CN106386869A - Trichoderma seed coating as well as preparation method and application thereof - Google Patents
Trichoderma seed coating as well as preparation method and application thereof Download PDFInfo
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
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Abstract
The invention discloses a trichoderma seed coating as well as a preparation method and an application thereof. The seed coating is prepared from trichoderma chlamydospore fermentation broth, a film-forming agent, chitosan oligosaccharide, phenazino-1-carboxylic acid and humic acid, wherein each liter of trichoderma chlamydospore fermentation broth contains 13%-17%(W/V) of the film-forming agent, 0.35%-0.45%(W/V) of the chitosan oligosaccharide, 0.5%-1.5% mu g/ml of phenazino-1-carboxylic acid and 0.05%-0.1%(W/V) of the humic acid. The seed coating is convenient to prepare, high in efficiency, low in toxicity and relatively safe to environment, performs coating easily and has good control effect on soil-borne diseases of corn.
Description
Technical field
The present invention relates to a kind of Trichoderma seed coat agent and its preparation method and application, especially with regard to the life of biological and ecological methods to prevent plant disease, pests, and erosion type Trichoderma
Thing seed coat agent preparation method and the application technology in corn stalk rot disease, banded sclerotial blight, root rot preventing and treating.
Background technology
Trichoderma is that a kind of habit of generally existing in soil and the plant rhizosphere ecosystem occupies fungi.Trichoderma is public in the world
The beneficial microbe recognized, mainly passes through the mechanism such as nutrition and competition for space, antibiosis, hyperparasitism and induction of resistance and prevents and treats
The soil-borne disease of all kinds of crops, including crop seedling diseases, root rot and droop etc..Trichoderma not only has the work(of controlling disease
Can, also there is promotion plant root growth simultaneously, improve nutritional utilization efficiency, improve crop yield and resistance, rehabilitating soil
Etc. function, it is that a kind of disease-preventing and yield-increasing acts on significant multifunctional microbial.
In recent years, Maize Soil Borne Disease, such as corn stalk rot disease and banded sclerotial blight occur to increase year by year, some areas generally time
The incidence of disease be 15-20%, serious time incidence reaches more than 40%, causes serious production loss, and has become complete
The chief threat of state's corn producing region good quality and high output.Moreover, because the high rate of corn stalk rot disease has caused serious jade
Rice lodging phenomenon, limits the popularization of corn mechanization production technology.Main with regard to Trichoderma biocontrol agent both at home and abroad for many years
Applied with forms such as compost, granules, seed coat agent is prepared with Trichoderma and successfully reports less, U.S.'s Biowork trichoderma
Bacterium biological seed coating agent is also only applied to the preventing and treating to corn seedling diseases and improves corn yield, and seldom has research to be used for preventing and treating corn
Stem rot (Fusarium spp., Pythium spp.) and banded sclerotial blight (Rhizoctonia solani) that late growth stage occurs.
Due to Corn stalk rot pathogens and sheath blight fungus, to infect period longer, all can infect, and aobvious disease is usually to exist from seedling stage to Adult plant
After pustulation period, therefore, during this requires biological control, microorganism colonizes to soil and root of the crop cording long period
Ability and the function of system induction plant resistance to environment stress, thus can prevent and treat later growth period disease after realizing microbiological treatment seed
Generation.
Content of the invention
For defect of the prior art, it is an object of the invention to provide a kind of Trichoderma seed coat agent and preparation method thereof and
Application, effective for preventing and treating corn stalk rot disease, banded sclerotial blight and root rot etc..The present invention is screening from substantial amounts of Trichoderma resource
The antagonistic Trichoderma bacterial strain obtaining, has very strong competitiveness in soil, the breeding with root table germ effectively in suppression soil
With colonize, the bacterial strain that what is more important is screened has Systemic induction resistance, that is, Trichoderma with seed sprouting colonize in
Maize root system, produces, in root system, the system expression that a series of exciton to induce defense genes in root system, and holds
The effect phase can reach whole corn growth stage (Wu Xiao etc. 2015), thus realizing the preventing and treating to the later stage disease such as stem rot, banded sclerotial blight.
Therefore, develop and can prevent and treat the Bio-control Trichoderma seed coat agent of Maize Soil Borne Disease disease and be promoted the use of to corn in the whole nation
Green prevention and control tool be of great significance, have a extensive future.
The purpose of the present invention is achieved by the following technical solution：
In a first aspect, the present invention provides a kind of Trichoderma seed coat agent, described seed coat agent includes trichoderma harzianum chlamydospore fermentation
Liquid, film forming agent, chitosan oligosaccharide, shenqinmycin, humic acid；
Wherein, 13-17% (W/V) film forming agent, 0.35-0.45% (W/V) are contained in every liter of trichoderma harzianum chlamydospore zymotic fluid
Chitosan oligosaccharide, 0.5-1.5ug/ml shenqinmycin, 0.05-0.1% (W/V) humic acid.
Preferably, shenqinmycin containing 1.0ug/ml, 0.06% (W/V) humic in every liter of trichoderma harzianum chlamydospore zymotic fluid
Acid.
Preferably, described film forming agent includes 853 film forming agents.
Preferably, described Trichoderma is Trichoderma harzianum SH2303, preserving number：CTCCSJ-A-QT11266.
Preferably, the preparation of described trichoderma harzianum chlamydospore zymotic fluid include as follows：Will with the inoculum concentration of 0.5% (V/V)
First class inoculum is inoculated in trichoderma liquid fermentation medium, 28 DEG C of fermented and cultured 7 days, obtains final product；
Wherein, described first class inoculum be by Trichoderma bacteria cake with the inoculum concentration of 3-5 piece/triangular flask in PD culture medium,
180r/min, under the conditions of 28 DEG C culture obtain within 5 days.
Preferably, the composition of described trichoderma liquid fermentation medium includes：Every 1000 liters of Trichoderma liquid fermentation and culture
40kg containing corn flour, magnesium sulfate 400g, ferrous sulfate 6.0g, manganese sulfate 2.0g, zinc sulfate 1.6g, potassium dihydrogen sulfate in base
3.06kg, sodium nitrate 1.13kg, ammonium nitrate 880g, calcium chloride 800g, balance of water, initial pH is adjusted to 4-5.
PD culture medium of the present invention and trichoderma liquid fermentation medium are before use all through 121 DEG C of autoclavings
25min.
Preferably, the tinning amount of described fermented and cultured controls：200L liquid fermentation medium loads in 300L fermentation tank.
Second aspect, the present invention provides a kind of preparation method of described seed coat agent, comprises the steps：Will in described ratio
Shitosan, shenqinmycin, humic acid, the mixing of trichoderma harzianum chlamydospore zymotic fluid, are subsequently adding film forming agent mixing, obtain final product kind of a clothing
Agent.
The third aspect, the present invention provides a kind of described Trichoderma seed coat agent controlling Maize Soil Borne Disease as biological prevention and control agent
In application.
Preferably, described soil-borne disease includes preventing and treating corn stalk rot disease, banded sclerotial blight, root rot etc..
Fourth aspect, the present invention provides a kind of user in controlling Maize Soil Borne Disease for described Trichoderma seed coat agent
Method, is 1 specially by described Trichoderma seed coat agent and corn seed in mass ratio:(10-15) uniformly mix, you can.
Through Trichoderma primary dcreening operation, secondary screening is stored in teat glass inclined-plane the present invention after purification.In corn flour nutrient solution
Carrying out liquid fermentation makes it produce trichoderma harzianum chlamydospore zymotic fluid, will be containing chlamydosporic zymotic fluid (100,000,000 CFU/ in a large number
ML) it is mixed and made into Trichoderma with film forming agent, chitosan oligosaccharide, shenqinmycin, humic acid according to the optimum proportioning that the design of response face obtains
Seed coat agent, Maize Seeds Coated (1:10-1:15W/W), sowed with field in greenhouse.Compared with the control, preventing and treating corn soil
Pass disease effect is significant, average out to more than 60%.
Compared with prior art, it is an advantage of the current invention that：
1. in this Trichoderma compound bio seed coat agent, do not only have Trichoderma thickness spore zymotic fluid, also chitosan oligosaccharide and Shen simultaneously
Two kinds of biogenic pesticide compositions of piperazine mycin (phenazine-1-carboxylic acid).Research shows, trichoderma fermented liquid has suppression Fusariumsp
(Fusarium spp), Rhizoctonia solani Kuhn (Rhizoctonia solani), the effect of pythium spp (Pythium spp) lure simultaneously
Lead corn resistance, chitosan oligosaccharide (Chitosan oligosaccharide) has the function of luring corn resistance, and shenqinmycin tool
There is the function of suppression sheath blight fungus.
2. Trichoderma biological seed coating agent is to be assembled by three kinds of biological source optimizing components to form, and functionally has complementary advantages, is one
Plant the new seed coat agent preventing and treating Maize Soil Borne Disease；Not only to corn stalk rot disease, banded sclerotial blight etc. has obvious preventive effect, also simultaneously
The growth indexes of plant are had by significant facilitation, improve stress resistance of plant, increases crop yield.
3. the Trichoderma biological seed coating agent providing according to the present invention, requires Maize Seeds Coated according to a certain percentage and broadcasts
Kind, pollution-free to soil.On Sichuan Nanchong, Yinchuan of Ningxia Province, Hebei Shijiazhuang, zunyi, guizhou, Dandong, Nanjing and other places
Field test, Trichoderma biological seed coating agent all can reach more than 60% to corn stalk rot disease, banded sclerotial blight preventive effect, corn yield increasing 10%
More than.
4. the present invention is applied to the trichoderma strain that great majority have good biocontrol effect, and preparation method is simple, low cost,
It is suitable for different scales to produce, wide market.
Brief description
The detailed description with reference to the following drawings, non-limiting example made by reading, the further feature of the present invention,
Objects and advantages will become more apparent upon：
Fig. 1 is film forming agent species the selection result schematic diagram；
Fig. 2 is seed coat agent shelf life measurement result schematic diagram of the present invention.
Specific embodiment
With reference to specific embodiment, the present invention is described in detail.Following examples will be helpful to the technology of this area
Personnel further understand the present invention, but the invention is not limited in any way.It should be pointed out that the ordinary skill to this area
For personnel, without departing from the inventive concept of the premise, some deformation can also be made and improve.These broadly fall into the present invention
Protection domain.
Embodiment 1
1st, flat board actication of culture：Trichoderma harzianum SH2303 is inoculated in PDA culture dish, in 28 DEG C of incubators, is inverted culture
4-5 days.
PDA culture medium：The upper strata juice filtering after 200g potato boiling, 20g glucose, 20g agar powder, pure water is fixed
Hold to 1L, every 250mL dispenses to 500mL triangular flask, 121 DEG C of autoclaving 25min.
2nd, the preparation of first class inoculum：200g potato, 20g glucose, pure water 1L, every 250mL dispenses to 500mL triangle
Bottle, 121 DEG C of autoclaving 25min.Access flat board bacterial classification, be placed in 180r/min, Shaking culture 5 days in 28 DEG C of shaking tables, breathed out
Thatch trichoderma SH2303 liquid fermentation bacterial classification.
3rd, trichoderma liquid fermentation：Corn flour 8kg, magnesium sulfate 80g, ferrous sulfate 1.2g, manganese sulfate 0.4g, zinc sulfate
0.32g, potassium dihydrogen sulfate 0.61kg, sodium nitrate 0.23kg, ammonium nitrate 176g, calcium chloride 160g, water supplies 200L, loads 300L
Fermentation tank.The initial pH of culture medium is adjusted to 4-5.High-temperature sterilization 40-60min at 121 DEG C.Access weight than the trichoderma for 0.5%
Bacteria liquid fermented bacterium, fermentation temperature is 28 DEG C, fermentation time 7 days, as final trichoderma harzianum chlamydospore zymotic fluid.
4th, Trichoderma biological seed coating agent preparation：Trichoderma fermented liquid 1L, shitosan 4g, shenqinmycin 1mg, humic acid
0.6g, is sufficiently mixed.Above-mentioned mixed liquor is mixed with 13% (W/V) film forming agent, as Trichoderma biological seed coating agent.
5th, Trichoderma biological seed coating agent promotes growth of maize effect
Greenhouse pot culture experimental result shows, compared with the control, seeding ratio after seed coat agent Cotton seeds, plant plant height and
Main root length has a certain degree of increase, and increment rate is respectively 6.32%, 9.84% and 3.34%, but does not all show with contrast difference
Write (p>0.05)；Other each growth indexes are notable (table 3) with contrast difference.
The impact to corn growth for the table 3 seed coat agent Cotton seeds
Embodiment 2
1st, flat board actication of culture：Trichoderma harzianum SH2303 is inoculated in PDA culture dish, in 28 DEG C of incubators, is inverted training
Foster 4-5 days.
PDA culture medium：The upper strata juice filtering after 200g potato boiling, 20g glucose, 20g agar powder, pure water is fixed
Hold to 1L, every 250mL dispenses to 500mL triangular flask, 121 DEG C of autoclaving 25min.
2nd, the preparation of first class inoculum：200g potato, 20g glucose, pure water 1L, every 250mL dispenses to 500mL triangle
Bottle, 121 DEG C of autoclaving 25min.Access flat board bacterial classification, be placed in 180r/min, Shaking culture 5 days in 28 DEG C of shaking tables, obtain wood
Mould liquid fermentation bacterial classification.
3rd, trichoderma liquid fermentation：Corn flour 8kg, magnesium sulfate 80g, ferrous sulfate 1.2g, manganese sulfate 0.4g, zinc sulfate
0.32g, potassium dihydrogen sulfate 0.61kg, sodium nitrate 0.23kg, ammonium nitrate 176g, calcium chloride 160g, water supplies 200L, loads 300L
Fermentation tank.The initial pH of culture medium is adjusted to 4-5.High-temperature sterilization 40-60min at 121 DEG C.Access weight than the trichoderma for 0.5%
Bacteria liquid fermented bacterium, fermentation temperature is 28 DEG C, fermentation time 7 days, as final trichoderma harzianum chlamydospore zymotic fluid.
4th, the preparation of Trichoderma biological seed coating agent：Trichoderma fermented liquid 1L, shitosan 4g, shenqinmycin 1mg, humic acid
0.6g, is sufficiently mixed.Above-mentioned mixed liquor is mixed with 17% (W/V) 853 film forming agent, as Trichoderma biological seed coating agent.By pharmacopoeia
Ratio 1:10 (W/W) dresses seed.With blank seed coat agent (only containing film forming agent) for comparison.
5th, biological seed coating agent community trial：Stem rot Fusariumsp or sheath blight fungus wheat medium culture 10 will be incubated at
My god, in corn seeding, it is seeded in around seed, every cave 2g wheat inoculum, each processes inoculation 50 caves, in triplicate, right
According to the blank seed coat agent of coating, treat that corn enters the pustulation period, investigate the incidence to corn stalk rot disease, calculate preventive effect.
Incidence of disease %=diseased plant number/investigation total strain number × 100%
Preventive effect=[(incidence of disease in the incidence of disease-process field in comparison field)/comparison field incidence of disease] × 100% community trial table
Bright：After biological seed coating agent Cotton seeds, disease index drops to 45.83 by 70 compareing；Dead leaf number, early ageing rate are all than comparison
Low, difference all reaches the pole level of signifiance (p<0.01)；Lodging rate is less than comparison, and difference all reaches the level of signifiance (p<0.05)；Fringe hangs down
Rate is also below comparison, but the not notable (p of difference>0.05).The generation feelings of Causal Organism of Maize Basal Stalk can be mitigated after showing seed coat agent coating
Condition (table 4).Investigation shows further：Trichoderma biological seed coating agent is obvious to stem rot and banded sclerotial blight preventive effect, to banded sclerotial blight preventive effect
Reach 57.35%, 75% has been reached to the preventive effect of stem rot.
The preventive effect to corn stalk rot disease for table 4 seed coat agent
The preventive effect to banded sclerotial blight and stem rot for table 5 seed coat agent
| Process | Banded sclerotial blight diseased plant rate (%) | Preventive effect (%) | Stem rot diseased plant rate (%) | Preventive effect (%) |
| Repeat 1 | 13.50 | 55.00 | 3.75 | 75.00 |
| Repeat 2 | 11.25 | 62.50 | 3.75 | 75.00 |
| Repeat 3 | 13.65 | 54.55 | 3.75 | 75.00 |
| CK | 30.00 | ---- | 15.00 | ---- |
Embodiment 3
1st, flat board actication of culture：It is inoculated in breathing out thatch green trichoderma SH2303 in PDA culture dish, in 28 DEG C of incubators, be inverted training
Foster 4-5 days.
PDA culture medium：The upper strata juice filtering after 200g potato boiling, 20g glucose, 20g agar powder, pure water is fixed
Hold to 1L, dispense to 250mL triangular flask, 121 DEG C of autoclaving 25min.
2nd, the preparation of first class inoculum：200g potato, 20g glucose, pure water 1L, dispense to 250mL triangular flask, 121
DEG C autoclaving 25min.Access flat board bacterial classification, be placed in 180r/min, in 28 DEG C of shaking tables, Shaking culture 5 days, obtains Trichoderma harzianum
SH2303 liquid fermentation bacterial classification.
3rd, trichoderma liquid fermentation：Corn flour 8kg, magnesium sulfate 80g, ferrous sulfate 1.2g, manganese sulfate 0.4g, zinc sulfate
0.32g, potassium dihydrogen sulfate 0.61kg, sodium nitrate 0.23kg, ammonium nitrate 176g, calcium chloride 160g, water supplies 200L, loads 300L
Fermentation tank.The initial pH of culture medium is adjusted to 4-5.High-temperature sterilization 40-60min at 121 DEG C.Access weight than the trichoderma for 0.5%
Bacteria liquid fermented bacterium, fermentation temperature is 28 DEG C, fermentation time 7 days, as final trichoderma harzianum chlamydospore zymotic fluid.
4th, Trichoderma biological seed coating agent preparation：Trichoderma fermented liquid 1L, shitosan 4g, shenqinmycin 1mg, humic acid
0.6g, is sufficiently mixed.Above-mentioned mixed liquor is mixed with 14% (W/V) 853 film forming agent, as Trichoderma biological seed coating agent.By pharmacopoeia
Ratio 1：15 (W/W) dresses seed.
5th, Field information test：Arrange as zunyi, guizhou, Sichuan Nanchong, Hebei Shijiazhuang and other places for examination base.Work as landlord
The corn variety planted, seed is uniform, germination percentage is normal.Selection generation over the years pest and disease damage is heavier, and physical features is flat, is difficult ponding, no
The plot that drought and waterlogging easily occur is for examination field block；Experimental design adopts random district's groups arrangement.Result shows：Trichoderma biological seed coating
Agent has preferable preventive effect (table 6) to these three diseases.
Table 6 Trichoderma biological seed coating agent prevents and treats soil-borne disease effect
Seed coat agent component of the present invention has different degrees of suppression and antagonism to various pathogenic bacteria.This test is passed through not
Show with concentration proportioning comparative study, chitosan concentration is 3.5-4.5mg/mL, piperazine mycin concentration is that both 1ug/mL are jointly right
Corn sickle-like bacteria has more obvious inhibitory action, and inhibiting rate is 53.3%.This concentration proportioning is with Trichoderma thickness wall zymotic fluid altogether
Reach 62.9% with to the inhibition of sickle-like bacteria.And on this basis, the humic acid of addition different proportion, to pathogen
Inhibitory action has apparent difference.Content of humic acid in the range of 0.05-0.1% (W/V), prevent to pathogen by combination matching
Effect is extremely notable compared with the control, the same significant difference than the proportioning preventive effect being added without humic acid simultaneously, and preventive effect significantly improves, and reaches
73.5%.And when humic acid concentration is less than 0.05% (W/V) or is higher than 0.1% (W/V) scope, preventive effect is decreased obviously, or even
The addition of humic acid has certain side effect to the suppression germ effect of the former proportioning.Therefore, in strict accordance with 0.05-0.1% (W/
V make compound Trichoderma seed coat agent in ratio addition proportioning) has extremely obvious preventive effect to pathogen.(table 7a, table 7b)
Meanwhile, seed coat agent component of the present invention has certain facilitation to the growth of corn.Chitosan concentration is 3.5-
4.5mg/mL, piperazine mycin concentration are that both 1ug/mL are common has facilitation to maize radicel and plumule growth, but facilitation
Inconspicuous；And after this concentration proportioning is combined with Trichoderma thickness wall zymotic fluid, the growth to maize germ and radicle has significantly
Facilitation；The increment of plumule reaches the twice of comparison, radicle elongation 35.2%.Above-mentioned compound proportioning adds certain than
After the humic acid of example, synergy is extremely notable, and radicle elongation increases by 52.6%, and plumule elongation increases by 42.3%.And add corruption
The amount growing acid is outside 0.05-0.1% (W/V), inconspicuous to the growth-promoting effect of maize radicel and plumule, or even has suppression
Growth tendency.This explanation, there is synergy with Trichoderma growth promotion mechanism in humic acid within the specific limits.(table 8a, table 8b)
By optimizing triple combination formula, and it is mixed and made into biological seed coating agent with carriers such as film forming agents, preparation method is easy,
Low cost, is suitable to different scales production and application.
After the present invention is filled a prescription by optimum organization, it is mixed and made into biological seed coating agent according to a certain percentage with film forming agent.Through
Test, the difference of different film forming agents and its content all has a certain impact to the protection effect of seed coat agent.Through screening, select one
Plant the film forming agent (Fig. 1) of the growth no impact on Bio-control Trichoderma.Test shows, film forming agent 853 is no secondary to trichoderma bacteria growing to be made
With, or even have certain promotion growth.
Formula after to optimum organization is mixed by the present invention with variable concentrations 853 film forming agent, to sickle-like bacteria and vertical withered
Rhizoctonia carries out the mensure (table 9) of bacteriostasis rate.Result shows, 853 film forming agents of variable concentrations with above-mentioned proportion optimizing formula
After mixing, there is obvious impact to the inhibitory action of germ.When film forming agent concentration range is at 13-17% (W/V), bacteriostasis rate is
68%, difference is extremely notable.And concentration range is when below or above this scope, the significance of difference is not compared with the control for antibacterial radius
Substantially even not notable.Formula after this explanation finite concentration film forming agent (acrylic amide modified synthetic polymer) and above-mentioned optimization
Composition is likely to be of synergy, the biological and ecological methods to prevent plant disease, pests, and erosion bacteriostasis promoting certain material or promoting Trichoderma, thus the suppression to pathogen
Rate processed significantly improves.
Seed coat agent of the present invention carries out proportioning with corn seed according to different proportion, observes different ratio and processes to Maize at Seedling Stage
The impact of the prevention effect of soil-borne disease, determines pesticide-seeds ratio.From following table, it is poor to exist between different pesticide-seeds ratio Cotton seeds
Different.Pesticide-seeds ratio 1 is to Maize at Seedling Stage stem rot and banded sclerotial blight:10 Cotton seeds preventive effect highest, relative control effect is respectively
78% and 67.59%.Trichoderma is individually coated preventive effect between various dose process and differs greatly, with pesticide-seeds ratio 1:When 20, relative control effect is
Height, preventive effect is not strong with dosage correlation.Wherein seed coat agent pesticide-seeds ratio 1:After 10 process, the disease index of banded sclerotial blight and stem rot is remote
Process far below comparison and chemical pesticide, respectively 17.50 and 16.00, relative control effect (67.59%) (78.00%) is highest.
Comprehensive analysis, after Trichoderma Bio-seed Coating agent Cotton seeds, it is possible to decrease Maize at Seedling Stage disease index, optimum proportioning is 1:10,
Growth and resistance against diseases for the later stage are had laid a good foundation.
Note：Process 1. -3. be respectively seed coat agent press 1:10、1:20、1:40 pesticide-seeds ratio capsuled seeds；4. -6. solid with trichoderma
Body fermentate presses 1:10、1:20、1:40 pesticide-seeds ratio capsuled seeds；7. it is chemical pesticide hymexazol；8. blank (CK).“-”
For not existing.
Embodiment 4
1st, flat board actication of culture：It is inoculated in breathing out thatch green trichoderma SH2303 in PDA culture dish, in 28 DEG C of incubators, be inverted training
Foster 4-5 days.
PDA culture medium：The upper strata juice filtering after 200g potato boiling, 20g glucose, 20g agar powder, pure water is fixed
Hold to 1L, dispense to 250mL triangular flask, 121 DEG C of autoclaving 25min.
2nd, the preparation of first class inoculum：200g potato, 20g glucose, pure water 1L, dispense to 250mL triangular flask, 121
DEG C autoclaving 25min.Access flat board bacterial classification, be placed in 180r/min, in 28 DEG C of shaking tables, Shaking culture 5 days, obtains Trichoderma harzianum
SH2303 liquid fermentation bacterial classification.
3rd, trichoderma liquid fermentation：Corn flour 8kg, magnesium sulfate 80g, ferrous sulfate 1.2g, manganese sulfate 0.4g, zinc sulfate
0.32g, potassium dihydrogen sulfate 0.61kg, sodium nitrate 0.23kg, ammonium nitrate 176, calcium chloride 160g, water supplies 200L, loads 300L
Fermentation tank.The initial pH of culture medium is adjusted to 4-5.High-temperature sterilization 40-60min at 121 DEG C.Access weight than the trichoderma for 0.5%
Bacteria liquid fermented bacterium, fermentation temperature is 28 DEG C, fermentation time 7 days, as final trichoderma harzianum chlamydospore zymotic fluid.
4th, Trichoderma biological seed coating agent preparation：Trichoderma fermented liquid 1L, shitosan 4g, shenqinmycin 1mg, humic acid
0.6g, is sufficiently mixed.Above-mentioned mixed liquor is mixed with 14% (W/V) film forming agent, as Trichoderma biological seed coating agent.
5th, above-mentioned biological seed coating agent is carried out with the mensure of shelf life.Spore is initially contained in trichoderma harzianum chlamydospore zymotic fluid
Measure as 2.5 × 108CFU/mL, is compareed with being not added with shitosan for seed coat agent.Result shows, shitosan not only can in seed coat agent
There is inhibitory action to pathogen, as a kind of glucide, protectant one kind, provide for the Trichoderma spore in seed coat agent and hold
, so as to shelf life extension, after 6 months, spore amount is up to 1.56 × 10 for continuous nutrient content8CFU/g, is shown in Fig. 2.
Above the specific embodiment of the present invention is described.It is to be appreciated that the invention is not limited in above-mentioned
Particular implementation, those skilled in the art can make various modifications or modification within the scope of the claims, this not shadow
Ring the flesh and blood of the present invention.
Claims (10)
1. a kind of Trichoderma seed coat agent it is characterised in that described seed coat agent include trichoderma harzianum chlamydospore zymotic fluid, film forming agent,
Chitosan oligosaccharide, shenqinmycin, humic acid；
Wherein, film forming agent containing 13-17%, 0.35-0.45% chitosan oligosaccharide, 0.5- in every liter of trichoderma harzianum chlamydospore zymotic fluid
1.5ug/ml shenqinmycin, 0.05-0.1% humic acid, described percentage is mass volume ratio.
2. Trichoderma seed coat agent according to claim 1 is it is characterised in that described film forming agent includes 853 film forming agents.
3. Trichoderma seed coat agent according to claim 1 is it is characterised in that described Trichoderma is Trichoderma harzianum SH2303.
4. Trichoderma seed coat agent according to claim 1 is it is characterised in that the system of described trichoderma harzianum chlamydospore zymotic fluid
Standby inclusion is as follows：With 0.5% inoculum concentration, first class inoculum is inoculated in trichoderma liquid fermentation medium, 28 DEG C of fermented and cultured
7 days, obtain final product；Described percentage is volume ratio；
Wherein, described first class inoculum be by Trichoderma bacteria cake with the inoculum concentration of 3-5 piece/triangular flask in PD culture medium, 180r/
Min, under the conditions of 28 DEG C culture obtain within 5 days.
5. Trichoderma seed coat agent according to claim 4 is it is characterised in that the group of described trichoderma liquid fermentation medium
Become to include：Every 1000L rises 40kg containing corn flour, magnesium sulfate 400g, ferrous sulfate 6.0g, sulphur in Trichoderma liquid fermentation medium
Sour manganese 2.0g, zinc sulfate 1.6g, potassium dihydrogen sulfate 3.06kg, sodium nitrate 1.13kg, ammonium nitrate 880g, calcium chloride 800g, surplus
For water, initial pH is adjusted to 4-5.
6. Trichoderma seed coat agent according to claim 4 is it is characterised in that the tinning amount of described fermented and cultured controls：
200L liquid fermentation medium loads in 300L fermentation tank.
7. the preparation method of seed coat agent described in a kind of any one according to claim 1 to 6 is it is characterised in that comprise the steps：
In described ratio by shitosan, shenqinmycin, humic acid, the mixing of trichoderma harzianum chlamydospore zymotic fluid, it is subsequently adding coalescing agents
Mixing, obtains final product seed coat agent.
8. seed coat agent described in a kind of any one according to claim 1 to 6 is controlling in Maize Soil Borne Disease as biological prevention and control agent
Application.
9. application according to claim 8 it is characterised in that described soil-borne disease include preventing and treating corn stalk rot disease, line withered
Disease, root rot.
10. user in controlling Maize Soil Borne Disease for the Trichoderma seed coat agent described in a kind of any one according to claim 1 to 6
Method is it is characterised in that being specially described Trichoderma seed coat agent and corn seed is 1 in mass ratio：(10-15) uniformly mix,
?.
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| CN106879585A (en) * | 2017-03-10 | 2017-06-23 | 上海交通大学 | Trichoderma brassin lactones wettable powder and its preparation method and application |
| CN108359631A (en) * | 2018-05-25 | 2018-08-03 | 上海交通大学 | A kind of production method of trichoderma chlamydospore |
| CN108739862A (en) * | 2018-06-29 | 2018-11-06 | 上海农乐生物制品股份有限公司 | A kind of plant rooting promoter |
| CN108935080A (en) * | 2018-08-20 | 2018-12-07 | 云南北玉种子科技有限公司 | Corn breeding method |
| CN109892327A (en) * | 2019-04-15 | 2019-06-18 | 微生物肥料技术研究推广中心 | A kind of preparation method and applications of microorganism seed coating |
| CN111466415A (en) * | 2020-04-24 | 2020-07-31 | 上海交通大学 | Method for inducing corn to resist fusarium ear rot by trichoderma agent |
| ES2872648A1 (en) * | 2020-04-29 | 2021-11-02 | Univ Leon | SEED COATING CONTAINING A BIOCONTROL AGENT AND SARGASSUM MUTICUM (Machine-translation by Google Translate, not legally binding) |
| ES2872599A1 (en) * | 2020-04-28 | 2021-11-02 | Univ Leon | SEED COATING INCLUDING A BIOCONTROL AGENT AND HOP CONES (Machine-translation by Google Translate, not legally binding) |
| CN114532363A (en) * | 2022-02-18 | 2022-05-27 | 南京思农生物有机肥研究院有限公司 | Application method and application of coating containing trichoderma seed |
| CN115152773A (en) * | 2022-06-17 | 2022-10-11 | 安徽久易农业股份有限公司 | Composition of prothioconazole and fludioxonil for preventing and treating basal stem rot of wheat |
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| CN109892327A (en) * | 2019-04-15 | 2019-06-18 | 微生物肥料技术研究推广中心 | A kind of preparation method and applications of microorganism seed coating |
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| WO2021219915A1 (en) * | 2020-04-28 | 2021-11-04 | Universidad De León | Seed coating comprising a biocontrol agent and hop cones |
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| WO2021219916A1 (en) * | 2020-04-29 | 2021-11-04 | Universidad De León | Seed coating containing a biocontrol agent and sargasssum muticum |
| CN114532363A (en) * | 2022-02-18 | 2022-05-27 | 南京思农生物有机肥研究院有限公司 | Application method and application of coating containing trichoderma seed |
| CN114532363B (en) * | 2022-02-18 | 2024-03-08 | 南京思农生物有机肥研究院有限公司 | Use method and application of trichoderma seed coating |
| CN115152773A (en) * | 2022-06-17 | 2022-10-11 | 安徽久易农业股份有限公司 | Composition of prothioconazole and fludioxonil for preventing and treating basal stem rot of wheat |
| CN115152773B (en) * | 2022-06-17 | 2023-12-29 | 安徽久易农业股份有限公司 | Composition of prothioconazole and fludioxonil for preventing and treating wheat stem rot |
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