CN1596658A - Antidisease induction agent containing yeast cell extract for plant - Google Patents
Antidisease induction agent containing yeast cell extract for plant Download PDFInfo
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Abstract
A disease resistant inducing agent for plants contains beta-1,3 glocosan, extract of yeast cell, wetting agent and disperser.
Description
Technical field
The present invention relates to a kind of yeast cells extract, a kind of more precisely plant disease resistance inductor and its preparation and application that contains fungi polysaccharide.
Background technology
The generation of various diseases can make crop production reduction 30～40%.The present plant disease mainly method of preventing and treating is to utilize chemical pesticide and disease-resistant variety, though chemical pesticide can fast and effeciently kill pathogen, exist as a series of problems such as environmental pollution, residue of pesticide, pathogen generation resistance and the destruction ecological balances.Disease-resistant variety is the effective measures of preventing and treating disease, especially develop rapidly along with engineered, increased the means that disease-resistant variety is cultivated, but this method cycle is long, expense is high, and much diseases can't solve by disease-resistant variety, so the exploitation of biopesticide comes into one's own day by day.The biological control of plant disease mainly comprises two aspects: utilize antagonistic microbe 1.; 2. the expression of inducing plant self disease resistance.Modern age, plant pathology was thought, the disease-resistant variety of plant still is that susceptible variety all exists a large amount of disease-resistant genes, what these disease-resistant genes had under nature can directly express, then needing of having induces and could express by suitable mode, thereby make the resistance of plant acquisition, i.e. induction of resistance to pathogen.The phenomenon of induction of resistance has just had been found that as far back as earlier 1900s initial people call acquisition resistance or acquired immunity to this phenomenon.Inducing anti-disease is defined as " utilize chemistry or the biological method of physics, anticipate plant, thereby change disease reaction, make original susceptible reaction produce resistance local or system ".Because inducing anti-disease is that plant is by reacting to exogenous stimulation, transfer the function of self vivo immuning system, stop pathogen to invade harm, and derivant itself does not have direct killing effect to pathogen, so it is harmless to animals and plants, can not cause biological strain to morph, pathogen also is difficult for producing resistance, can not destroy the ecological balance etc.Therefore, inducing anti-disease become gain great popularity in the numerous resistance types of plant disease and plant disease control in have a kind of method of application potential most.
Plant in the long-term evolution process owing to need constantly resist the infringement of microorganism; therefore a series of complexity have been formed gradually and effective system of defense; when plant is subjected to infecting; induce various biochemical reactions, control even remove pathogen and reach the purpose of protection self.The factor that can inducing plant produces disease resistance is called elicitor or exciton, be divided into biotic factor and abiotic component two classes, biotic factor comprises fungi, bacterium and nontoxic pathogen, the pathogen culturing filtrate of virus and some biotic factor that is derived from pathogen, handle wheat, barley seedling as Stenzel with the culture fluid of a bacterial strain of bacillus, energy inducing wheat and barley mildew-resistance bacterium are infected, not forming but do not influence powdery mildew conidia germination and appresorium, mainly is the formation that suppresses haustorium and bacterium colony.Abiotic component comprises oxalate, benzene thiocarbamide, sodium selenite, two chlorcyclopropanes, salicylic acid, 2,6-dichloro-isonicotinic acid (INA), benzothiazolethio methyl acetate (BTH) etc.Especially BTH, behind the foliage-spray, can evoking tobacco and arabidopsis various viruses, bacterium and fungus-caused disease are produced resistance, seedling stage, processings can make the paddy rice whole growth season rice blast be had resistant effect, can induce the resistance to rust, powdery mildew and leaf spot after the processing wheat.The success and the registration of BTH development are used, and are to use the induction of resistance protective plant for many years from studying to an important breakthrough of putting into practice.
The mechanism of induction of resistance is very complicated, and it relates to plant and aspects such as pathogen and interaction thereof.1, the reaction of host's lignification is one of main mechanism of inducing plant generation resistance.Lignin is the main component of plant cell wall, and the lignification reaction is one of effective means of plant opposing pathogen infection.Studies show that, the wheat leaf is inoculated in spite of wound that a near the lignification rapidly inoculation point can make Mai Ye avoid infection process with the Botrytis cinerea of non-pathogenic.2, protective plant protecting agent increases.The deposition of protective plant protecting agent can suppress growth of pathogenic bacteria and infect, thereby reaches disease-resistant purpose.3, the accumulation of aldehydes matter.Aldehydes matter can be oxidized to the quinone to the toxic effect of pathogen, is again simultaneously the precursor that lignin forms.4, pathogenic associated protein (Pathogenesis-related proteins, accumulation PRs).A little less than the associated protein of causing a disease does not exist in healthy plant or shows, when being induced, produce accumulation rapidly by elicitor, as inducing the tobacco plant that downy mildew is produced resistance, find that the quantity that produces pathogenic associated protein increases after inoculating 2 days or 4 days with electrophoresis methods analyst process.Have now found that tens plant species can be induced pathogenic associated protein.In the PR protein family, PR-3 is accredited as chitinase at present, and PR-2 is accredited as beta-1,3-glucanase.Ubiquity chitinase and β-1,3 dextranase in the higher plant exists but find no chitin so far, and β-1,3 glucan also only exists in plant corpus part cell on a small quantity.Because the main component of a lot of plant pathogenic fungi cell walls is chitin and β-1,3 glucan, so the chitinase in the plant and β-1,3 dextranase can be cleared up the cell wall of pathogen.Under the normal condition, chitinase and β-1,3 dextranase have only low-level constructive expression in the plant corpus, but with making it produce high-caliber chitinase and beta-1,3-glucanase after the exciton processing.5, the variation of defensive ferment system.Mainly comprise phenylalnine ammonialyase (PAL), peroxidase (POD) and polyphenol oxidase (PPO) etc.PAL is the key enzyme and the rate-limiting enzyme of shikimic acid metabolism, it is the synthetic key enzyme of aldehydes matter, protective plant protecting agent and lignin, POD and isodynamic enzyme thereof play a major role in above-mentioned antibacterial material building-up process and influence the metabolism of phenols, and the major function of PPO is that aldehydes matter is oxidized to the quinones substance stronger to pathogen toxicity.
Summary of the invention
The purpose of this invention is to provide a kind of method of extracting the yeast cells extract, its main component is β-1,3 glucan.
Another object of the present invention provides the method that a kind of preparation contains the plant disease resistance inductor of yeast cells extract, inducing plant self energy that enhances disease resistance.
Last purpose of the present invention provides the application of this plant disease resistance inductor aspect plant disease-resistant.
This plant disease resistance inductor that contains the yeast cells extract, be that a kind of main active is β-1, the aqueous suspension agent of 3 glucans, it is characterized in that: it contains yeast cells extract, wetting agent, dispersant, thickener, sagging inhibitor, defoamer, antifreezing agent, and their percentage by weights therein are as follows:
Yeast cells extract 40-60%
Wetting agent 2.0-4.0%
Dispersant 1.0-3.5%
Thickener 0.05-0.15%
Sagging inhibitor 1.0-4.0%
Defoamer 0.2-1.0%
Antifreezing agent 1.0-4.0%
Water 23.35-54.75%
After 500mg/L contained the plant disease resistance inductor spraying processing of yeast cells extract, the disease-resistant active substance level improved in the plant bodies such as cucumber, and sick leaf rate and disease index all obviously descend.
Beneficial effect of the present invention:
Compare with the general chemistry bactericide, plant disease resistance inductor has that disease-resistant spectrum is wide, the lasting period is long and the characteristics of system's resistance, and safety non-pollution, is a kind of pest control method that has much application potential.This yeast cells extract is a polysaccharose substance, and through being accredited as β-1,3 glucan, this polysaccharide does not have direct killing effect to phytopathogen, but can inducing cucumber plant body in phenylalnine ammonialyase, the raising of defensive ferment activity such as polyphenol oxidase; β-1, the raising of pathogenic associated protein activity such as 3-dextranase and chitinase, and the raising of content of lignin, can strengthen the resistance of plant to diseases such as powdery mildew of cucumber, downy mildew, gray mold and cladosporium leaf and fruit mould of tomato, be a kind of effective plant disease resistance inductor therefore.
Description of drawings
No accompanying drawing
Specific embodiments
This plant disease resistance inductor that contains the yeast cells extract, be that a kind of main active is β-1, the aqueous suspension agent of 3 glucans, it contains yeast cells extract, wetting agent, dispersant, thickener, sagging inhibitor, defoamer, antifreezing agent, and their percentage by weights therein are as follows:
Yeast cells extract 40-60%
Wetting agent 2.0-4.0%
Dispersant 1.0-3.5%
Thickener 0.05-0.15%
Sagging inhibitor 1.0-4.0%
Defoamer 0.2-1.0%
Antifreezing agent 1.0-4.0%
Water 23.35-54.75%
Containing active component main in the plant disease resistance inductor of yeast cells extract is β-1, the 3-glucan.
Wetting agent can be a farming breast 0204;
Dispersant can be sodium lignin sulfonate or calcium lignosulfonate;
Thickener can be an xanthans;
Sagging inhibitor can be a carclazyte;
Defoamer can be an isoamyl alcohol;
Antifreezing agent can be a glycerine.
The yeast cells preparation method of extract comprises the steps:
1) yeast cells is dissolved in 1mol/L NaOH (W: V=1: 2), 75 ℃ the reaction 6 hours, repeat once to be washed to neutrality, obtain the extract of brown thickness;
2) this extract is dissolved in the HCl (W: V=1: 1) of 1mol/L, 30 ℃ were reacted 2 hours, be washed to neutrality, obtain light brown crude extract, ethanol decolorization degreasing, ether dehydrate, obtain white powder, contain the needed yeast cells extract of plant disease resistance inductor of yeast cells extract for preparation.
This application that contains the plant disease resistance inductor of yeast cells extract, contain the plant disease resistance inductor spraying processing of yeast cells extract through 500mg/L after, the phenylalnine ammonialyase activity begins to improve in the cucumber plant body, in the time of 14 days, enzymic activity reaches maximum, comparison begins to descend according to exceeding 3 times more than afterwards, and treated plant enzyme is lived and still is higher than contrast in the time of 28 days;
Contain the plant disease resistance inductor spraying processing of yeast cells extract through 500mg/L after, polyphenol oxidase activity improves in the cucumber plant body, and activity peaks in the time of 7 days, slowly descend afterwards, and it is flat to keep certain enzyme running water in a long time.
Through the plant disease resistance inductor that 500mg/L contains the yeast cells extract induce handle after 3 days, β-1,3 dextranase activity begins to improve in the cucumber plant body, reach maximum in the time of 15 days, enzyme is lived comparison according to improving more than 1 times, begins in the time of 20 days to descend, and it is flat still to maintain high enzyme running water in the time of 25 days.
After derivant is handled 3 days, the chitinase activity began to improve in the cucumber plant body, reaches maximum in the time of 7 days, began afterwards to descend, and in the time of 25 days, its enzyme is lived and still is higher than contrast.
After derivant was handled, content of lignin began slow raising in the cucumber plant body, reaches maximum in the time of 15 days, begins slow decline afterwards, in the time of 30 days still a little more than contrast.
The inducing action of this derivant has conductivity, when cucumber seedling grows to 2 cotyledons, 2 leaf periods, handle 3 blades in bottom of cucumber seedling with the plant disease resistance inductor that contains the yeast cells extract, and in the time of 7 days, measure the phenylalnine ammonialyase activity of the 4th blade and newborn blade, find about 2.6 times of the 4th leaf enzyme raising alive, the blade enzyme that newly grows is lived also slight raising, shows that the inducing action of derivant has conductivity, and can conduct to newborn blade.
After derivant was handled, sick leaf rate of cucumber and tomato plant and disease index all obviously descended.
In the greenhouse after 500mg/L and 1000mg/L derivant are handled, cucumber plant improves the resistance of gray mold, plant sick leaf rate in 14 days is compared respectively according to having descended 37.9～57.9 and 38.4～59.0, best relative immunity effect reaches 90.9% and 92.1%, and the two each in period the relative immunity effect similar.
After derivant was handled, plant was obviously improved the resistance of cladosporium leaf and fruit mould of tomato, and sick leaf rate is starkly lower than blank, and the relative immunity effect has reached 76.8%.
After derivant was handled, sick leaf rate of cucumber downy mildew and disease index comparison were according to all there being obvious decline, and after derivant was handled, the relative immunity effect reached 61.4%.
Further illustrate the preparation method of the disease-resistant derivant of the present invention below by embodiment.
[embodiment 1]
1. the separation and Extraction of yeast cells extract
1) yeast cells is dissolved in 1mol/L NaOH (W: V=1: 2), 75 ℃ the reaction 6 hours, repeat once to be washed to neutrality, obtain the extract of brown thickness;
2) this extract is dissolved in the HCl (W: V=1: 1) of 1mol/L, 30 ℃ were reacted 2 hours, be washed to neutrality, obtain light brown crude extract, ethanol decolorization degreasing, ether dehydrate, obtain white powder, contain the needed yeast cells extract of plant disease resistance inductor of yeast cells extract for preparation.
2. prepared yeast cells extract is prepared into the aqueous suspension agent-plant disease resistance inductor that contains 40% yeast cells extract in following ratio:
Yeast cells extract 40%
Farming breast 0,204 3.6%
Sodium lignin sulfonate (or calcium lignosulfonate) 3%
Xanthans 0.15%
Carclazyte 2.4-3.6%
Isoamyl alcohol 0.6%
Glycerine 2.4-3.6%
Water 45.45-47.85%
[embodiment 2]
1. the separation and Extraction of yeast cells extract is with [embodiment 1]
2. prepared yeast cells extract is prepared into the aqueous suspension agent-plant disease resistance inductor that contains 50% yeast cells extract in following ratio:
Yeast cells extract 50%
Farming breast 0,204 3%
Sodium lignin sulfonate (or calcium lignosulfonate) 2.5%
Xanthans 0.10%
Carclazyte 2-3%
Isoamyl alcohol 0.5%
Glycerine 2-3%
Water 37.90-39.90%
[embodiment 3]
1. the separation and Extraction of yeast cells extract is with [embodiment 1]
2. prepared yeast cells extract is prepared into the aqueous suspension agent-plant disease resistance inductor that contains 60% yeast cells extract in following ratio:
Yeast cells extract 60%
Farming breast 0,204 2.4%
Sodium lignin sulfonate (or calcium lignosulfonate) 2%
Xanthans 0.05%
Carclazyte 1.6-2.4%
Isoamyl alcohol 0.4%
Glycerine 1.6-2.4%
Water 30.35-31.95%
Further set forth the present invention below by experimental example.
[experimental example 1] this disease-resistant derivant is to the influence of disease fungus spore germination
Influence to spore germination: glucose preparation B.cinerea with 2%, the spore suspension of F.oxysporum f.sp.Cucumerinum and F.fulva, mix with the yeast cells extract aqueous suspension agent equal-volume of variable concentrations, making the conidium concentration that contains 100mg/L, 500mg/L and 1000mg/L yeast cells extract aqueous suspension agent is the soup spore suspension in 20-30 spore/visual field under 10 * 10 times of mirrors, get 0.1mL respectively and be added drop-wise on the slide, establish corresponding solvent and be contrast.Preserve moisture respectively at 22 ℃ and 25 ℃ of dark and to cultivate 24 hours, check the spore germination situation, the result shows, the spore of botrytis cinerea pers, cucumber fusarium axysporum and cladosporium leaf and fruit mould of tomato bacterium all can normally be sprouted and be grown in the yeast cells extract aqueous suspension agent solution of 100,500 and 1000 (removing) mg/L, promptly yeast cells extract aqueous suspension agent to the spore germination of botrytis cinerea pers, wilt and cladosporium leaf and fruit mould of tomato bacterium all less than influence.
[experimental example 2] this disease-resistant derivant is to the influence of pathogen mycelial growth
Adopt face-off to cultivate determination method, supply the examination pathogen in the inoculation of PDA culture medium flat plate center, after placing the interior 48h of cultivation of 25 ℃ of insulating boxs, with the diameter of sterilization is that the circular filter paper sheet of 6mm places the culture dish edge, and drips this aqueous suspension agent solution of 10mL variable concentrations, makes filter paper moistening fully, if corresponding solvent and clear water are contrast, continuation is cultivated 48h in 25 ℃ of incubators, measure antibacterial distance, and promptly the filter paper edge is to the distance at fungus colony edge.Six plates of every processing, repeat for three times, the result shows, botrytis cinerea pers, cucumber fusarium axysporum and cladosporium leaf and fruit mould of tomato bacterium 100,500 and the PDA flat board of 1000mg/L yeast cells extract aqueous suspension agent on all can normal growth, do not find any anomaly, so this aqueous suspension agent is to the not influence of mycelial growth of botrytis cinerea pers, cucumber fusarium axysporum and cladosporium leaf and fruit mould of tomato bacterium.
The disease-resistant Study on Mechanism of [experimental example 3] this disease-resistant derivant
It is the solution of 500mg/L that this disease-resistant derivant water is mixed with by the active ingredient calculating concentration, after cucumber emerged 3 days, handle with this disease-resistant derivant spraying of 500mg/L, measure defensive ferment PAL and PPO associated protein chitinase alive, pathogenic and β-1,3 dextranase activity and content of lignin in spraying back different time.
The extraction of PAL and mensuration: through inducing the cucumber plant 5g of processing, add the borate buffer that 10mL contains the 5mM mercaptoethanol (0.025M, PH8.8), 0.5g polyvinylpyrrolidone (PVP) grinds in the ice bath, the centrifugal 15min of 10000g, supernatant are the PAL crude extract.1ml enzyme liquid, 1mL 0.02M L-phenyl alanine, 2mL distilled water, cumulative volume 4mL.Contrast replaces substrate with 1mL distilled water; Reactant liquor places 30 ℃ of waters bath with thermostatic control to be incubated, and measures light absorption value (OD value) with ultraviolet specrophotometer under the 290nm wavelength behind the 30min, and it is enzyme unit alive that the OD value changes 0.01.The result shows that after the spraying of 500mg/L cell wall polysaccharides of yeast was handled, the PAL enzymic activity began to improve in the plant body, and in the time of 14 days, enzymic activity reaches maximum, and comparison begins to descend according to exceeding 3 times more than afterwards, and treated plant enzyme is lived and still is higher than contrast in the time of 28 days.
The extraction of PPO and mensuration: through inducing the cucumber plant 5g of processing, (0.05M pH7.0), grinds in the ice bath, and the centrifugal 5min of 10000g, supernatant are the PPO crude extract to add the 10mL phosphate buffer.1.5mL the catechol solution of 0.02M adds 1.4mL 0.05M phosphate buffer, 0.1mL enzyme liquid, and contrast is with 0.1mL distilled water replacement enzyme liquid, and 30 ℃ of reaction 2min measure light absorption value under the 398nm wavelength, and it is enzyme unit alive that the OD value changes 0.01.The result shows that after the spraying of 500mg/L cell wall polysaccharides of yeast was handled, the PPO activity improved in the cucumber plant body, and activity peaks in the time of 7 days, slowly descends afterwards, and maintains the flat relative long period of certain enzyme running water.
The extraction and the mensuration of β-1,3 dextranase:, add 8mL0.1mol/L citric acid-0.2mol/L sodium hydrogen phosphate buffer solution (pH4.8) through inducing the cucumber plant flesh tissue 2g of processing, grind in the ice bath, the centrifugal 15min of 10000g, supernatant are β-1,3 dextranase crude enzyme liquid.With reference to people such as Abeles (1970) method
[7], zyme extract 1mL, 0.1mol/L citric acid-0.2mol/L sodium hydrogen phosphate buffer solution (pH4.8) 2.5mL, 1% (w/v) laminariose 0.5mL, 30 ℃ are incubated 1 hour, and anthrone method is measured sugared growing amount, is a unit of activity per hour to form 1mg glucose.The result shows that the 500mg/L cell wall polysaccharides of yeast is induced and handled back 3 days, and the beta-1,3-glucanase activity begins to improve in the cucumber plant body, reaches maximum in the time of 15 days, and enzyme is lived comparison according to improving more than 1 times; Begin in the time of 20 days to descend, it is flat still to maintain high enzyme running water in the time of 25 days.
The extraction of chitinase and mensuration: through inducing the cucumber plant 5g of processing, add 7.5mL 0.1mol/L sodium citrate buffer solution (pH5.0), the centrifugal 15min of 10000g, supernatant are the chitinase crude extract.Enzyme extract 200 μ L, 10m mol/L sodium phosphate buffer (pH6.4), the gluey chitin of 10mg/mL.Mixing behind 37 ℃ of water bath with thermostatic control 2h, places 100 ℃ of waters bath with thermostatic control to boil 3min, the centrifugal 10min of 6000g.Get supernatant 400 μ L, add 20 μ L 1mol/L sodium phosphate buffers (pH7.1), 80 μ L 1% (w/v) glusulases, behind 37 ℃ of water bath with thermostatic control 30min, the colorimetric method of pressing Reissiget al. is measured the N-acetylglucosamine.Get the 0.5mL sample, add 0.1mL 0.8mol/L potassium borate (pH9.1), boiling water bath 3min, cooling adds 3mL DMAB, mixes immediately, places 36-38 ℃ of 20min, cooling, 585nm measures the OD value.The result shows that after this disease-resistant derivant was handled, the chitinase activity began to improve in the plant body, reaches maximum in the time of 7 days, begins afterwards to descend, and in the time of 25 days, its enzyme is lived and still is higher than contrast.
The extraction of lignin and mensuration: dry rapidly through inducing the cucumber plant 5g of processing, get dry substance 0.1g, handle with 1% acetic acid earlier and isolate sugar, organic acid and other soluble compounds; Use acetone treatment, isolate chlorophyll, fat and other fat-soluble compounds; The concentrated sulfuric acid of adding 72% is isolated cellulose and hemicellulose; The centrifugal 5min of 6000g, the distilled water wash precipitation, sulfuric acid exists down, and with the lignin in the potassium dichromate oxidation hydrolysate, the potassium bichromate that iodometric determination is excessive adds the 1mL starch indicator before the terminal point, be titrated to green.The result shows that after this disease-resistant derivant was handled, content of lignin began slow raising in the plant body, reaches maximum in the time of 15 days, begins slow decline afterwards, still is slightly higher than contrast in the time of 30 days.
The systematicness test of [experimental example 4] disease-resistant derivant
When cucumber seedling grows to 2 cotyledons, 2 leaf periods, be 3 blades in bottom that the disease-resistant derivant of 500mg/L is handled cucumber seedling with active ingredient concentration, get the 4th blade and newborn blade during respectively at 7 days, measure the PAL enzymic activity.The result shows, during after this disease-resistant derivant is handled the 3rd blade of cucumber plant 7 days, get the 4th leaf and newborn measuring blade PAL activity respectively, the result shows: spraying was handled after 7 days, handle the 4th leaf enzyme of plant and live comparison according to improving about 2.6 times, handle the newborn blade enzyme of plant and live comparison according to also slight raising being arranged, promptly disease-resistant derivant has conductivity to the inducing action of enzymic activity, and can conduct to newborn blade (table 1).
The PAL active unit of the different blades of table 1 cucumber plant: U/g.min
The 4th newborn blade of leaf
The control treatment control treatment
0.046 0.166 0.051 0.056
[experimental example 5] disease-resistant derivant is to the preventive and therapeutic effect of gray mold
Cucumber seeds is dipped in respectively in the disease-resistant derivant solution of variable concentrations, carries out vernalization after 4 hours, be seeded in during seed sprouting in the vermiculite, place 25 ± 1 ℃ of greenhouses, make its natural occurrence, handle 20 strains at every turn, repeat investigation incidence in the time of after planting 7,14 and 30 days for 3 times.The result shows, the sick leaf rate of handling through disease-resistant derivant of plant all is lower than contrast, but different using dosages induce difference on effect very significantly (table 2).Not obvious through sick leaf rate of plant and contrast difference that the disease-resistant derivant of 100mg/L is handled, show that disease-resistant derivant dosage is too small, induce the effect of plant generation disease resistance not obvious.And be greatly improved through the plant disease resistance of 500mg/L and the disease-resistant derivant processing of 1000mg/L, after inducing 14 days, sick leaf rate is compared respectively according to having descended 37.9～57.9 and 38.4～59.0 percentage points, but prolongation along with the time, after handling one month, the sick leaf rate of plant through inducing processing and adjoining tree has all reached 100%.The sick leaf rate of the plant that 500mg/L and 1000mg/L handle all differed not quite in each period, illustrate that this disease-resistant derivant using dosage reaches finite concentration after, the inducing anti-disease effect is no longer with the dosage increasing and improve proportionally; Through the plant that the disease-resistant derivant of 100mg/L is handled, the relative immunity effect is relatively poor, and the plant of handling through 500mg/L and 1000mg/L, and best relative immunity effect reaches 90.9% and 92.1%, and the two each in period the relative immunity effect similar.And all along with the prolongation of induction time, the relative immunity effect descends in each processing, but the immune effect that the disease-resistant derivant of higher dosage is handled still obviously is better than contrast (table 3).
The disease-resistant derivant of table 2 is induced the sick leaf rate (%) of back gray mold of cucumber
Induce back time (fate) 100mg/L 500mg/L 1000mg/L contrast
7 38.5 3.8 3.3 41.7
14 51.3 11.1 10.0 69.0
21 100 100 100 100
The disease-resistant derivant of table 3 is to the relative immunity effect of gray mold of cucumber
Induce back time (fate) survey item 100mg/L 500mg/L 1000mg/L contrast
Disease index 38.5 3.8 3.3 41.7
7
Relative immunity effect (%) 7.9 90.9 92.1
Disease index 34.6 8.9 8.8 44.4
14
Relative immunity effect (%) 22.1 80.0 80.2
Disease index 81.9 51.9 51.3 83.1
21
Relative immunity effect (%) 1.4 37.5 38.3
[experimental example 6] disease-resistant derivant is to the preventive and therapeutic effect of cladosporium leaf and fruit mould of tomato
Spring calendar year 2001, after Xin Kou town green house of vegetables sprays this aqueous suspension agent 22 days, cladosporium leaf and fruit mould of tomato takes place, spray derivant and the agricultural chemicals lucky star missible oil that active ingredient concentration is 500mg/L in their early stage, 7 days investigation incidences after the dispenser, the result shows: after derivant is handled, obviously improved the resistance of plant to cladosporium leaf and fruit mould of tomato, sick leaf rate is starkly lower than blank, but be higher than agricultural chemicals lucky star missible oil, but disease index and lucky star missible oil are more or less the same, but well below blank, the relative immunity effect has reached 76.8% (table 4).
The disease-resistant derivant aqueous suspension agent of table 4 is to the preventive and therapeutic effect of cladosporium leaf and fruit mould of tomato
Sick leaf rate (%) the disease index relative immunity effect (%) of processing mode
Derivant 13.1 2.6 76.8
Lucky star missible oil 4.5 1.2 89.3
Blank 27.1 11.2
[experimental example 7] disease-resistant derivant is to the preventive and therapeutic effect of cucumber downy mildew
Autumn in 2002,42 days generation downy mildew after the blue or green town of willow green house of vegetables sprays this aqueous suspension agent, spraying derivant and the agricultural chemicals gram that active ingredient concentration is 500mg/L in back 7 days in morbidity reveals, 7 days investigation incidences after the dispenser, the result shows: after derivant was handled, sick leaf rate of plant and disease index were all compared illumination and are shown decline.But because this disease a run of wet weather when taking place is unfavorable to disease control, and the control time is late excessively, so the preventive effect of derivant only is 61.4%, and the preventive effect of agricultural chemicals gram dew has also only reached 74.2% (table 5).
The disease-resistant derivant of table 5 is to the preventive and therapeutic effect of cucumber downy mildew
Sick leaf rate (%) the disease index relative immunity effect (%) of processing mode
Derivant 21.4 9.1 61.4
Gram reveals 15.6 6.1 74.2
Blank 48.9 23.6
Claims (14)
1, a kind of plant disease resistance inductor that contains the yeast cells extract, be that a kind of its main active is β-1, the aqueous suspension agent of 3 glucans, it is characterized in that: it contains yeast cells extract, wetting agent, dispersant, thickener, sagging inhibitor, defoamer, antifreezing agent, and their percentage by weights therein are as follows:
Yeast cells extract 40-60%
Wetting agent 2.0-4.0%
Dispersant 1.0-3.5%
Thickener 0.05-0.15%
Sagging inhibitor 1.0-4.0%
Defoamer 0.2-1.0%
Antifreezing agent 1.0-4.0%
Water 23.35-54.75%
2, the plant disease resistance inductor that contains the yeast cells extract according to claim 1 is characterized in that: main active component is β-1 in the yeast cells extract, the 3-glucan.
3, the plant disease resistance inductor that contains the yeast cells extract according to claim 1 is characterized in that:
Wetting agent can be a farming breast 0204;
Dispersant can be sodium lignin sulfonate or calcium lignosulfonate;
Thickener can be an xanthans;
Sagging inhibitor can be a carclazyte;
Defoamer can be an isoamyl alcohol;
Antifreezing agent can be a glycerine.
4, the plant disease resistance inductor that contains the yeast cells extract according to claim 2, it is characterized in that: the yeast cells preparation method of extract comprises the steps:
1) yeast cells is dissolved in 1mol/L NaOH (W: V=1: 2), 75 ℃ the reaction 6 hours, repeat once to be washed to neutrality, obtain the extract of brown thickness;
2) this extract is dissolved in 1mol/L HCl (W: V=1: 1), 30 ℃ the reaction 2 hours, be washed to neutrality,
To light brown crude extract, ethanol decolorization degreasing, ether dehydrate and obtain white powder, contain the needed yeast cells extract of plant disease resistance inductor of yeast cells extract for preparation.
5. application that contains the plant disease resistance inductor of yeast cells extract, it is characterized in that: contain the plant disease resistance inductor spraying processing of yeast cells extract through 500mg/L after, the phenylalnine ammonialyase activity begins to improve in the cucumber plant body, in the time of 14 days, enzymic activity reaches maximum, comparison begins to descend according to exceeding 3 times more than afterwards, and treated plant enzyme is lived and still is higher than contrast in the time of 28 days.
6, the application that contains the plant disease resistance inductor of yeast cells extract according to claim 5, it is characterized in that: contain the plant disease resistance inductor spraying processing of yeast cells extract through 500mg/L after, polyphenol oxidase activity improves in the cucumber plant body, activity peaks in the time of 7 days, slowly descend afterwards, and it is flat to keep certain enzyme running water in a long time.
7, the application that contains the plant disease resistance inductor of yeast cells extract according to claim 5, it is characterized in that: through the plant disease resistance inductor that 500mg/L contains the yeast cells extract induce handle after 3 days, β-1 in the cucumber plant body, 3 dextranase activities begin to improve, reach maximum in the time of 15 days, enzyme is lived comparison according to improving more than 1 times, begins in the time of 20 days to descend, and it is flat still to maintain high enzyme running water in the time of 25 days.
8, the application that contains the plant disease resistance inductor of yeast cells extract according to claim 5, it is characterized in that: after derivant is handled 3 days, the chitinase activity begins to improve in the cucumber plant body, reach maximum in the time of 7 days, begin afterwards to descend, in the time of 25 days, its enzyme is lived and still is higher than contrast.
9, the application that contains the plant disease resistance inductor of yeast cells extract according to claim 5, it is characterized in that: after derivant was handled, content of lignin began slow raising in the cucumber plant body, reaches maximum in the time of 15 days, begin slow decline afterwards, in the time of 30 days still a little more than contrast.
10. the application that contains the plant disease resistance inductor of yeast cells extract according to claim 5, it is characterized in that: the inducing action of this derivant has conductivity, when cucumber seedling grows to 2 cotyledons, during 2 leaf periods, handle 3 blades in bottom of cucumber seedling with the plant disease resistance inductor that contains the yeast cells extract, and in the time of 7 days, measure the phenylalnine ammonialyase activity of the 4th blade and newborn blade, find about 2.6 times of the 4th leaf enzyme raising alive, the blade enzyme that newly grows is lived also slight raising, the inducing action that shows derivant has conductivity, and can conduct to newborn blade.
11. the application that contains the plant disease resistance inductor of yeast cells extract according to claim 5 is characterized in that: after derivant was handled, sick leaf rate of cucumber and tomato plant and disease index all obviously descended.
12. the application that contains the plant disease resistance inductor of yeast cells extract according to claim 5, it is characterized in that: gray mold of cucumber is had preventive and therapeutic effect, after 500mg/L and the processing of 1000mg/L derivant, plant sick leaf rate in 14 days is compared respectively according to having descended 37.9～57.9 and 38.4～59.0, best relative immunity effect reaches 90.9% and 92.1%, and the two each in period the relative immunity effect similar.
13. the application that contains the plant disease resistance inductor of yeast cells extract according to claim 5, it is characterized in that: the cladosporium leaf and fruit mould of tomato disease is had preventive and therapeutic effect, after derivant is handled, plant is obviously improved the resistance of cladosporium leaf and fruit mould of tomato, sick leaf rate is starkly lower than blank, and the relative immunity effect has reached 76.8%.
14. the application that contains the plant disease resistance inductor of yeast cells extract according to claim 5, it is characterized in that: cucumber downy mildew is had preventive and therapeutic effect, after derivant is handled, sick leaf rate of cucumber downy mildew and disease index comparison are according to obvious decline is all arranged, after derivant was handled, the relative immunity effect reached 61.4%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100200998A CN1313006C (en) | 2004-07-23 | 2004-07-23 | Antidisease induction agent containing yeast cell extract for plant |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100200998A CN1313006C (en) | 2004-07-23 | 2004-07-23 | Antidisease induction agent containing yeast cell extract for plant |
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| Publication Number | Publication Date |
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| CN1596658A true CN1596658A (en) | 2005-03-23 |
| CN1313006C CN1313006C (en) | 2007-05-02 |
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| Application Number | Title | Priority Date | Filing Date |
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| CNB2004100200998A Expired - Fee Related CN1313006C (en) | 2004-07-23 | 2004-07-23 | Antidisease induction agent containing yeast cell extract for plant |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2241186A1 (en) * | 2008-01-11 | 2010-10-20 | Ajinomoto Co., Inc. | Disease resistance enhancer for plants and method of controlling plant disease by using the same |
| CN101374417B (en) * | 2005-12-21 | 2013-06-05 | 乐斯福公司 | Protection of plants against their pathogenic agents |
| CN103468627A (en) * | 2013-04-11 | 2013-12-25 | 浙江大学 | Method for improving fruit disease control effectiveness of antagonisitic yeast |
| CN107118016A (en) * | 2017-05-08 | 2017-09-01 | 安徽新荣久农业科技有限公司 | A kind of tea seed breed breeding soak |
| CN107484643A (en) * | 2017-08-03 | 2017-12-19 | 安徽省雷氏农业科技有限公司 | A kind of peony seeds breed breeding culture medium |
| CN107593766A (en) * | 2017-09-30 | 2018-01-19 | 武汉理工大学 | A kind of efficient and environment-friendly type agent for coating seed of watermelon and preparation method thereof |
| CN109645107A (en) * | 2019-01-06 | 2019-04-19 | 浙江大学 | The fruit disease control method of resistant activity is lured based on rhodosporidium toruloides cell wall (1 → 3)-callose |
| CN111631219A (en) * | 2020-06-10 | 2020-09-08 | 菏泽学院 | Method and medicament for preventing and treating greenhouse cucumber leaf blight and resisting low temperature |
| CN118085124A (en) * | 2024-04-25 | 2024-05-28 | 山东劲脉植物细胞信息技术有限公司 | Preparation method of beta-1, 3-glucan and application of beta-1, 3-glucan in biologically inducing disease prevention and disease resistance of plants |
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2004
- 2004-07-23 CN CNB2004100200998A patent/CN1313006C/en not_active Expired - Fee Related
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101374417B (en) * | 2005-12-21 | 2013-06-05 | 乐斯福公司 | Protection of plants against their pathogenic agents |
| EP2241186A1 (en) * | 2008-01-11 | 2010-10-20 | Ajinomoto Co., Inc. | Disease resistance enhancer for plants and method of controlling plant disease by using the same |
| EP2241186A4 (en) * | 2008-01-11 | 2013-08-21 | Ajinomoto Kk | Disease resistance enhancer for plants and method of controlling plant disease by using the same |
| US9173407B2 (en) | 2008-01-11 | 2015-11-03 | Ajinomoto Co., Inc. | Disease resistance enhancer for plants and method of controlling plant disease by using the same |
| CN103468627A (en) * | 2013-04-11 | 2013-12-25 | 浙江大学 | Method for improving fruit disease control effectiveness of antagonisitic yeast |
| CN103468627B (en) * | 2013-04-11 | 2015-06-03 | 浙江大学 | Method for improving fruit disease control effectiveness of antagonisitic yeast |
| CN107118016A (en) * | 2017-05-08 | 2017-09-01 | 安徽新荣久农业科技有限公司 | A kind of tea seed breed breeding soak |
| CN107484643A (en) * | 2017-08-03 | 2017-12-19 | 安徽省雷氏农业科技有限公司 | A kind of peony seeds breed breeding culture medium |
| CN107593766A (en) * | 2017-09-30 | 2018-01-19 | 武汉理工大学 | A kind of efficient and environment-friendly type agent for coating seed of watermelon and preparation method thereof |
| CN109645107A (en) * | 2019-01-06 | 2019-04-19 | 浙江大学 | The fruit disease control method of resistant activity is lured based on rhodosporidium toruloides cell wall (1 → 3)-callose |
| CN111631219A (en) * | 2020-06-10 | 2020-09-08 | 菏泽学院 | Method and medicament for preventing and treating greenhouse cucumber leaf blight and resisting low temperature |
| CN118085124A (en) * | 2024-04-25 | 2024-05-28 | 山东劲脉植物细胞信息技术有限公司 | Preparation method of beta-1, 3-glucan and application of beta-1, 3-glucan in biologically inducing disease prevention and disease resistance of plants |
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