CN1250084C - Microbial preparation for crop disease control and production increase - Google Patents
Microbial preparation for crop disease control and production increase Download PDFInfo
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- CN1250084C CN1250084C CN 03135663 CN03135663A CN1250084C CN 1250084 C CN1250084 C CN 1250084C CN 03135663 CN03135663 CN 03135663 CN 03135663 A CN03135663 A CN 03135663A CN 1250084 C CN1250084 C CN 1250084C
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Abstract
The present invention discloses microbial preparation for preventing diseases and increasing yield for crops, which has following main components and weight distribution ratio: 12 to 16 shares of Trichoderma harzianum bacterial strain T23 CGMCC No. 0975, and 84 to 88 shares of crushing materials of plants. The crushing materials is composed of corn flour or wheat bran or the mixture of corn flour and wheat bran, rice bran, powder of corn stalks, powder of cotton seed shells, and one kind of wood flour or the mixture of at least two kinds of wood flour. The microbial preparation of the present invention has the advantages of short production period, low cost, wide source of raw material, low price, large sporulation quantity, convenient operation, high safety, wide applicable crops, multiple disease preventing objects, and obvious function of promoting yield, reduces pollution and provides technical assurance of good-quality nuisanceless agricultural products.
Description
Affiliated technical field
The present invention relates to microorganism formulation that is used for the crop disease prevention volume increase and preparation method thereof.
Background technology
Soil-borne diseases such as crop pest such as sclerotinia rot of colza, peanut sclerotium rolfsii, eggplant verticillium wilt, capsicum epidemic disease, red sage root root rot, safflower fusarium wilt, cotton seedling blight and fusarium wilt not only have a strong impact on the yield and quality of crop, and the pathogen of disease can survive in soil and propagate with soil.For the control of these diseases, common chemical medicament not only control efficiency is poor, and the abuse chemical pesticide also can cause serious agricultural residual pollution problem.At present, domestic and international biological control at crop pests such as gray mold, damping off, southern blight, banded sclerotial blight, fusarium wilts, separate some and had the trichoderma strain of certain protection effect, and carried out corresponding enlarged culture research, but also there are some problems, as the preparation applicable crops of producing is few, the diseases prevention narrow range; Perhaps the source of raw materials for production is limited by the region; Perhaps production process complexity, production cost height; Perhaps large usage quantity, often every mu needs with several kilograms to tens kilograms.As Chinese patent application " a kind of microbial bactericide and preparation method thereof ", publication number CN1337164A, this microbial bactericide is only applicable to tobacco, only effective to tobacco damping off, damping off, balck shank, rust, the source region of its raw materials for production offal is strong, and needs to use the special equipment fermentation tank in the production process.Chinese patent application " quick-acting disease-resistant organic fungus fertilizer and production method thereof " and for example, publication number CN1283599, this disease-resistant fungi fertilizer large usage quantity, every mu of need are used 40kg-50kg.
Summary of the invention
The purpose of this invention is to provide a kind of microorganism formulation that is used for crop disease prevention volume increase, this kind microorganism formulation: with short production cycle, cost is low; Raw material sources are wide, cheap; Sporulation quantity is big; Easy to use, safe; Applicable crops is wide, the diseases prevention object is many, the short production-increasing function of giving birth to is remarkable; But also pollution abatement provides technical guarantee for producing and developing the high-quality non-polluted farm product.
Another purpose of the present invention provides a kind of above-mentioned method that is used for the microorganism formulation of crop disease prevention volume increase for preparing.
The technical solution adopted for the present invention to solve the technical problems is: a kind of microorganism formulation that is used for the crop disease prevention volume increase, component that it contains and quality proportioning are: 12～16 parts Trichoderma harzianum (Trichoderma harzianum) bacterial strain T23 CGMCC No.0975, the spore content of this bacterial strain T23 reaches 3 * 10
10Individual/gram; 84～88 parts plant comminution of material, the consisting of of this plant comminution of material: the mixture of a kind of in corn flour or wheat bran or the mixture of the two and rice bran, milled powders of cornstalk, cotton seed hulls powder, the wood chip or at least two kinds.
Prepare above-mentioned a kind of method that is used for the microorganism formulation of crop disease prevention volume increase, its steps in sequence is:
(1) the bacterial classification test tube is cultivated: adopt potato glucose agar-agar solid culture medium (PDA), trichoderma harzianum strain T23 is inoculated in the solid culture medium (PDA) that contains streptomycin 40～70ppm, under 18 ℃～30 ℃ temperature, cultivated 1～7 day;
(2) liquid fermentation and culture: corn flour or wheat bran or the mixture of the two and water are loaded in the triangular flask by 1: 300～500 mass ratio mixing, and the high-temperature pressurizing sterilization is 20～30 minutes under 121 ℃, 15 pounds, makes culture fluid; The trichoderma harzianum strain T23 that after the cooling (1) step is turned out in the test tube is by being seeded in the triangular flask, the mass ratio of bacterial strain and culture fluid is 1: 400～500, and be 20 ℃～30 ℃ in temperature, Light To Dark Ratio is 1: in the alternation of light and darkness environment of 0.8-1.2, and constant-temperature shaking culture 3～5 days;
(3) solid fermentation is cultivated: with the mixture of a kind of in rice bran, milled powders of cornstalk, cotton seed hulls powder, the wood chip or at least two kinds and water in 1: 0.8～1.2 ratio mixing, be loaded in the heatproof plastic sack, the high-temperature pressurizing sterilization is 120～150 minutes under 121 ℃, 15 pounds, pour into after the cooling in sterilized culture bed, form solid culture medium; Liquid fermentation and culture thing and this solid culture medium that (2) step makes are inoculated the liquid fermentation culture medium by 1: 8～10 mass ratio, in the culturing room of 18 ℃～32 ℃, relative moisture more than 90%, under the natural lighting, sterilization, ventilate to cultivating and promptly made solid fermentation culture in 1～3 day.
(4) preparation of preparation: with the solid fermentation culture natural air drying to water content 8%～10%, pulverize with cracker again, cross 100 mesh sieves after, measure spore content, bacterial strain T23 spore content reaches 3 * 10
10Individual/gram is certified products.
The trichoderma harzianum strain T23 that the present invention adopts is from the traditional Chinese medicine red sage root rhizosphere soil of Jianyang, Sichuan, utilizes pathogen to make the bait separation screening and gets.Promptly adopt nylon mesh sheet to make carrier, earlier the nylon mesh sheet of sterilization is put into the culture dish that is connected to pathogen, after covering with thalline on the nylon mesh sheet, nylon mesh sheet is placed the soil sample surface, cultivate after 3-5 days, the antagonism Trichoderma can be parasitic to the pathogen of nylon mesh sheet, takes nylon mesh sheet off and be placed on potato glucose agar-agar solid (PDA) medium, choose the trichoderma strain purifying and cultivate the trichoderma strain that just can obtain to have antagonism; By antagonistic effect, therefrom filter out the significant trichoderma harzianum strain T23 of antagonism to these bacterial strains and multiple pathogen.
This bacterial strain is to 1 grade of the antagonism index reached of pathogens such as the Rhizoctonia solani Kuhn that causes diseases such as the dead seedling of crops, fusarium wilt, root rot and southern blight, Fusarium oxysporum, Sclerotium rolfsii, to the pathogen mycelial growth inhibition rate all more than 85%.
The trichoderma harzianum strain T23 that the present invention adopts has following Microbiological Characteristics:
1, morphological characteristic: sporophore clump Shu Shusong, annular arrangement, main branch is tree-shaped, and secondary branch is multistage, and normal 2～3 one group, the right angle stretches out; Spore sphere, subsphaeroidal or obovate.
2, cultivate to learn characteristic: on the PDA medium, bacterium colony be white in 1～2 day, transferred dirty-green to after aging.
This bacterial strain is on 07 03rd, 2003, and in the common micro-organisms center preservation of Chinese microorganism strain administration committee, its preservation registration number is CGMCC NO.0975.
Compared with prior art, the invention has the beneficial effects as follows:
(1) with short production cycle, operate simple and easy.Production whole process from the spawn culture to the preparation only needs 5～11 days.
(2) production cost is low: the required composts or fertilisers of cultivating of liquid and solid fermentation is respectively corn flour, rice bran etc., and its source is very abundant, and is cheap.
(3) sporulation quantity is big: the Trichoderma spore content in the liquid fermentation and culture thing of producing according to this method is 0.5 * 10
8～2 * 10
8Individual/ml; In the microorganism formulation finished product of making, the Trichoderma spore content is 3 * 10
10～9 * 10
10Individual/gram, the percentage by weight of Trichoderma conidial powder accounts for 12%～16%, and plant comminution of material such as rice bran, corn flour account for 84%～88%.
(4) applicable crops is wide, and the diseases prevention object is many, and the short production-increasing function of giving birth to is remarkable.Applicable crops comprises grain and oil crop, vegetables, traditional Chinese medicine, the diseases prevention object relates to the soil-borne disease that chemical pesticides such as the fusarium wilt that caused by cause of disease mattresses such as sickle-like bacteria, Rhizoctonia solani Kuhn, big beautiful Verticillium dahliae, sclerotium bacteria, vaccine and Sclerotium rolfsiis, damping off, stalk break, southern blight are difficult to prevent and treat, control efficiency is remarkable, and crop is significantly increased production.
(5) adopt the inventive method to prepare above-mentioned microorganism formulation, not only can diseases prevention, volume increase, reach the purpose that improves planting benefit, but also pollution abatement, for producing and development high-quality non-polluted farm product provides technical guarantee.
Microorganism formulation of the present invention is proved by following contrast experiment the effect of various crop disease-preventing and yield-increasing:
The test that is used for rape shows, the soil mentioned microorganism preparation 0.2kg/ mu of spreading manuer in holes when rape is transplanted, can alleviate the generation of sclerotinia rot of colza effectively, the control efficiency of sclerotinia rot of colza is reached 62.9%, better than the control efficiency (55.7%) of present common chemical pesticide carbendazim.And the thousand kernel weight of rape increases by 5.6% than blank, rape yield increasing 5.3%.
The test that is used for peanut shows, the soil mentioned microorganism preparation 0.25kg/ mu of spreading manuer in holes during peanut seeding, peanut emerges soon, neat, to lack nest few.Peanut was emerged early 1-2 days, and emergence rate improves 7 percentage points, lacked the nest rate and descended 7 percentage points, production ratio blank volume increase 10.2%.
The test that is used for capsicum shows, the soil mentioned microorganism preparation 0.2kg/ mu of spreading manuer in holes can make capsicum volume increase 14.0% when capsicum was transplanted.
The test that is used for eggplant shows, the soil mentioned microorganism preparation 0.2kg/ mu of spreading manuer in holes when eggplant is transplanted, can alleviate the generation of eggplant verticillium wilt effectively, control efficiency to its verticillium wilt is 55.9%, control efficiency (40.4%) than present common chemical pesticide carbendazim is good, and makes eggplant volume increase 12.9%.
The test that is used for the red sage root shows, the soil mentioned microorganism preparation 0.2kg/ mu of spreading manuer in holes is 55.7% to the red sage root root rot control efficiency during red sage root sowing, and is suitable with the control efficiency (57.5%) of carbendazim; The red sage root emerged facilitation, and the emergence rate after the processing is 83.8%, be significantly higher than carbendazim 71.3% and blank 64.4%; And make the red sage root increase production 8.2%.
The test that is used for Flos Carthami shows, Flos Carthami when sowing soil mentioned microorganism preparation 0.2kg/ mu of spreading manuer in holes, can effectively control the generation of Flos Carthami fusarium wilt, be 70.4% to the control efficiency of its fusarium wilt, is higher than the control efficiency (60.9%) of chemical agent carbendazim.And promote to emerge and growth of seedlings, the Flos Carthami emergence rate after the processing is 87.5%, is significantly higher than 73.9% of blank; Single-strain fresh weight and dry weight are respectively 3.23g and 0.46g, all are significantly higher than the 2.70g and the 0.41g of blank.
The test that is used for cotton shows, with behind the above-mentioned microorganism formulation seed dressing processing cotton (preparation: cotton seed is 1: 100), can effectively reduce the generation of dead seedling of cotton and fusarium wilt during sowing cotton seed.Control efficiency to cotton seedling blight is 70.0%, can reach 60.4% to the preventive effect of the fusarium wilt of cotton, is higher than or is equivalent to the control efficiency (being respectively 54.6% and 61.2%) of chemical agent carbendazim; Simultaneously, be 80.9% with the cotton emergence rate after this microorganism formulation processing, be significantly higher than the emergence rate (51.6%) of blank.
The present invention is further described below in conjunction with embodiment.
Embodiment:
Embodiment one
The consisting of of plant comminution of material in this routine microorganism formulation: corn flour and rice bran.The steps in sequence of its preparation method is:
(1) the bacterial classification test tube is cultivated: adopt potato glucose agar-agar solid culture medium (PDA), trichoderma harzianum strain T23 is inoculated in the solid culture medium (PDA) that contains streptomycin 50ppm, in temperature is under 25 ℃, cultivates 1～3 day, and concrete incubation time is covered with media surface with the mattress silk and is as the criterion.
Add the purpose of streptomycin in the PDA medium, be to prevent the growth of bacterium and other mould.
(2) liquid fermentation and culture: employing corn flour and water are loaded in the triangular flask by 1: 400 mass ratio mixing, and high-temperature pressurizing sterilization 25min makes culture fluid under 121 ℃, 15 pounds; After the cooling trichoderma harzianum strain T23 in the test tube is seeded in the triangular flask, the mass ratio of Trichoderma harzianum and culture fluid is 1: 450, places 25 ℃ of temperature, and Light To Dark Ratio is in 1: 1 the alternation of light and darkness environment, shaken cultivation 3～5 days is cultivated and is made the liquid fermentation and culture thing.Concrete incubation time reaches 0.5 * 10 can guarantee the bacterial strain T23 spore content in the liquid fermentation and culture thing
8～2 * 10
8Individual/ml is advisable.
(3) solid fermentation is cultivated: rice bran and water are pressed 1: 1 mass ratio mixing, be loaded in the heatproof plastic sack, high-temperature pressurizing sterilization 130min under 121 ℃, 15 pounds, pour into after the cooling in sterilized culture bed, by the liquid fermentation and culture thing: solid culture medium is 1: 9 a mass ratio inoculation liquid fermentation culture medium, more than 90%, natural lighting was made solid fermentation culture in 1～3 day in the cultivation room ventilation cultivation of sterilization in 25 ℃ of temperature, relative moisture.Concrete incubation time should can reach 3 * 10 with the bacterial strain T23 spore content in the solid fermentation culture
10～9 * 10
10Individual/gram is advisable.
(4) preparation of preparation: with the solid fermentation culture natural air drying to water content after 8%～10%, pulverize with cracker, cross 100 mesh sieves after, carry out spore content and measure, the bacterial strain T23 spore content in the microorganism formulation reaches 3 * 10
10Individual/gram is certified products.
Embodiment two
The consisting of of plant comminution of material in this routine microorganism formulation: wheat bran and milled powders of cornstalk.The steps in sequence of its preparation method is:
(1) the bacterial classification test tube is cultivated: adopt potato glucose agar-agar solid culture medium (PDA), trichoderma harzianum strain T23 is inoculated in the solid culture medium (PDA) that contains streptomycin 40ppm, in temperature is under 22 ℃, cultivates 1～3 day, and concrete incubation time is covered with media surface with mycelia and is as the criterion.
(2) liquid fermentation and culture: employing wheat bran and water are loaded in the triangular flask by 1: 500 mass ratio mixing, and high-temperature pressurizing sterilization 30min makes culture fluid under 121 ℃, 15 pounds; After the cooling trichoderma harzianum strain T23 in the test tube is seeded in the triangular flask, the mass ratio of trichoderma harzianum strain T23 and culture fluid is 1: 500, places 23 ℃ of temperature, and Light To Dark Ratio is in 1: 1.2 the alternation of light and darkness environment, shaken cultivation 3～5 days is cultivated and is made the liquid fermentation and culture thing.Concrete incubation time reaches 0.5 * 10 can guarantee the bacterial strain T23 spore content in the liquid fermentation and culture thing
8～2 * 10
8Individual/ml is advisable.
(3) solid fermentation is cultivated: milled powders of cornstalk and water are pressed 1: 0.8 mass ratio mixing, be loaded in the heatproof plastic sack, high-temperature pressurizing sterilization 150min under 121 ℃, 15 pounds, pour into after the cooling in sterilized culture bed, by the liquid fermentation and culture thing: solid culture medium is 1: 10 a mass ratio inoculation liquid fermentation culture medium, more than 90%, natural lighting was made solid fermentation culture in 1～3 day in the cultivation room ventilation cultivation of sterilization in 28 ℃ of temperature, relative moisture.Concrete incubation time can reach 3 * 10 with the bacterial strain T23 spore content in the solid fermentation culture
10～9 * 10
10Individual/gram is advisable.
(4) preparation of preparation: with embodiment one.
Embodiment three
The consisting of of plant comminution of material in this routine microorganism formulation: wheat bran and rice bran.The steps in sequence of its preparation method is:
(1) the bacterial classification test tube is cultivated: adopt potato glucose agar-agar solid culture medium (PDA), trichoderma harzianum strain T23 is inoculated in the solid culture medium (PDA) that contains streptomycin 70ppm, in temperature is under 28 ℃, cultivates 1～3 day, and concrete incubation time is covered with media surface with mycelia and is as the criterion.
(2) liquid fermentation and culture: employing wheat bran and water are loaded in the triangular flask by 1: 300 mass ratio mixing, and high-temperature pressurizing sterilization 20min makes culture fluid under 121 ℃, 15 pounds; After the cooling trichoderma harzianum strain T23 in the test tube is seeded in the triangular flask, the mass ratio of trichoderma harzianum strain T23 and culture fluid is 1: 400, places 27 ℃ of temperature, and Light To Dark Ratio is in 1: 0.8 the alternation of light and darkness environment, shaken cultivation 3～5 days is cultivated and is made the liquid fermentation and culture thing.Concrete incubation time reaches 0.5 * 10 can guarantee the bacterial strain T23 spore content in the liquid fermentation and culture thing
8～2 * 10
8Individual/ml is advisable.
(3) solid fermentation is cultivated: rice bran and water are pressed 1: 1.2 mass ratio mixing, be loaded in the heatproof plastic sack, high-temperature pressurizing sterilization 120min under 121 ℃, 15 pounds, pour into after the cooling in sterilized culture bed, by the liquid fermentation and culture thing: solid culture medium is 1: 8 a mass ratio inoculation liquid fermentation culture medium, more than 90%, natural lighting was made solid fermentation culture in 1～3 day in the cultivation room ventilation cultivation of sterilization in 22 ℃ of temperature, relative moisture.Concrete incubation time can reach 3 * 10 with the bacterial strain T23 spore content in the solid fermentation culture
10～9 * 10
10Individual/gram is advisable.
(4) preparation of preparation: with embodiment one.
Embodiment four
The consisting of of plant comminution of material in this routine microorganism formulation: the mixture of corn flour, wheat bran and cotton seed hulls powder.The steps in sequence of its preparation method is:
(1) the bacterial classification test tube is cultivated: adopt potato glucose agar-agar solid culture medium (PDA), trichoderma harzianum strain T23 is inoculated in the solid culture medium (PDA) that contains streptomycin 60ppm, in temperature is under 18 ℃, cultivates 1～7 day, and concrete incubation time is covered with media surface with mycelia and is as the criterion.
(2) liquid fermentation and culture: the mixture of employing corn flour and wheat bran and water are loaded in the triangular flask by 1: 450 mass ratio mixing, and high-temperature pressurizing sterilization 30min makes culture fluid under 121 ℃, 15 pounds; After the cooling trichoderma harzianum strain T23 in the test tube is seeded in the triangular flask, the mass ratio of trichoderma harzianum strain T23 and culture fluid is 1: 450, places 20 ℃, and Light To Dark Ratio is in 1: 0.9 the alternation of light and darkness environment, shaken cultivation 3～5 days is cultivated and is made the liquid fermentation and culture thing.Concrete incubation time reaches 0.5 * 10 can guarantee the bacterial strain T23 spore content in the liquid fermentation and culture thing
8～2 * 10
8Individual/ml is advisable.
(3) solid fermentation is cultivated: cotton seed hulls powder and water are pressed 1: 0.9 mass ratio mixing, be loaded in the heatproof plastic sack, high-temperature pressurizing sterilization 150min under 121 ℃, 15 pounds, pour into after the cooling in sterilized culture bed, by the liquid fermentation and culture thing: solid culture medium is 1: 10 a mass ratio inoculation liquid fermentation culture medium, more than 90%, natural lighting was made solid fermentation culture in 1～3 day in the cultivation room ventilation cultivation of sterilization in 18 ℃ of temperature, relative moisture.Concrete incubation time can reach 3 * 10 with the bacterial strain T23 spore content in the solid fermentation culture
10～9 * 10
10Individual/gram is advisable.
(4) preparation of preparation: with embodiment one.
Embodiment five
The consisting of of plant comminution of material in this routine microorganism formulation: the mixture of corn flour, wheat bran and wood chip.The steps in sequence of its preparation method is:
(1) the bacterial classification test tube is cultivated: adopt potato glucose agar-agar solid culture medium (PDA), trichoderma harzianum strain T23 is inoculated in the solid culture medium (PDA) that contains streptomycin 70ppm, in temperature is under 30 ℃, cultivates 1～3 day, and concrete incubation time is covered with media surface with mycelia and is as the criterion.
(2) liquid fermentation and culture: the mixture of employing corn flour, wheat bran and water are loaded in the triangular flask by 1: 450 mass ratio mixing, and high-temperature pressurizing sterilization 20min makes culture fluid under 121 ℃, 15 pounds; After the cooling trichoderma harzianum strain T23 in the test tube is seeded in the triangular flask, the mass ratio of trichoderma harzianum strain T23 and culture fluid is 1: 450, places 30 ℃, and Light To Dark Ratio is in 1: 1 the alternation of light and darkness environment, shaken cultivation 3～5 days is cultivated and is made the liquid fermentation and culture thing.Concrete incubation time reaches 0.5 * 10 can guarantee the bacterial strain T23 spore content in the liquid fermentation and culture thing
8～2 * 10
8Individual/ml is advisable.
(3) solid fermentation is cultivated: wood chip and water are pressed 1: 1.1 mass ratio mixing, be loaded in the heatproof plastic sack, high-temperature pressurizing sterilization 120min under 121 ℃, 15 pounds, pour into after the cooling in sterilized culture bed, by the liquid fermentation and culture thing: solid culture medium is 1: 8 a mass ratio inoculation liquid fermentation culture medium, more than 90%, natural lighting was made solid fermentation culture in 1～3 day in the cultivation room ventilation cultivation of sterilization in 32 ℃ of temperature, relative moisture.Concrete incubation time can reach 3 * 10 with the bacterial strain T23 spore content in the solid fermentation culture
10～9 * 10
10Individual/gram is advisable.
(4) preparation of preparation: with embodiment one.
The microorganism formulation of the various embodiments described above all contains the trichoderma harzianum strain T23 of 12-16 part quality, the plant flour material of 84-88 part quality.
The concrete mode of the container that uses during the solid fermentation of the various embodiments described above is cultivated and culturing room's sterilization is: adopts fumigation, fumigates 10～12 hours with potassium permanganate 1g, formaldehyde 10ml by every square metre, and standby.
The plant comminution of material that solid fermentation used when cultivating both can be above-mentioned for each embodiment chooses a kind of in rice bran, corn straw part, cotton seed hulls part or the wood chip like that, also can choose at least two kinds mixture in them.
Microorganism formulation of the present invention is employed vegetable material when liquid culture, solid culture, and also available other suitable existing vegetable material of cultivating trichoderma harzianum substitutes.
In the preparation process of microorganism formulation of the present invention:
1, the preference temperature of bacterial classification test tube cultivation is 22 ℃～28 ℃, and 1～3 day mycelia can be covered with media surface under this temperature condition; Below 18 ℃, mycelial growth is slower, needs 6 days just can be covered with media surface, and more than 30 ℃ the time, mycelia needed 7 days just can be covered with media surface.
2, the preference temperature of liquid fermentation and culture is 23 ℃～27 ℃, and its incubation time is generally and can meets the demands in 84-96 hour; Support temperature below 20 ℃, bacterial strain T23 spore content is less than 0.5 * 10
8Individual/ml, more than 30 ℃ the time, bacterial strain T23 spore content is less than 0.4 * 10
8Individual/ml, in the time of 23 ℃～27 ℃, bacterial strain T23 spore content is 0.5 * 10
8Individual～2 * 10
8Individual/ml, spore content reaches and is up to 2 * 10 in the time of 25 ℃
8Individual/ml.
3, the preference temperature of solid fermentation cultivation is 22 ℃-28 ℃, incubation time was generally 36-60 hour, below 18 ℃ the time, need to cultivate more than 4 days, more than 32 ℃ the time, wooden mould growth obviously is obstructed, also need cultivate more than 4 days, need to cultivate 3.5 days mycelia at 22 ℃ and just can be covered with solid fermentation thing surface, in the time of 25 ℃, mycelia can be covered with solid fermentation thing surface in 1 day.
Claims (2)
1, a kind of microorganism formulation that is used for the crop disease prevention volume increase, component that it contains and quality proportioning are: 12～16 parts Trichoderma harzianum (Trichoderma harzianum) bacterial strain T23 CGMCC No.0975, the spore content of this bacterial strain T23 reaches 3 * 10
10Individual/gram; 84～88 parts plant comminution of material, the consisting of of this plant comminution of material: the mixture of a kind of in corn flour or wheat bran or the mixture of the two and rice bran, milled powders of cornstalk, cotton seed hulls powder, the wood chip or at least two kinds.
2, a kind of method for preparing the described microorganism formulation of claim 1, its steps in sequence is:
(1) the bacterial classification test tube is cultivated: adopt potato glucose agar-agar solid culture medium (PDA), trichoderma harzianum strain T23 is inoculated in the solid culture medium (PDA) that contains streptomycin 40～70ppm, under 18 ℃～30 ℃ temperature, cultivated 1～7 day;
(2) liquid fermentation and culture: corn flour or wheat bran or the mixture of the two and water are loaded in the triangular flask by 1: 300～500 mass ratio mixing, and the high-temperature pressurizing sterilization is 20～30 minutes under 121 ℃, 15 pounds, makes culture fluid; After the cooling trichoderma harzianum strain T23 that turns out in (1) step test tube is seeded in the triangular flask, the mass ratio of bacterial strain and culture fluid is 1: 400～500, and be 20 ℃～30 ℃ in temperature, Light To Dark Ratio is 1: in the alternation of light and darkness environment of 0.8-1.2, and constant-temperature shaking culture 3～5 days;
(3) solid fermentation is cultivated: the mixture and the water a kind of or at least two kinds of rice bran, milled powders of cornstalk, cotton seed hulls powder, wood chip are pressed 1: 0.8～1.2 mass ratio mixing, be loaded in the heatproof plastic sack, the high-temperature pressurizing sterilization is 120～150 minutes under 121 ℃, 15 pounds, pour into after the cooling in sterilized culture bed, form solid culture medium; Liquid fermentation and culture thing and this solid culture medium that (2) step makes are inoculated the liquid fermentation culture medium by 1: 8～10 mass ratio, in the culturing room of 18 ℃～32 ℃, relative moisture more than 90%, under the natural lighting, sterilization, ventilate to cultivating and promptly made solid fermentation culture in 1～3 day.
(4) preparation of preparation: with the solid fermentation culture natural air drying to water content 8%～10%, pulverize with cracker again, cross 100 mesh sieves after, measure spore content, bacterial strain T23 spore content reaches 3 * 10
10Individual/gram is certified products.
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Cited By (2)
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| CN101485259B (en) * | 2009-03-04 | 2011-04-20 | 四川省农业科学院经济作物育种栽培研究所 | Disease-controlling and seedling-promoting non-pollution processing method for sprout of Ligusticum wallichii |
| CN103952321A (en) * | 2014-04-28 | 2014-07-30 | 唐山金土生物有机肥有限公司 | Trichoderma harzianum and applications thereof in preventing and controlling turfgrass brown patch disease |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101485336B (en) * | 2009-03-04 | 2011-10-12 | 四川省农业科学院经济作物育种栽培研究所 | Trichoderma sp. preparation for preventing and treating soil-borne disease of crop |
| CN101897346A (en) * | 2010-08-05 | 2010-12-01 | 宁夏农林科学院 | Trichoderma granules for preventing and controlling blight of watermelons growing on gravel-covered land |
| CN102382791A (en) * | 2011-11-21 | 2012-03-21 | 滨州职业学院 | Fermentation process of trichoderma |
| CN109337829B (en) * | 2018-12-18 | 2020-11-03 | 山东五福生生态工程有限公司 | Trichoderma harzianum solid fermentation method, solid fermentation product, trichoderma harzianum microbial agent, and preparation method and application thereof |
-
2003
- 2003-08-25 CN CN 03135663 patent/CN1250084C/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101485259B (en) * | 2009-03-04 | 2011-04-20 | 四川省农业科学院经济作物育种栽培研究所 | Disease-controlling and seedling-promoting non-pollution processing method for sprout of Ligusticum wallichii |
| CN103952321A (en) * | 2014-04-28 | 2014-07-30 | 唐山金土生物有机肥有限公司 | Trichoderma harzianum and applications thereof in preventing and controlling turfgrass brown patch disease |
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