CN109645107A - The fruit disease control method of resistant activity is lured based on rhodosporidium toruloides cell wall (1 → 3)-callose - Google Patents
The fruit disease control method of resistant activity is lured based on rhodosporidium toruloides cell wall (1 → 3)-callose Download PDFInfo
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- CN109645107A CN109645107A CN201910010053.4A CN201910010053A CN109645107A CN 109645107 A CN109645107 A CN 109645107A CN 201910010053 A CN201910010053 A CN 201910010053A CN 109645107 A CN109645107 A CN 109645107A
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- fruit
- cell wall
- callose
- rhodosporidium toruloides
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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
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Materials Engineering (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Sustainable Development (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The present invention proposes a kind of fruit disease control preparation that resistant activity is lured based on rhodosporidium toruloides cell wall (1 → 3)-callose: containing 1~5g rhodosporidium toruloides cell wall (1 → 3)-callose in every liter of preparation, surplus is water.The present invention also propose it is a kind of utilize above-mentioned preparation carry out fruit disease control method, step are as follows: preparation is inoculated in the wound of fruit, make it that fruit wound be completely covered, after the preparation natural air drying of fruit wound, which is put into container and is saved under sealing state.The present invention is conducive to change the fruit fungal disease control method based on dependence chemical bactericide for a long time, and food-safe, environmental protection and growth of agricultural efficiency, increasing peasant income and economic society sustainable development etc. are of great significance.
Description
Technical field
The present invention relates to postharvest diseases of fruit Prevention Technique field, it is especially a kind of by rhodosporidium toruloides cell wall (1 →
3)-callose induction fruit resistance ability biological way of keeping fresh.
Background technique
Fruit is the important sources of vitamin, minerals and dietary fiber, is that people maintain health, augment nutritional must
Indispensable Main Foods are the agricultural product for being only second to grain in the world.But huge, root is lost caused by fruit postharvest diseases
According to generally conservative estimation, loss is 20%~25% or so after China's fruit is adopted, serious to affect its market supply and agriculture
The economic well-being of workers and staff of the people.There are many factor lost after causing fruit to be adopted, and rot to be that fruit is caused to adopt as caused by pathogen infection
After the main reason for losing, wherein mould is fruit Main postharvest pathogens.Cause the fungal species to rot during fruit storage
Very much, but the pathogenic bacteria of every kind of fruit dominance generally have one kind.For example, apple postharvest decay is mainly penicillium expansum
Penicilliosis caused by (Penicillium expansum) and the gray mold as caused by grey grape (Botrytis cinerea);Mandarin orange
Tangerine postharvest decay is mainly penicilliosis and Penicillium digitatum caused by Italian mould (Penicillium italicum)
Green mould caused by (Penicillium digitatum);Strawberry postharvest decay is mainly rhizopus stolonifer (Rhizopus
Stolonifer gray mold caused by head mold disease caused by) and Botrytis cinerea (Botrytis cinerea) etc..These pathogens
It relies primarily on the natural openings such as fruit wound or stomata, hole skin and infects.Moreover, disease fungus is to adopting the harm of rear fruit not only
Be heavy losses caused by it will lead to fruit quantitatively, and due to many disease fungus can secrete generate it is many secondary
Metabolite is so as to cause serious food-safety problem, the stick generated such as the aflatoxin that Aspergillus generates, penicillium expansum
Aspergillin, the botrydial etc. that Botrytis cinerea generates.
The main method of control postharvest diseases of fruit has cryopreservation and uses chemical bactericide at present.Cryopreservation technology
Though can extend fruits and vegetables shelf life to a certain extent, the cold chain technology in China's transport is very not perfect, and fruit is refrigerating
Phenomenon easily is damaged to plants caused by sudden drop in temperature in the process, seriously affects the fresh-keeping effect of fruits and vegetables.For a long time, both at home and abroad to the control master of postharvest disease
To depend on chemical bactericide.Chemical bactericide has the clear mechanism of action, potency height and effect stability, and can control and adopt preceding dive
The advantages that volt infection, so being up to now still the main method for controlling postharvest disease.But chemical bactericide long-term and
It is a large amount of to use, environment is seriously polluted, human health is impaired.On the other hand, single chemical bactericide, pathogen is used for a long time
May evolve and form drug resistance, directly result in allow at present using some fungicide to the control efficiency of disease worse and worse,
Such as thiabendazole and imazalil class fungicide.Therefore, it is low to find toxicity, preventive effect height, the fungicide of low-residual and environmental protection
Substitute it is very urgent.
Plant forms a set of complicated and effective defence machine during common evolutionary long-term with pathogen
System, when encountering pest infestation, this defense mechanism can be induced, and then a series of defence for starting elaborates is anti-
It answers, the generation of harm is mitigated or eliminated, shows as resistance.Disease resistance of plant can not only be induced by biotic factor, and can
It is induced by some chemokines and physical agent.Wherein, Chemical elicitor production, transport, storage are convenient, at low cost, induction
Easy to operate, effect stability, application is relatively broad." a kind of pyridyl-pyrimidine alcohol compound is in plant inducer for Chinese invention patent
Application in impedance " application number: 201310535851.1, it provides a kind of pyridyl-pyrimidine alcohol compound and is induced in plant
Application in resistance, the compound can significantly improve rapidly the ability of plant resistant pathogen infection.Chinese invention patent
" a method of excitation rice Induced insect resistance ", application publication number: CN105145572A is disclosed a kind of by fluorobenzene oxygen second
Acid improves rice and flies the resistance of wind to rice to mitigate rice and fly wind to the method for the harm of rice.A kind of Chinese invention patent " use
The plant extracts to powder mildew resistance is generated in evoking tobacco ", application number: 201610544509.1, it discloses one kind and is used for
Evoking tobacco generate to the plant extracts of powder mildew resistance (Ginger P.E, soapberry extract, extra large coconut extractive,
Loquat extract) preparation method.
Beta glucan is a kind of polysaccharide being widely present in microorganism, mushroom and plant, it is composition higher plant, ferment
One of female bacterium and the structural macromolecules of fungal cell wall.Have compared with strong biological activity beta glucan important sources first is that ferment
Mother accounts for the 35-60% of cell wall dry weight.Yeast dextran have extensive physiological function, including strengthen immunity, it is anti-infective,
Anti-radiation and reduction blood lipid etc..A kind of Chinese invention patent " side that alkali-insoluble glucan is extracted from beer waste yeast residue
Method " a kind of method that alkali-insoluble glucan is extracted from beer waste yeast residue is disclosed, it is opened for the comprehensive utilization of waste yeast
A new approach has been warded off, has realized appreciating and enlarging for beer waste yeast.A kind of Chinese invention patent " anti-Ichemia reperfusion injury
Drug ", application number 01114983.3, provide it is a kind of using (1 → 3)-callose as the anti-ischemic of effective component with fill again
Infuse damage medicine and application thereof.Chinese invention patent " a kind of raising immunity, the health food of reducing blood lipid and preparation method thereof ",
Application number 201210516557.1 discloses a kind of having containing yeast beta-dextran and improves immunity, effect for reducing blood fat
Health food.Chinese invention patent " peracid beverage products and the method for extending probiotic stability ", application number
201080053351.9, it discloses the beverage comprising at least one fruit juice, at least one sweetener, probiotics and beta glucan and produces
Product, wherein beta glucan can increase the shelf life of the product." a kind of reduction cholesterol is antiviral for Chinese invention granted patent
Feed addictive and its production technology and application ", publication number CN101637220 discloses a kind of reduction gallbladder containing beta glucan
Sterol anti-virus feed additive and its production technology and application.Chinese invention patent " the plant containing yeast cell extract
Plant elicitors ", application number 200410020099.8 discloses a kind of plant disease-resistant induction containing yeast cell extract
Agent, main component are (1 → 3)-callose, are that a kind of disease-resistant spectrum is wide, the lasting period is long, safety non-pollution, have much application
The plant disease resistance inductor of potentiality.
Summary of the invention
Under conditions of not using chemical bactericide the technical problem to be solved in the present invention is to provide one kind, using chemi-excitation
Son inhibits the preservation technology of postharvest diseases of fruit.
In order to solve the above-mentioned technical problem, the present invention proposes a kind of based on rhodosporidium toruloides cell wall (1 → 3) Portugal-β-D-
Glycan lures the fruit disease of resistant activity to control preparation:
Contain 1~5g rhodosporidium toruloides cell wall (1 → 3)-callose in every liter of preparation, surplus is water.
Note: preparation of the invention is suspension.
As the present invention is based on rhodosporidium toruloides cell wall (1 → 3)-calloses, and the fruit disease of resistant activity to be lured to control
The improvement of preparation:
Contain 2g rhodosporidium toruloides cell wall (1 → 3)-callose in every liter of preparation, surplus is water.
As the present invention is based on rhodosporidium toruloides cell wall (1 → 3)-calloses, and the fruit disease of resistant activity to be lured to control
The further improvement of preparation:
Rhodosporidium toruloides cell wall (1 → the 3)-callose the preparation method is as follows:
S1, preparation rhodosporidium toruloides cell wall: rhodosporidium toruloides cell is subjected to Mechanical Crushing, obtains rhodosporidium toruloides cell
Wall;
S2, preparation rhodosporidium toruloides cell wall (1 → 3)-callose, comprising the following steps:
2.1, by rhodosporidium toruloides cell wall yeast cell wall obtained by step S1 according to the ratio of 1g:48~52mL be added to
Concentration is, in 70~80 DEG C of 4~8h of reaction, to shake up once during reaction every 30 ± 5min in the NaOH solution of 3% (g/mL),
Obtain reactant;
2.2, step 2.1 gained reactant is centrifuged 40~50min under the conditions of 7500~8000g, obtains sediment;
2.3, the sediment after cleaning is added according to the solid-liquid ratio of 1g:48~52mL for 2.2 gained sediment of cleaning step
The CH of 0.5mol/L3In COOH solution, 2~4h, centrifugal filtration are hydrolyzed in 85~95 DEG C of water-bath;Repeat above-mentioned steps
1~2 time, it will be freeze-dried that (vacuum degree 0.2Pa, -40 DEG C of temperature, the time is after the cleaning of final centrifugal filtration products therefrom
36h), (1 → 3)-callose powder is obtained.
Note: above-mentioned rhodosporidium toruloides (Rhodosporidium paludigenum&Fell Tallman) are preserved in Britain
International fungal studies institute's International Agriculture and Bio-Centers genetic resources collection (International Mycological
Institute, CABI Genetic Resource Collection), deposit number: IMI394084, in Patent No.
Preservation is submitted to prove in 200610155209.0 patent of invention " a kind of the bio-preservative of fruit and vegetable and preparation method thereof ".
As the present invention is based on rhodosporidium toruloides cell wall (1 → 3)-calloses, and the fruit disease of resistant activity to be lured to control
The further improvement of preparation:
The step S2 prepares rhodosporidium toruloides cell wall (1 → 3)-callose, comprising the following steps:
2.1, rhodosporidium toruloides cell wall yeast cell wall is added according to the ratio of 1g:50mL to concentration is 3% (g/
ML it in NaOH solution), in 75 DEG C of reaction 6h, is shaken up once during reaction every 30min;
2.2, step 2.1 gained reactant is centrifuged 45min under the conditions of 7800g, obtains sediment;
2.3, by step 2.2 gained sediment 3 times wash with distilled water;It will be after cleaning according to the solid-liquid ratio of 1g:50mL
The CH of sediment addition 0.5mol/L3In COOH solution, 3h, centrifugal filtration are hydrolyzed in 90 DEG C of water-bath;Repeat above-mentioned
Step 1 time is freeze-dried (vacuum degree after being neutrality to pH wash with distilled water for final centrifugal filtration products therefrom
0.2Pa, -40 DEG C of temperature, time 36h), obtain (1 → 3)-callose powder.
Note: filtering gained precipitating is replaced into step 2.2 gained sediment, it is primary to repeat above-mentioned steps, that is, will filter
Gained precipitates the CH that isometric 0.5mol/L is added again3In COOH solution, 3h is hydrolyzed in 90 DEG C of water-bath, was centrifuged
Filter;
In order to solve the above-mentioned technical problem, the present invention utilizes above-mentioned preparation, proposes a kind of based on rhodosporidium toruloides cell wall
(1 → 3)-callose lures the fruit disease control method of resistant activity, comprising the following steps:
Preparation is inoculated in the wound of fruit, makes it that fruit wound be completely covered, to fruit wound preparation natural wind
After dry, which is put into container and is saved under sealing state.
Note: general pathogen infection fruit is since the edge of fruit wound, therefore the principle of above-mentioned inoculation is preparation
Fruit wound can be completely covered, that is, when fruit wound diameter is 5mm, inoculum concentration is 30 μ L when depth 2mm.When fruit wound
Diameter is 3mm, and inoculum concentration is 20 μ L when depth 2mm.
Above-mentioned by the fruit is exactly to carry out induction of resistance response, specific embodiment party of the present invention to it under state in being sealed
In 25 DEG C in formula, the lower induction of relative humidity 90% is to have optimal induction time when for 24 hours in order to illustrate the present invention for 24 hours.
As the present invention is based on rhodosporidium toruloides cell wall (1 → 3)-calloses, and the fruit disease of resistant activity to be lured to control
The improvement of method:
After the preparation natural air drying of fruit wound, which is put into container after being sealed using preservative film, in room temperature
It is saved under (15~35 DEG C).
As the present invention is based on rhodosporidium toruloides cell wall (1 → 3)-calloses, and the fruit disease of resistant activity to be lured to control
Further improvements in methods:
The fruit is pear fruit.
Compared with prior art, the present invention having following technical advantage:
(1) rhodosporidium toruloides cell wall (1 → 3)-callose employed in the present invention is resourceful, at low cost
It is honest and clean, cost performance is high, be easy production, transport, have biodegradability;
(2) induction of resistance is a kind of natural reaction generated after plant is infected by pest and disease damage, can generate and hold to pest and disease damage
Long and system resistance of wide spectrum;
(3) postharvest disease that the present invention can significantly reduce fruit has to environmental and human health impacts without any toxic action
The features such as economical and practical, safe and efficient, environmental-friendly, has good social benefit and ecological benefits.
Detailed description of the invention
Specific embodiments of the present invention will be described in further detail with reference to the accompanying drawing.
Fig. 1 is that various concentration rhodosporidium toruloides cell wall (1 → 3)-callose is anti-to the induction of pear fruit anti-penicilliosis
The influence schematic diagram of property;As a result it is obtained for detection in the 3rd day after access pathogen.Wherein figure (a) is disease incidence, and figure (b) is scab
Diameter;Different letters represent the significance of difference (P=0.05).
Fig. 2 is influence signal of various concentration oat (1 → the 3)-callose to the anti-penicilliosis induction of resistance of pear fruit
Figure;As a result it is obtained for detection in the 3rd day after access pathogen.Wherein figure (a) is disease incidence, and figure (b) is lesion diameter;Different letters
Represent the significance of difference (P=0.05).
Fig. 3 is various concentration brewing yeast cell wall (1 → 3)-callose to the anti-penicilliosis induction of resistance of pear fruit
Influence schematic diagram;As a result it is obtained for detection in the 3rd day after access pathogen.Wherein figure (a) is disease incidence, and figure (b) is that scab is straight
Diameter;Different letters represent the significance of difference (P=0.05).
Fig. 4 is various concentration rhodosporidium toruloides cell (1 → 3)-callose to the anti-penicilliosis induction of resistance of pear fruit
Influence schematic diagram;As a result it is obtained for detection in the 3rd day after access pathogen.Wherein figure (a) is disease incidence, and figure (b) is that scab is straight
Diameter;Different letters represent the significance of difference (P=0.05).
Specific embodiment
The present invention is described further combined with specific embodiments below, but protection scope of the present invention is not limited in
This.
Embodiment 1 lures the fruit disease of resistant activity to control system based on rhodosporidium toruloides cell wall (1 → 3)-callose
Agent (hereinafter referred to as preparation), the preparation method is as follows:
1), rhodosporidium toruloides cell prepares and (belongs to the prior art):
Deposit number is that the rhodosporidium toruloides of IMI394084 are stored in NYDA culture medium (beef extract under low temperature (4 DEG C)
8g, yeast extract 5g, glucose 10g, agar 20g are settled to 1000mL with water, and (lower 121 DEG C of 0.1MPa go out high pressure steam sterilization
Bacterium 20min) in, when activation, takes out;The rhodosporidium toruloides are cultivated 48h (25 DEG C) in NYDA culture medium, repeats secondary culture 2
After secondary, activated rhodosporidium toruloides are inoculated into NYDB with oese, after being cultivated for 24 hours under the conditions of 200rpm, 28 DEG C, are received
Collect culture solution;
Gained culture solution is collected into rhodosporidium toruloides thallus in 3000g (centrifugal force) centrifugation 10min, and uses sterile distilled water
3 rhodosporidium toruloides thallus are cleaned to remove remaining culture medium, obtain rhodosporidium toruloides cell.
2) rhodosporidium toruloides cell wall, is prepared: preparation rhodosporidium toruloides cell wall, the specific steps are as follows:
Rhodosporidium toruloides cell obtained by step 1) is resuspended in the phosphate buffer of 0.2mol/L (PBS, pH 8.0) to match
It is made 1 × 108The bacteria suspension of cell/mL.0.5g pickling glass pearl (Sigma company, partial size are added into the bacteria suspension of 0.5mL
425-600 μm), smudge cells are ground in (frequency 70Hz) on automatic sample grinder, 3min/ circulation handles 5 circulations,
4000rpm is centrifuged 10min in refrigerated centrifuge (4 DEG C) after broken, with sterile water washing 8 times, collects precipitating, and in 121 DEG C
Sterilize 20min, and freeze-drying (vacuum degree 0.2Pa, -40 DEG C of temperature, time 36h) is afterwards up to rhodosporidium toruloides cell wall.
3) rhodosporidium toruloides cell wall (1 → 3)-callose, is prepared:
3.1, rhodosporidium toruloides cell wall obtained by step 2) is added according to the ratio of 1:50 (g/mL) to concentration is 3%
(g/mL) it in NaOH solution, in 75 DEG C of reaction 6h, is shaken up once during reaction every 30min, obtains reactant.
3.2, step 3.1 gained reactant is centrifuged 45min under the conditions of 7800g, obtains sediment;
3.3, by step 3.2 gained sediment 3 times wash with distilled water;After being cleaned according to the solid-liquid ratio of 1:50 (g/mL)
Sediment be added to the CH of 0.5mol/L3In COOH solution, 3h, centrifugal filtration are handled in 90 DEG C of water-bath;
It is primary that precipitating obtained by centrifugal filtration is repeated into above-mentioned steps, that is, the precipitating is used into corresponding volume again
The CH of 0.5mol/L3COOH solution brings up again once centrifugal filtration;
(vacuum degree is freeze-dried after being neutrality to pH wash with distilled water for products therefrom after final centrifugal filtration
0.2Pa, -40 DEG C of temperature, time 36h), (1 → 3)-callose powder is obtained, is saved backup in -20 DEG C.
4) it, is prepared using (1 → 3)-callose powder obtained by step 3.3 and sterile water and obtains preparation, is i.e. configuration pair
Answer the bacteria suspension of (1 → 3)-callose of concentration as preparation.
5g (1 → 3)-callose powder is taken in the present embodiment, sterile distilled water is added to be diluted to 1L, and being configured to concentration is
0.5% (1 → 3)-callose solution.
Test 1, influence of (1 → 3)-callose difference induced concentration to pear fruit penicilliosis resistance:
1, experimental material:
Fruit is pears, and kind is Shuijing Pear.
Pathogen: penicillium expansum (Penicillium expansum), 25 DEG C activation 7 days it is spare.
2, it handles:
(1) fruit pre-processes: it chooses that appearance is neat, no disease and pests harm, the fruit having no mechanical damage, is first cleaned with tap water,
Then it immerses again in 0.1% liquor natrii hypochloritis and sterilizes 2min, take out, then rinsed well with tap water, wash away remaining chlorine
Sour sodium dries spare.
(2) unified size (5mm) and depth as identical as possible are formed on the surface of each fruit with sterilized punch
Spend wound 6 of (2mm).Each wound be separately added into equivalent (30 μ L) water (control) and concentration be 0.01%,
0.1%, (1 → 3)-callose solution of 0.2%, 0.5%, 1% concentration, is being placed at room temperature for 2h, after evaporating moisture, in
It is induced for 24 hours under the conditions of 25 DEG C of constant temperature, constant humidity (relative humidity 90%).
Preparation in this experiment with water as a control group, respectively with 0.01%, 0.1%, 0.2%, 0.5%, 1% concentration
Preparation of (1 → 3)-callose solution as each concentration processing group.
(1 → 3)-callose solution of above-mentioned 0.5% concentration is that embodiment 1 prepares the Portugal resulting (1 → 3)-β-D-
Glycan solution, that is, indicate to contain 5g (1 → 3)-callose in every liter of (1 → 3)-callose solution, surplus is water.
Remaining processing group and so on.
(3) 1 × 10 is accessed in wound430 μ of spores/mL penicillium expansum Penicillium expansum spore suspension
L, (25 DEG C) storages at normal temperature after being disposed, and gone bail for wet process with PE plastic film seal, knot is observed and recorded after timing
Fruit is compared various concentration (1 → 3)-callose effect, as a result straight with average attack rate (%) and average scab
Diameter (mm) indicates.Choosing 9 fruits is one group of repetition, and 3 repetitions, experiment is repeated twice, and is subject to identical result.
Influence of (1 → 3)-callose difference induced concentration to pear fruit penicilliosis resistance is as shown in Figure 1.
3, result:
As shown in Figure 1, each concentration processing group of (1 → 3)-callose is compared with the control group, the hair of pear fruit penicilliosis
State of an illness condition obviously weakens, wherein the most obvious with the processing group effect that (1 → 3)-callose concentration is 0.2%.It is connecing
Enter after pathogen the 3rd day, compared with the control group, the disease incidence and lesion diameter of pear fruit wound reduce by 70.37% He respectively
8.22mm illustrates that (1 → 3)-callose effectively slows down the occurrence and development of penicilliosis.Although as (1 → 3)-β-
The increase of D- glucan concentration, the inhibitory effect of penicilliosis are declined, but compared with the control group, (1 → 3) Portugal-β-D- is poly-
The processing group disease incidence and lesion diameter that sugared concentration is 1% still decline significantly.
Therefore, (1 → 3)-callose of the present invention 0.2% is the most suitable concentration for the treatment of for inducing pear fruit resistance.
Comparative experiments 1, will experiment 1 in (1 → 3)-callose by " rhodosporidium toruloides cell wall preparation gained (1 →
3)-callose " is changed to " purchase poly- in oat (1 → 3) Portugal-β-D- of Hangzhou Zhong Zhikang mushroom Bioisystech Co., Ltd
Sugar ", remaining is equal to experiment 1.
As a result as shown in Figure 2.
Fig. 2 shows that (1 → 3)-callose from oat cannot be such that the mold decay of pear fruit is delayed
Solution, compared with the control group, no matter disease incidence or lesion diameter there are no significant changes.Illustrate from (1 → 3)-of oat
Callose not can induce the resistance of pear fruit.
(1 → 3)-callose in experiment 1 is changed to " saccharomyces cerevisiae by " rhodosporidium toruloides cell " by comparative experiments 2
Cell " cultivates brewing yeast cell according to embodiment 1 and extracts brewing yeast cell wall (1 → 3)-callose, remaining etc.
It is same as experiment 1.
As a result as shown in Figure 3.
Fig. 3 shows that brewing yeast cell wall (1 → 3)-callose concentration is in 0.2~1.0% range, Ke Yixian
It lands and inhibits pear fruit mold decay.It is and right when brewing yeast cell wall (1 → 3)-callose concentration is 0.2%
It is compared according to group, disease incidence and lesion diameter have dropped 44.44% and 6.03mm respectively, effectively slow down the generation of penicilliosis
And development.When concentration increases to 1.0%, disease incidence and lesion diameter reduce by 74.07% and 8.22mm respectively, and it is best to reach it
Inhibition concentration.
Comparative experiments 3, will experiment 1 in (1 → 3)-callose by " rhodosporidium toruloides cell wall preparation gained (1 →
3)-callose " is changed to by " rhodosporidium toruloides cell preparation gained (1 → 3)-callose ", remaining equivalent experiments
1。
Note: the method for rhodosporidium toruloides cell preparation gained (1 → 3)-callose is Patent No.
200410020099.8 patent of invention " plant disease resistance inductor containing yeast cell extract " proposed in yeast it is thin
Born of the same parents' extract (1 → 3)-callose preparation method, that is, replace beer yeast cells to mention in rhodosporidium toruloides cell
It takes.
As a result as shown in Figure 4.
After hot winter spore yeast cells (1 → 3)-callose induction processing for 24 hours, the disease incidence of pear fruit wound is sent out
It reduces to raw conspicuousness, when (1 → 3)-callose concentration is 0.2%, compared with the control group, disease incidence and scab are straight
Diameter reduces by 22.22% and 4.44mm respectively.When (1 → 3)-callose concentration increases to 1%, effect reaches most preferably, morbidity
Rate and lesion diameter reduce by 44.44% and 7.26mm respectively, restrained effectively pear fruit Postharvest Penicillium and rot.
In conclusion rhodosporidium toruloides cell wall (1 → 3)-callose, brewing yeast cell wall (1 → 3)-β-D-
Glucan, rhodosporidium toruloides (1 → 3)-callose can effectively inhibit the mold decay for adopting rear pear fruit, source
The putrefactive phenomenon of fruit wound cannot be reduced in (1 → 3)-callose of oat.But brewing yeast cell wall (1 →
3) biocontrol effect when-callose and rhodosporidium toruloides (1 → 3)-callose concentration are 0.2% is not so good as red winter spore
Yeast cell wall (1 → 3)-callose.
The above list is only a few specific embodiments of the present invention for finally, it should also be noted that.Obviously, this hair
Bright to be not limited to above embodiments, acceptable there are many deformations.Those skilled in the art can be from present disclosure
All deformations for directly exporting or associating, are considered as protection scope of the present invention.
Claims (6)
1. luring the fruit disease of resistant activity to control preparation, feature based on rhodosporidium toruloides cell wall (1 → 3)-callose
It is:
Contain 1~5g rhodosporidium toruloides cell wall (1 → 3)-callose in every liter of preparation, surplus is water;
Rhodosporidium toruloides cell wall (1 → the 3)-callose the preparation method is as follows:
S1, preparation rhodosporidium toruloides cell wall: rhodosporidium toruloides cell is subjected to Mechanical Crushing, obtains rhodosporidium toruloides cell wall;
S2, preparation rhodosporidium toruloides cell wall (1 → 3)-callose, comprising the following steps:
2.1, rhodosporidium toruloides cell wall yeast cell wall obtained by step S1 is added according to the ratio of 1g:48~52mL to concentration
In 70~80 DEG C of 4~8h of reaction, to shake up once, being reacted every 30 ± 5min during reaction in 3% NaOH solution
Object;
2.2, step 2.1 gained reactant is centrifuged 40~50min under the conditions of 7500~8000g, obtains sediment;
2.3,2.2 gained sediment of cleaning step;The sediment after cleaning is added according to the solid-liquid ratio of 1g:48~52mL
The CH of 0.5mol/L3In COOH solution, 2~4h, centrifugal filtration are handled in 85~95 DEG C of water-bath;Repeat above-mentioned steps
1~2 time, it will be freeze-dried after the cleaning of final centrifugal filtration products therefrom, obtain (1 → 3)-callose powder.
2. the fruit according to claim 1 for luring resistant activity based on rhodosporidium toruloides cell wall (1 → 3)-callose
Disease management preparation, it is characterised in that:
The step S2 prepares rhodosporidium toruloides cell wall (1 → 3)-callose, comprising the following steps:
2.1, rhodosporidium toruloides cell wall yeast cell wall is added according to the ratio of 1g:50mL to concentration molten for 3% NaOH
In liquid, in 75 DEG C of reaction 6h, shaken up once during reaction every 30min;
2.2, step 2.1 gained reactant is centrifuged 45min under the conditions of 7800g, obtains sediment;
2.3, by step 2.2 gained sediment 3 times wash with distilled water;According to the solid-liquid ratio of 1g:50mL by the precipitating after cleaning
The CH of object addition 0.5mol/L3In COOH solution, 3h, centrifugal filtration are hydrolyzed in 90 DEG C of water-bath;Repeat above-mentioned steps
Once, it is freeze-dried after being neutrality to pH wash with distilled water for final centrifugal filtration products therefrom, obtains (1 → 3)-β-
D- glucan powder.
3. the fruit according to claim 1 or 2 for luring resistant activity based on rhodosporidium toruloides cell wall (1 → 3)-callose
Real Disease management preparation, it is characterised in that:
Contain 2g rhodosporidium toruloides cell wall (1 → 3)-callose in every liter of preparation, surplus is water.
4. carrying out fruit disease control method using the preparation as described in claims 1 to 3 is any, feature includes following step
It is rapid:
Preparation is inoculated in the wound of fruit, makes it that fruit wound be completely covered, after the preparation natural air drying of fruit wound,
The fruit is put into container and is saved under sealing state.
5. the fruit according to claim 4 for luring resistant activity based on rhodosporidium toruloides cell wall (1 → 3)-callose
Disease management method, it is characterised in that:
After the preparation natural air drying of fruit wound, which is put into container using after preservative film sealing, is protected at room temperature
It deposits.
6. the fruit according to claim 4 or 5 for luring resistant activity based on rhodosporidium toruloides cell wall (1 → 3)-callose
Real Disease management method, it is characterised in that:
The fruit is pear fruit.
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CN110583762A (en) * | 2019-08-12 | 2019-12-20 | 天津科技大学 | Oligosaccharide preservative for inhibiting fruit softening and relieving mechanical damage and use method and application thereof |
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Application publication date: 20190419 |