CN102925367A - Trichoderma viride strain and application thereof - Google Patents
Trichoderma viride strain and application thereof Download PDFInfo
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Abstract
The invention discloses a Trichoderma viride strain and application thereof. The Trichoderma viride stain is WLTR35, and the collection number is CGMCC No.6673. The invention also discloses application of the strain or biological control preparation comprising the strain in controlling plant soil-borne disease. The experiment indicates that the Trichoderma viride strain WLTR35 has the characteristics of fast growth, large spore production quantity, wide action spectrum and the like, can quickly and massively implement colonization, and therefore, has wide application prospects. The biological control preparation prepared from the Trichoderma viride WLTR35 can efficiently control the plant soil-borne disease and effectively enhance the crop yield.
Description
Technical field
The invention belongs to biological control Plant diseases field.Particularly, the present invention relates to strain viride and an application thereof, and the biological control preparation that adopts this trichoderma viride strain preparation.
Background technology
Plant diseases has a strong impact on the farm-forestry crop growth, even causes that crop is dead, causes great financial loss.For the control of Plant diseases, the conventional method that adopts comprises traditional chemical prevention, cultivates disease-resistant variety and biological control.The disease-resistant variety cycle is long, and resistance is single.Chemical prevention makes that the with medicament time is long, usage quantity is large, causes pesticide residue, environmental pollution; Simultaneously, the life-time service of chemical pesticide so that pathogenic bacteria develops immunity to drugs to it, causes preventive effect to descend even failure.Utilize the beneficial microorganism controlling plant diseases to have broad prospects.
Wood mould (Trichoderma spp.) belongs to hyphomycetes, Moniliales, the Moniliaceae of imperfect fungi, be that a class is distributed widely in the fungi on soil, air, dry branches and fallen leaves and the various fermented product, all can be separated to from roots of plants circle, blade and seed, bulb surface.Wood is mould to produce the mechanism such as resistance by producing microbiotic, nutrient competition, superparasitism, cell wall degrading enzyme and inducing plant, has antagonistic action for the various plants pathogenic bacteria, can suppress plurality of plant diseases, be to study the most widely one of bacterial strain in the present biocontrol strain.The mould biological control mechanism to phytopathogen of wood is various and complicated, usually be the coefficient result of number of mechanisms, the special emphasis of different Biocontrol Activity mechanism is different, and type, agrotype and the envrionment conditions of its Biocontrol Effect effect and Strain type, pathogenic fungi are closely related.
Summary of the invention
The purpose of this invention is to provide a kind of new trichoderma strain for the high-efficiency prevention and control plurality of plant diseases---viride (Trichoderma viride) WLTR35.Trichoderma viride strain of the present invention has that fast growth, sporulation quantity are large, action spectrum is extensively and the characteristics such as can rapid, high volume surely growing, and therefore has a good application prospect.
Another object of the present invention provide a kind of adopt viride WLTR35 preparation can the high-efficiency prevention and control Plant diseases the biological control preparation.
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
The invention provides a kind of viride (Trichoderma viride) bacterial strain WLTR35, the deposit number of described bacterial strain is CGMCC No.6673.
Bacterial strain provided by the present invention is viride WLTR35, separate acquisition from the Wulian County mushroom, be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on October 12nd, 2012 and (be called for short CGMCC, address: Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica), preserving number is CGMCC No.6673.It has following biological characteristics: fast in the growth of PDA nutrient agar, cultivate 3d under 30 ℃ of dark conditions, colony diameter 70mm.Bacterium colony initial stage white is sparse, and the later stage forms yellow-green colour and produces the spore district.Reverse side is colourless.Mycelia is poly-every, branch.Conidiophore forms the branch profile of pine and cypress formula, the stigma Shu Sheng of branch end, to life, alternate or Dan Sheng, doleiform, 4.5-10 * 2.5-3.5 μ m.Conidium elliposoidal, single, closely colourless, be light green during gathering, wall is slightly coarse, 3.5-5 * 3-4 μ m.The rRNA gene sequencing result (ITS-5.8S-ITS2 district) of this bacterial strain is shown in SEQ-1.
Cultural method or the propagation method of trichoderma viride WLTR35 of the present invention comprise:
(1) common cultivation is preserved and is adopted the PDA substratum, and filling a prescription is: potato 200g, glucose 20g, agar 12g, distilled water 1000mL;
(2) laboratory fluids is cultivated and is adopted the PDB substratum, and filling a prescription is: potato 200g, glucose 20g, distilled water 1000mL.
(3) solid culture based formulas: Gu material and inorganic salt solution, the in mass ratio ratio of 1:1.8 preparation.Wherein said solid material is that corn cob and the wheat bran of 60:40 forms by mass ratio; By mass percentage, described inorganic salt solution comprises 3.5% potassium primary phosphate, 0.04% sal epsom, and 4% ammonium sulfate remains and is water.
(4) bulk fermentation is cultivated prescription: with solid culture based formulas in (3)
The present invention also provides a kind of biological control preparation for the control plant soil-borne diseases, and described biological control preparation comprises described viride WLTR35.
The preparation method of biological control preparation of the present invention may further comprise the steps:
(1) seed liquor of preparation viride WLTR35;
(2) seed liquor with step (1) preparation is inoculated into constant temperature culture in the solid medium;
(3) culture of step (2) being cultivated dilutes with sterilized water, filters, and filtrate is seeded to carries out fermentation culture in the bulk fermentation substratum.
In a specific embodiments of the present invention, described preparation method may further comprise the steps:
(1) spore of viride WLTR35 is transplanted in the PDB liquid nutrient medium, 28 ℃ of shaking table shaking culture 3～5d obtain seed liquor;
(2) seed liquor that step (1) is prepared in mass ratio 10% ratio is inoculated in the solid medium 28 ℃ of lower shaking culture 3～5d;
(3) culture of step (2) being cultivated with sterilized water in mass ratio the ratio of 1:15 mix, filter, with filtrate by volume the ratio of 1:6 be seeded to the bulk fermentation substratum, fermentation culture 8～9d in 28 ℃ of room temperatures, the proving room of relative humidity more than 85%.
The present invention also provides a kind of biological control method for the control plant soil-borne diseases, and described biological control method comprises to the plant with soil-borne disease uses above-mentioned viride WLTR35 or above-mentioned biological control preparation.
The present invention also provides above-mentioned trichoderma viride WLTR35 or the purposes of above-mentioned biological control preparation in the control plant soil-borne diseases.
With regard to above-mentioned biological control preparation, biological control method and purposes, described plant soil-borne diseases can be selected from cucumber anthracnose, walnut anthrax, apple anthracnose, bitter rot or anthracnose of grape, fruit white rot of grape, watermelon anthrax, winter jujube black spot, Cure of Coryneum kunzei, pomegranate dry rot etc. one or more.
Experiment shows, trichoderma viride strain WLTR35 of the present invention has that fast growth, sporulation quantity are large, action spectrum is extensively and the characteristics such as can rapid, high volume surely growing, and therefore has a good application prospect.By the biological control preparation of this viride WLTR35 preparation, not only can prevent and treat efficiently plant soil-borne diseases, can also the Effective Raise crop yield, be a kind of biological control preparation that has application prospect.This biological control preparation can be used as biological pesticide or bio-feritlizer, prevent and treat multiple soil and pass Plant diseases, comprise cucumber anthracnose, walnut anthrax, apple anthracnose, bitter rot or anthracnose of grape, fruit white rot of grape, watermelon anthrax, winter jujube black spot, Cure of Coryneum kunzei, pomegranate dry rot etc. one or more.
Embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage and disadvantage of the present invention will be more clear along with description.But these embodiment only are exemplary, scope of the present invention are not consisted of any restriction.It will be understood by those skilled in the art that lower without departing from the spirit and scope of the present invention and can make amendment or replace the details of technical solution of the present invention and form, but these modifications and replacing all fall within the scope of protection of the present invention.
Embodiment 1
1, isolation and purification trichoderma viride (T.viride) WLTR35(CGMCC No.6673)
Viride of the present invention (T.viride) WLTR35(CGMCC No.6673) adopt plate streak to separate from the Wulian County mushroom and obtain, separation method is:
⑴ the separation of trichoderma strain: get the mushroom bacteria bag of being injured serious, use first 75% alcohol Local treatment 1min, again after processing 20min with ultraviolet lamp on the Bechtop, cut treatment sites with the sterilization blade, with a small amount of Trichoderma spore of inoculating needle picking, rule at the PDA flat board, regularly observe the colony growth situation.Then adopt plate streak, the purifying trichoderma strain goes to the PDA test tube slant and saves backup.
⑵ the screening of the efficient antagonistic Trichoderma bacterial strain of cucumber anthracnose
1. primary dcreening operation: adopt the face-off culture method, preparation PDA is dull and stereotyped, buys the bacterium cake that cut-off directly is 5mm with punch tool at Trichoderma, cucumber anthracnose, walnut anthrax bacteria edge, is implanted in respectively dull and stereotyped relative both sides central, 25 ℃ of constant temperature culture are observed Trichoderma day by day to the restraining effect of pathogenic bacteria.
2. sieve again: the trichoderma strain with efficient antagonistic activity that will screen carries out multiple sieve, mainly is through temperature tolerance, resistance to acids and bases, drug-resistant test, screens preferably trichoderma strain of patience, carries out potted plant control test and field test.
Wherein, the PDA culture medium prescription: potato 200g(peeling), glucose 20g, agar 12g, distilled water 1000mL.
The inventor obtains trichoderma viride (T.viride) the WLTR35(CGMCC No.6673 that a strain can the high-efficiency prevention and control plurality of plant diseases by a large amount of screening operations).Experiment showed, that the former powder of this viride all demonstrates very efficiently prevention effect in control cucumber anthracnose, walnut anthrax, so that farm crop are significantly increased production.Thereby viride of the present invention is the new bacterial strain of viride with wide application prospect, can be for the preparation of the biological control preparation of preventing and treating plurality of plant diseases.
2, identification of strains
(1) Microbiological Characteristics: fast in the growth of PDA nutrient agar, cultivate 3d under 30 ℃ of dark conditions, colony diameter 70mm.Bacterium colony initial stage white is sparse, and the later stage forms yellow-green colour and produces the spore district.Reverse side is colourless.Mycelia is poly-every, branch.Conidiophore forms the branch profile of pine and cypress formula, the stigma Shu Sheng of branch end, to life, alternate or Dan Sheng, doleiform, 4.5-10 * 2.5-3.5 μ m.Conidium elliposoidal, single, closely colourless, be light green during gathering, wall is slightly coarse, 3.5-5 * 3-4 μ m.
(2) molecular biological characteristic
The rRNA gene sequencing result (ITS-5.8S-ITS2 district) following (SEQ-1) of this bacterial strain:
CTTTCTTGGCACAATCGTGTCCGACAATCCTGTTCTCAGTCTTGCCAATTT
TTCTCGCGTCGTCACACCCCGCTCTACCTGTCTACCCCTCCTTTGGCACA
GCAGAAATTTTCTGGCTGCCTTGTTTGGCTTTTAGTGGGGTGCCAATTTTT
TTTTGGCAACAACCCCGCTATCGCCGTTGTCCCTCATCCATCGTCCCAAC
AATTTGATCTCACTCAATCACATCGTCTTCTGGTTCATTGTGTCAAAGATT
CGATGATTCACTGAATTCTGCAATTCACATTACTTATCGCATTTCGCTGCG
TTCTTCATCGATGCCAGAACCAAGAGATCCGTTGTTGAATGGGTTCTTGA
CAAGCTCAAGGCCGGCGTGAGCGTGGTATCACCATCGACATTGCCCTCTG
GAAGTTCGAGACTCCCAAGTACTATGTCACCGTCATTGGTATGTTTTGTG
TTTCCTCATTGACCCCCGGAATCCTTATTCTAACGTGCCGCTCTACAGACG
CTCCCGGTCACGTGATTTCATCAAGAACATGATCACTGGTACTTCCCAGG
CTGACTGCGTATCCTGATTATCGCTGCCGGTACTGGTGAGTTCGAGGCTG
GTATCTCCAAGGATGGCCAGACCCGTGAGCACGCTCTGCTCGCCTACACC
CTGGGTGTCAAGCAGCTCATCGTTGCCATCAACAAGATGGACACTGCCA
ACTGGGCCGAGGCTCGTTACCTTGAGATCATCAAGGAGACTCCAACTTCA
TCAAGAAGGTCGGCTTCAACCCCAAGTCCGTCGCCGCGCTCCCGGTGCG
AGTGTGCAACTACTGCGCAGGAGAGGCTGCGGCGAGACCAACCCCTGG
AAGGGCT?GGGAGAAGGAGACCAAGGCT?GGCAAGAGCA?GCG
Embodiment 2
1, trichoderma viride fermenting process
The PDB culture medium prescription: potato 200g(peeling), glucose 20g, distilled water 1000mL.
A large amount of solid fermentation culture medium prescriptions (quality percentage composition):
Gu material: corn cob 60%, wheat bran 40%
Inorganic salt solution: potassium primary phosphate 3.5%, sal epsom 0.04%, ammonium sulfate 4% remains and is water.
Solid-to-liquid ratio is the 1:1.8(mass ratio)
Viride (T.viride) WLTR35(CGMCC No.6673) a large amount of solid fermentation process:
1. the bacterial classification seed liquor is cultivated: with a small amount of spore of trichoderma viride (T.viride) WLTR35CGMCC No.6673 picking from the test tube slant, move in the PDB liquid nutrient medium, and 28 ℃ of shaking table shaking culture 3～5d, this is seed liquor.
2. the cultivation of solids manufacture bacterial classification: seed liquor is inoculated in the solid medium (500mL triangular flask) 28 ℃ of constant temperature culture 3～5d, middle multiple oscillation by 10% mass ratio.
3. a large amount of solid fermentations: the culture of solid culture press the 1:15 dilution proportion with sterilized water in will be 2., and filters with sterile gauze, goes out thick slag, is production bacterium liquid, and inoculative proportion is inoculated in the bulk fermentation substratum by 1:6.The raw material of inoculation is placed proving room (28 ℃ and relative humidity more than 85%) fermentation culture 8～9d, can obtain the former powder of Trichoderma.
Embodiment 3
The present embodiment provides viride (T.viride) WLTR35(CGMCC No.6673) former powder is to the related experiment of cucumber anthracnose disease-preventing and yield-increasing.
1, reagent agent
The former powder of viride WLTR35 (preparation among the embodiment 2), 70% mancozeb wettable powder (commercially available).
2, for studying thing and controlling object
Supplying to study thing is cucumber, and kind is Chang Chun Mi Ci;
Controlling object is anthrax.
3, situation experimental field
Experimental field be located at cucumber greenhouse gardening base, Shouguang, Shandong Province, this base plastic tent cucumber anthrax occured seriously in recent years, and control difficulty, test is in 600 square metres of (0.9 mu) cucumber booths, soil property is light loam, and organic content is that 1.09%, pH value is 6.7, all experimental plot cultivation conditions and control measures are consistent, and cucumber anthracnose is in their early stage during dispenser.
4, test design and arrangement
200 times, 400 times, 600 times in the former powder of trichoderma viride WLTR35,800 times of liquid of 70% mancozeb wettable powder are established in this test and not dispenser clear water compares totally 5 processing, repeat 4 times, and totally 20 residential quarters, each residential quarter random alignment, the residential quarter area is 20m
2Respectively spray medicine once on June 9th, 2012 and June 16, with plant leaf spray up and down evenly, thoughtful, block with plastic cloth during the spray medicine, mu using liquid 60kg, medicinal sprayer tool is workers and peasants-16 type atomizer.
5, pilot survey and method of calculation
(1) meteorological conditions
The 1st dispenser (on June 9th, 2012) was fine the same day, and 2 grades of wind-force, the highest temperature are 29 ℃, and the lowest temperature is 15 ℃, and relative humidity is 65%.The 2nd dispenser (on June 16th, 2012), the same day is fine, and 3 grades of wind-force, the highest temperature are 25 ℃, and the lowest temperature is 18 ℃, and relative humidity is 75%; Because of the plastic greenhouse envrionment conditions relatively stable, so this weather is on not impact of drug effect.
(2) drug effect and security survey
Efficacy survey: 14d investigates after the front and last dispenser of the 1st dispenser.4 point samplings are adopted in every residential quarter, look into 2 strains at every, investigate whole blades, Investigate incidence, and the record disease index also calculates disease index.
Security survey: 7d and 14d observe the security of cucumber after the dispenser first time, occur if any poisoning, describe symptom of chemical damage in detail and press poisoning grading standard poisoning degree.
Stage division (take blade as unit):
0 grade: without scab
1 grade: lesion area accounts for below 5% of whole leaf area;
3 grades: lesion area accounts for the 6%-10% of whole leaf area;
5 grades: lesion area accounts for the 11%-25% of whole leaf area;
7 grades: lesion area accounts for the 26%-30% of whole leaf area;
9 grades: lesion area accounts for more than 50% of whole leaf area;
(3) control time and number of times
Investigation state of an illness radix before the dispenser, 14d investigation prevention result before the next dispenser and after the last dispenser.
(4) drug effect method of calculation
Drug effect is calculated by formula (1), (2):
(5) on the direct impact of crop
Observe medicament crop is had or not poisoning, type and the degree of record poisoning.
6, result
(1) reagent agent is to the prevention effect of cucumber anthracnose
Table 1 result shows, disease index rate of increase proportion by subtraction is 188.68%, 229.50%, 278.71% after 200 times, 400 times, 600 times processing of the former powder of trichoderma viride WLTR35, the disease index rate of increase of contrast medicament is 306.56%, all is lower than the disease index rate of increase 1155.05% of contrast.The correcting controling effect that the former powder of trichoderma viride WLTR35 is 200 times, 400 times is respectively 83.66%, 80.13%, and there was no significant difference on P＜0.05 level all is higher than the correcting controling effect of other processing.600 times in the former powder of trichoderma viride WLTR35 and prevention effect on P＜0.05 level the there was no significant difference of 70% mancozeb wettable powder to cucumber anthracnose are respectively 75.87%, 73.46%.This shows that the former powder of trichoderma viride WLTR35 can effectively control the harm of cucumber anthracnose.
Each processes disease index and the prevention effect of rear cucumber anthracnose table 1
(2) cucumber security survey: observe through dispenser 7d and 14d, each chemicals treatment district compares with the check plot, and cucumber growth is normal, produces without poisoning, illustrates that the former powder of trichoderma viride WLTR35 is supplying examination concentration to cucumber safety.
(3) the former powder of viride is to the cucumber production promoting effect
The former powder of table 2 trichoderma viride WLTR35 is to cucumber production promoting effect analysis table
Annotate: residential quarter area 33.4m
2Processing 1 is 200 times in the former powder of trichoderma viride WLTR35, and processing 2 is 400 times in the former powder of trichoderma viride WLTR35, and processing 3 is 600 times in the former powder of trichoderma viride WLTR35, and processing 4 is control group.Different letter representation significant difference (P＜0.01) behind the Duncan multiple comparisons after the data in the table.
By table 2 result as can be known, each processes the contrast check plot at the former powder of cucumber sprinkling irrigation trichoderma viride WLTR35, and stimulation ratio is respectively 27.24%, 25.52% and 20.44%; Effect is fairly obvious.
Therefore, from disease index rate of increase and prevention effect, the former powder of trichoderma viride WLTR35 has preferably prevention effect to cucumber anthracnose, and prevention effect can reach more than 75%, significantly better than contrast medicament, significant difference.From the cucumber production promoting effect, the former powder of trichoderma viride WLTR35 has the significantly short effect of increasing production of giving birth to cucumber, and stimulation ratio reaches more than 20% after 2 dispensers.
Embodiment 4
The present embodiment provides the related experiment of the former powder of trichoderma viride WLTR35 to walnut anthrax prevention effect.
1) reagent agent
The former powder of trichoderma viride WLTR35 (preparation among the embodiment 2); 70% mancozeb wettable powder (commercially available).
2) for studying thing and controlling object:
Supplying to study thing is walnut, and kind is the fragrant tinkling of pieces of jade;
Controlling object: anthrax.
3) experimental field situation, test design and arrangement
Experimental field be located at walnut plantation, Tai’an, Shandong Province, soil is sand ground.Walnut is 10 years fragrant tinkling of pieces of jade of life, spacing in the rows 3m * 4m, and tree vigo(u)r is medium on the weak side, and disease occurs serious.
200 times, 400 times, 600 times in the former powder of trichoderma viride WLTR35 is established in this test, and 800 times of liquid of 70% mancozeb wettable powder and not dispenser clear water compare totally 5 processing, repeat 4 times.3 all medications after walnut is bloomed every 15d dispenser 1 time, amount to 7 times.The front 5d that gathers investigates, and each processes 300 fruits of random searching, calculates diseased fruit rate, prevention effect, whole pilot survey 1 time.With plant leaf spray up and down evenly, thoughtful, block with plastic cloth during the spray medicine, it is workers and peasants-16 type atomizer that mu is used liquid 60kg, medicinal sprayer tool.
5) pilot survey and method of calculation
(1) meteorological conditions
The 1st dispenser (on April 19th, 2012) was fine the same day, and gentle breeze, the highest temperature are 23 ℃, and the lowest temperature is 12 ℃, and relative humidity is 75%; The 2nd dispenser (on May 4th, 2011), the same day is fine, and 2 grades of wind-force, the highest temperature are 29 ℃, and the lowest temperature is 15 ℃, and relative humidity is 70%; The 3rd dispenser (on May 19th, 2011), the same day is fine, and 2 grades of wind-force, the highest temperature are 30 ℃, and the lowest temperature is 17 ℃, and relative humidity is 65%; The 4th dispenser (on June 3rd, 2011), the same day is fine, and 2 grades of wind-force, the highest temperature are 30 ℃, and the lowest temperature is 19 ℃, and relative humidity is 60%; The 5th dispenser (on June 18th, 2011), the same day is clear to cloudy, and 4 grades of wind-force, the highest temperature are 34 ℃, and the lowest temperature is 21 ℃, and relative humidity is 75%; The 6th dispenser (on July 3rd, 2012), the same day is fine, and 2 grades of wind-force, the highest temperature are 34 ℃, and the lowest temperature is 24 ℃, and relative humidity is 85%; The 7th dispenser (on July 18th, 2011), the same day is overcast to cloudy, and 4 grades of wind-force, the highest temperature are 33 ℃, and the lowest temperature is 24 ℃, and relative humidity is 85%.
(2) drug effect and security survey
Efficacy survey: the front 5d investigation of gathering, each processes 300 fruits of random searching, calculates diseased fruit rate, prevention effect
Security survey: 7d observes the security of walnut after each dispenser.
(3) drug effect method of calculation
Drug effect is calculated by formula (1), (2):
(4) on the direct impact of crop
Observe medicament crop is had or not poisoning, type and the degree of record poisoning.And record is to the other influences of crop.
6) result
(1) reagent agent is to the prevention effect of walnut anthrax
The front 5d that gathers investigates diseased fruit rate, table 3 result demonstration, and the diseased fruit rate of clear water contrast is up to 5.366%, and the diseased fruit rate of each processing all is lower than the clear water contrast.Diseased fruit rate after 200 times, 400 times, 600 times processing of the former powder of trichoderma viride WLTR35 is respectively 8.63%, 9.98%, 12.36%, all is lower than the diseased fruit rate of contrast medicament 18.69%; The prevention effect of former each concentration of powder of trichoderma viride WLTR35 is respectively 83.92%, 81.40%, 76.97%, prevention effect there was no significant difference on P＜0.05 level of 200 times and 400 times wherein, 70% mancozeb wettable powder is 65.17% to the prevention effect of walnut anthrax.The former powder of trichoderma viride WLTR35 can effectively be controlled the harm of walnut anthrax.
Each processes sickness rate and the prevention effect of rear walnut anthrax table 3
Annotate: with there being same letter to represent behind the Duncan multiple comparisons difference not significantly (P＜0.05) behind the data line.
(2) walnut security survey: through observing, each chemicals treatment district compares with the check plot, and the walnut growth is normal, produces without poisoning, illustrates that the former powder of trichoderma viride WLTR35 is supplying examination concentration to walnut safety.
Therefore, from sickness rate and prevention effect, trichoderma viride WLTR35 has preferably prevention effect to walnut anthrax, and prevention effect can reach more than 75% after the dispenser, significantly better than contrast medicament, significant difference.
Claims (8)
1. a viride (Trichoderma viride) bacterial strain WLTR35, the deposit number of described bacterial strain is CGMCC No.6673.
2. trichoderma viride strain WLTR35 claimed in claim 1 is in the purposes of control in the plant soil-borne diseases.
3. a biological control preparation comprises trichoderma viride strain WLTR35 claimed in claim 1.
4. a method for preparing biological control preparation claimed in claim 3 is characterized in that, may further comprise the steps:
(1) seed liquor of preparation trichoderma viride strain WLTR35;
(2) seed liquor with step (1) preparation is inoculated into constant temperature culture in the solid medium;
(3) culture of step (2) being cultivated dilutes with sterilized water, filters, and filtrate is seeded to carries out fermentation culture in the bulk fermentation substratum.
5. preparation method as claimed in claim 4 is characterized in that,
(1) spore of trichoderma viride strain WLTR35 is transplanted in the PDB liquid nutrient medium, 28 ℃ of shaking table shaking culture 3～5d obtain seed liquor;
(2) seed liquor that step (1) is prepared in mass ratio 10% ratio is inoculated in the solid medium 28 ℃ of lower shaking culture 3～5d;
(3) culture of step (2) being cultivated with sterilized water in mass ratio the ratio of 1:15 mix, filter, with filtrate by volume the ratio of 1:6 be seeded to the bulk fermentation substratum, fermentation culture 8～9d in 28 ℃ of room temperatures, the proving room of relative humidity more than 85%;
Described PDB substratum is: potato 200g, glucose 20g, distilled water 1000mL; Described solid medium is: Gu material and inorganic salt solution, the in mass ratio ratio of 1:1.8 preparation; Wherein said solid material is that corn cob and the wheat bran of 60:40 forms by mass ratio; Described inorganic salt solution comprises 3.5% potassium primary phosphate by mass percentage, 0.04% sal epsom, and 4% ammonium sulfate remains and is water; The same solid medium of described bulk fermentation substratum.
6. biological control preparation claimed in claim 3 is in the purposes of control in the plant soil-borne diseases.
7. purposes as claimed in claim 6 is characterized in that, described plant soil-borne diseases is cucumber anthracnose, walnut anthrax, apple anthracnose, bitter rot or anthracnose of grape, fruit white rot of grape, watermelon anthrax, winter jujube black spot, Cure of Coryneum kunzei or pomegranate dry rot.
8. the biological control method of a plant soil-borne diseases is characterized in that, uses the trichoderma viride strain WLTR35 of claim 1 or the biological control preparation of claim 3 to the plant with soil-borne disease.
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| CN108587922A (en) * | 2018-04-17 | 2018-09-28 | 山东省林业科学研究院 | One plant of pick up the ears trichoderma and its application in preventing winter jujube anthracnose |
| CN109042742A (en) * | 2018-08-18 | 2018-12-21 | 杭州渔森农业技术开发有限公司 | Compound formulation is used to prevent and treat soil and passes the purposes fallen ill |
| CN113881578A (en) * | 2021-11-15 | 2022-01-04 | 中国农业科学院特产研究所 | Soil conditioner for trichoderma viride and panax plants and application thereof |
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| CN103131642A (en) * | 2013-03-18 | 2013-06-05 | 济南巨微生物技术有限公司 | Trichoderma viride and culture method and application thereof |
| CN103131642B (en) * | 2013-03-18 | 2014-07-23 | 赵钦刚 | Trichoderma viride and culture method and application thereof |
| CN105567573A (en) * | 2016-01-13 | 2016-05-11 | 西南林业大学 | Biocontrol strain for preventing and treating colletotrichum gloeosporium penz and screening method and application thereof |
| CN106857682A (en) * | 2017-01-26 | 2017-06-20 | 新疆农垦科学院 | A kind of biological control method of dense planting jujube garden black spot |
| CN107118971A (en) * | 2017-06-08 | 2017-09-01 | 黄河三角洲京博化工研究院有限公司 | A kind of Trichoderma viride and its cultural method |
| CN108587922A (en) * | 2018-04-17 | 2018-09-28 | 山东省林业科学研究院 | One plant of pick up the ears trichoderma and its application in preventing winter jujube anthracnose |
| CN108587922B (en) * | 2018-04-17 | 2020-03-27 | 山东省林业科学研究院 | Trichoderma aureoviride and application thereof in preventing and treating winter jujube anthracnose |
| CN109042742A (en) * | 2018-08-18 | 2018-12-21 | 杭州渔森农业技术开发有限公司 | Compound formulation is used to prevent and treat soil and passes the purposes fallen ill |
| CN113881578A (en) * | 2021-11-15 | 2022-01-04 | 中国农业科学院特产研究所 | Soil conditioner for trichoderma viride and panax plants and application thereof |
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