CN103814754A - Method for preparing AMF (Arbuscular Mycorrhizal Fungus) capsule fungicide - Google Patents
Method for preparing AMF (Arbuscular Mycorrhizal Fungus) capsule fungicide Download PDFInfo
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- CN103814754A CN103814754A CN201410073976.1A CN201410073976A CN103814754A CN 103814754 A CN103814754 A CN 103814754A CN 201410073976 A CN201410073976 A CN 201410073976A CN 103814754 A CN103814754 A CN 103814754A
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Abstract
The invention provides a method for preparing AMF (Arbuscular Mycorrhizal Fungus) capsule fungicide. The method is characterized in that sodium alga acid is used as a basic carrier, a mixture is obtained by matching and mixing multiple fillers such as chitosan, gelatin, glycerol, rice, peat, coal cinder and malt sprout in the basic carrier, a fungicide mixture can be obtained by embedding the fungicide in the mixture after the mixture is sterilized, a spherical object can be prepared by dripping the fungicide mixture into calcium chloride solution in a physical extrusion way, and the AMF capsule fungicide can be obtained after the spherical object is solidified and cleaned. According to the method disclosed by the invention, spores, mycorrhiza and hypha of inner glomus intraradices fungus are embedded in the sodium alga acid, a complete infection process can be completed when the spores, the mycorrhiza and the hypha are seeded in unpasteurized soil, and the infection rate can be increased with the prolonging of the infection time. The method disclosed by the invention has the advantages of simpleness, low cost and low toxicity; an embedding material of the AMF capsule fungicide prepared by the invention can be easily decomposed by microorganism, the preservation time is longer, and the AMF capsule fungicide can be simultaneously and directly seeded in a big field.
Description
Technical field
The invention belongs to capsule microbial inocula preparing technical field, relate in particular to a kind of preparation method of arbuscular mycorrhizal fungi capsule microbial inocula.
Background technology
Arbuscular mycorrhiza (Arbuscularmycorrhizalfungus, arbuscular mycorrhizal fungi) fungi is the endotrophic mycorrhiza fungi being extensively present in soil, and it can form syntaxial system with 80% plant on the earth.Research shows, arbuscular mycorrhizal fungi can be improved water metabolism in plant corpus, improves drought-resistant, salt alkali ability; Strengthen plant disease-resistant ability; Promote absorption and the utilization of plant to various mineral elements, thereby promote growth, improve quality.The carbohydrate that arbuscular mycorrhizal fungi utilizes host plant to provide maintains self Growth and reproduction.Arbuscular mycorrhizal fungi is obligate symbiosis fungi, self can not synthetic DNA, therefore also cannot as external fungi, carry out so far pure culture and then industrial fermentation, and can only interdependent plant root and its common breeding growth.
Liquid bacterial agent is produced simple, with low cost, easy to use, but transportation is difficult, is unfavorable for the Growth and reproduction of fungi, and the storage time is unsuitable long.Solid fungicide is on liquid bacterial agent basis, adds suitable material and bacterial classification to produce, make simple, transportation and store conveniently, shortcoming is that the microbial inoculum bacteria containing amount of equal quality and volume is lower than liquid bacterial agent.Pulvis is that spore is sneaked into the microbial inoculum of making in inserts, stores, transports and use more for convenience, but in microbial inoculum, only having spore, the bad result of use that can affect microbial inoculum of spore-germination.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of arbuscular mycorrhizal fungi capsule microbial inocula, be intended to the problem solving.
The present invention is achieved in that a kind of preparation method of arbuscular mycorrhizal fungi capsule microbial inocula, comprises the following steps:
(1) take sodium alginate as underlying carrier, in described underlying carrier, add filler to mix, by described mixture sterilizing, and arbuscular mycorrhiza liquid bacterial agent is evenly embedded in described mixture, obtain microbial inoculum mixture; Wherein, described filler be shitosan, gelatin, glycerine, peat, cinder, malt root and rice meal wherein at least one; In described microbial inoculum mixture, the mass percent of sodium alginate, filler and arbuscular mycorrhiza is respectively 2～3%, 2～9% and 1～2%;
(2) described microbial inoculum mixture being splashed into mass concentration ratio by physics fashion of extrusion is in 1.5～4.5% calcium chloride solutions, under stirring, the thing that splashes into of described microbial inoculum mixture forms sphere in calcium chloride solution, after solidifying, takes out described sphere, and by surface chlorination calcium with drying after sterile water wash, obtain arbuscular mycorrhizal fungi capsule microbial inocula.
Preferably, in step (1), described filler is to account for shitosan that microbial inoculum mixture quality percent value is 2%, 3% gelatin, 5% glycerine, 3～5% peat, 7～9% cinder, 2～5% malt root or 2～5% rice meal.
Preferably, in step (1), described filler grinds evenly before use;
In step (1), after described liquid bacterial agent and sterilizing, the temperature of mixture is all lower than 40 ℃, and described sterilizing is steam sterilizing 20min under 121 ℃, 0.12MPa pressure;
In step (1), the temperature of described embedding is 20～40 ℃.
Preferably, in step (1), arbuscular mycorrhiza liquid bacterial agent is prepared by following steps:
A, arbuscular mycorrhiza is carried out to dual cultivation with carrot excised root, obtain arbuscular mycorrhizal fungi culture;
B, described arbuscular mycorrhizal fungi culture is blended after, cross 400 mesh sieves and screen out impurity with sterile water, leave and take the arbuscular mycorrhiza liquid bacterial agent that contains mycelia, spore and mycorhiza.
Preferably, described arbuscular mycorrhiza is Glomus intraradices bacterium (Glomusintraradices).
Preferably, in step (2), described physics fashion of extrusion is for to push described microbial inoculum mixture with constant flow pump or syringe;
In step (2), the diameter of described sphere is 3～6mm;
In step (2), the speed of described stirring is not more than 30rpm/min;
In step (2), the described curing time is 8～16h.
For overcoming the deficiencies in the prior art and shortcoming, a kind of preparation method of arbuscular mycorrhizal fungi capsule microbial inocula is provided in the present invention, by take sodium alginate as underlying carrier, coordinate shitosan, gelatin, glycerine, rice, peat, cinder, the multiple filler of malt root to be mixed to get mixture, after mixture sterilizing, microbial inoculum is embedded into and in mixture, obtains microbial inoculum mixture, microbial inoculum mixture is splashed in calcium chloride solution and made sphere by physics fashion of extrusion, after sphere solidifies cleaning, obtain arbuscular mycorrhizal fungi capsule microbial inocula.In the present invention, spore, mycorhiza and the mycelia of the mould fungi of interior sacculus are embedded in sodium alginate, while sowing in unpasteurized soil, can complete complete infection processs, and with the prolongation of time of infection, infection rate all has lifting.Preparation method of the present invention is simple, with low cost, toxicity is little, and arbuscular mycorrhizal fungi capsule microbial inocula embedded material of the present invention is easy to be decomposed by microorganism, and the holding time is longer, can directly be seeded in large Tanaka simultaneously.
Accompanying drawing explanation
Fig. 1 is the profile observed result figure of arbuscular mycorrhizal fungi capsule microbial inocula 1～8 in effect embodiment of the present invention;
Fig. 2 is the result comparison diagram of the infection rate of different microbial inoculums after 25 days in effect embodiment of the present invention;
Fig. 3 be in effect embodiment of the present invention different microbial inoculums along with under incubation time infection rate change schematic diagram;
Fig. 4 is that in effect embodiment of the present invention, arbuscular mycorrhizal fungi capsule microbial inocula 5 changes schematic diagram to the infection rate of Chinese sorghum after storing different time;
Fig. 5 is that in effect embodiment of the present invention, arbuscular mycorrhizal fungi capsule microbial inocula 1,2 and 4 freeze dryings front and back infection rates change schematic diagram.
Embodiment
In order to make object of the present invention, technical scheme and advantage clearer, below in conjunction with drawings and Examples, the present invention is further elaborated.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not intended to limit the present invention.
The production of embodiment arbuscular mycorrhizal fungi capsule microbial inocula
(1) preparation of liquid bacterial agent
Get Glomus intraradices bacterium (Glomusintraradices) solid fungicide (the concrete preparation method of this microbial inoculum is by name referring to publication number CN101565689A: the patent of invention of " the purebred production method of high density of arbuscular mycorrhizal fungi spore ") that the dual cultivation of Ri-TDNA Carrot Roots obtains, putting into pulper adds a small amount of water to blend, after taking-up, in 400 object sieves, repeatedly sieve and wash with sterile water, remove agar medium, what retain is concentration for (3～6) × 10
2the arbuscular mycorrhizal fungi liquid bacterial agent of CFU/ml, contains mycorhiza, spore and mycelia that height infects.
(2) preparation of underlying carrier and filler
Be that sodium alginate and filler mix by underlying carrier, wherein, the mass ratio of underlying carrier and filler is 2-3:2～9, kind and the consumption of filler are as shown in table 1, after mixing, add water and be settled to 200ml, mixture is carried out to sterilization treatment 20min in 121 ℃, the high-pressure sterilizing pot of 0.12MPa, be cooled to lower than 40 ℃, for subsequent use.
(3) preparation of capsule microbial inocula
Under aseptic condition by 50ml(500 spore/ml of arbuscular mycorrhizal fungi liquid bacterial agent) at 20～40 ℃, be embedded in the mixture going out in step (2) after mixed, mix, make microbial inoculum mixture.
(4) take syringe or constant flow pump at the uniform velocity splashes into mass concentration in 1.5～4.5% aseptic calcium chloride solution 500ml by described microbial inoculum mixture, with magnetic stirring apparatus to stir lower than 30rpm/min rotating speed, in calcium chloride solution, to form diameter be 3～6mm sphere to the thing that splashes into of described microbial inoculum mixture, after solidifying 8～16h, takes out described sphere, and by surface chlorination calcium with drying after sterile water wash, obtain arbuscular mycorrhizal fungi capsule microbial inocula.
In embodiment of the present invention step (2), the arbuscular mycorrhizal fungi capsule microbial inocula of filler used, consumption and corresponding preparation thereof is as shown in table 1 below:
The profile of effect embodiment arbuscular mycorrhizal fungi capsule microbial inocula is observed and is infected active detection
1, the profile of arbuscular mycorrhizal fungi capsule microbial inocula 1～8 is observed
The arbuscular mycorrhizal fungi capsule microbial inocula 1～7 that above-described embodiment is prepared is observed after taking pictures under the microscope, result as shown in Figure 1, HG is arbuscular mycorrhizal fungi capsule microbial inocula 1 prepared by sodium alginate and glycerine, HK is arbuscular mycorrhizal fungi capsule microbial inocula 2 prepared by sodium alginate-chitosan, H is arbuscular mycorrhizal fungi capsule microbial inocula 3 prepared by sodium alginate-rice, HM is arbuscular mycorrhizal fungi capsule microbial inocula 4 prepared by sodium alginate-gelatin, HN is arbuscular mycorrhizal fungi capsule microbial inocula 5 prepared by sodium alginate-peat, H-malt root is arbuscular mycorrhizal fungi capsule microbial inocula 6 prepared by sodium alginate-maltose, H-cinder is arbuscular mycorrhizal fungi capsule microbial inocula 7 prepared by sodium alginate-cinder, sodium alginate is the arbuscular mycorrhizal fungi capsule microbial inocula 8 that does not add the blank group of any filler.
2, arbuscular mycorrhizal fungi capsule microbial inocula 1～7 infect active detection
Take unpasteurized soil as culture matrix, taper iron sheet barrel is culture vessel, and Sorghum Seedlings is host plant, and Chinese sorghum is added to co-incubation in soil with arbuscular mycorrhizal fungi capsule microbial inocula 1～7 respectively.Regularly detect Chinese sorghum root infection rate.
(1) the infection rate situation to Chinese sorghum
Result as shown in Figure 2, as can be seen from Figure 2, after adding filler, microbial inoculum infection rate is had to impact in various degree, arbuscular mycorrhizal fungi capsule microbial inocula 3(H take rice as filler) infection rate to plant after 25 days is the highest, reach 42.9%, next is the arbuscular mycorrhizal fungi capsule microbial inocula 1(HG take glycerine as filler) infection rate reaches 39.5%, take the arbuscular mycorrhizal fungi capsule microbial inocula 4(HM as filler of gelatin) and be only second to 36.6% of liquid bacterial agent, reach 36.5% infection rate.
(2) arbuscular mycorrhiza (Arbuscularmycorrhizalfungus) liquid bacterial agent, arbuscular mycorrhizal fungi capsule microbial inocula 1,2 and 4(HG, HK, HM) with the altogether variation of incubation time infection rate of Chinese sorghum
Result as shown in Figure 3, as can be seen from Figure 3, three kinds of capsule microbial inoculas and liquid bacterial agent are all the increases along with incubation time, infection rate also constantly increases, but still the infection rate of liquid bacterial agent is higher on the whole, and this is due in liquid bacterial agent, and AM fungi directly contacts with Chinese sorghum root, other three kinds of microbial inoculums can first discharge from the capsule of embedding, and then infect Chinese sorghum.
Shitosan, gelatin, glycerine, rice, peat, cinder, malt root difference are filled adding of nutriment, and the infection rate of microbial inoculum is had nothing in common with each other.Although capsule microbial inocula of the present invention is a little less than the effect that infects of liquid bacterial agent, but the inconvenience of liquid bacterial agent storage and transport, then inconvenient user's large-scale application, and the present invention is conducive to storage and transport, in the situation that infection rate is more or less the same, the present invention has more the significant market advantage.
(3) inquire into the infection rate storing after different time as an example of arbuscular mycorrhizal fungi capsule microbial inocula 5 example
As shown in Figure 4, as can be seen from Figure 4, arbuscular mycorrhizal fungi capsule microbial inocula 5(HN) preserve 1 month at 4 ℃, after 2 months, Chinese sorghum is infected and still has higher infection rate.Other capsule microbial inocula also has similar preservation effect.
(4) arbuscular mycorrhizal fungi capsule microbial inocula 1,2 and 4(HG, HK, HM) comparison of infection rate before and after freeze drying
As shown in Figure 5, as can be seen from Figure 5, arbuscular mycorrhizal fungi capsule microbial inocula freeze drying of the present invention still has stronger infection rate after preserving.
Compare and the shortcoming and defect of prior art, the present invention has following beneficial effect:
(1) in the present invention, in unpasteurized soil, after planting can complete complete infection processs, infection rate is high.
(2) preparation method of the present invention is simple, with low cost, toxicity is little.
(3) arbuscular mycorrhizal fungi capsule microbial inocula embedded material material source of the present invention extensively, be easy to be decomposed by microorganism, the holding time is longer, can suit measures to local conditions to produce, and can be directly in large Tanaka's sowing.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any modifications of doing within the spirit and principles in the present invention, be equal to and replace and improvement etc., within all should being included in protection scope of the present invention.
Claims (6)
1. a preparation method for arbuscular mycorrhizal fungi capsule microbial inocula, is characterized in that comprising the following steps:
(1) take sodium alginate as underlying carrier, in described underlying carrier, add filler to mix, by described mixture sterilizing, and arbuscular mycorrhiza liquid bacterial agent is evenly embedded in described mixture, obtain microbial inoculum mixture; Wherein, described filler be shitosan, gelatin, glycerine, peat, cinder, malt root and rice meal wherein at least one; In described microbial inoculum mixture, the mass percent of sodium alginate, filler and arbuscular mycorrhiza is respectively 2～3%, 2～9% and 1～2%;
(2) described microbial inoculum mixture being splashed into mass concentration by physics fashion of extrusion is in 1.5～4.5% calcium chloride solutions, under stirring, the thing that splashes into of described microbial inoculum mixture forms sphere in calcium chloride solution, after solidifying, takes out described sphere, and by surface chlorination calcium with drying after sterile water wash, obtain arbuscular mycorrhizal fungi capsule microbial inocula.
2. the preparation method of arbuscular mycorrhizal fungi capsule microbial inocula as claimed in claim 1, it is characterized in that, in step (1), described filler is to account for shitosan that microbial inoculum mixture quality percent value is 2%, 3% gelatin, 5% glycerine, 3～5% peat, 7～9% cinder, 2～5% malt root or 2～5% rice meal.
3. the preparation method of described arbuscular mycorrhizal fungi capsule microbial inocula as claimed in claim 1, is characterized in that, in step (1), described filler grinds evenly before use;
In step (1), after described liquid bacterial agent and sterilizing, the temperature of mixture is all lower than 40 ℃, and described sterilizing is steam sterilizing 20min under 121 ℃, 0.12MPa pressure;
In step (1), the temperature of described embedding is 20～40 ℃.
4. the preparation method of arbuscular mycorrhizal fungi capsule microbial inocula as claimed in claim 3, is characterized in that, in step (1), arbuscular mycorrhiza liquid bacterial agent is prepared by following steps:
A, arbuscular mycorrhiza is carried out to dual cultivation with carrot excised root, obtain arbuscular mycorrhizal fungi culture;
B, described arbuscular mycorrhizal fungi culture is blended after, cross 400 mesh sieves and screen out impurity with sterile water, leave and take the arbuscular mycorrhiza liquid bacterial agent that contains mycelia, spore and mycorhiza.
5. the preparation method of arbuscular mycorrhizal fungi capsule microbial inocula as claimed in claim 4, is characterized in that, described arbuscular mycorrhiza is Glomus intraradices bacterium (Glomusintraradices).
6. the preparation method of arbuscular mycorrhizal fungi capsule microbial inocula as claimed in claim 5, is characterized in that, in step (2), described physics fashion of extrusion is for to push described microbial inoculum mixture with constant flow pump or syringe;
In step (2), the diameter of described sphere is 3～6mm;
In step (2), the speed of described stirring is not more than 30rpm/min;
In step (2), the described curing time is 8～16h.
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| CN110668876A (en) * | 2019-11-05 | 2020-01-10 | 内蒙古农业大学 | Composite mycorrhiza biological fertilizer and preparation method and application thereof |
| CN112385507A (en) * | 2020-11-16 | 2021-02-23 | 南京农业大学 | Seedling culture medium containing arbuscular mycorrhiza and preparation method and application thereof |
| CN113249224A (en) * | 2020-02-11 | 2021-08-13 | 江苏财经职业技术学院 | Portable arbuscular mycorrhizal fungus application method |
| CN115530035A (en) * | 2022-09-30 | 2022-12-30 | 西南林业大学 | Paris polyphylla planting and cultivating method |
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| CN113249224A (en) * | 2020-02-11 | 2021-08-13 | 江苏财经职业技术学院 | Portable arbuscular mycorrhizal fungus application method |
| CN112385507A (en) * | 2020-11-16 | 2021-02-23 | 南京农业大学 | Seedling culture medium containing arbuscular mycorrhiza and preparation method and application thereof |
| CN112385507B (en) * | 2020-11-16 | 2021-07-23 | 南京农业大学 | Seedling culture medium containing arbuscular mycorrhiza and preparation method and application thereof |
| CN115530035A (en) * | 2022-09-30 | 2022-12-30 | 西南林业大学 | Paris polyphylla planting and cultivating method |
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Application publication date: 20140528 |