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Method for making alfalfa rhizobium inoculant and rhizobium inoculant made through same and application thereof

An alfalfa rhizobia inoculum and alfalfa rhizobia technology are applied in the field of agricultural microorganisms, which can solve the problems that rhizobia are not suitable for industrialized production and processing, the shelf life of inoculum products is short, and the nitrogen-fixing efficiency of rhizobia is low, so as to achieve high-efficiency survival and good mobility. , the effect of high effective number of viable bacteria

Inactive Publication Date: 2016-04-06
克劳沃(北京)生态科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0029] What the present invention aims to solve is the following one or more technical problems that conventional meliloti rhizobia exist: (1) the nitrogen fixation efficiency of the rhizobia is low; (2) the rhizobia is not suitable for industrialized production and processing; (3) the bacterial agent product has poor stability; (4) The effectiveness of bacterial agent products is low; (5) The shelf life of bacterial agent products is short

Method used

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  • Method for making alfalfa rhizobium inoculant and rhizobium inoculant made through same and application thereof
  • Method for making alfalfa rhizobium inoculant and rhizobium inoculant made through same and application thereof
  • Method for making alfalfa rhizobium inoculant and rhizobium inoculant made through same and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1 strain culture

[0067] The present embodiment adopts YMA agar slant culture method, and concrete method and culture medium formula are as follows:

[0068] (1) Production of YMA agar medium: Weigh potassium dihydrogen phosphate (KH 2 PO 4 ) 0.25g; Magnesium sulfate heptahydrate (MgSO 4 ·7H 2 O) 0.1g; Sodium chloride (NaCl) 0.05g; Mannitol (C 6 h 14 o 6 ) 5g; yeast powder 0.4g; agar 10g; add 500ml of distilled water and stir well, heat until boiling, so that the medium is fully dissolved. Transfer the culture medium to the test tube while it is hot through the funnel (the filling volume accounts for about 1 / 4 of the test tube height). After the packaging is completed, seal it with a silica gel stopper, then tie 7 test tubes into a bundle, and wrap the silica gel stopper of the test tube with parafilm and kraft paper.

[0069] (2) Sterilization of YMA agar medium: Sterilize in a sterilizing pot at 121.5°C for half an hour. During the sterilization, a...

Embodiment 2

[0072] Embodiment 2 Shake flask culture

[0073] The present embodiment adopts yeast-glycerin medium culture method, and specific method and medium formula are as follows:

[0074] (1) prepare yeast-glycerol medium, formula is glycerol 20g; Yeast powder 2.2g; Dipotassium hydrogen phosphate (K 2 HPO 4 ) 1.2g; diammonium hydrogen phosphate [(NH 4 )HPO 4 ] 1.0g; Magnesium sulfate heptahydrate (MgSO 4 ·7H 2 O) 0.7g; Calcium chloride dihydrate (CaCl 2H 2 O) 0.16g; Ferric chloride (FeCl 3 ) 0.0083g; manganese sulfate heptahydrate (MnSO 4 ·7H 2 O) 0.008g; Boric acid (H 3 BO 5 ) 0.01g; Sodium molybdate (Na 2 MoO 4 2H 2 O) 0.01g; defoamer (tributyl phosphate) 0.05ml; water 1000ml, fully stir evenly, transfer the solution to a 500ml triangular flask with a beaker, 200ml for each triangular flask, use sealing film and kraft paper after subpackaging Bandage.

[0075] (2) Yeast-glycerin medium is sterilized, put the yeast-glycerin medium into the Erlenmeyer flask for sterili...

Embodiment 3 1

[0078] Example 3 primary culture

[0079] In this embodiment, a fermenter is used to carry out primary culture, and the specific steps are as follows:

[0080] (1) Sterilize the empty fermenter.

[0081] (2) Prepare glycerol medium: take by weighing respectively with balance: glycerol 400g, yeast powder 44g, dipotassium hydrogen phosphate (K 2 HPO 4 ) 24g, diammonium hydrogen phosphate [(NH 4 )HPO 4 ] 20g, magnesium sulfate heptahydrate (MgSO 4 ·7H 2 O) 14g, calcium chloride dihydrate (CaCl 2H 2 O) 3.2g, ferric chloride (FeCl 3 ) 0.166g, manganese sulfate heptahydrate (MnSO 4 ·7H 2 O) 0.16g, boric acid (H 3 BO 5 ) 0.2g, sodium molybdate (Na 2 MoO 4 2H 2 O) 0.2g, defoamer (tributyl phosphate) 1mL, demineralized water 20L, fully stir and dissolve the above medium, and add it to a 30L fermenter.

[0082] (3) Sterilization of the fermenter culture medium: the fermenter was sterilized at 121.5° C. for half an hour, and then cooled.

[0083] (4) Culture medium inocul...

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Abstract

The invention provides a method for making alfalfa rhizobium inoculant and rhizobium inoculant made through the method and application thereof. The method includes the following steps that S1, bacterium liquid of alfalfa rhizobia is acquired; S2, powder containing kaolin powder and bentonite powder is acquired, and the particle size of the kaolin powder and the bentonite powder ranges from 20 microns to 100 microns; S3, the bacterium liquid and the powder are mixed, dried and smashed so that powder where the alfalfa rhizobium inoculant is adsorbed to can be acquired. The alfalfa rhizobium inoculant has the advantages that the number of effective living bacteria is high, the storage life is long, and the alfalfa rhizobium inoculant is environmentally friendly and suitable for commercial and industrial production.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganisms, in particular, the invention relates to a highly stable alfalfa rhizobia agent and its production method and application. Background technique [0002] It is an indisputable topic that rhizobia can form symbiotic nitrogen fixers with legumes, but what kind of rhizobia can form symbionts with legumes will change with the expansion of the range of rhizobia and the increase of the diversity of rhizobia. increasingly complex. Leguminous forages can symbiotically fix nitrogen with rhizobia, and only when there is a certain amount of bacteria specific to a certain leguminous forage in the soil, this kind of root nodule can form. The bacteria that can form nodules on the roots of legume grasses are called rhizobium bacteria. Rhizobia can fix free nitrogen from the air and transform it into nitrogenous compounds that legumes can absorb and utilize for the growth of legumes. For leg...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F11/08C12R1/41
Inventor 王跃栋王伟苏爱莲刘自学范龙刘艺杉侯湃王圣乾
Owner 克劳沃(北京)生态科技有限公司
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