CN107868759A - One plant of grape wine that can effectively prevent and treat postharvest disease of fruits and vegetables has Hanseniaspora valbyensis and its preparation and application method - Google Patents
One plant of grape wine that can effectively prevent and treat postharvest disease of fruits and vegetables has Hanseniaspora valbyensis and its preparation and application method 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 preventing and treating is wide, grape wine of effect stability has Hanseniaspora valbyensis Hanseniaspora vineae BY19 and its application method.The bacterial strain is CGMCC No.14903 in the preservation strain numbering of China Committee for Culture Collection of Microorganisms's common micro-organisms center.The grape wine has the application method of Hanseniaspora valbyensis：Bacterial strain is activated, with YPD fermented and cultureds, centrifugation, thalline is made into 1 × 10 with sterilized water8Cell/mL bacteria suspension；The fruits and vegetables such as apple, pears, 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 grape wine has Hanseniaspora valbyensis bacterial strain to control green mold of apple and ash arrhizus bacteria simultaneously, pears penicilliosis and ash arrhizus bacteria, grape grey mould, aspergillosis, black spot, fusarium mildew and anthracnose, grey mould fruit rot of strawberry, Penicillium italicum, and cherry tomato gray mold and aspergillosis, loss caused by reducing postharvest disease, there is 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 grape wine controlled has a Hanseniaspora valbyensis (Hanseniaspora vineae), the bacterial strain to apple, pears, grape, strawberry, citrus,
The main postharvest disease of cherry tomato is respectively provided with significant prevention effect.
Background technology
The quality deterioration of fresh fruit of vegetables product is influenceed by factors, but rotten caused by disease especially fungal disease
Rotten go bad is the factor of most serious in being lost after fruit is adopted.Although postharvest disease of fruits and vegetables can by cultural control, physical control,
Many approach such as chemical prevention and biological control are prevented and treated, but the major measure of control fruit postharvest diseases is that chemistry is anti-at present
Control (Eckert＆Ogawa, 1985,1988).But long-term use of chemical pesticide not only causes pathogen to produce drug resistance and reduce and kill
Bacterium effect (Prusky et al., 1985；As et al., 1991；Holmes＆Eckert, 1999), and continually make
The persticide residue on fruit, the health of serious threat people are also increased with the chemical agent of high concentration, and causes environment dirty
Contaminate (Gullion＆Kuijipers, 1994).Therefore, safe efficient, nontoxic, low resistance is developed, non-harmful control fruit is adopted
Afterwards disease new technology turn into our times various countries research emphasis (Falik et al., 1995；Tian et al., 2001；
Kulakiotuet al., 2004).It is to be proved to safe and effective at present wherein to prevent and treat postharvest diseases of fruit using antagonist
New method (Wilson＆Wisniewski, 1989；Janisiewicz＆Koersten, 2002).
So far, scientists from all over the world have filtered out many has obvious fungistatic effect to fruit postharvest pathogenic fungi
Bacterium, saccharomycete and small-sized filamentous fungi, wherein using antagonistic yeast prevent and treat postharvest diseases of fruit be currently to have been demonstrated
Efficient, less toxic, low resistance new technology (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 because 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 spectrum is wide, the grape wine of effect stability has Hanseniaspora valbyensis BY19 and its application method and purposes.This kind is used for postharvest fruit and vegetable disease
The grape wine of evil biological control has Hanseniaspora valbyensis BY19, in China Committee for Culture Collection of Microorganisms's common micro-organisms
The preservation strain numbering of the heart is CGMCC No.14903.
The grape wine has the method that Hanseniaspora valbyensis bacterial strain carries out postharvest disease of fruits and vegetables preventing and treating and preservation and freshness, according to following
Step is carried out：Grape wine there is into Hanseniaspora valbyensis activation, with YPD solution culture fermentation cultures, centrifugation obtains thalline, by thalline
Concentration is configured to as 1 × 10 with sterilized water8Cell/mL bacteria suspension；By apple, pears, grape, strawberry, citrus and cherry tomato etc.
Fruits and vegetables are put into bacteria suspension, and immersion is taken out after 30 seconds, are air-dried；It is put into crisper, after sealing, is put into room temperature storage.
The preparation method that the grape wine has Hanseniaspora valbyensis BY19 is that bacterial strain is taken out from -80 DEG C of refrigerators, is cultivated with YPDA
Base is activated, and picking single bacterium is dropped down onto in YPD fluid nutrient mediums, and 24h, 4000rpm centrifugations are cultivated under the conditions of 26 DEG C, 200r/min
5min collects thalline, is 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.
Described grape wine has Hanseniaspora valbyensis bacterial strain to be used to control green mold of apple and ash arrhizus bacteria, pear fruit simultaneously
Penicilliosis and ash arrhizus bacteria, grape grey mould, aspergillosis, black spot, fusarium mildew and anthracnose, grey mould fruit rot of strawberry and aspergillus
Disease, 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, pears,
There is the grape wine of notable prevention effect to have Hanseniaspora valbyensis BY19 for grape, strawberry, citrus, cherry tomato postharvest disease, can be extensive
For the preventing and treating of postharvest disease of fruits and vegetables, loss caused by reducing postharvest disease, there is good application prospect.
Advantages of the present invention：(1) it is this laboratory from grape that grape wine used in the present invention, which has Hanseniaspora valbyensis BY19,
Screening obtains in wine fermentation mash, and its is harmless, safe.(2) grape wine used in the present invention has the spore Chinese to give birth to ferment
Female BY19 antimicrobial spectrums are wide, can control green mold of apple and ash arrhizus bacteria, pear fruit penicilliosis and ash arrhizus bacteria, light grey and pinkish simultaneously
Mildew, aspergillosis, black spot, fusarium mildew and anthracnose, grey mould fruit rot of strawberry and aspergillosis, Penicillium italicum, and cherry tomato
Gray mold and aspergillosis.(3) grape wine used in the present invention has Hanseniaspora valbyensis BY19 well-growns in YPD culture mediums,
It is easy to cultivate, character is stable, and the finite concentration bacteria suspension is used alone just can effectively reduce a variety of postharvest fruit and vegetable fungal diseases
Incidence, use cost is low, wide market.(4) present invention has Hanseniaspora valbyensis BY19 to replace using grape wine
Chemical bactericide prevents and treats fruit postharvest diseases, avoids using harm of the chemical bactericide to people, and free from environmental pollution, society and life
State remarkable benefit.
Pass through the more detailed explanation present invention of following embodiment.Following embodiment is merely illustrative, the present invention
Do not limited by these embodiments.
Brief description of the drawings
Fig. 1 is the 26S rDNA D1/ that grape wine of the present invention has Hanseniaspora valbyensis Hanseniaspora vineae BY19
D2 areas Evolution of Nucleic Acid Sequences graph of a relation.
Fig. 2 is that grape wine has Hanseniaspora valbyensis BY19 to green mold of apple and the inhibition of gray mold.Note：Control：
Sterilized water, i.e. control group；Hv：1×108Cell/mL grape wine has Hanseniaspora valbyensis BY19 bacteria suspensions.It is poor that different letters represent
Different notable (P ＜ 0.05).
Fig. 3 is that grape wine has Hanseniaspora valbyensis BY19 to have Hanseniaspora valbyensis to grape wine to pear fruit penicilliosis and gray mold
Inhibition.Note：Control：Sterilized water, i.e. control group；Hv：1×108Cell/mL grape wine has Hanseniaspora valbyensis
BY19 bacteria suspensions.Different letters represent significant difference (P ＜ 0.05).
Fig. 4 is that grape wine has Hanseniaspora valbyensis BY19 to grape grey mould, aspergillosis, black spot, fusarium mildew and anthrax
The inhibition of disease.Note：Control：Sterilized water, i.e. control group；Hv：1×108Cell/mL grape wine has Hanseniaspora valbyensis
BY19 bacteria suspensions.Different letters represent significant difference (P ＜ 0.05).
Fig. 5 is that grape wine has Hanseniaspora valbyensis BY19 to grey mould fruit rot of strawberry and the inhibition of aspergillosis.Note：Control：
Sterilized water, i.e. control group；Hv：1×108Cell/mL grape wine has Hanseniaspora valbyensis BY19 bacteria suspensions.It is poor that different letters represent
Different notable (P ＜ 0.05).
Fig. 6 is that grape wine has inhibitions of the Hanseniaspora valbyensis BY19 to Penicillium italicum.Note：Control：Sterilized water,
That is control group；Hv：1×108Cell/mL grape wine has Hanseniaspora valbyensis BY19 bacteria suspensions.Different letters represent significant difference
(P ＜ 0.05).
Fig. 7 is that grape wine has Hanseniaspora valbyensis BY19 to cherry tomato gray mold and the inhibition of aspergillosis.Note：
Control：Sterilized water, i.e. control group；Hv：1×108Cell/mL grape wine has Hanseniaspora valbyensis BY19 bacteria suspensions.Different words
Mother represents significant difference (P ＜ 0.05).
Embodiment
Embodiment 1：Grape wine has Hanseniaspora valbyensis Hanseniaspora vineae BY19 biological characteristics
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 ') enter performing PCR amplification yeast 26S rDNA D1/D2 areas nucleotide sequence, by the survey of PCR primer
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, it is determined that the bacterial strain screened, which is grape wine, Hanseniaspora valbyensis (Hanseniaspora
vineae)。
The grape wine of the present invention has Hanseniaspora valbyensis BY19 to be preserved in city of BeiJing, China Chaoyang District North Star West Road 1 institute 3
Number Institute of Microorganism, Academia Sinica China Microbiological bacterial strain preservation committee common micro-organisms center, the preservation time is
On November 15th, 2017, deposit number are CGMCC No.14903, it is proposed that Classification And Nomenclature have Hanseniaspora valbyensis for grape wine
Hanseniaspora vineae。
The grape wine of embodiment 2 has Hanseniaspora valbyensis BY19 to green mold of apple and the inhibition of gray mold
1. experimental program
There is Hanseniaspora valbyensis BY19 to be taken out from -80 DEG C of refrigerators grape wine, with YPDA culture mediums (yeast extract 10g, albumen
Peptone 20g, glucose 20g, agar 18g, deionized water 1000ml, natural pH, 121 DEG C of sterilizing 30min) activation, picking single bacterium colony
To YPD fluid nutrient mediums, 24h is cultivated under the conditions of 26 DEG C, 200r/min, 4000rpm centrifugation 5min, abandons supernatant, collection
Thalline is cleaned 3 times repeatedly with sterilized water, and blood counting chamber, which counts to be made into, is configured to concentration as 1 × 108Cell/mL grape wine has
Hanseniaspora valbyensis BY19 bacteria suspensions.
Mould germ (Penicillium expansum) or ash arrhizus bacteria (Botrytis cinerea) are cultivated in PDA
Activated on base flat board, 7~14d is cultivated at 26 DEG C, scrapes appropriate spore, concentration is configured to as 5 × 10 with sterilized water4Cell/mL
Mould germ or ash arrhizus bacteria spore suspension.By health 2% hypochlorite disinfectant 5min of lossless Apple, spend
Ionized water rinses, and dries, makes a call to 5 holes 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)1×108Cell/mL grape wine has Hanseniaspora valbyensis BY19 bacterium to hang
Liquid；(2) sterile distilled water.20 μ L moulds germs or ash arrhizus bacteria spore suspension are inoculated with after 4h.After drying, fruit is put into modeling
Hopper, relative humidity 95% is kept, room temperature (25 DEG C) is placed in and the fruit incidence of disease is recorded after 4 days, evaluating grape wine with this has the spore Chinese
Raw yeast BY19 fungistatic effect.The calculation formula of the incidence of disease is：The fruit of the incidence of disease (%)=morbidity/fruit sum ×
100%.
2. result of the test
Tested according to above-mentioned steps, the incidence of disease result for counting apple is as follows：
(1) grape wine has inhibitions of the Hanseniaspora valbyensis BY19 to green mold of apple
As shown in Fig. 2 the control group green mold of apple incidence of disease is 100%, have by grape wine at Hanseniaspora valbyensis BY19
The green mold of apple incidence of disease of reason is 6%, therefore grape wine has Hanseniaspora valbyensis BY19 effectively to control green mold of apple sick
Evil.
(2) grape wine has inhibitions of the Hanseniaspora valbyensis BY19 to apple gray mold
As shown in Fig. 2 the incidence of disease of control group apple gray mold is 100%, there is Hanseniaspora valbyensis BY19 by grape wine
The apple gray mold incidence of disease of processing is 0, therefore grape wine has Hanseniaspora valbyensis BY19 effectively to control apple gray mold sick
Evil.
The grape wine of embodiment 3 has Hanseniaspora valbyensis BY19 to pear fruit penicilliosis and the inhibition of gray mold
1. experimental program
There is Hanseniaspora valbyensis BY19 to be taken out from -80 DEG C of refrigerators grape wine, with YPDA culture mediums (yeast extract 10g, albumen
Peptone 20g, glucose 20g, agar 18g, deionized water 1000ml, natural pH, 121 DEG C of sterilizing 30min) activation, picking single bacterium colony
To YPD fluid nutrient mediums, 24h is cultivated under the conditions of 26 DEG C, 200r/min, 4000rpm centrifugation 5min, abandons supernatant, collection
Thalline is cleaned 3 times repeatedly with sterilized water, and blood counting chamber, which counts to be made into, is configured to concentration as 1 × 108Cell/mL grape wine has
Hanseniaspora valbyensis BY19 bacteria suspensions.
Mould germ (Penicillium expansum) or ash arrhizus bacteria (Botrytis cinerea) are cultivated in PDA
Activated on base flat board, 7~14d is cultivated at 26 DEG C, scrapes appropriate spore, concentration is configured to as 5 × 10 with sterilized water4Cell/mL
Mould germ or ash arrhizus bacteria spore suspension.By the lossless pear fruit of health with 2% hypochlorite disinfectant 5min, spend from
Sub- water rinses, and dries, makes a call to 5 holes at fruit equator with aseptic card punch, superficial cut is 2mm (diameter) × 2mm (depth).Often
Individual wound equivalent adds the 20 following treatment fluids of μ L：(1)1×108Cell/mL grape wine has Hanseniaspora valbyensis BY19 bacterium to hang
Liquid；(2) sterile distilled water.20 μ L moulds germs or ash arrhizus bacteria spore suspension are inoculated with after 4h.After drying, fruit is put into modeling
Hopper, relative humidity 95% is kept, room temperature (25 DEG C) is placed in and the fruit incidence of disease is recorded after 4 days, evaluating grape wine with this has the spore Chinese
Raw yeast BY19 fungistatic effect.The calculation formula of the incidence of disease is：The fruit of the incidence of disease (%)=morbidity/fruit sum ×
100%.
2. result of the test
Tested according to above-mentioned steps, the incidence of disease result for counting pear fruit is as follows：
(1) grape wine has inhibitions of the Hanseniaspora valbyensis BY19 to pear fruit penicilliosis
As shown in figure 3, the control group pear fruit penicilliosis incidence of disease is 100%, there is Hanseniaspora valbyensis BY19 by grape wine
The pear fruit penicilliosis incidence of disease of processing is 33%, therefore grape wine has Hanseniaspora valbyensis BY19 effectively to control pear fruit blue or green
Mildew disease.
(2) grape wine has inhibitions of the Hanseniaspora valbyensis BY19 to pear fruit gray mold
As shown in figure 3, the incidence of disease of control group pear fruit gray mold is 100%, there is Hanseniaspora valbyensis by grape wine
The pear fruit gray mold incidence of disease of BY19 processing is 6%, therefore grape wine has Hanseniaspora valbyensis BY19 effectively to control the operatic circle
Real gray mold disease.
The grape wine of embodiment 4 has inhibitions of the Hanseniaspora valbyensis BY19 to grape postharvest disease
1. experimental program
There is Hanseniaspora valbyensis BY19 to be taken out from -80 DEG C of refrigerators grape wine, with YPDA culture mediums (yeast extract 10g, albumen
Peptone 20g, glucose 20g, agar 18g, deionized water 1000ml, natural pH, 121 DEG C of sterilizing 30min) activation, picking single bacterium colony
To YPD fluid nutrient mediums, 24h is cultivated under the conditions of 26 DEG C, 200r/min, 4000rpm centrifugation 5min, abandons supernatant, collection
Thalline is cleaned 3 times repeatedly with sterilized water, and blood counting chamber, which counts to be made into, is configured to concentration as 1 × 108Cell/mL grape wine has
Hanseniaspora valbyensis BY19 bacteria suspensions.
By botrytis cinerea (Botrytis porri), aspergillus echinulatus (Aspergillus aculeatus), alternaric bacteria
(Alternaria eichhorniae), fragrant Fusariumsp (Fusarium redolens) or fruit life anthrax-bacilus
(Colletotrichum fructicola) is activated on PDA culture medium flat board, and 7~14d is cultivated at 26 DEG C, scrapes appropriate spore
Son, concentration is configured to as 5 × 10 with sterilized water4Cell/mL botrytis cinerea, aspergillus echinulatus, alternaric bacteria, fragrant Fusariumsp or fruit
Raw anthrax spores suspension.
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)1×108Cell/mL grape wine has Hanseniaspora valbyensis BY19 bacteria suspensions；(2) sterile distilled water.4h is followed by
20 μ L botrytis cinereas of kind, aspergillus echinulatus, alternaric bacteria, fragrant Fusariumsp or fruit life anthrax spores suspension.After drying, by fruit
Plastic box is put into, relative humidity 95% is kept, is placed in room temperature (25 DEG C) and the fruit incidence of disease is recorded after 4 days, grape wine is evaluated with this
There is Hanseniaspora valbyensis BY19 fungistatic effect.The calculation formula of the incidence of disease is：The fruit of the incidence of disease (%)=morbidity/fruit sum
× 100%.
2. result of the test
Tested according to above-mentioned steps, the incidence of disease result for counting grape fruit is as follows：
(1) grape wine has inhibitions of the Hanseniaspora valbyensis BY19 to grape fruit gray mold
As shown in figure 4, the control group grape fruit gray mold incidence of disease is 100%, there is Hanseniaspora valbyensis by grape wine
The grape fruit gray mold incidence of disease of BY19 processing is 0, therefore grape wine has Hanseniaspora valbyensis BY19 effectively to control grape
Fruit gray mold disease.
(2) grape wine has inhibitions of the spore Chinese life ferment BY19 to grape fruit aspergillosis
As shown in figure 4, the incidence of disease of control group grape fruit aspergillosis is 100%, there is Hanseniaspora valbyensis by grape wine
The grape fruit aspergillosis incidence of disease of BY19 processing is 33%, therefore grape wine has Hanseniaspora valbyensis BY19 effectively to control Portugal
Grape fruit aspergillosis disease.
(3) grape wine has inhibitions of the Hanseniaspora valbyensis BY19 to grape fruit black spot
As shown in figure 4, the incidence of disease of control group grape fruit black spot is 87%, there is Hanseniaspora valbyensis by grape wine
The BY19 grape fruit black spot incidence of disease is 67%, therefore grape wine has Hanseniaspora valbyensis BY19 to control uva solid black
Pinta disease.
(4) grape wine has inhibitions of the Hanseniaspora valbyensis BY19 to grape fruit fusarium mildew
As shown in figure 4, the incidence of disease of control group grape fruit fusarium mildew is 100%, there is the spore Chinese to give birth to ferment by grape wine
The grape fruit fusarium mildew incidence of disease of female BY19 processing is 73%, therefore grape wine has Hanseniaspora valbyensis BY19 effectively to control
Grape fruit fusarium mildew disease processed.
(5) grape wine has inhibitions of the Hanseniaspora valbyensis BY19 to grape fruit anthracnose
As shown in figure 4, the incidence of disease of control group grape fruit anthracnose is 100%, there is Hanseniaspora valbyensis by grape wine
The grape fruit anthracnose incidence of disease of BY19 processing is 80%, therefore grape wine has Hanseniaspora valbyensis BY19 effectively to control Portugal
Grape Fruit Anthracnose does harm to.
The grape wine of embodiment 5 has Hanseniaspora valbyensis BY19 to strawberry fruit gray mold and the inhibition of aspergillosis
1. experimental program
There is Hanseniaspora valbyensis BY19 to be taken out from -80 DEG C of refrigerators grape wine, with YPDA culture mediums (yeast extract 10g, albumen
Peptone 20g, glucose 20g, agar 18g, deionized water 1000ml, natural pH, 121 DEG C of sterilizing 30min) activation, picking single bacterium colony
To YPD fluid nutrient mediums, 24h is cultivated under the conditions of 26 DEG C, 200r/min, 4000rpm centrifugation 5min, abandons supernatant, collection
Thalline is cleaned 3 times repeatedly with sterilized water, and blood counting chamber, which counts to be made into, is configured to concentration as 1 × 108Cell/mL grape wine has
Hanseniaspora valbyensis BY19 bacteria suspensions.
Ash arrhizus bacteria (Botrytis porri) or aspergillus echinulatus (Aspergillus aculeatus) are cultivated in PDA
Activated on base flat board, 7~14d is cultivated at 26 DEG C, scrapes appropriate spore, concentration is configured to as 5 × 10 with sterilized 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)1×108Cell/mL grape wine has Hanseniaspora valbyensis BY19 bacterium to hang
Liquid；(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 modeling
Hopper, relative humidity 95% is kept, room temperature (25 DEG C) is placed in and the fruit incidence of disease is recorded after 4 days, evaluating grape wine with this has the spore Chinese
Raw yeast BY19 fungistatic effect.The calculation formula of the incidence of disease is：The fruit of the incidence of disease (%)=morbidity/fruit sum ×
100%.
2. result of the test
Tested according to above-mentioned steps, the incidence of disease result for counting strawberry fruit is as follows：
(1) grape wine has inhibitions of the Hanseniaspora valbyensis BY19 to strawberry fruit gray mold
As shown in Figure 5, the control group strawberry fruit gray mold incidence of disease is 80%, has Hanseniaspora valbyensis by grape wine
The strawberry fruit gray mold incidence of disease of BY19 processing is 30%, therefore grape wine has Hanseniaspora valbyensis effectively to control strawberry fruit
Real gray mold disease.
(2) grape wine has inhibitions of the Hanseniaspora valbyensis BY19 to strawberry fruit aspergillosis
As shown in Figure 5, the incidence of disease of control group strawberry fruit aspergillosis is 60%, has Hanseniaspora valbyensis by grape wine
The strawberry fruit aspergillosis incidence of disease of BY19 processing is 12%, therefore grape wine has Hanseniaspora valbyensis effectively to control strawberry fruit
Real aspergillosis disease.
The grape wine of embodiment 6 has inhibitions of the Hanseniaspora valbyensis BY19 to Penicillium italicum
1. experimental program
There is Hanseniaspora valbyensis BY19 to be taken out from -80 DEG C of refrigerators grape wine, with YPDA culture mediums (yeast extract 10g, albumen
Peptone 20g, glucose 20g, agar 18g, deionized water 1000ml, natural pH, 121 DEG C of sterilizing 30min) activation, picking single bacterium colony
To YPD fluid nutrient mediums, 24h is cultivated under the conditions of 26 DEG C, 200r/min, 4000rpm centrifugation 5min, abandons supernatant, collection
Thalline is cleaned 3 times repeatedly with sterilized water, and blood counting chamber, which counts to be made into, is configured to concentration as 1 × 108Cell/mL grape wine has
Hanseniaspora valbyensis BY19 bacteria suspensions.
Mould germ (Penicillium italicum) is activated on PDA culture medium flat board, at 26 DEG C cultivate 7~
14d, appropriate spore is scraped, concentration is configured to as 5 × 10 with sterilized water4Cell/mL mould germ spore suspension.
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)1×108Cell/mL grape wine has Hanseniaspora valbyensis BY19 bacteria suspensions；(2) sterile distilled water.After 4h
It is inoculated with 20 μ 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) record the fruit incidence of disease after 4 days, evaluating grape wine with this has Hanseniaspora valbyensis BY19 fungistatic effect.The meter of the incidence of disease
Calculating formula is：The fruit of the incidence of disease (%)=morbidity/fruit sum × 100%.
2. result of the test
Tested according to above-mentioned steps, count the incidence of disease result of citrusfruit as shown in fig. 6, control group citrusfruit mould
The sick incidence of disease is 73%, and the citrusfruit penicilliosis incidence of disease for having Hanseniaspora valbyensis BY19 to handle by grape wine is 24.3%,
Therefore grape wine has Hanseniaspora valbyensis BY19 effectively to control citrusfruit penicilliosis disease.
The grape wine of embodiment 7 has Hanseniaspora valbyensis BY19 to cherry tomato gray mold and the inhibition of aspergillosis
1. experimental program
There is Hanseniaspora valbyensis BY19 to be taken out from -80 DEG C of refrigerators grape wine, with YPDA culture mediums (yeast extract 10g, albumen
Peptone 20g, glucose 20g, agar 18g, deionized water 1000ml, natural pH, 121 DEG C of sterilizing 30min) activation, picking single bacterium colony
To YPD fluid nutrient mediums, 24h is cultivated under the conditions of 26 DEG C, 200r/min, 4000rpm centrifugation 5min, abandons supernatant, collection
Thalline is cleaned 3 times repeatedly with sterilized water, and blood counting chamber, which counts to be made into, is configured to concentration as 1 × 108Cell/mL grape wine has
Hanseniaspora valbyensis BY19 bacteria suspensions.
Ash arrhizus bacteria (Botrytis porri) or aspergillus echinulatus (Aspergillus aculeatus) are cultivated in PDA
Activated on base flat board, 7~14d is cultivated at 26 DEG C, scrapes appropriate spore, concentration is configured to as 5 × 10 with sterilized 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
Individual wound equivalent adds the 20 following treatment fluids of μ L：(1)1×108Cell/mL grape wine has Hanseniaspora valbyensis BY19 bacterium to hang
Liquid；(2) sterile distilled water.20 μ L ash arrhizus bacterias or spore suspension are inoculated with after 4h.After drying, fruit is put into plastic box, protected
Relative humidity 95% is held, room temperature (25 DEG C) is placed in and the fruit incidence of disease is recorded after 4 days, evaluating grape wine with this has Hanseniaspora valbyensis
BY19 fungistatic effect.The calculation formula of the incidence of disease is：The fruit of the incidence of disease (%)=morbidity/fruit sum × 100%.
2. result of the test
Tested according to above-mentioned steps, statistics cherry tomato incidence of disease result is as follows：
(1) grape wine has inhibitions of the Hanseniaspora valbyensis BY19 to cherry tomato gray mold
As shown in Figure 7, the control group cherry tomato gray mold incidence of disease is 100%, has Hanseniaspora valbyensis by grape wine
The cherry tomato gray mold incidence of disease of BY19 processing is 66%, therefore grape wine has Hanseniaspora valbyensis BY19 effectively to control holy girl
Fruit gray mold disease.
(2) grape wine has inhibitions of the Hanseniaspora valbyensis BY19 to cherry tomato aspergillosis
As shown in Figure 7, the incidence of disease of control group cherry tomato aspergillosis is 90%, has Hanseniaspora valbyensis by grape wine
The cherry tomato aspergillosis incidence of disease of BY19 processing is 0%, therefore grape wine has Hanseniaspora valbyensis BY19 effectively to control holy girl
Fruit aspergillosis disease.
Claims (6)
1. one plant of grape wine for being used for diseases biological control of postharvest fruits and vegetables has Hanseniaspora valbyensis Hanseniaspora vineae
BY19 and its application method, it is in the preservation strain numbering of China Committee for Culture Collection of Microorganisms's common micro-organisms center
CGMCC No.14903。
2. the grape wine described in claim 1 has the side that Hanseniaspora valbyensis BY19 carries out postharvest disease of fruits and vegetables preventing and treating and preservation and freshness
Method, it is characterised in that follow the steps below：Grape wine there is into Hanseniaspora valbyensis activation, trained with YPD solution culture fermentations
Support, centrifugation obtains thalline, and thalline is configured into concentration for 1 × 10 with sterilized water8Cell/mL bacteria suspension；Fruits and vegetables are put into bacterium
In suspension, immersion is taken out after 30 seconds, is air-dried；It is put into crisper, after sealing, is put into room temperature storage.
3. postharvest disease of fruits and vegetables preventing and treating according to claim 2 and the method for preservation and freshness, it is characterised in that wherein described
Fruits and vegetables are apple, pears, grape, strawberry, citrus and cherry tomato.
4. grape wine according to claim 1 has Hanseniaspora valbyensis BY19 preparation method, it is characterised in that by bacterial strain
Take out, activated with YPDA culture mediums, picking single bacterium is dropped down onto in YPD fluid nutrient mediums, in 26 DEG C, 200r/min from -80 DEG C of refrigerators
Under the conditions of cultivate 24h, 4000rpm centrifugations 5min collects thalline, with sterile washing 3 times.
5. grape wine according to claim 4 has Hanseniaspora valbyensis BY19 preparation method, it is characterised in that the YPDA
Culture medium is：Yeast extract 10g, peptone 20g, glucose 20g, agar 18g, deionized water 1000ml, natural pH, 121 DEG C
Sterilize 30min.
6. a kind of grape wine as claimed in claim 1 has spore Chinese life ferment BY19 purposes, it is characterised in that：It is used for same time control
Green mold of apple processed and ash arrhizus bacteria, pear fruit penicilliosis and ash arrhizus bacteria, grape grey mould, aspergillosis, black spot, fusarium
Disease and anthracnose, grey mould fruit rot of strawberry and aspergillosis, Penicillium italicum, and cherry tomato gray mold and aspergillosis.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110628655A (en) * | 2019-10-21 | 2019-12-31 | 福州大学 | Yeast Lg-3 for controlling postharvest diseases of fruits and vegetables and using method thereof |
| CN113273604A (en) * | 2021-05-10 | 2021-08-20 | 江苏大学 | Application of abnormal yeast Weikehan in preventing and treating postharvest diseases of tomatoes and storing and refreshing |
| CN114451450A (en) * | 2022-02-24 | 2022-05-10 | 兰州大学 | Hansenula polymorpha capable of antagonizing botrytis cinerea and application thereof |
| CN114600950A (en) * | 2022-01-26 | 2022-06-10 | 四川农业大学 | Preparation method and application of hansenula polymorpha antistaling agent |
| CN114752513A (en) * | 2022-04-19 | 2022-07-15 | 青岛农业大学 | Hansenula polymorpha MP261 in vineyard for inhibiting phytopathogens and application thereof |
| CN114874914A (en) * | 2022-01-18 | 2022-08-09 | 吉林大学 | Broad-spectrum bacteriostatic self-flocculating non-saccharomyces cerevisiae strain CC-P5 and application thereof |
| CN117264789A (en) * | 2023-09-05 | 2023-12-22 | 云南大学 | Hansenula polymorpha strain and application thereof in preparation of blueberry fruit wine |
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| CN103740604A (en) * | 2014-01-26 | 2014-04-23 | 北京绿环国际科技有限公司 | Hanseniaspora uvarum and application thereof |
| CN105875292A (en) * | 2016-04-12 | 2016-08-24 | 江苏大学 | Method for preventing and treating postharvest diseases of citrus reticulata blanco by combining phosphatidylcholine with hanseniaspora uvarum |
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| CN103740604A (en) * | 2014-01-26 | 2014-04-23 | 北京绿环国际科技有限公司 | Hanseniaspora uvarum and application thereof |
| CN105875292A (en) * | 2016-04-12 | 2016-08-24 | 江苏大学 | Method for preventing and treating postharvest diseases of citrus reticulata blanco by combining phosphatidylcholine with hanseniaspora uvarum |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110628655A (en) * | 2019-10-21 | 2019-12-31 | 福州大学 | Yeast Lg-3 for controlling postharvest diseases of fruits and vegetables and using method thereof |
| CN113273604A (en) * | 2021-05-10 | 2021-08-20 | 江苏大学 | Application of abnormal yeast Weikehan in preventing and treating postharvest diseases of tomatoes and storing and refreshing |
| CN114874914A (en) * | 2022-01-18 | 2022-08-09 | 吉林大学 | Broad-spectrum bacteriostatic self-flocculating non-saccharomyces cerevisiae strain CC-P5 and application thereof |
| CN114874914B (en) * | 2022-01-18 | 2024-01-26 | 吉林大学 | Broad-spectrum antibacterial self-flocculating non-saccharomyces cerevisiae strain CC-P5 and application thereof |
| CN114600950A (en) * | 2022-01-26 | 2022-06-10 | 四川农业大学 | Preparation method and application of hansenula polymorpha antistaling agent |
| CN114451450A (en) * | 2022-02-24 | 2022-05-10 | 兰州大学 | Hansenula polymorpha capable of antagonizing botrytis cinerea and application thereof |
| CN114752513A (en) * | 2022-04-19 | 2022-07-15 | 青岛农业大学 | Hansenula polymorpha MP261 in vineyard for inhibiting phytopathogens and application thereof |
| CN117264789A (en) * | 2023-09-05 | 2023-12-22 | 云南大学 | Hansenula polymorpha strain and application thereof in preparation of blueberry fruit wine |
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