CN104312930B - A kind of preparation method of the type that the sprays composite bacteria agent capable for rape stalk and sclerotium decomposition - Google Patents

A kind of preparation method of the type that the sprays composite bacteria agent capable for rape stalk and sclerotium decomposition Download PDF

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CN104312930B
CN104312930B CN201410584321.0A CN201410584321A CN104312930B CN 104312930 B CN104312930 B CN 104312930B CN 201410584321 A CN201410584321 A CN 201410584321A CN 104312930 B CN104312930 B CN 104312930B
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microbial inoculum
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trichoderma harzianum
aspergillus aculeatus
bulbus allii
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CN104312930A (en
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谢立华
廖星
胡小加
余常兵
李银水
秦路
张银波
胡磊
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Oil Crops Research Institute of Chinese Academy of Agriculture Sciences
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Abstract

The present invention relates to a kind of type that sprays composite bacteria agent capable preparation method for rape stalk and sclerotium decomposition.The preparation method of a kind of type that sprays composite bacteria agent capable for rape stalk and sclerotium decomposition, it is characterised in that comprise the steps: 1) preparation of Trichoderma harzianum microbial inoculum；2) preparation of microorganism Aspergillus aculeatus microbial inoculum；3) by Trichoderma harzianum microbial inoculum: the mass ratio of microorganism Aspergillus aculeatus microbial inoculum is 1～2:1, Trichoderma harzianum microbial inoculum and microorganism Aspergillus aculeatus microbial inoculum are mixed pack, obtains spraying type composite bacteria agent capable.The living bacteria count of the composite bacteria agent capable that the method obtains is high, can efficiently decomposition rape stalk and sclerotium, the method low cost.

Description

A kind of preparation method of the type that the sprays composite bacteria agent capable for rape stalk and sclerotium decomposition
Technical field
The present invention relates to a kind of type that sprays composite bacteria agent capable preparation method for rape stalk and sclerotium decomposition.
Background technology
In recent years, China's Brassica campestris L entire mechanization progress faster, along with the increase of Brassica campestris L mechanical harvest area, adjoint phenomenon is Substantial amounts of rape straw direct returning to farmland, but produce to land for growing field crops succession crop and bring two major issues, one be rape straw how Can quick composting, neither affect succession crop growth of seedling, the nutrient demand of succession crop seedling can be quickly provided again, two be as Sclerotium in the parasitic rape stalk of what degraded, reduces the harm to Brassica campestris L of lower season of the soil-borne disease sclerotinia rot of colza.So, field Biology in situ converts, the type that the sprays composite bacteria agent capable of the simple and effective rape stalk returned crop stocks to the field and sclerotium decomposition (and preparation method thereof) Use be support mechanization Rape-seed production important channel.
Summary of the invention
It is an object of the invention to provide the preparation method of a kind of type that sprays composite bacteria agent capable for rape stalk and sclerotium decomposition, should The living bacteria count of the composite bacteria agent capable that method obtains is high, can efficiently decomposition rape stalk and sclerotium, the method low cost.
To achieve these goals, the technical solution used in the present invention is: a kind of spraying for rape stalk and sclerotium decomposition The preparation method of type composite bacteria agent capable, it is characterised in that comprise the steps:
1) preparation of Trichoderma harzianum microbial inoculum:
1. inoculated fungi: be seeded in the enamel tray containing V8+S culture medium by Trichoderma harzianum thalline, covers clean double-layer sand cloth, Cultivating 5 days for 28 DEG C, then 35 DEG C air-dry 2 days；
Trichoderma harzianum is first configured to spore suspension, and inoculum concentration is that per kilogram V8+S culture medium adds 10 milliliters of spore suspension, Every milliliter of spore amount is >=5 × 104Individual；
2. as shared by each component, percetage by weight is: soluble starch 85.0%, glucose 13.5%, fluorescent whitening agent 0.3%, Fungal spore 1.2%, chooses soluble starch, glucose, fluorescent whitening agent and step the most postvaccinal Trichoderma harzianum fungal spore； After being sufficiently mixed with the fungal spore being dried with soluble starch, glucose and fluorescent whitening agent, crossing 100 eye mesh screens, 35 DEG C again Air-dry 1 day, obtain Trichoderma harzianum microbial inoculum；
2) preparation of microorganism Aspergillus aculeatus microbial inoculum:
1. inoculated fungi: be seeded in the enamel tray containing V8+S culture medium by microorganism Aspergillus aculeatus thalline, covers clean double-layer sand cloth, Cultivating 5 days for 28 DEG C, then 35 DEG C air-dry 2 days；
Microorganism Aspergillus aculeatus is first configured to spore suspension, and inoculum concentration is that per kilogram V8+S culture medium adds 10 milliliters of spore suspension, Every milliliter of spore amount is >=5 × 104Individual；
2. as shared by each component, percetage by weight is: soluble starch 85.0%, glucose 13.5%, fluorescent whitening agent 0.3%, Fungal spore 1.2%, chooses soluble starch, glucose, fluorescent whitening agent and step the most postvaccinal microorganism Aspergillus aculeatus fungal spore； After being sufficiently mixed with the fungal spore being dried with soluble starch, glucose and fluorescent whitening agent, crossing 100 eye mesh screens, 35 DEG C again Air-dry 1 day, obtain microorganism Aspergillus aculeatus microbial inoculum；
3) by Trichoderma harzianum microbial inoculum: the mass ratio of microorganism Aspergillus aculeatus microbial inoculum is 1～2:1, by Trichoderma harzianum microbial inoculum and microorganism Aspergillus aculeatus bacterium Agent mixing pack, obtains spraying type composite bacteria agent capable.
Step 3) in by every bag of 15 grams of subpackages (15 grams every bag have 10 grams for Trichoderma harzianum microbial inoculum, another 5 grams is microorganism Aspergillus aculeatus Microbial inoculum), amount of application is 15 grams every mu；Finally every gram is 10 containing fungal spore amount11Individual/gram more than.
Described V8+S culture medium is: Rhizoma Solani tuber osi 30wt%, Fructus Lycopersici esculenti 20wt%, Radix Dauci Sativae 10wt%, Bulbus Allii Cepae 10wt%, Capsicum annuum L. 10wt%, Herba Apii graveolentis 5wt%, Herba Spinaciae 5wt%, Bulbus Allii 5wt%, sucrose 5wt%.
The preparation of above-mentioned V8+S culture medium:
1) percetage by weight as shared by each component: Rhizoma Solani tuber osi 30%, Fructus Lycopersici esculenti 20%, Radix Dauci Sativae 10%, Bulbus Allii Cepae 10%, Capsicum annuum L. 10%, Herba Apii graveolentis 5%, Herba Spinaciae 5%, Bulbus Allii 5%, sucrose 5%, choose Rhizoma Solani tuber osi, Fructus Lycopersici esculenti, Radix Dauci Sativae, Bulbus Allii Cepae, green grass or young crops Green pepper, Herba Apii graveolentis, Herba Spinaciae, Bulbus Allii and sucrose；
2) Rhizoma Solani tuber osi, Fructus Lycopersici esculenti, Radix Dauci Sativae, Bulbus Allii Cepae, Capsicum annuum L., Herba Apii graveolentis, Herba Spinaciae, Bulbus Allii are cut into small pieces respectively, and mix Uniformly, pulverize, add sucrose mix homogeneously, obtain V8+S culture medium.
Wherein, V8 represents 8 kinds of vegetables, and S represents sucrose.
The present invention is with V8+S culture medium culturing and produces a large amount of fungal spores, and soluble starch is carrier, adsorbing spores；This sprays After type composite bacteria agent capable sprays, glucose can shorten the time that spore is sprouted the growth accelerating mycelia, decomposition rape stalk and sclerotium； Fluorescent whitening agent can defend sun middle-ultraviolet lamp to fungal spore and the injury of mycelia effectively, it is ensured that lasting high bacterium number and prolongation The microbial survival time, sustainable development form local superiority's monoid on straw and sclerotium.It has resistance to environmental condition, effectively Viable count is high, can preserve for a long time in room temperature, and using (spraying) injury that is the most affected by environment little and that bring ultraviolet has necessarily Protection, can in situ quickly decomposition rape stalk and sclerotium in field effectively.
The invention has the beneficial effects as follows:
1, the highest effective bacterium number held by the microbial inoculum of the present invention, exceedes the living bacteria count of current all bacterial manure.Room temperature storage 12 After individual month, fungal spore remains at 1011Individual/gram more than.
2, the resistance to environmental condition of the microbial inoculum of the present invention, it is not necessary to sterile working and aseptic packaging, production technology is easy, low cost is (single Position usage amount reduces, and reduces production and use cost).
3, the microbial inoculum of the present invention, it is achieved rape stalk field biology in situ converts also field and soil-borne disease (sclerotinia rot of colza etc.) Biological control, promote second stubble crop growth.
4, the microbial inoculum of the present invention, uses conveniently, can machinery also can artificial spraying, the fluorescent whitening agent in said preparation can be effective Ground defence sun middle-ultraviolet lamp is to fungal spore and the injury of mycelia, it is ensured that lasting high bacterium number and prolongation microbial survival time, Sustainable development form local superiority's monoid on straw and sclerotium.
The type that the sprays composite bacteria agent capable of the present invention is suitable for during rape harvest using, it is achieved rape stalk field biology in situ convert also field and Pass the Biological control of disease (sclerotinia rot of colza etc.).
Accompanying drawing explanation
Fig. 1 is the powder figure that the present invention sprays type composite bacteria agent capable.
Detailed description of the invention
In order to be more fully understood that the present invention, it is further elucidated with present disclosure below in conjunction with embodiment, but present disclosure is not It is limited only to the following examples.
In following embodiment: V8+S culture medium (being made up of 8 kinds of vegetables, this culture medium is beneficial to fungus and forms spore) is: horse Bell potato 30wt%, Fructus Lycopersici esculenti 20wt%, Radix Dauci Sativae 10wt%, Bulbus Allii Cepae 10wt%, Capsicum annuum L. 10wt%, Herba Apii graveolentis 5wt%, spinach Dish 5wt%, Bulbus Allii 5wt%, sucrose 5wt%.
The preparation of above-mentioned V8+S culture medium:
1) percetage by weight as shared by each component: Rhizoma Solani tuber osi 30%, Fructus Lycopersici esculenti 20%, Radix Dauci Sativae 10%, Bulbus Allii Cepae 10%, Capsicum annuum L. 10%, Herba Apii graveolentis 5%, Herba Spinaciae 5%, Bulbus Allii 5%, sucrose 5%, choose Rhizoma Solani tuber osi, Fructus Lycopersici esculenti, Radix Dauci Sativae, Bulbus Allii Cepae, green grass or young crops Green pepper, Herba Apii graveolentis, Herba Spinaciae, Bulbus Allii and sucrose；
2) Rhizoma Solani tuber osi, Fructus Lycopersici esculenti, Radix Dauci Sativae, Bulbus Allii Cepae, Capsicum annuum L., Herba Apii graveolentis, Herba Spinaciae, Bulbus Allii (8 kinds of vegetables) are cut into respectively Surface is the fritter of 1-2 square centimeter, and mix homogeneously, then smashes with pressure-even pulp crusher, adds sucrose mix homogeneously, keep flat into In enamel tray, height, less than 0.5 centimetre, obtains V8+S culture medium (i.e. can be used for inoculated fungi thalline), standby.
Embodiment 1:
The preparation method of a kind of type that sprays composite bacteria agent capable for rape stalk and sclerotium decomposition, comprises the steps:
1) preparation of Trichoderma harzianum microbial inoculum:
1. inoculated fungi: be seeded in the enamel tray containing V8+S culture medium by Trichoderma harzianum thalline, covers clean double-layer sand cloth, Cultivating 5 days for 28 DEG C, then 35 DEG C air-dry 2 days；
Trichoderma harzianum is first configured to spore suspension, and inoculum concentration is that per kilogram V8+S culture medium adds 10 milliliters of spore suspension, Every milliliter of spore amount is >=5 × 104Individual；
2. as shared by each component, percetage by weight is: soluble starch 85.0%, glucose 13.5%, fluorescent whitening agent 0.3%, Fungal spore 1.2%, chooses soluble starch, glucose, fluorescent whitening agent and step the most postvaccinal Trichoderma harzianum fungal spore； After being sufficiently mixed with the fungal spore being dried with soluble starch, glucose and fluorescent whitening agent, crossing 100 eye mesh screens, 35 DEG C again Air-dry 1 day, obtain Trichoderma harzianum microbial inoculum；
2) preparation of microorganism Aspergillus aculeatus microbial inoculum:
1. inoculated fungi: be seeded in the enamel tray containing V8+S culture medium by microorganism Aspergillus aculeatus thalline, covers clean double-layer sand cloth, Cultivating 5 days for 28 DEG C, then 35 DEG C air-dry 2 days；
Microorganism Aspergillus aculeatus is first configured to spore suspension, and inoculum concentration is that per kilogram V8+S culture medium adds 10 milliliters of spore suspension, Every milliliter of spore amount is >=5 × 104Individual；
2. as shared by each component, percetage by weight is: soluble starch 85.0%, glucose 13.5%, fluorescent whitening agent 0.3%, Fungal spore 1.2%, chooses soluble starch, glucose, fluorescent whitening agent and step the most postvaccinal microorganism Aspergillus aculeatus fungal spore； After being sufficiently mixed with the fungal spore being dried with soluble starch, glucose and fluorescent whitening agent, crossing 100 eye mesh screens, 35 DEG C again Air-dry 1 day, obtain microorganism Aspergillus aculeatus microbial inoculum；
3) by Trichoderma harzianum microbial inoculum: the mass ratio of microorganism Aspergillus aculeatus microbial inoculum is 2:1, Trichoderma harzianum microbial inoculum and microorganism Aspergillus aculeatus microbial inoculum are mixed Attach together bag,
By every bag of 15 grams of subpackages (having 10 grams in 15 grams every bag is Trichoderma harzianum microbial inoculum, and another 5 grams is microorganism Aspergillus aculeatus microbial inoculum), execute Consumption is 15 grams every mu；Finally every gram is 10 containing fungal spore amount11Individual/gram more than, obtain spraying type composite bacteria agent capable.
Trichoderma harzianum and the subpackage ratio of microorganism Aspergillus aculeatus is determined according to the sclerotinia rot of colza disease index that field then is investigated.Brassica campestris L During sclerotiniose disease index≤20, the subpackage ratio of Trichoderma harzianum and microorganism Aspergillus aculeatus is 2:1.The ability of Trichoderma harzianum decomposition straw It is better than microorganism Aspergillus aculeatus, but the ability of microorganism Aspergillus aculeatus decomposition Brassica campestris L sclerotium is better than Trichoderma harzianum.Its object is to guarantee Brassica campestris L straw Stalk and sclerotium energy field simultaneously the most effectively decomposition.
Application effect: investigation in 30 days, compares with not spraying " spraying type composite bacteria agent capable ", and rape stalk decomposition rate improves 16.5%；Sclerotium decomposition rate improves 38.1%.
Embodiment 2:
The preparation method of a kind of type that sprays composite bacteria agent capable for rape stalk and sclerotium decomposition, comprises the steps:
1) preparation of Trichoderma harzianum microbial inoculum:
1. inoculated fungi: be seeded in the enamel tray containing V8+S culture medium by Trichoderma harzianum thalline, covers clean double-layer sand cloth, Cultivating 5 days for 28 DEG C, then 35 DEG C air-dry 2 days；
Trichoderma harzianum is first configured to spore suspension, and inoculum concentration is that per kilogram V8+S culture medium adds 10 milliliters of spore suspension, Every milliliter of spore amount is >=5 × 104Individual；
2. as shared by each component, percetage by weight is: soluble starch 85.0%, glucose 13.5%, fluorescent whitening agent 0.3%, Fungal spore 1.2%, chooses soluble starch, glucose, fluorescent whitening agent and step the most postvaccinal Trichoderma harzianum fungal spore； After being sufficiently mixed with the fungal spore being dried with soluble starch, glucose and fluorescent whitening agent, crossing 100 eye mesh screens, 35 DEG C again Air-dry 1 day, obtain Trichoderma harzianum microbial inoculum；
2) preparation of microorganism Aspergillus aculeatus microbial inoculum:
1. inoculated fungi: be seeded in the enamel tray containing V8+S culture medium by microorganism Aspergillus aculeatus thalline, covers clean double-layer sand cloth, Cultivating 5 days for 28 DEG C, then 35 DEG C air-dry 2 days；
Microorganism Aspergillus aculeatus is first configured to spore suspension, and inoculum concentration is that per kilogram V8+S culture medium adds 10 milliliters of spore suspension, Every milliliter of spore amount is >=5 × 104Individual；
2. as shared by each component, percetage by weight is: soluble starch 85.0%, glucose 13.5%, fluorescent whitening agent 0.3%, Fungal spore 1.2%, chooses soluble starch, glucose, fluorescent whitening agent and step the most postvaccinal microorganism Aspergillus aculeatus fungal spore； After being sufficiently mixed with the fungal spore being dried with soluble starch, glucose and fluorescent whitening agent, crossing 100 eye mesh screens, 35 DEG C again Air-dry 1 day, obtain microorganism Aspergillus aculeatus microbial inoculum；
3) by Trichoderma harzianum microbial inoculum: the mass ratio of microorganism Aspergillus aculeatus microbial inoculum is 1:1, Trichoderma harzianum microbial inoculum and microorganism Aspergillus aculeatus microbial inoculum are mixed Attach together bag,
By every bag of 15 grams of subpackages (having 7.5 grams in 15 grams every bag is Trichoderma harzianum microbial inoculum, and another 7.5 grams is microorganism Aspergillus aculeatus microbial inoculum), Amount of application is 15 grams every mu；Finally every gram is 10 containing fungal spore amount11Individual/gram more than, obtain spraying type composite bacteria agent capable.
Trichoderma harzianum and the subpackage ratio of microorganism Aspergillus aculeatus is determined according to the sclerotinia rot of colza disease index that field then is investigated.Brassica campestris L During sclerotiniose disease index ＞ 20, the subpackage ratio of Trichoderma harzianum and microorganism Aspergillus aculeatus is 1:1.The ability of Trichoderma harzianum decomposition straw It is better than microorganism Aspergillus aculeatus, but the ability of microorganism Aspergillus aculeatus decomposition Brassica campestris L sclerotium is better than Trichoderma harzianum.Its object is to guarantee Brassica campestris L straw Stalk and sclerotium energy field simultaneously the most effectively decomposition.
Application effect: investigation in 30 days, compares with not spraying " spraying type composite bacteria agent capable ", and rape stalk decomposition rate improves 23.9%； Sclerotium decomposition rate improves 31.4%.

Claims (3)

1. the preparation method of the type that the sprays composite bacteria agent capable being used for rape stalk and sclerotium decomposition, it is characterised in that include walking as follows Rapid:
1) preparation of Trichoderma harzianum microbial inoculum:
1. inoculated fungi: be seeded in the enamel tray containing V8+S culture medium by Trichoderma harzianum thalline, covers clean double-layer sand cloth, Cultivating 5 days for 28 DEG C, then 35 DEG C air-dry 2 days；
Trichoderma harzianum is first configured to spore suspension, and inoculum concentration is that per kilogram V8+S culture medium adds 10 milliliters of spore suspension, Every milliliter of spore amount is >=5 × 104Individual；
Described V8+S culture medium is: Rhizoma Solani tuber osi 30wt%, Fructus Lycopersici esculenti 20wt%, Radix Dauci Sativae 10wt%, Bulbus Allii Cepae 10wt%, Capsicum annuum L. 10wt%, Herba Apii graveolentis 5wt%, Herba Spinaciae 5wt%, Bulbus Allii 5wt%, sucrose 5wt%；
2. as shared by each component, percetage by weight is: soluble starch 85.0%, glucose 13.5%, fluorescent whitening agent 0.3%, Fungal spore 1.2%, chooses soluble starch, glucose, fluorescent whitening agent and step the most postvaccinal Trichoderma harzianum fungal spore； After being sufficiently mixed with the fungal spore being dried with soluble starch, glucose and fluorescent whitening agent, crossing 100 eye mesh screens, 35 DEG C again Air-dry 1 day, obtain Trichoderma harzianum microbial inoculum；
2) preparation of microorganism Aspergillus aculeatus microbial inoculum:
1. inoculated fungi: be seeded in the enamel tray containing V8+S culture medium by microorganism Aspergillus aculeatus thalline, covers clean double-layer sand cloth, Cultivating 5 days for 28 DEG C, then 35 DEG C air-dry 2 days；
Microorganism Aspergillus aculeatus is first configured to spore suspension, and inoculum concentration is that per kilogram V8+S culture medium adds 10 milliliters of spore suspension, Every milliliter of spore amount is >=5 × 104Individual；
Described V8+S culture medium is: Rhizoma Solani tuber osi 30wt%, Fructus Lycopersici esculenti 20wt%, Radix Dauci Sativae 10wt%, Bulbus Allii Cepae 10wt%, Capsicum annuum L. 10wt%, Herba Apii graveolentis 5wt%, Herba Spinaciae 5wt%, Bulbus Allii 5wt%, sucrose 5wt%；
2. as shared by each component, percetage by weight is: soluble starch 85.0%, glucose 13.5%, fluorescent whitening agent 0.3%, Fungal spore 1.2%, chooses soluble starch, glucose, fluorescent whitening agent and step the most postvaccinal microorganism Aspergillus aculeatus fungal spore； After being sufficiently mixed with the fungal spore being dried with soluble starch, glucose and fluorescent whitening agent, crossing 100 eye mesh screens, 35 DEG C again Air-dry 1 day, obtain microorganism Aspergillus aculeatus microbial inoculum；
3) by Trichoderma harzianum microbial inoculum: the mass ratio of microorganism Aspergillus aculeatus microbial inoculum is 1～2:1, by Trichoderma harzianum microbial inoculum and microorganism Aspergillus aculeatus bacterium Agent mixing pack, obtains spraying type composite bacteria agent capable.
The preparation method of a kind of type that sprays composite bacteria agent capable for rape stalk and sclerotium decomposition the most according to claim 1, It is characterized in that: step 3) in by every bag of 15 grams of subpackages, every gram of amount Han fungal spore is 1011Individual/gram more than.
The preparation method of a kind of type that sprays composite bacteria agent capable for rape stalk and sclerotium decomposition the most according to claim 1, It is characterized in that: the preparation of described V8+S culture medium:
1) percetage by weight as shared by each component: Rhizoma Solani tuber osi 30%, Fructus Lycopersici esculenti 20%, Radix Dauci Sativae 10%, Bulbus Allii Cepae 10%, Capsicum annuum L. 10%, Herba Apii graveolentis 5%, Herba Spinaciae 5%, Bulbus Allii 5%, sucrose 5%, choose Rhizoma Solani tuber osi, Fructus Lycopersici esculenti, Radix Dauci Sativae, Bulbus Allii Cepae, green grass or young crops Green pepper, Herba Apii graveolentis, Herba Spinaciae, Bulbus Allii and sucrose；
2) Rhizoma Solani tuber osi, Fructus Lycopersici esculenti, Radix Dauci Sativae, Bulbus Allii Cepae, Capsicum annuum L., Herba Apii graveolentis, Herba Spinaciae, Bulbus Allii are cut into small pieces respectively, and mix Uniformly, pulverize, add sucrose mix homogeneously, obtain V8+S culture medium.
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