CN105794954B - The method of bamboo-leaves flavones combination Cali Bick Pichia pastoris prevention and treatment Biocontrol of Postharvest Diseases in Apple Fruits - Google Patents
The method of bamboo-leaves flavones combination Cali Bick Pichia pastoris prevention and treatment Biocontrol of Postharvest Diseases in Apple Fruits Download PDFInfo
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- CN105794954B CN105794954B CN201610143541.9A CN201610143541A CN105794954B CN 105794954 B CN105794954 B CN 105794954B CN 201610143541 A CN201610143541 A CN 201610143541A CN 105794954 B CN105794954 B CN 105794954B
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/14—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
- A23B7/153—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
- A23B7/154—Organic compounds; Microorganisms; Enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/14—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
- A23B7/153—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
- A23B7/154—Organic compounds; Microorganisms; Enzymes
- A23B7/155—Microorganisms; Enzymes; Antibiotics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses the methods of bamboo-leaves flavones combination Cali Bick Pichia pastoris prevention and treatment Biocontrol of Postharvest Diseases in Apple Fruits, belong to biologically controlling postharvest disease of fruit field.Cali's Bick Pichia pastoris is activated, is inoculated into NYDB culture medium and cultivates 20h, centrifugation obtains thallus, and Cali's Bick Pichia pastoris thallus is diluted to 1 × 10 with the bamboo-leaves flavones of twice of final concentration8 The bacteria suspension of cells/mL (between the final concentration of 100 μ g/mL-500 μ g/mL of bamboo-leaves flavones)；Bacteria suspension is uniformly sprayed on apple, natural air drying；It is placed in plastic crate, after being sealed with preservative film, is placed under the conditions of room temperature (20 DEG C) and stores.The present invention is used in combination using bamboo-leaves flavones and Cali's Bick Pichia pastoris and improves its effect to Biocontrol of Postharvest Diseases in Apple Fruits biological control, and using simple, easy to operate, effect is good, at low cost.
Description
Technical field
The present invention relates to a kind of bamboo-leaves flavones (BLF) to prevent and treat in conjunction with Cali's Bick Pichia pastoris (Pichia caribbica)
The method of Biocontrol of Postharvest Diseases in Apple Fruits belongs to biologically controlling postharvest disease of fruit field.
Background technique
Apple is one of big fruit in the world four, is the most important fruit tree species of world Temperate Region in China cultivation, in world fruit
Product occupy an important position in market.The nutritional ingredient very abundant of apple.Contain protein, fat, carbohydrate, dimension life
The organic acids such as plain C, vitamin B1, vitamin B2, carrotene and calcium, phosphorus, iron, citric acid, tartaric acid and pectin, fiber
Element etc..Currently, whole world apple production state (area) has 80 or so, China and South America are that Apple development is most fast in the past 20 years
Countries and regions.568.5 ten thousand hectares of world's apple area in 2001, yield 6.3 × 1010Kilogram, wherein China's apple area
240.1 ten thousand hectares, yield 2.4 × 1010Kilograms ten thousand tons, the 42.2% and 38.2% of world's apple gross area and total output is accounted for,
It ranks the first in the world.
Loss is huge caused by Biocontrol of Postharvest Diseases in Apple Fruits.According to generally conservative estimation, loss is after China's apple is adopted
10%-30% or so.The cause of loss and deterioration is known as very much, including since moisture and nutrient consumption are made after causing apple to be adopted
At weightlessness and the qualities fission such as withered；As adopting the imbalance of physiological metabolism caused by rear adverse circumstance；It is caused due to adopting after ripening aging
Fruit tissue's structure collapse and rot caused by being infected as pathogen etc..Wherein rotted as caused by pathogen infection
It is the main reason for loss after causing fruit to be adopted.Cause the fungal species to rot during fruit storage very much, but every kind of fruit accounts for
The pathogenic bacteria of advantage generally have 2-3 kind.For example, apple adopt after by fungus-caused disease be mainly penicillium expansum
Penicilliosis caused by (Penicillium expansum) and the gray mold as caused by Botrytis cinerea (Botrytis cinerea).
Disease fungus is not only in that it leads to the heavy losses of fruit quantitatively, and many disease fungus to the harm for adopting rear fruit
The many secondary metabolites of generation, which can be secreted, can cause serious food-safety problem, the main pathogenic bacteria penicillium expansum on apple
(P.expansum) clavacin (patulin) can be generated.Patulin is also known as clavacin, is Penicillium, aspergillus, silk clothes
The mould toxic fungus metabolite of one kind for the moulds metabolism such as belonging to and generating, has even lethal etc. adverse effects of carcinogenic, aberration inducing.
For a long time, the major measure for controlling postharvest disease of fruits and vegetables is using fungicide.It is counted, is sterilized according to latest data
Agent accounts for about 26% share in crop disease control market, and consumption sum is up to 13,300,000,000 dollars within 2011.Chemical bactericide has
Good, the quick feature of economy, bactericidal effect, the common chemical bactericide for controlling fruit postharvest diseases mainly have imazalil
(Imazzlii), benomyl (Benomyl), Guanoctine (Guazatine).But more than ten years in past, the public and scientific circles are for food
The concern of product and Environmental Pesticide (especially fungicide) residual hazard problem increasingly increases.The long-term and a large amount of use of fungicide, seriously
Environment is polluted, human health is harmful to.The report (1987) of National Academy of Sciences (NAS) points out that the application of fungicide is to people
Class health causes harm；In the sales volume of the 9 kinds of neoplastic compounds in the whole America, fungicide accounts for 90% or so, for the complete of food
In portion's pesticide, the tumorigenesis that fungicide constitutes 60% is dangerous.In addition, high-frequency can make pathogen using the fungicide of single kind
Develop drug resistance problem, substantially reduces the control efficiency of fungicide.
Therefore, the disease for studying and preventing and treating using the substitute of chemical bactericide plant especially fruits and vegetables, is facing mankind
A urgent task.Biological bactericide can mitigate harmful microorganism and fruits and vegetables are infected and protected with environment, be production safety
The effective ways of property food.Thus research natural biological fungicide replaces the prevention and treatment of pesticide progress postharvest disease of fruits and vegetables, to reduction
The residual of Pesticide protects environment, it is ensured that human health increases fruits and vegetables outlet, all has deep meaning.
Currently increasingly attention carries out the research of biologically controlling postharvest disease of fruit with beneficial microbe in the world, and thinks
This is one of the direction for being most hopeful substituted chemistry fungicide.Saccharomycete is due to inheritance stability, and antimicrobial spectrum is wide, potency is high, generally
It does not generate to people and the harmful metabolite of host plant, highly-safe and low to nutritional requirement, growth is fast, and to a variety of sides of body
Urgent, adverse circumstance has stronger tolerance, insensitive to most of fungicide, can be compatible to other chemically and physically handle, because
Antagonism yeast is as fruit postharvest diseases biocontrol microorganisms in recent years for this, by international extensive concern.
Since nineteen fifty-three, Gutter etc. report for the first time bacillus subtilis to citrus fruit pathogen have antagonism with
Come, made breakthrough progress at present using the biological control that Antagonistic Fungi carries out fruit postharvest diseases, it is considered to be most have
The method for wishing to substitute chemical bactericide.Such as in the U.S., prevented from Environmental Protection Agency approval in 1979 using first biology
Since microorganism is controlled as biocontrol microorganisms, by the end of 2004, existing 26 microorganisms (14 kinds of bacteriums and 12 kinds of fungies) were by the mechanism
Approve and registers.Pichia pastoris (Pichia guilliermondii) is to be reported that fruit cause of disease can be inhibited true in the world earliest
The biological control yeast of bacterium, and Candida oleophilia isolate I-182 is in nineteen ninety-five as biological bactericide
It is registered at Environmental Protection Agency (EPA), and via U.S.'s Ecogen company trade.In addition, short of money after one plant of South Africa is adopted
Anti- yeast Cryptococcus albidus (Cryptococcus albidus) has also been commercialized, trade name Yield Plus.
Up to the present, the research of postharvest fruit and vegetable biological control has semicentennial history, but is applied to produce
Antagonistic Fungi type it is few, a few launch such as only Biosave and Aspire, and China is at present still without Antagonistic Fungi
Applied to actual production.Main reason is that Antagonistic Fungi reported in the literature is used to prevent and treat the pathogen of fruits and vegetables wound mostly, and
It is not the pathogen invasion for being used directly in fruits and vegetables epidermis and preventing and treating normal fruits and vegetables；On the other hand, under the conditions of commodity production, due to ring
The uncertainty of border condition, preventive effect or does not reach requirement or unstable, and compared with chemical bactericide control cost compared with
It is high；For Antagonistic Fungi as antistaling agent, usage mode is convenient not as good as chemical bactericide.In addition, antagonism yeast needs under higher concentration
Very big economic cost pressure could will be also generated the characteristics of useful effect.These factors affects Antagonistic Fungi is protected as fruits and vegetables
Fresh dose of business in production uses.Therefore the effect of antagonism yeast biological control must be further improved and improve.
Biocontrol effect is an important indicator for determining Antagonistic Fungi good job, and determines that can microorganism Antagonistic Fungi be applied to
The foundation of actual production.Recent years studies have shown that can from it is following it is several in terms of improve the biocontrol effect of Antagonistic Fungi.1.
Being used in mixed way Antagonistic Fungi using the mixture of Antagonistic Fungi has three big advantages: (1) can widen sphere of action, prevent and treat a variety of cause of diseases
Bacterium；(2) validity can be enhanced, reduce frequency of use；(3) it allows to combine different biological control features.2. and chemical sterilization
Agent is used in combination biological agent and is effectively combined with chemical bactericide, so that with sterilization more less than conventional antimicrobial agent dose
Agent control disease is possibly realized, so that the usage amount of fungicide is reduced to the safe dose of human body level below, is reduced
The pesticide residue of fruits and vegetables.3. antagonistic microbe group number can be increased using certain adjuvants using adjuvant and enhance its survival rate,
And then increase the biological and ecological methods to prevent plant disease, pests, and erosion ability of Antagonistic Fungi.4. being used in mixed way the 5. many researchs of effect for being combined other measures with certain metal ions
Show the methods of to be heat-treated, refrigerate to be used in combination with biocontrol bacterium, the fungistatic effect of Antagonistic Fungi can be reinforced.
Bamboo-leaves flavones are the biological flavones with physiological activity extracted from the leaf of bamboo, are a kind of efficient biologiesIt is anti- Oxidant(Anti-Oxidant), modem animal toxicological experiment sufficiently demonstrates the security reliability of bamboo-leaves flavones.As the leaf of bamboo
Function extract, bamboo-leaves flavones are rich in Flavonoid substances, are widely used in food additives.The leaf of bamboo contain natural activity at
Point, there is various biological function effects and extraordinary radicals scavenging effect.In addition, as food additives, research
Show that bamboo-leaves flavones are used in combination as antioxidant and antagonism yeast, activities of antioxidant enzymes and antagonism yeast can be improved to oxygen
The tolerance for changing stress, to improve its biological control effect to fruit postharvest diseases.
Cali's Bick Pichia pastoris has been proved to possess antagonistic potencies, and the present invention is finished using bamboo-leaves flavones and Cali's Bick
Red yeast, which is used in combination, improves its effect to biologically controlling postharvest disease of fruit, to reduce the rotting rate of fruit, Jin Erda
The purpose fresh-keeping to fruit storage.
Summary of the invention
The present invention is used in combination using bamboo-leaves flavones (BLF) and Cali's Bick Pichia pastoris, is improved the yeast and is adopted to fruit
The biological control effect of disease afterwards.Compared with Cali, Bick Pichia pastoris is individually handled, bamboo-leaves flavones and Cali's Bick finish red ferment
Mother is used in combination, and can significantly enhance the yeast to the inhibiting effect of green mold of apple, prevent and treat its postharvest decay.Present invention tool
Have safety, efficiently, it is at low cost the advantages that, can be widely used for during the biological control of fruit postharvest diseases, reduce fruit
It is lost caused by postharvest disease.
The technical solution adopted by the present invention
The present invention provides the methods of bamboo-leaves flavones combination Cali Bick Pichia pastoris prevention and treatment Biocontrol of Postharvest Diseases in Apple Fruits, under
It states step progress: Cali's Bick Pichia pastoris (Pichia caribbica) being activated, with NYDB culture medium culture Cali Bick
Pichia pastoris, centrifugation obtain thallus, prepare 1 × 10 containing bamboo-leaves flavones8The yeast bacteria suspension of cells/mL；Bacteria suspension is uniform
It is sprayed on fruit, natural air drying；It is placed in plastic crate, after being sealed with preservative film, is placed under the conditions of room temperature (20 DEG C) and stores.
The wherein NYDB culture medium are as follows: beef extract 8g, yeast extract 5g, glucose 10g, distilled water 1000ml, pH
It is natural.
Wherein the preparation containing bamboo-leaves flavones 1 × 108Cells/mL yeast-like fungi suspension method, as steps described below into
Row: the dilution of thallus sterile water is prepared into 2 × 108Cells/mL bacteria suspension is the bamboo-leaves flavones of twice of final concentration with concentration
1:1 is mixed by volume, and the ultimate density of bamboo-leaves flavones is 100 μ g/mL-500 μ g/mL.
Advantages of the present invention:
(1) Cali's Bick Pichia pastoris (Pichia caribbica) used in the present invention is this laboratory screening,
Antagonistic potencies are strong, harmless.
(2) bamboo-leaves flavones (BLF) used in the present invention are from a wealth of sources, cheap, nontoxic.
(3) present invention is used in combination using bamboo-leaves flavones and Cali's Bick Pichia pastoris, at exclusive use bamboo-leaves flavones
Reason or the processing of Cali's Bick Pichia pastoris are compared, and the antagonistic activity of Cali's Bick Pichia pastoris can be significantly improved, to reduce
The rotting rate of fruit, and then achieve the purpose that fruit storage is fresh-keeping.
(4) present invention is used in combination using bamboo-leaves flavones and Cali's Bick Pichia pastoris, and it is anti-to can replace chemical bactericide
Fruit postharvest diseases are controlled, the harm using chemical bactericide to people is avoided, there is significant economic benefit and social benefit.
Detailed description of the invention
Fig. 1 is that the control effect to green mold of apple is used in combination with Cali's Bick Pichia pastoris for bamboo-leaves flavones.Note: CK:
Sterile distilled water, i.e. control group；Pc:1 × 108The P.carribbica yeast bacteria suspension of cells/ml；0.01%BLF:100 μ
The bamboo-leaves flavones (BLF) of g/mL；The bamboo-leaves flavones (BLF) of 0.025%BLF:250 μ g/mL；The bamboo of 0.05%BLF:500 μ g/mL
Leaf flavones (BLF)；0.01%BLF+Pc:1 × 108The P.carribbica yeast bacteria suspension of cells/ml (contains final concentration of 100
The BLF of μ g/mL)；0.025%BLF+Pc:1 × 108The P.carribbica yeast bacteria suspension of cells/ml is (containing final concentration of
The BLF of 250 μ g/mL)；0.05%BLF+Pc:1 × 108The P.carribbica yeast bacteria suspension of cells/ml is (containing final concentration of
The BLF of 500 μ g/mL).Different letters represent significant difference (P < 0.05).
The influence rotted naturally to apple is used in combination with Cali's Bick Pichia pastoris for bamboo-leaves flavones in Fig. 2.Note: CK: nothing
Bacterium distilled water, i.e. control group；The bamboo-leaves flavones of 0.01%BLF:100 μ g/mL；Pc:1 × 108Cells/ml's
P.carribbica yeast bacteria suspension；0.01%BLF+Pc:1 × 108The P.carribbica yeast bacteria suspension of cells/ml (contains
The BLF of final concentration of 100 μ g/mL).Different letters represent significant difference (P < 0.05).
Specific embodiment
By the way that the present invention will be explained in more detail by following embodiment.Following embodiment is merely illustrative, this
Invention is not limited by these embodiments.
This laboratory screening of embodiment 1 Cali Bick Pichia pastoris (P.carribbica) system obtains, and applies for Chinese invention
Patent simultaneously authorizes, patent No. ZL201010198119.6, patent name Cali Bick Pichia pastoris, in fruit storage is fresh-keeping
Using and application method.It is protected in NYDA (adding 2% agar (quality meter) on the basis of NYDB culture medium) 4 DEG C of low temperature of culture medium
It deposits.Yeast extract 5g in NYDB culture medium, beef extract 8g, glucose 10g, purified water l000ml, natural pH.
It cultivates program are as follows: (1) solid activates: Cali's Bick Pichia pastoris is inoculated in NYDA culture medium, trained at 28 DEG C
Support 48h；(2) Liquid Culture: it is packed into the NYDB seed culture medium of 50ml in the triangular flask of 250ml, accesses a ring with oese
Activated Cali's Bick Pichia pastoris, cultivates 20h under the conditions of 180rpm, 28 DEG C；(3) it is centrifugated settling flux: above-mentioned ferment
Female culture mix is centrifuged 10min under the conditions of 8000g, and is washed 2 times with sterile distilled water, to remove culture medium, with nothing
Bacterium distilled water settling flux again, and adjusting barm cell concentration with blood counting chamber is 2 × 108Cells/ml, then with sterilized
Distilled water or bamboo-leaves flavones (BLF) solution be diluted to required concentration.
The inhibitory effect to apple Postharvest Penicillium is used in combination with Cali's Bick Pichia pastoris for 2 bamboo-leaves flavones of embodiment
One) testing program
Pass through fine selection for the fruit of examination, as far as possible selection fruit uniformity, surface is without scratch！Machinery wound and disease pest wound
Evil.Apple is punched with aseptic card punch, superficial cut is 5mm (diameter) × 3mm (depth).30 μ l are added in each wound equivalent
Following treatment fluid: (1) 1 × 108Cali's Bick Pichia pastoris suspension (containing each concentration bamboo-leaves flavones) of cells/mL；(2)1×
108Cali's Bick Pichia pastoris bacteria suspension of cells/mL；(3) sterile water.After 3h, then to fruit wound it is inoculated with 30 μ L respectively
Following pathogenic bacteria spore suspension: the wound Xiang Pingguo inoculation 5 × 104Penicillium expansum (the Penicillium of spores/mL
Expanusm) spore suspension.After natural drying, fruit is put into plastic crate and is sealed with preservative film, trained in constant incubator
It supports (humidity 95%), is placed in 20 DEG C.Every processing is repeated 3 times, and repeats 12 fruits every time.Entire test is repeated 2 times.After 8 days, note
Fruit disease incidence is recorded, with the fungistatic effect of this evaluation card Libiee gram Pichia pastoris.
The calculation formula of disease incidence is as follows:
Disease incidence (%)=morbidity fruit/fruit sum × 100%
Two) test result
It is tested according to above-mentioned steps, the rotting rate result for counting fruit is as follows:
As shown in fig. 1, the disease incidence of control group is about 91.7%, and individual yeast processing group disease incidence is 22.2%,
Therefore P.caribbica can effectively control the postharvest disease that apple is caused by penicillium expansum at 20 DEG C.At the same time, with card
Libiee gram Pichia pastoris individually compare by processing group, and bamboo-leaves flavones can improve the biocontrol effect of yeast significantly.Wherein 100ug/ml
BLF combination Cali Bick Pichia pastoris processing group has best anti-corrosion effect, and green mold of apple disease disease incidence is only
5.6%, i.e. effect is the most significant.
3 bamboo-leaves flavones of embodiment are used in combination to naturally rotten and storage quality the shadow of apple with Cali's Bick Pichia pastoris
It rings
One) testing program
Following treatment fluid is uniformly sprayed on fruit surface, naturally dry: (1) 1 × 108Cali's Bick of cells/mL is finished red
Yeast bacteria suspension (BLF concentration is 100 μ g/mL)；(2)1×108Cali's Bick Pichia pastoris bacteria suspension of cells/mL；(3)
BLF (concentration is 100 μ g/mL) and (4) sterile water.Then, the fruit handled well is put into plastic crate to be sealed with preservative film.20℃
(RH95%) it is stored 35 days in constant incubator.Rot fruit number is counted every 5 days taking-up fruit since the 10th day, and
Observe Decay.After 35 days, fruit storage quality is surveyed.Every processing is repeated 3 times, and entire experiment is repeated 2 times.Bamboo-leaves flavones and card
The combined use of Libiee gram Pichia pastoris indicates the inhibitory effect that fruit rots naturally with disease incidence.
1, the calculation formula of disease incidence is as follows:
Disease incidence (%)=morbidity fruit/fruit sum × 100%
2, quality determination method is as follows:
(1) total dissoluble solid content: being measured with hand-held saccharometer, be measured using saccharometer, is measured
As a result it is expressed as g/100g sample.
(2) brown stain degree: weighing 20g pulp from 6 fruits, 40ml distilled water added to grind rapidly, 10000 × g, 4 DEG C of items
It is centrifuged 10mim under part, 10mim is centrifuged under the conditions of taking supernatant 10ml that 15ml, 10000 × g, 4 DEG C of 95% ethyl alcohol is added, takes supernatant
Liquid measures absorbance value in 420nm, indicates browning degree with this.3 repetitions of every processing.
(3) titrable acidity: using titration, samples from 5 fruit, weighs 10g sample, distilled water is added to grind rapidly,
It is settled to 100mL, is filtered, is taken 10mL filtrate, carried out being titrated to pH 8.1 with 0.1mol/L NaOH, be calculated after titration, is tied
Fruit is indicated with the percentage containing malic acid.
(4) Vc content: 2,6- dichloropheno-lindophenol methods.5g sample is taken to add the grinding homogenate of 2% oxalic acid of 5mL from 5 fruit,
100mL volumetric flask is poured into, washs residue with 1% oxalic acid, adds 30% zinc sulfate of 1mL, shakes up.Again plus 15% ferrocyanide of 1mL
Potassium (depigmentation) shakes up filtering with 1% oxalic acid constant volume.Filtered fluid 4mL is taken, 2,6- dichloropheno-lindophenol dye then is added to each pipe
Expect solution, dimethylbenzene 5mL is added immediately, shakes about 0.5min rapidly, dimethylbenzene is separated with water layer after standing, by upper layer dimethylbenzene
Extract liquor measures absorbance under 500nm wavelength.Standard curve is looked into according to the OD value of measurement liquid, finds out corresponding vitamin
As a result the content of C (Vc) is indicated with milligram Vc/100g fruit.
Two) test result
1, it is tested according to above-mentioned steps, the natural rotting rate result of statistics is as follows:
Figure it is seen that after 35 days, the independent processing group of BLF, the independent processing group of Cali's Bick Pichia pastoris, Cali's ratio
Gram Pichia pastoris and the Apple disease incidence of BLF combination processing are substantially less than control group (79.4%).Bamboo-leaves flavones and card
Libiee gram Pichia pastoris combination processing group fruit rotting rate is minimum, and only 8.3%, this, which is substantially less than, is used alone BLF and Cali
The natural rotting rate of Bick Pichia pastoris.Illustrate that BLF (concentration is 100 μ g/mL) can significantly improve Cali's Bick Pichia pastoris
To the control efficiency of Biocontrol of Postharvest Diseases in Apple Fruits.
2, it is tested according to above-mentioned steps, the storage quality result of statistics is as follows:
The influence to Apples quality is used in combination with Cali's Bick Pichia pastoris for 1 bamboo-leaves flavones of table
Note: CK: sterile distilled water, i.e. control group；The bamboo-leaves flavones of 0.01%BLF:100 μ g/mL；Pc:1 ×
108The P.carribbica yeast bacteria suspension of cells/ml；0.01%BLF+Pc:1 × 108The P.carribbica of cells/ml
Yeast bacteria suspension.Different letters represent significant difference (P < 0.05).
As it can be seen from table 1 after 35 days, compared with the control group, the apple through BLF Yu Cali's Bick Pichia pastoris combination processing
Fruit its Vc content, titratable acid and brown stain degree are significantly reduced without significant difference, soluble solid, not to every index of quality
Generate adverse effect.
Claims (1)
1. the method that bamboo-leaves flavones combination Cali Bick Pichia pastoris prevents and treats Biocontrol of Postharvest Diseases in Apple Fruits carries out as steps described below: will
Cali's Bick Pichia pastoris (Pichia caribbica) activation, with NYDB culture medium culture Cali Bick Pichia pastoris, centrifugation
Thallus is obtained, 1 × 10 containing bamboo-leaves flavones is prepared8The yeast bacteria suspension of cells/mL；Bacteria suspension is uniformly sprayed on fruit,
Natural air drying；It is placed in plastic crate, after being sealed with preservative film, puts and store at 20 °C；
The wherein NYDB culture medium are as follows: 8 g of beef extract, 5 g of yeast extract, 10 g of glucose, distilled water 1000 ml, pH
It is natural；
Wherein the preparation containing bamboo-leaves flavones 1 × 108 Cells/mL saccharomycete suspension method carries out as steps described below:
The dilution of thallus sterile distilled water is prepared into 2 × 108 Cells/mL bacteria suspension is the bamboo-leaves flavones of twice of final concentration with concentration
1:1 is mixed by volume, and the ultimate density of bamboo-leaves flavones is 100 μ g/mL；
Wherein the fruit is apple.
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