CN102919280A - Trichoderma asperellum wettable powder and applications thereof - Google Patents
Trichoderma asperellum wettable powder and applications thereof Download PDFInfo
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- CN102919280A CN102919280A CN2012104926823A CN201210492682A CN102919280A CN 102919280 A CN102919280 A CN 102919280A CN 2012104926823 A CN2012104926823 A CN 2012104926823A CN 201210492682 A CN201210492682 A CN 201210492682A CN 102919280 A CN102919280 A CN 102919280A
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Abstract
The invention discloses trichoderma asperellum wettable powder and applications thereof, and relates to a biological pesticide and applications thereof. The trichoderma fungus is a biological control pathogenic bacterium with extensive application prospects; and the trichoderma asperellum wettable powder is prepared from the following components in percentage by weight: 10 to 30 percent of trichoderma asperellum, 2 to 10 percent of a wetting agent, 10 to 15 percent of a dispersing agent, 0.1 to 0.5 percent of an ultraviolet protective agent, and the balance of a carrier. The trichoderma asperellum wettable powder can be used for controlling agriculture and forestry pathogenic fungus and bacterial diseases, and has the advantages of high activity, stable effect, long lasting period, small environmental influence and convenience in use.
Description
Technical field
The present invention relates to a kind of biopesticide and application thereof, be specifically related to wetting powder of a kind of Trichoderma and uses thereof.
Background technology
Trichoderma (Trichoderma spp) belongs to filamentous fungi, and saprophytic property is strong, and wide adaptability can be prevented and treated multiple soil-borne disease, and 18 genus more than 29 kind disease funguses and Various Diseases indigenous bacteria are had antagonism, is considered to one of the most promising BIOLOGICAL CONTROL material.At present more than the 100 kind of biologic product take Trichoderma as main component arranged in the world, and successfully apply to the greenhouse bactericide.The mechanism of action of Trichoderma comprise the solubilising of the competition in hyperparasitism, antibiosis, nutrition or space, the growth compression resistance by strengthening root system and plant, inorganic nutrient salt and seal up for safekeeping, the complex processes such as inactivation of induction of resistance, pathogens chitinase.
Summary of the invention
Be a kind of biological and ecological methods to prevent plant disease, pests, and erosion pathogen with wide application prospect for the mould fungi of wood, the invention provides a kind of active height, effect stability, the lasting period is long, environmental influence is little, easy to use trichoderma asperellum bacterium wetting powder.
Trichoderma asperellum bacterium wetting powder of the present invention is made by following composition: 10～30wt.% trichoderma asperellum, 2～10%wt.% wetting agent, 7～20%wt.% dispersant, 0.1～0.5wt.% uv-protector, surplus are carrier.
Described pulvis is made by following composition: 10～30wt.% trichoderma asperellum, 2～10%wt.% wetting agent, 7～20%wt.% dispersant, 0.1～0.5wt.% uv-protector, 40～80wt.% carrier.
Described pulvis is made by following composition: 10～30wt.% trichoderma asperellum, 2～10%wt.% wetting agent, 10～15%wt.% dispersant, 0.1～0.5wt.% uv-protector, 45～78wt.% carrier.
The quality of described trichoderma asperellum is 7,000,000,000/g.
Described wetting agent is TERWET1010, WLNO, Morwet EFW, SDS, MF-5, pull open one or more the mixture in the powder; Described dispersant is one or more the mixture in calcium lignosulfonate, TERSPERSE 2020, NNO, Morwet D425, the sodium lignin sulfonate; Described carrier is one or more the mixture in attapulgite, kaolin, diatomite, bentonite, the white carbon; Described uv-protector is one or more the mixture in dextrin, xanthans, fluorescein sodium, the VC nano zine oxide.
Described wetting agent is TERWET1010 and Morwet EFW; Described dispersant is calcium lignosulfonate and Morwet D425; Described carrier is bentonite; Described uv-protector is the VC nano zine oxide.
Described pulvis is made by following composition: 10wt.% trichoderma asperellum, 0.5wt.%MorwetEFW, 1.5wt.%TERWET1010,2wt.%Morwet D425,5wt.% calcium lignosulfonate, 0.1wt.% nano zine oxide, 80.9wt.% bentonite.
Described pulvis is made by following composition: 20wt.% trichoderma asperellum, 1wt.%Morwet EFW, 5wt.%TERWET1010,5wt.%Morwet D425,7wt.% calcium lignosulfonate, 0.3wt.% nano zine oxide, 61.7wt.% bentonite.
Described pulvis is made by following composition: 30wt.% trichoderma asperellum, 2wt.%Morwet EFW, 8wt.%TERWET1010,10wt.%Morwet D425,10wt.% calcium lignosulfonate, 0.5wt.% nano zine oxide, 39.5wt.% bentonite.
Trichoderma is trichoderma asperellum (accession number: JX270638) by morphology and ITS-rDNA Sequence Identification in this test.The present invention is processed into wetting powder with the trichoderma asperellum bacterium, in order to prevent and treat agricultural pathogenic fungus and bacteriosis.Compared with prior art, the present invention has following outstanding advantage:
1, on auxiliary agent is selected, pay attention to itself and the biocompatibility of trichoderma asperellum, the auxiliary agent of selection is on trichoderma asperellum conidia germination and mycelia day growth amount and produce not impact or affect littlely of spore, is desirable auxiliary agent.
2, product performance index is good, the spore germination rate of trichoderma asperellum 〉=88.57%; Wetability＜1min; The quality suspensibility is 75.79%, and the spore suspensibility is 80.12%, and moisture is 3.52%; Fineness (crossing 325 mesh standard sieves) 〉=98%; The pH value is 6.7; Be qualified trichoderma asperellum bacterium wetting powder.
3, the present invention is trichoderma asperellum bacterium wetting powder, and this formulation has prolonged period of storage and lasting period, has enlarged the control spectrum, provides new material for carrying out the non-environmental pollution control agriculture and forestry injurious insect.
Description of drawings
Fig. 1 is trichoderma asperellum bacterium wetting powder particle diameter distribution map.
Embodiment
Embodiment one: the trichoderma asperellum wetting powder of present embodiment is made by following composition: 10～30wt.% trichoderma asperellum, 2～10%wt.% wetting agent, 7～20%wt.% dispersant, 0.1～0.5wt.% uv-protector, 40～80wt.% carrier.
In the present embodiment, described wood is mould to be trichoderma asperellum (Trichoderma asperellum), and quality is 7,000,000,000/g.
In the present embodiment, described wetting agent is TERWET1010, WLNO, MorwetEFW, SDS, MF-5, pull open one or more the mixture in the powder.
In the present embodiment, described dispersant is one or more the mixture in calcium lignosulfonate, TERSPERSE 2020, NNO, Morwet D425, the sodium lignin sulfonate.
In the present embodiment, described carrier is one or more the mixture in attapulgite, kaolin, diatomite, bentonite, the white carbon.
In the present embodiment, described uv-protector is one or more the mixture in dextrin, xanthans, fluorescein sodium, the VC nano zine oxide.
Embodiment two: what present embodiment and embodiment one were different is that described wetting powder is made by following composition: 10～30wt.% trichoderma asperellum, 2～10%wt.% wetting agent, 7～20%wt.% dispersant, 0.1～0.5wt.% uv-protector, 45～78wt.% bentonite.
Embodiment three: what present embodiment and embodiment one were different is, described wetting powder is made by following composition: 10～30wt.% trichoderma asperellum, 0.5～2wt.%Morwet EFW, 1.5～8wt.%TERWET1010,2～10wt.%Morwet D425,5～10wt.% calcium lignosulfonate, 0.1～0.5wt.% nano zine oxide, 45～78wt.% bentonite, and the preparation method is as follows:
(1) trichoderma asperellum, Morwet EFW, TERWET1010, Morwet D425, calcium lignosulfonate, nano zine oxide and bentonite are mixed according to the above ratio;
(2) 325 mesh standard sieves are crossed in raw material precomminution;
(3) quality inspection, the germination rate of Trichoderma 〉=88.57%, wetability＜1min, quality suspensibility 75.79%, spore suspensibility 80.12%, moisture is 3.52%, fineness 〉=98% (is crossed 325 mesh standard sieves, particle mean size is 27.00 μ m), the pH value is 6.7, is qualified trichoderma asperellum bacterium wetting powder.
Embodiment four: the trichoderma asperellum bacterium wetting powder of present embodiment, the raw material composition sees Table 1.
Table 1 raw material proportioning unit: mass percent %
| Component | Formula I | Formula I I | The prescription III | The prescription IV |
| The trichoderma asperellum conidial powder | 10 | 15 | 20 | 30 |
| Morwet?EFW | 0.5 | 0.8 | 1 | 2 |
| TERWET1010 | 1.5 | 3 | 5 | 8 |
| Morwet?D425 | 2 | 4 | 5 | 10 |
| Calcium lignosulfonate | 5 | 6 | 7 | 10 |
| Nano zine oxide | 0.1 | 0.2 | 0.3 | 0.5 |
| Bentonite | 80.9 | 71.0 | 61.7 | 39.5 |
Embodiment five: present embodiment prepares trichoderma asperellum bacterium wetting powder by the following method:
One, materials and methods
1, test material
(1) conidial powder: thorn spore Trichoderma conidial powder 7,000,000,000/gram;
(2) trichoderma asperellum pathogen: Northeast Forestry University's Pathology Lab is preserved bacterial classification;
(3) carrier: attapulgite, kaolin, diatomite, bentonite, white carbon;
(4) wetting agent: Morwet EFW, pull open powder, MF-5, SDS, TERWET1010, WLNO;
(5) dispersant: sodium lignin sulfonate, calcium lignosulfonate, TERSPERSE 2020, Morwet D-425, NNO;
(6) uv-protector: dextrin, xanthans, fluorescein sodium, VC nano zine oxide;
(7) chemical pesticide of contrast: 70% tpn wetting powder, 25% Fluoxastrobin SC.
2, test method
2.1 the antagonism of trichoderma asperellum and pathogen
On culture dish (d=90mm), access respectively diameter and be 5mm PDA medium culture 3d etc. Trichoderma and the pathogen of quality, spacing distance is 3cm; Contrast is for accessing respectively Trichoderma and the pathogen that diameter is 5mm, and experiment repeats 3 times, places 25 ℃ of incubator dark culturing, and every 8h measures bacterium colony trend radius, calculates suppressed rate and calculates antagonistic effect with relative bacteriostasis rate.
Suppressed rate=[(cultivating separately colony radius)-(bacterium colony trend radius)]/cultivate separately colony radius
The relative suppressed rate of the suppressed rate/Antagonistic Fungi of bacteriostasis rate=pathogen
2.2 the biocompatibility of carrier, wetting agent, dispersant screening
(1) will join respectively among the PDA of thawing for 1000mg/ml carrier, 120 μ g/ml wetting agents, the 250 μ g/ml dispersants of examination, flat board is made in sterilization.Get the spore suspension (10 of 0.05ml
4Individual spore/ml) evenly coat on the mixed culture medium flat board, with improvement PDA in contrast, each processes 4 repetitions, is positioned over 28 ± 1 ℃ of dark culturing, calculates clump count in 48h.
(2) in above-mentioned mixed culture medium, get the Trichoderma mycelia piece that growth is vigorous, cell age is identical with card punch (d=5mm), be inoculated in the dull and stereotyped center of mixed culture medium, with the PDA culture medium flat plate in contrast, each processes 3 repetitions, and interval 6h surveys a colony diameter.Colony diameter daily growth amount (mm/d)=diameter/growth fate.
2.3 the selection of wetting agent and dispersant
(1) at first test wetting agent and dispersant are carried out primary dcreening operation, selective wetting time＜1min, the dispersant that the suspension situation is good, then carrying out biocompatibility measures, wherein wetting time is carried out according to the GB/T5451-2001 standard, disperse situation by visual observations, the dispersion situation is divided into 3 grades:
1. scattered: as to be cloud in the water and automatically to disperse, sink without visible particle;
2. disperse general: in water, automatically disperse, have particle to sink, jetsam can slowly disperse or jog after disperse;
3. poor dispersion: in water, can not automatically disperse, be graininess and sink or cotton-shaped sinking, after acutely shaking, could disperse.
Filter out preferably dispersant of dispersive property, then add respectively 20% Trichoderma conidial powder, 5% wetting agent, 10% dispersant and 65% carrier, be processed into wetting powder, measure wetting time and suspensibility.
(2) screening of wetting agent and dispersant optimal proportion: selected auxiliary agent kind and ratio are carried out orthogonal design, determine the best usage ratio of the two.
(3) screening of uv-protector: will add in the Trichoderma spore suspension for the uv-protector of examination, take do not add uv-protector without ultra violet lamp for contrast one, not adding uv-protector, through ultra violet lamp for contrasting two, 100 μ l10
3Individual/ml trichoderma asperellum spore is applied on the PDA flat board, and each is processed and repeats 3 times, behind uviol lamp (30W, light intensity 120lx) irradiation 2min, places 28 ± 1 ℃ of incubator dark culturing 48h to calculate clump count.
2.4 the processing of formulation
The various auxiliary agents of selecting through testing sieve are crossed respectively 400 mesh standard sieves, join in the carrier in proportion to stir, and 7,000,000,000/g Trichoderma conidial powder of then drying being crossed joins in the mixture, is processed into wetting powder after mixing.
2.5 properties of product are measured
Spore germination rate is measured, and the Trichoderma spore suspension is inoculated in the PDA nutrient solution, and after cultivating 20h under 28 ± 1 ℃ of temperature and the 160r/min rotating speed, microscope inspection statistics spore count alive calculates the spore rate of living.
Wetting time press GB/T5451-2001 measure (≤1min).Suspensibility is pressed GB/T14825-2006 and is measured (〉=75%).The pH value is pressed GB/T1600-93 and is measured, and fineness is measured by the laser particle distribution instrument.
2.6 the test of pesticide effectiveness
(1) isolated test
Trichoderma wettable powder is diluted to respectively 40,80,120,160 times of conidial suspensions, draw respectively each spore suspension 1mL and inject the sterilization culture dish, add again 9mL and be cooled to 45 ° of PDA medium about C, shake gently and make it mixing, make flat board, then picking pathogen bacterium piece is transplanted to dull and stereotyped central authorities respectively, every processing repeats 3 times, replace the bacterial strain spore suspension in contrast with sterile water, behind constant temperature culture 3d, measure the pathogen colony diameter, calculate and respectively process inhibiting rate.
Wherein, 4mm is the diameter of access bacterium cake.
(2) pot experiment
Respectively botrytis cinerea, rape sclerotium and three kinds of pathogens of rice sheath blight disease are gone to the dull and stereotyped activation of PDA 3d, go to again in the PD liquid nutrient medium, every bottle turns 5, cultivate 6d, beat mycelia, survey the light transmittance spray inoculation, moisturizing is cultivated cucumber seedling to morbidity, and three kinds of pathogen light transmittances are respectively 1.1%, 3.4% and 9.8%; Trichoderma wettable powder is diluted respectively 400 times, 800 times and 1600 times of liquid, and spraying control pathogen checks control efficiency behind the 7d, according to the sick level index criteria for classifying (table 2), calculates control efficiency according to following formula.
The sick grade index criteria for classifying of table 2
| Sick level | The state of an illness |
| 0 | Without any symptom |
| 1 | Lesion area accounts for below 5% of one-piece blade area |
| 3 | Lesion area accounts for the 6-10% of one-piece blade area |
| 5 | Lesion area accounts for the 11-25% of one-piece blade area |
| 7 | Lesion area accounts for the 26-50% of one-piece blade area |
| 9 | Lesion area accounts for more than 50% of one-piece blade area |
Disease index=(∑ (sick value of series * this disease level number of sheets) * 100)/(sick level peak * investigation number of sheets)
Control efficiency (%)=(CK disease index-processing disease index) * 100/CK disease index
3, data statistic analysis
Experimental data adopts SPSS 16.0 statistical softwares to carry out variance analysis.Adopt the otherness (P=0.05) between Duncan ' s duncan's new multiple range method comparison process, digital back lowercase is different in the table, is illustrated in significant difference on 95% level.
Two, results and analysis
1, antagonism interpretation of result
Trichoderma shows the antagonistic effect of Rhizoctonia solani Kuhn: the trichoderma asperellum bacterium has inhibitory action to Rhizoctonia solani Kuhn.Because the growth rate of Rhizoctonia solani Kuhn is fast than Trichoderma, Rhizoctonia solani Kuhn contacts with the mycelia of Trichoderma behind the 1d, Rhizoctonia solani Kuhn stops growing behind the 2d, and aerial hyphae is cleared up, the bacterium colony attenuation is through the continued growth of microscopy Trichoderma, and Trichoderma is covered with whole culture dish behind the 5d, has covered the Rhizoctonia solani Kuhn bacterium colony, the atrophy of Rhizoctonia solani Kuhn bacterium colony, and produce a large amount of green spores.
Trichoderma shows the antagonistic effect of Fusarium oxysporum: when Trichoderma and Fusarium oxysporum are cultivated in the face-off of PDA flat board, the growth of Trichoderma is rapid, Trichoderma contacts with the Fusarium oxysporum silk behind the 2d, Trichoderma begins to surround Fusarium oxysporum and falls behind the 3d, the Fusarium oxysporum of contact falls to beginning atrophy, stops growing.Position in contact behind the 4d produces the antagonism line, and this moment, Trichoderma was covered with whole flat board.Trichoderma produces a large amount of green spores at most advanced and sophisticated spore bacterium colony behind the 5d.
Trichoderma shows willow skin rot antagonistic effect: when Trichoderma and willow skin rot are cultivated in the face-off of PDA flat board, the growth rate of Trichoderma falls faster than Cytospora chrysosperma, Trichoderma contacts with the Cytospora chrysosperma silk behind the 2d, Trichoderma begins to surround Cytospora chrysosperma and falls behind the 3d, Trichoderma is further invaded the pathogen bacterium colony, finally occupy the pathogen growth and be subject to severe inhibition, pathogen face-off radius is much smaller compared with the control, and Trichoderma is covered with full ware and produces a large amount of spores willow skin rot pathogen when cultivating 4d.In the face-off incubation, to wilt with the pathogen mycelia of trichoderma asperellum bacterium face-off, bacterium colony is uneven.
Trichoderma shows the antagonistic effect of leaf blight: when Trichoderma and leaf blight are cultivated in the face-off of PDA flat board, the growth rate of Trichoderma falls faster than leaf spoting bacteria, 1.5d rear trichoderma asperellum bacterium contacts with leaf blight, the edge pathogen colony growth of contact is subject to obvious inhibition, the growth radius of its bacterium colony is starkly lower than the contrast radius, and wilting appears in the edge, the pigment of leaf blight is thin out simultaneously, the recessed low injustice of bacterium colony, behind the 2d Trichoderma from around surround the former bacterium of leaf blight, an antibacterial band appears in Trichoderma and leaf blight after cultivating 4d, observes simultaneously Trichoderma and grows in leaf blight, and produce a large amount of green spores.
Trichoderma shows the antagonistic effect of gray mold: when trichoderma asperellum bacterium and gray mold are cultivated in the face-off of PDA flat board, 1.5d rear trichoderma asperellum bacterium contacts with the gray mold mycelia, the trichoderma asperellum bacterium begins to surround the gray mold bacterium colony behind the 3d, the pathogen growth is suppressed, and no longer to the mould Directional Extension of inoculation wood, the trichoderma asperellum mycelia further expands again invades the pathogen bacterium colony, is wilted by the position bacterium colony that Trichoderma is invaded, and the pathogen bacterium colony is uneven.Trichoderma is covered with full ware and produces a large amount of spores in gray mold when cultivating 4d, the mycelia of picking two bacterium contact, microscopy observe trichoderma asperellum to the grey mold mycelia by winding, hyperparasitism, infect, dissolve, Antagonizing destroys gray mold to form attachment filament and wear out etc., and then the growth of inhibition botrytis cinerea mycelia.
Table 3 trichoderma asperellum bacterium is to the face-off result of each pathogen
As shown in Table 3: the trichoderma asperellum bacterial strain has inhibitory action to 4 kinds of pathogens, inhibiting rate is followed successively by leaf blight from high to low〉Fusarium oxysporum〉the mashed skin of poplar〉gray mold〉Rhizoctonia solani Kuhn, because when Antagonistic Fungus suppressed pathogen, itself was also suppressed by pathogen, for this reason, when calculating antagonistic effect, adopt relative fungistatic effect, what wherein inhibition was best is leaf blight, secondly the willow skin rot, Fusarium oxysporum takes second place, and is Rhizoctonia solani Kuhn at last.The quality of inhibition illustrates that also Trichoderma has certain selectivity to the Competition of plant pathogenic fungi.
2, the screening of carrier
Table 4 carrier is on the impact of Trichoderma spore germination and growth rate
| Carrier | Working concentration (mg/ml) | Bacterium colony (individual/ware) | Colony diameter daily growth amount (mm/d) |
| Attapulgite | 1000 | 41c | 20.08cd?CD |
| White carbon | 1000 | 43c | 24.25aA |
| Kaolin | 1000 | 38c | 18.92dD |
| Diatomite | 1000 | 54ab | 21.33bc?BC |
| Bentonite | 1000 | 51b | 21.75bB |
| Contrast | 1000 | 58a | 22.5bB |
Annotate: lowercase alphabet shows 0.05 horizontal significant difference in the table, and capitalization represents 0.01 horizontal significant difference (together lower).
Table 4 shows for 5 kinds of carriers of examination from the conidia germination impact, except diatomite and contrast without (P〉0.05) significant difference, all the other 4 kinds of carriers and contrast difference obvious (P＜0.05); From on the mycelial growth, white carbon promotes mycelial growth, than contrast difference significantly (P＜0.05), yet as spore as the main body that infects, and then consider carrier from the angle of spore germination, diatomite, bentonite and contrast difference be significantly (P＜0.05) not; Available conidial powder as trichoderma wettable powder.
3, the screening of wetting agent and dispersant
Table 5 wetting agent wetting time relatively
| Wetting agent | Wetting time (s) |
| TERWET1010 | 11.00±1.00cCD |
| WLNO | 48.67±0.58aA |
| Morwet?EFW | 6.67±0.58dD |
| SDS | 13.33±0.58cC |
| MF-5 | 13.33±0.58cC |
| Pull open powder | 42.33±2.52bB |
Annotate: the wetting agent working concentration is 5%, and each experiment repeats 5 times.
It is 5% that the usage amount of each wetting agent is selected in this test, and to carrying out scalping for the examination wetting agent, wetting agent wetting time comparative result sees Table 5, and the result shows that the wetting power of different wetting agent there are differences, and TERWET1010, SDS, MF-5 difference are not remarkable.The wetting agent (TERWET1010, Morwet EFW, SDS, MF-5) that filters out wetting time 30s is taken into account simultaneously biocompatibility test and is seen Table 6.The result shows: Morwet EFW, TERWET 1010, MF-5 all can be considered as its wetting agent to Trichoderma conidia germination and contrast difference not significantly (P＜0.01).
Table 6 wetting agent is on the impact of Trichoderma spore germination and growth rate
| Wetting agent | Concentration (μ g/mL) | Bacterium colony (individual/ware) | Colony diameter daily growth amount (mm/d) |
| Morwet?EFW | 120 | 52ab?AB | 19.00bB |
| MF-5 | 120 | 45c?B | 23.42aA |
| SDS | 120 | 30d?C | 12.58cC |
| TERWET?1010 | 120 | 49b?B | 12.67cC |
| Contrast | - | 58a?AB | 22.5aA |
Table 7 dispersant is on the impact of Trichoderma spore germination and growth rate
Filter out preferably dispersant of dispersiveness through range estimation, can find out through biocompatibility test (table 7): on the impact of Trichoderma conidia germination, except TERSPERSE 2020, NNO, sodium lignin sulfonate and contrast difference significantly (P＜0.05), the dispersant shown in all the other and contrast difference not remarkable (P〉0.05).On the day growth amount to colony diameter, 250 μ g/mL calcium lignosulfonates and NNO play a driving role to the growth of Trichoderma bacterium colony, with contrast difference remarkable (P＜0.05), than contrast difference not significantly (P〉0.05), 2020 pairs of colony growths of TERSPERSE play obvious inhibitory action to colony growth for sodium lignin sulfonate and Morwet D425.One or several mix dispersant as Trichoderma conidium wetting powder can to consider calcium lignosulfonate, Morwet D425, sodium lignin sulfonate in conjunction with both.Selected wetting agent, dispersant are added in the Trichoderma conidial powder according to 5%, 10% ratio respectively, and carrier complements to 100%, is processed into wetting powder, surveys its wetting time and suspensibility (table 8).Select that suspensibility is higher, wetting time shorter one is as quadrature factor, different auxiliary agents the results are shown in Table 9 by 3 levels of mass fraction consumption design commonly used.By orthogonal, obtained experimental result is in table 10.His-and-hers watches 10 data are carried out the results of analysis of variance in table 11.Can be drawn by table 9 and 10, the more excellent prescription of trichoderma wettable powder is A1B2C2D3, i.e. Morwet EFW1%, TERWET 10105%, and Morwet D425 5% calcium lignosulfonate 7%, by variance analysis, influence factor is followed successively by D＞B＞C＞A.
Wetting powder wetting time and the suspensibility of the agent of table 8 different wetting and dispersant preparation
Factor and the level of table 9 wetting agent and dispersant screening
Annotate: data are the mass fraction that is of 5 factors.
Table 10 orthogonal experiments
Table 11 variance analysis
| The source | SS | DF | MS | F | P |
| A (error E rror) | 43.60 | 2 | 21.80 | ? | ? |
| B | 95.76 | 2 | 47.88 | 2.20 | ? |
| C | 86.66 | 2 | 43.33 | 1.99 | ? |
| D | 974.52 | 2 | 487.26 | 22.35 | <0.05 |
4, the screening of uv-protector
The different uv-protectors of table 12 are on the impact of Trichoderma spore germination
| Protectant | Clump count/cfu |
| VC | 125±7.00bA |
| Xanthans | 119±4.04bB |
| Nano zine oxide | 126±8.89bA |
| Fluorescein sodium | 88±9.00cC |
| Contrast 1 | 144±8.62aA |
| Contrast 2 | 79±5.51cC |
Annotate: working concentration is 0.2%.
The nano zine oxide of table 13 variable concentrations is to the protective effect of spore
Result of the test is (table 12) as can be known, and selected four kinds of uv-protectors are different to the conidial protective effect of Trichoderma, and it is not obvious to the conidial protective effect of Trichoderma to remove fluorescein sodium, not remarkable than contrast difference; Though all the other 3 kinds with contrast 1 significant difference, 2 compare with contrast, obvious to the protective effect of spore, reach utmost point significance level (P＜0.01), secondly the protection best results of nano zine oxide is VC, xanthans.Therefore select nano zine oxide as the uv-protector of trichoderma wettable powder.Through further screening (table 13), be 0.3% from economical and protection effect selection optium concentration.
5, properties of product detect
According to above-mentioned result of study preparation trichoderma wettable powder (formula I II), carry out property indices and measure.The germination rate of Trichoderma 〉=88.57%, wetability＜1min, quality suspensibility 75.79%, spore suspensibility 80.12%, moisture 3.52%, fineness (cross 325 mesh standard sieves, particle mean size is 27.00m) 〉=98%, the pH value is 6.7.The performance indications of the wetting powder of preparing meet the requirement of commodity preparation.
6, the test of pesticide effectiveness
This test selects 10 kinds of agricultural pathogens to carry out stripped control experiment (table 14), the result shows that trichoderma asperellum bacterium wetting powder has preferably inhibiting rate to the cercospora brown spot of peanut, ring rot of apple, early blight of tomato, rice sheath blight disease, and 1200 times of liquid of the wetting powder of this Trichoderma are better than the drug effect of contrast to the inhibiting rate of ring rot of apple, and to the cercospora brown spot of peanut, early blight of tomato significantly better than the contrast drug effect; Though to the too late contrast of the control drug effect of sclerotinia rot of colza, its inhibiting rate is also very considerable, the inhibiting rate of low dosage reaches 81.8%; And the wetting powder of this Trichoderma is withered to cucumber, the control of gibberella saubinetii, gray mold of cucumber, potato late blight is not as good as the contrast drug effect, but the inhibiting rate that high dose is processed cucumber fusarium axysporum, gray mold of cucumber, potato late blight also reaches about 70%.
Table 14 trichoderma asperellum bacterium wetting powder is to the inhibitory action of each pathogen
Trichoderma wettable powder shows (table 15) to gray mold of cucumber, rape sclerotium and rice banded sclerotial blight control efficiency, and 400 times, 800 times and 1600 times liquid of trichoderma wettable powder all are lower than contrast 50ppm tpn and 250ppm bactericide Fluoxastrobin to three kinds of pathogen control efficiency; Wherein, best to the rice sheath blight disease controlling effect, be sclerotinia rot of colza secondly, control efficiency all reaches more than 50%, and the poorest to the gray mold of cucumber control efficiency, and this experimental result is consistent with the inhibition experiment of exsomatizing.The trichoderma wettable powder of the present invention's development can be used for preventing and treating the formulation of electing of rape sclerotium and rice sheath blight disease.
Table 15 trichoderma asperellum bacterium wetting powder is to the control efficiency of each pathogen
Claims (10)
1. trichoderma asperellum bacterium wetting powder, it is characterized in that described pulvis made by following composition: 10～30wt.% trichoderma asperellum, 2～10%wt.% wetting agent, 7～20%wt.% dispersant, 0.1～0.5wt.% uv-protector, surplus are carrier.
2. trichoderma asperellum bacterium wetting powder according to claim 1 is characterized in that described pulvis made by following composition: 10～30wt.% trichoderma asperellum, 2～10%wt.% wetting agent, 7～20%wt.% dispersant, 0.1～0.5wt.% uv-protector, 40～80wt.% carrier.
3. trichoderma asperellum bacterium wetting powder according to claim 1 is characterized in that described pulvis made by following composition: 10～30wt.% trichoderma asperellum, 2～10%wt.% wetting agent, 10～15%wt.% dispersant, 0.1～0.5wt.% uv-protector, 45～78wt.% carrier.
4. according to claim 1,2 or 3 described trichoderma asperellum bacterium wetting powders, the quality that it is characterized in that described trichoderma asperellum is 7,000,000,000/g.
5. according to claim 1,2 or 3 described trichoderma asperellum bacterium wetting powders, it is characterized in that described wetting agent is TERWET1010, WLNO, Morwet EFW, SDS, MF-5, pulls open one or more the mixture in the powder; Described dispersant is one or more the mixture in calcium lignosulfonate, TERSPERSE 2020, NNO, Morwet D425, the sodium lignin sulfonate; Described carrier is one or more the mixture in attapulgite, kaolin, diatomite, bentonite, the white carbon; Described uv-protector is one or more the mixture in dextrin, xanthans, fluorescein sodium, the VC nano zine oxide.
6. trichoderma asperellum bacterium wetting powder according to claim 5 is characterized in that described wetting agent is TERWET1010 and Morwet EFW; Described dispersant is calcium lignosulfonate and Morwet D425; Described carrier is bentonite; Described uv-protector is the VC nano zine oxide.
7. trichoderma asperellum bacterium wetting powder according to claim 6 is characterized in that described pulvis made by following composition: 10wt.% trichoderma asperellum, 0.5wt.% Morwet EFW, 1.5wt.%TERWET1010,2wt.%Morwet D425,5wt.% calcium lignosulfonate, 0.1wt.% nano zine oxide, 80.9wt.% bentonite.
8. trichoderma asperellum bacterium wetting powder according to claim 6 is characterized in that described pulvis made by following composition: 20wt.% trichoderma asperellum, 1wt.% Morwet EFW, 5wt.%TERWET1010,5wt.%Morwet D425,7wt.% calcium lignosulfonate, 0.3wt.% nano zine oxide, 61.7wt.% bentonite.
9. trichoderma asperellum bacterium wetting powder according to claim 6 is characterized in that described pulvis made by following composition: 30wt.% trichoderma asperellum, 2wt.% Morwet EFW, 8wt.%TERWET1010,10wt.%Morwet D425,10wt.% calcium lignosulfonate, 0.5wt.% nano zine oxide, 39.5wt.% bentonite.
10. the described trichoderma asperellum bacterium of the arbitrary claim of claim 1-9 wetting powder is as the application of control agricultural pathogenic fungus and bacteriosis.
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| CN103710270A (en) * | 2013-12-19 | 2014-04-09 | 广西壮族自治区林业科学研究院 | Decomposition agent for degrading litters of pine trees |
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| CN105794855A (en) * | 2014-12-30 | 2016-07-27 | 姜汉军 | Control method of potato cyst nematode |
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| CN105941484A (en) * | 2016-05-11 | 2016-09-21 | 江西正邦生物化工有限责任公司 | Trichoderma asperellum spore and phenazino-1-carboxylic acid compound sterilization composition |
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| CN107058126A (en) * | 2017-03-28 | 2017-08-18 | 东北林业大学 | One plant of trichoderma asperellum and its application |
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| CN110343621B (en) * | 2019-07-10 | 2021-04-30 | 贵州省植物保护研究所 | Trichoderma asperellum strain and application thereof |
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