effectively control strawberry fruit gray mold disease.
(2) inhibitions of the Bayer Zygosaccharomyces BY39 to strawberry fruit aspergillosis
Shown in Fig. 4, control group strawberry fruit aspergillosis incidence is 60%, the grass handled by Bayer Zygosaccharomyces BY39
Certain kind of berries fruit aspergillosis incidence is 0, therefore Bayer Zygosaccharomyces can effectively control strawberry fruit aspergillosis disease.
Inhibitions of the 5 Bayer Zygosaccharomyces BY39 of embodiment to Penicillium italicum
1. experimental program
Bayer Zygosaccharomyces BY39 is taken out from -80 DEG C of refrigerators, with YPDA culture mediums (yeast extract 10g, peptone 20g,
Glucose 20g, agar 18g, deionized water 1000ml, natural pH, 121 DEG C of sterilizing 30min) activation, picking single bacterium drops down onto YPD liquid
In body culture medium, 24h is cultivated under the conditions of 26 DEG C, 200r/min, 4000rpm centrifugation 5min, abandon supernatant, the thalline nothing of collection
Bacterium water cleans 3 times repeatedly, and blood counting chamber, which counts to be made into, is configured to concentration as 1 × 108The Bayer Zygosaccharomyces BY39 of cell/mL
Bacteria suspension.
Mould germ (Penicillium italicum) is activated on PDA culture medium tablet, at 26 DEG C cultivate 7~
14d, scrapes appropriate spore, and concentration is configured to as 5 × 10 with sterile water4Mould germ (the spore suspension of cell/mL.
By health 2% hypochlorite disinfectant 5min of lossless strawberry fruit, with deionized water rinsing, dry, use is sterile
Card punch makes a call to 1 hole at fruit equator, and superficial cut is 2mm (diameter) × 2mm (depth).Each wound equivalent adds 20 μ L
Following treatment fluid：(1)1×108The Bayer Zygosaccharomyces BY39 bacteria suspensions of cell/mL；(2) sterile distilled water.20 μ are inoculated with after 4h
L mould germ spore suspensions.After drying, fruit is put into plastic box, relative humidity 95% is kept, is placed in room temperature (25 DEG C) 4
Fruit incidence is recorded after it, the fungistatic effect of Bayer Zygosaccharomyces BY39 is evaluated with this.The calculation formula of incidence is：Morbidity
The fruit of rate (%)=morbidity/fruit sum × 100%.
2. result of the test
Tested according to above-mentioned steps, counting the incidence of citrusfruit, the results are shown in Figure 5, control group citrusfruit mould
Sick incidence is 73%, and the citrusfruit penicilliosis incidence handled by Bayer Zygosaccharomyces BY39 is 60%, therefore Bayer
Zygosaccharomyces BY39 can effectively control citrusfruit penicilliosis disease.
Inhibitions of the 6 Bayer Zygosaccharomyces BY39 of embodiment to cherry tomato gray mold and aspergillosis
1. experimental program
Bayer Zygosaccharomyces BY39 is taken out from -80 DEG C of refrigerators, with YPDA culture mediums (yeast extract 10g, peptone 20g,
Glucose 20g, agar 18g, deionized water 1000ml, natural pH, 121 DEG C of sterilizing 30min) activation, picking single bacterium drops down onto YPD liquid
In body culture medium, 24h is cultivated under the conditions of 26 DEG C, 200r/min, 4000rpm centrifugation 5min, abandon supernatant, the thalline nothing of collection
Bacterium water cleans 3 times repeatedly, and blood counting chamber, which counts to be made into, is configured to concentration as 1 × 108The Bayer Zygosaccharomyces BY39 of cell/mL
Bacteria suspension.
Ash arrhizus bacteria (Botrytis porri) or aspergillus echinulatus (Aspergillus aculeatus) are cultivated in PDA
Activated on base tablet, 7~14d is cultivated at 26 DEG C, scrapes appropriate spore, concentration is configured to as 5 × 10 with sterile water4Cell/mL
Ash arrhizus bacteria or aspergillus echinulatus spore suspension.By the lossless cherry tomato of health with 2% hypochlorite disinfectant 5min, spend from
Sub- water rinses, and dries, makes a call to 1 hole at fruit equator with aseptic card punch, superficial cut is 2mm (diameter) × 2mm (depth).Often
A wound equivalent adds the 20 following treatment fluids of μ L：(1)1×108The Bayer Zygosaccharomyces BY39 bacteria suspensions of cell/mL；(2) nothing
Bacterium distilled water.20 μ L ash arrhizus bacterias or aspergillus echinulatus spore suspension are inoculated with after 4h.After drying, fruit is put into plastic box, is protected
Relative humidity 95% is held, room temperature (25 DEG C) is placed in and fruit incidence is recorded after 4 days, Bayer Zygosaccharomyces bacterial strain BY39 is evaluated with this
Fungistatic effect.The calculation formula of incidence is：The fruit of incidence (%)=morbidity/fruit sum × 100%.
2. result of the test
Tested according to above-mentioned steps, statistics cherry tomato incidence result is as follows：
(1) inhibitions of the Bayer Zygosaccharomyces BY39 to cherry tomato gray mold
As shown in fig. 6, control group cherry tomato gray mold incidence is 100%, handled by Bayer Zygosaccharomyces BY39
Cherry tomato gray mold incidence is 60%, therefore Bayer Zygosaccharomyces BY39 can effectively control the holy girl as caused by ash arrhizus bacteria
Fruit gray mold disease.
(2) inhibitions of the Bayer Zygosaccharomyces BY39 to cherry tomato aspergillosis
As shown in Figure 6, the incidence of control group cherry tomato aspergillosis is 90%, by Bayer Zygosaccharomyces BY39 processing
Cherry tomato aspergillosis incidence be 20%, therefore Bayer Zygosaccharomyces BY39 can effectively control cherry tomato aspergillosis disease.
Claims (6)
- A kind of 1. Bayer Zygosaccharomyces Zygosaccharomyces bailii bacterial strains for diseases biological control of postharvest fruits and vegetables BY39 and its preparation and application method, in the preservation strain of China Committee for Culture Collection of Microorganisms's common micro-organisms center Numbering is CGMCC No.14912.
- 2. the method that the Bayer Zygosaccharomyces BY39 described in claim 1 carries out postharvest disease of fruits and vegetables prevention and preservation and freshness, its It is characterized in that following the steps below：Pichia pastoris is activated, with YPD solution culture fermentation cultures, centrifugation obtains thalline, Thalline is configured to concentration as 1 × 10 with sterile water8The bacteria suspension of cell/mL；Fruits and vegetables are put into bacteria suspension, after soaking 30 seconds Take out, air-dry；It is put into crisper, after sealing, is put into room temperature storage.
- 3. postharvest disease of fruits and vegetables prevention according to claim 2 and the method for preservation and freshness, it is characterised in that wherein described Fruits and vegetables are apple, grape, strawberry, citrus and cherry tomato.
- 4. the preparation method of Bayer Zygosaccharomyces BY39 according to claim 1, it is characterised in that by bacterial strain from -80 DEG C Refrigerator takes out, and is activated with YPDA culture mediums, and picking single bacterium is dropped down onto in YPD fluid nutrient mediums, trained under the conditions of 26 DEG C, 200r/min 24h is supported, 4000rpm centrifugations 5min collects thalline, washed 3 times with sterile.
- 5. the preparation method of Bayer Zygosaccharomyces BY39 according to claim 4, it is characterised in that the YPDA culture mediums It is：Yeast extract 10g, peptone 20g, glucose 20g, agar 18g, deionized water 1000ml, natural pH, 121 DEG C of sterilizings 30min。
- A kind of 6. purposes of Bayer Zygosaccharomyces BY39 as claimed in claim 1, it is characterised in that：It is used to control apple at the same time Fruit black spot, grape grey mould, aspergillosis and fusarium mildew, grey mould fruit rot of strawberry and aspergillosis, Penicillium italicum, and cherry tomato Gray mold and aspergillosis.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114317299A (en) * | 2021-11-30 | 2022-04-12 | 浙江中烟工业有限责任公司 | Zygosaccharomyces bailii and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1308506A1 (en) * | 2001-11-06 | 2003-05-07 | Eidgenössische Technische Hochschule Zürich | Mixtures of Propionibacterium jensenii and Lactobacillus sp. with antimicrobial activities for use as a natural preservation system |
| CN101892167A (en) * | 2010-06-10 | 2010-11-24 | 江苏大学 | Rhodotorula mucilaginosa, application thereof in preventing and curing postharvest disease of fruit and use method thereof |
| CN102834520A (en) * | 2009-10-30 | 2012-12-19 | 生物基因创新有限责任公司 | Use of methylsulfonylmethane (MSM) to modulate microbial activity |
| EP2717691A1 (en) * | 2011-06-10 | 2014-04-16 | IMD Natural Solutions GmbH | Long chain glycolipids useful to avoid perishing or microbial contamination of materials |
| CN103918723A (en) * | 2014-04-18 | 2014-07-16 | 安徽省淮北市农药厂 | Method for preparing biological agent by radix euphorbiae ebracteolatae |
-
2017
- 2017-12-15 CN CN201711373890.0A patent/CN107937290B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1308506A1 (en) * | 2001-11-06 | 2003-05-07 | Eidgenössische Technische Hochschule Zürich | Mixtures of Propionibacterium jensenii and Lactobacillus sp. with antimicrobial activities for use as a natural preservation system |
| CN102834520A (en) * | 2009-10-30 | 2012-12-19 | 生物基因创新有限责任公司 | Use of methylsulfonylmethane (MSM) to modulate microbial activity |
| CN101892167A (en) * | 2010-06-10 | 2010-11-24 | 江苏大学 | Rhodotorula mucilaginosa, application thereof in preventing and curing postharvest disease of fruit and use method thereof |
| EP2717691A1 (en) * | 2011-06-10 | 2014-04-16 | IMD Natural Solutions GmbH | Long chain glycolipids useful to avoid perishing or microbial contamination of materials |
| CN103918723A (en) * | 2014-04-18 | 2014-07-16 | 安徽省淮北市农药厂 | Method for preparing biological agent by radix euphorbiae ebracteolatae |
Non-Patent Citations (7)
| Title |
|---|
| FERDINAND RADLER 等: "Investigation of a killer strain of Zygosaccharomyces bailii", 《JOURNAL OF GENERAL MICROBIOLOGY》 * |
| W.J.JANISIEWICZ 等: "Yeasts associated with nectarines and their potential for biological control of brown rot", 《YEAST》 * |
| WOJCIECH J.JANISIEWICZ 等: "Yeasts associated with plums and their potential for controlling brown rot after harvest", 《YEAST》 * |
| 彭景贤: "酵母菌对柑橘采后病害的生物防治及防治机理的研究", 《中国优秀硕士学位论文全文数据库（电子期刊）农业科技辑》 * |
| 徐占利: "梨果采后病害新型拮抗菌分离、筛选及安全性研究", 《中国优秀硕士学位论文全文数据库（电子期刊）农业科技辑》 * |
| 李喜宏 等: "《果蔬经营与商品化处理技术》", 31 March 2009, 天津科学出版社 * |
| 王祥红 等: "嗜杀酵母的生物学功能及其应用", 《生物技术通报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114317299A (en) * | 2021-11-30 | 2022-04-12 | 浙江中烟工业有限责任公司 | Zygosaccharomyces bailii and application thereof |
| CN114317299B (en) * | 2021-11-30 | 2024-04-09 | 浙江中烟工业有限责任公司 | Zygosaccharomyces bailii strain and application thereof |
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