CN105087414A - High-concentration compound rhizobium inoculant for symbiotic nitrogen fixation and preparation method thereof - Google Patents
High-concentration compound rhizobium inoculant for symbiotic nitrogen fixation and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a high-concentration compound rhizobium inoculant for symbiotic nitrogen fixation and a preparation method thereof. The high-concentration compound rhizobium inoculant is microbial inoculum compound liquid prepared by, by weight, 10-30 parts of pea rhizobium inoculant, 10-30 parts of cowpea rhizobium inoculant, 40-50 parts of milk vetch rhizobium inoculant and 10-30 parts of slow-growing soybean rhizobium inoculant. The preparation method includes: mixing the pea rhizobium inoculant, the cowpea rhizobium inoculant, the milk vetch rhizobium inoculant and the slow-growing soybean rhizobium inoculant according to the above proportion to obtain mixed liquid; performing three-stage enlarged cultivation, namely shaking table liquid cultivation, seed tank liquid cultivation and fermentation liquid tank enlarged cultivation to obtain a high-concentration finished product with microbial count reaching 4-5 billion per ml. The high-concentration compound rhizobium inoculant has good seed soaking, seed mixing and seed fertilizer treating effect on crops, yield of grain and oil crops like peanut, soybean, sweet potato and potato can be increased by 15-30%, yield of commercial crops like ginger, garlic, sweet potato and rhizome medicinal materials can be increased by 20-40%, and the high-concentration compound rhizobium inoculant has the advantages of low investment and high benefit.
Description
Technical field
The present invention relates to a kind of root nodule bacterium fertilizer, particularly relate to a kind of symbiotic nitrogen fixation high density compound nitragin and preparation method thereof.
Background technology
Root nodule bacterium and a kind of root nodule structure of root system of plant organization formation, set up syntaxial system.In the multiple root crop syntaxial systems such as root nodule bacterium and soybean, peanut, cowpea and Herba Astragali Melilotoidis (Herba Astragali Sinici), the crops such as pulse family provide photosynthate to be the energy of nitrogen fixation to root nodule bacterium, and the nitrogen that the root nodule bacterium in root nodule are fixed is transported to object from nodule cell, for plant provides nutrition, form the Symbiotic association of Soybean and Other Crops and root nodule bacterium, production-increasing function is played to leguminous crop and root crop.The infectivity of root nodule bacterium, specificity and validity are had nothing in common with each other.Infectivity is bred wherein and generates root nodule after referring to that root nodule bacterium invade leguminous plants root.
Specificity refer to root nodule bacterium immerse leguminous plants root system time be have certain scope.As rhizobium leguminosarum only infect pea, broad bean, sweet potato, mountain smoke beans, Lens culinaris, garbanzo.Cowpea rhizobium infects cowpea, peanut, mung bean, red bean, pig urine beans, Stem or leaf of Shrub Lespedeza and soybean.Rhizobium of Milk Vetch is high to astragali, Astragalus and prairie milk vetch infection rate.Rihizobium japonicum infection, Glycine and wild soybean crop.
Validity refers to that can root nodule bacterium carry out nitrogen fixation.The root nodule with nitrogen fixation claims effective root nodule, though some bacterial strain can form root nodule, can not fixed nitrogen, and this root nodule is called ineffetive nodulation.Effective root nodule volume is general larger, inner pinkiness, and ineffetive nodulation is smaller, inner in shallow white or light green.
Root nodule strain excellent is functional very strong, and root nodule fixes 75-100 kilogram of nitrogen in soybean per hectare whole breeding time, can provide the 50%-66% of soybean nitrogen requirement throughout one's life.Nodule nitrogen fixation amount and soil, soybean varieties, fertilizer and water condition are relevant.Black earth root nodule bacterium amount of nitrogen fixation is per hectare 105 kg, can meet soybean to 62% of nitrogen demand.Atteration per hectare nodule nitrogen fixation amount is 115.5 kilograms, can meet soybean to 72% of nitrogen demand.Grouan soil per hectare nodule nitrogen fixation amount is 139.5 kilograms, can meet soybean to 68% of nitrogen demand.
The nitrogen-fixing efficiency of Symbiotic association is high more than the nitrogen-fixing efficiency of spontaneous fixed nitrogen system, and reason is syntaxial system is an entirety on physiology, is conducive to the carrying out of nitrogen fixation.
In microbial fertilizer, comparatively early, wherein soybean, peanut, Herba Astragali Melilotoidis (Herba Astragali Sinici) and leguminous forage vaccination area are maximum, and effect of increasing production is obvious in rhizobia fertilizer application.Use Rhizobium of Milk Vetch in the area of not planting Herba Astragali Melilotoidis (Herba Astragali Sinici), grass yield can be doubled and redoubled; Soybean inoculation root nodule bacterium per hectare volume increase 225-300 kilogram; Peanut Rhizobium Inoculation volume increase 10%-50%, the nitrogen that leguminous crop obtains from root nodule bacterium is the 30%-80% of required nitrogen in its all one's life.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of root nodule bacterium fertilizer, especially provides a kind of symbiotic nitrogen fixation high density compound nitragin and preparation method thereof.
Technical problem to be solved by this invention is achieved through the following technical solutions:
A kind of symbiotic nitrogen fixation high density compound nitragin is the microbial inoculum complex liquid be prepared from by following four bacterial classification microbial inoculums; Described four bacterial classification microbial inoculums are following parts by weight of component:
Pea root nodule microbial inoculum 10-30; Cowpea rhizobium agent 10-30; Astragalus Nodules microbial inoculum 40-50; Slow-growing soybean rhizobia agent 10-30;
Described pea root nodule microbial inoculum is Rhizobiumleguminosarum (Frank1879) Frank1889 rhizobium leguminosarum ACCC-16104 ← the Chinese Academy of Agricultural Sciences clay fertilizer institute Liu Hui qin, Ningguo praises and adopted knurl separation from herding in 1986, former numbering CP8636, with pea, broad bean, sweet potato symbiotic nitrogen fixation; 28 ~ 30 DEG C, cultivate and obtain;
Described cowpea rhizobium agent is Bradyrhizobiumvigna cowpea rhizobium ACCC14010 ← clay fertilizer institute of Chinese Academy of Agricultural Sciences separation 1010 from mung bean root nodule, with peanut symbiotic nitrogen fixation; Substratum: 53; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains;
Described Astragalus Nodules microbial inoculum is Rhizobiumastragula Rhizobium of Milk Vetch ACCC13004 ← China Agriculture Academe Fertilizer Institute ← Institute of Micro-biology of Zhejiang academy of agricultural sciences; Original number: 38D; With Herba Astragali Melilotoidis (Herba Astragali Sinici) symbiotic nitrogen fixation nitrogenous fertilizer; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains;
Described Slow-growing soybean rhizobia is Bradyrhizobiumjaponicum (Kirchner) Jordan1982 Slow-growing soybean rhizobia ACCC15005 ← Chinese Academy of Agricultural Sciences clay fertilizer 2005 ← U.S. root nodule bacterium company (NitrogenCompany-Inc) 3I1b39; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains; This ratio is conducive to various bacterium and occupies equilibrium amount at finished product, improves the broad spectrum effect of this microbial inoculum to crop pest kind.
Described symbiotic nitrogen fixation high density compound nitragin, the preferred following parts by weight of component of described four bacterial classification microbial inoculums: pea root nodule microbial inoculum 20; Cowpea rhizobium agent 20; Astragalus Nodules microbial inoculum 30; Slow-growing soybean rhizobia agent 20.
Present invention also offers a kind of preparation method of described symbiotic nitrogen fixation high density compound nitragin, comprise the following steps:
(1) respectively with nutrition dross medium preparing pea root nodule microbial inoculum, cowpea rhizobium agent, Astragalus Nodules microbial inoculum account for, Slow-growing soybean rhizobia agent;
Described nutrition dross substratum is: potassium primary phosphate (KH
2pO
4) 0.22mg, Repone K (KCl) 140mg, magnesium sulfate (MgSO
47H
2o) 70mg, zinc sulfate (ZnSO
47H
2o) 0.25mg, copper sulfate (CuSO
45H
2o) 0.25mg, boric acid (H
3bO
5) solution 0.25mg, Sodium orthomolybdate (Ma
2moO
42H
2o) 0.05mg, ironic citrate (FeC
6h
5o
2) 50mg, nitric acid is received (NaNO
3) 60mg, agar 5-10g, water 1000mL, pH7.0.
(2) pea root nodule microbial inoculum, cowpea rhizobium agent, Astragalus Nodules microbial inoculum, Slow-growing soybean rhizobia agent are mixed by aforementioned proportion, obtain mixed solution;
(3) described mixed solution carries out three grades of enlarged culturing; Described three grades of enlarged culturing comprise shaking table liquid culture, seeding tank liquid culture, fermented liq tank enlarged culturing; Described three grades of inoculum sizes are 10%-20%; Bacterium number reaches the high density finished product of 40-50 hundred million/ml.
Described fermentation is aerobic fermentation, and in whole fermenting process, ventilation and stirring are the important method improving aerobic microbial bacteria number.So sterile air filters and plenum system is key prepared by high density nitragin, preferred fermentation condition: the clean air air flow 0.4-0.8L (m of sterilizing
3min); Stirring velocity 120-200r/min; Envrionment temperature 28-30 DEG C of degree; Cultivate 72-96 hour, compound concentrates finished product microbial inoculum qualified product.
(4) microscopy 5,000,000,000/ml; The compound of obtained enlarged culturing concentrates finished product microbial inoculum.
Present invention also offers a kind of described symbiotic nitrogen fixation high density compound root nodule solid fungicide, comprise the symbol composite solid preparation that following parts by weight of component is prepared from: described compound concentrates microbial inoculum 2-4, peat composed of rotten mosses solids 0.5-1.5.Preferred ingredient weight part: described compound concentrates microbial inoculum 3, peat composed of rotten mosses solids 1.
Present invention also offers a kind of preparation method of described symbiotic nitrogen fixation high density compound root nodule solid fungicide, comprise the following steps:
Mixed with peat composed of rotten mosses solids by described liquid concentrated formula, absorption, makes solid articles;
Described peat composed of rotten mosses water content is less than 10%; Described peat composed of rotten mosses fineness is greater than 80 orders;
The described peat composed of rotten mosses and microbial inoculum parts by weight of component are (2-4): 1, preferably 3: 1; After mixing, rapid stirring is even,
Described solid articles water content 30%-35%.
In order to improve the effect of increasing production of root nodule bacterium, preferably adding ammonium molybdate weight part 0.025-0.075, preferably adding weight part 0.05.
Beneficial effect of the present invention:
1. the root nodule bacterial classification of described four kinds of high infection rates that the present invention is selected, they are extremely strong to leguminous crop root system infectivity such as soybean, peanut, cowpea, Herba Astragali Melilotoidis (Herba Astragali Sinici), pea, broad beans, and dross rate reaches 89%; Through being combined with organic fertilizer, 15-30% being reached to radicant effect of increasing production such as Ipomoea batatas, potato, ginger, garlics, improving and infecting object and infection intensity, solving technical problem of the present invention.
2. farm crop are soaked seed with composite bacteria liquid of the present invention, dress seed, process seed manure, respond well.Particularly make coating agent for seed, better effects if.Generally can make the grain and oil crop volume increase 15-30% such as peanut, soybean, Ipomoea batatas, potato, and can make the cash crop volume increase 20-40% such as ginger, garlic, pachyrhizus, root medicinal material, realization such as drops into little, the effect of high efficiency.
specific implementation method
Embodiment 1
A kind of symbiotic nitrogen fixation high density compound nitragin is the microbial inoculum complex liquid (KG) be prepared from by following four bacterial classification microbial inoculums:
Pea root nodule microbial inoculum 10; Cowpea rhizobium agent 10; Astragalus Nodules microbial inoculum 40; Slow-growing soybean rhizobia agent 10;
Described pea root nodule microbial inoculum is Rhizobiumleguminosarum (Frank1879) Frank1889 rhizobium leguminosarum ACCC-16104 ← the Chinese Academy of Agricultural Sciences clay fertilizer institute Liu Hui qin, Ningguo praises and adopted knurl separation from herding in 1986, former numbering CP8636, with pea, broad bean, sweet potato symbiotic nitrogen fixation; 28 ~ 30 DEG C, cultivate and obtain;
Described cowpea rhizobium agent is separated, with peanut symbiotic nitrogen fixation for Bradyrhizobiumvigna cowpea rhizobium ACCC14010 ← Chinese Academy of Agricultural Sciences's clay fertilizer from mung bean root nodule; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains;
Described Astragalus Nodules microbial inoculum is Rhizobiumastragula Rhizobium of Milk Vetch ACCC13004 ← China Agriculture Academe Fertilizer Institute ← Institute of Micro-biology of Zhejiang academy of agricultural sciences; Original number: 38D; With Herba Astragali Melilotoidis (Herba Astragali Sinici) symbiotic nitrogen fixation nitrogenous fertilizer; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains;
Described Slow-growing soybean rhizobia is Bradyrhizobiumjaponicum (Kirchner) Jordan1982 Slow-growing soybean rhizobia ACCC15005 ← Chinese Academy of Agricultural Sciences clay fertilizer 2005 ← U.S. root nodule bacterium company (NitrogenCompany-Inc) 3I1b39; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains; This ratio is conducive to various bacterium and occupies equilibrium amount at finished product, improves the broad spectrum effect of this microbial inoculum to crop pest kind.
Described symbiotic nitrogen fixation high density compound nitragin, described four bacterial classification microbial inoculum components (KG): pea root nodule microbial inoculum 20; Cowpea rhizobium agent 20; Astragalus Nodules microbial inoculum 30; Slow-growing soybean rhizobia agent 20.
Preparation method:
(1) respectively with nutrition dross medium preparing pea root nodule microbial inoculum, cowpea rhizobium agent, Astragalus Nodules microbial inoculum account for, Slow-growing soybean rhizobia agent;
Described nutrition dross substratum is: potassium primary phosphate (KH
2pO
4) 0.22mg, Repone K (KCl) 140mg, magnesium sulfate (MgSO
47H
2o) 70mg, zinc sulfate (ZnSO
47H
2o) 0.25mg, copper sulfate (CuSO
45H
2o) 0.25mg, boric acid (H
3bO
5) solution 0.25mg, Sodium orthomolybdate (Ma
2moO
42H
2o) 0.05mg, ironic citrate (FeC
6h
5o
2) 50mg, nitric acid is received (NaNO
3) 60mg, agar 5-10g, water 1000mL, pH7.0.
(2) pea root nodule microbial inoculum, cowpea rhizobium agent, Astragalus Nodules microbial inoculum, Slow-growing soybean rhizobia agent are mixed by aforementioned proportion, obtain mixed solution;
(3) described mixed solution carries out three grades of enlarged culturing; Described three grades of enlarged culturing comprise shaking table liquid culture, seeding tank liquid culture, fermented liq tank enlarged culturing; Described three grades of inoculum sizes are 10%; Bacterium number reaches the high density finished product of 4,000,000,000/ml.
Described fermentation is aerobic fermentation, and in whole fermenting process, ventilation and stirring are the important method improving aerobic microbial bacteria number.So sterile air filters and plenum system is key prepared by high density nitragin, preferred fermentation condition: the clean air air flow 0.4L (m of sterilizing
3min); Stirring velocity 120r/min; Envrionment temperature 28 DEG C degree; Cultivate 72 hours, compound concentrates finished product microbial inoculum qualified product.
(4) microscopy, the compound of obtained enlarged culturing concentrates finished product microbial inoculum.
The present embodiment additionally provides described symbiotic nitrogen fixation high density compound root nodule solid fungicide, following component preparation (KG): described compound concentrates microbial inoculum 2, peat composed of rotten mosses solids 0.5; Described compound concentrates microbial inoculum 3, peat composed of rotten mosses solids 1.
The preparation method of described symbiotic nitrogen fixation high density compound root nodule solid fungicide:
Mixed with peat composed of rotten mosses solids by described liquid concentrated formula, absorption, makes solid articles; Described peat composed of rotten mosses water content is less than 10%; Described peat composed of rotten mosses fineness is greater than 80 orders; The described peat composed of rotten mosses and microbial inoculum parts by weight of component are 2: 1; After mixing, rapid stirring is even, described solid articles water content 30%%.
In order to improve the effect of increasing production of root nodule bacterium, preferably add ammonium molybdate 0.025KG.
The successful of the present embodiment root nodule bacterium high-concentration bacterial liquid in seed soaking, and application method is simple and easy, suitable popularization.With 30% bacterium liquid of grain weight, protect seed and close seed soaking 4-8 hour, sowed by seed airing, general effect of increasing production is about 15%.In the application must with leaching with, the seed after having soaked generally was sown into soil in 12 hours, otherwise will affect percentage of germination.This bacterium validity period more than 6 months.
Embodiment 2
A kind of symbiotic nitrogen fixation high density compound nitragin is the microbial inoculum complex liquid (KG) be prepared from by following four bacterial classification microbial inoculums:
Pea root nodule microbial inoculum 130; Cowpea rhizobium agent 30; Astragalus Nodules microbial inoculum 50; Slow-growing soybean rhizobia agent 30;
Described pea root nodule microbial inoculum is Rhizobiumleguminosarum (Frank1879) Frank1889 rhizobium leguminosarum ACCC-16104 ← the Chinese Academy of Agricultural Sciences clay fertilizer institute Liu Hui qin, Ningguo praises and adopted knurl separation from herding in 1986, former numbering CP8636, with pea, broad bean, sweet potato symbiotic nitrogen fixation; 28 ~ 30 DEG C, cultivate and obtain;
Described cowpea rhizobium agent is separated, with peanut symbiotic nitrogen fixation for Bradyrhizobiumvigna cowpea rhizobium ACCC14010 ← Chinese Academy of Agricultural Sciences's clay fertilizer from mung bean root nodule; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains;
Described Astragalus Nodules microbial inoculum is Rhizobiumastragula Rhizobium of Milk Vetch ACCC13004 ← China Agriculture Academe Fertilizer Institute ← Institute of Micro-biology of Zhejiang academy of agricultural sciences; Original number: 38D; With Herba Astragali Melilotoidis (Herba Astragali Sinici) symbiotic nitrogen fixation nitrogenous fertilizer; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains;
Described Slow-growing soybean rhizobia is Bradyrhizobiumjaponicum (Kirchner) Jordan1982 Slow-growing soybean rhizobia ACCC15005 ← Chinese Academy of Agricultural Sciences clay fertilizer 2005 ← U.S. root nodule bacterium company (NitrogenCompany-Inc) 3I1b39; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains; This ratio is conducive to various bacterium and occupies equilibrium amount at finished product, improves the broad spectrum effect of this microbial inoculum to crop pest kind.
Described symbiotic nitrogen fixation high density compound nitragin, the preferred following parts by weight of component of described four bacterial classification microbial inoculums: pea root nodule microbial inoculum 20; Cowpea rhizobium agent 20; Astragalus Nodules microbial inoculum 30; Slow-growing soybean rhizobia agent 20.
The preparation method of described symbiotic nitrogen fixation high density compound nitragin:
(1) respectively with nutrition dross medium preparing pea root nodule microbial inoculum, cowpea rhizobium agent, Astragalus Nodules microbial inoculum account for, Slow-growing soybean rhizobia agent;
Described nutrition dross substratum is: potassium primary phosphate (KH
2pO
4) 0.22m
g, Repone K (KCl) 140mg, magnesium sulfate (MgSO
47H
2o) 70mg, zinc sulfate (ZnSO
47H
2o) 0.25mg, copper sulfate (CuSO
45H
2o) 0.25mg, boric acid (H
3bO
5) solution 0.25mg, Sodium orthomolybdate (Ma
2moO
42H
2o) 0.05mg, ironic citrate (FeC
6h
5o
2) 50mg, nitric acid is received (NaNO
3) 60mg, agar 5-10g, water 1000mL, pH7.0.
(2) pea root nodule microbial inoculum, cowpea rhizobium agent, Astragalus Nodules microbial inoculum, Slow-growing soybean rhizobia agent are mixed by aforementioned proportion, obtain mixed solution;
(3) described mixed solution carries out three grades of enlarged culturing; Described three grades of enlarged culturing comprise shaking table liquid culture, seeding tank liquid culture, fermented liq tank enlarged culturing; Described three grades of inoculum sizes are 20%; Bacterium number reaches the high density finished product of 5,000,000,000/ml.
Described fermentation is aerobic fermentation, and in whole fermenting process, ventilation and stirring are the important method improving aerobic microbial bacteria number.So sterile air filters and plenum system is key prepared by high density nitragin, preferred fermentation condition: the clean air air flow 0.8L (m of sterilizing
3min); Stirring velocity 200r/min; Envrionment temperature 30 DEG C degree; Cultivate 96 hours, compound concentrates finished product microbial inoculum qualified product.
(4) microscopy 5,000,000,000/ml; The compound of obtained enlarged culturing concentrates finished product microbial inoculum.
The present embodiment additionally provides a kind of described symbiotic nitrogen fixation high density compound root nodule solid fungicide, comprises following component preparation (KG): described compound concentrates microbial inoculum 4, peat composed of rotten mosses solids 1.5.Preferred ingredient weight part: described compound concentrates microbial inoculum 3, peat composed of rotten mosses solids 1.
The preparation method of described symbiotic nitrogen fixation high density compound root nodule solid fungicide: described liquid concentrated formula is mixed with peat composed of rotten mosses solids, absorption, makes solid articles; Described peat composed of rotten mosses water content is less than 10%; Described peat composed of rotten mosses fineness is greater than 80 orders; The described peat composed of rotten mosses and microbial inoculum parts by weight of component are 4: 1; After mixing, rapid stirring is even, described solid articles water content 35%.
In order to improve the effect of increasing production of root nodule bacterium, add ammonium molybdate part 0.075KG.
The present embodiment high concentration liquid microbial inoculum and solid bacterium powder also can be used for the Dressing to seed, make increasing crop yield effect reach 10%-15%.Seed dressing method is: with grain weight 8%-10% solid fungicide, add a small amount of clear water, stir, seed and microbial inoculum are fully mixed, allow every seed speckle with microbial inoculum, sow after waiting airing.The seed mixed answers sowing in time, is sown into soil and is advisable, can not affect percentage of germination in general 2-3 days.Liquid seed dressing method is identical with Solid Method.
Example example 3
A kind of symbiotic nitrogen fixation high density compound nitragin is the microbial inoculum complex liquid (KG) be prepared from by following four bacterial classification microbial inoculums:
Pea root nodule microbial inoculum 20; Cowpea rhizobium agent 20; Astragalus Nodules microbial inoculum 45; Slow-growing soybean rhizobia agent 20;
Described pea root nodule microbial inoculum is Rhizobiumleguminosarum (Frank1879) Frank1889 rhizobium leguminosarum ACCC-16104 ← the Chinese Academy of Agricultural Sciences clay fertilizer institute Liu Hui qin, Ningguo praises and adopted knurl separation from herding in 1986, former numbering CP8636, with pea, broad bean, sweet potato symbiotic nitrogen fixation; 28 ~ 30 DEG C, cultivate and obtain;
Described cowpea rhizobium agent is separated, with peanut symbiotic nitrogen fixation for Bradyrhizobiumvigna cowpea rhizobium ACCC14010 ← Chinese Academy of Agricultural Sciences's clay fertilizer from mung bean root nodule; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains;
Described Astragalus Nodules microbial inoculum is Rhizobiumastragula Rhizobium of Milk Vetch ACCC13004 ← China Agriculture Academe Fertilizer Institute ← Institute of Micro-biology of Zhejiang academy of agricultural sciences; Original number: 38D; With Herba Astragali Melilotoidis (Herba Astragali Sinici) symbiotic nitrogen fixation nitrogenous fertilizer; Substratum: 54, culture temperature: 28 ~ 30 DEG C, cultivates and obtains;
Described Slow-growing soybean rhizobia is Bradyrhizobiumjaponicum (Kirchner) Jordan1982 Slow-growing soybean rhizobia ACCC15005 ← Chinese Academy of Agricultural Sciences clay fertilizer 2005 ← U.S. root nodule bacterium company (NitrogenCompany-Inc) 3I1b39; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains; This ratio is conducive to various bacterium and occupies equilibrium amount at finished product, improves the broad spectrum effect of this microbial inoculum to crop pest kind.
Described symbiotic nitrogen fixation high density compound nitragin, described four bacterial classification microbial inoculum components preparation (KG): pea root nodule microbial inoculum 20; Cowpea rhizobium agent 20; Astragalus Nodules microbial inoculum 30; Slow-growing soybean rhizobia agent 20.
The preparation method of described symbiotic nitrogen fixation high density compound nitragin:
(1) nutrition dross medium preparing pea root nodule microbial inoculum, cowpea rhizobium agent, Astragalus Nodules microbial inoculum, Slow-growing soybean rhizobia agent is used respectively;
Described nutrition dross substratum is that a kind of promotion dross is fast, and a kind of improved culture medium that nodule number is many, composition is: potassium primary phosphate (KH
2pO
4) 0.22mg, Repone K (KCl) 140mg, magnesium sulfate (MgSO
47H
2o) 70mg, zinc sulfate (ZnSO
47H
2o) 0.25mg, copper sulfate (CuSO
45H
2o) 0.25mg, boric acid (H
3bO
5) solution 0.25mg, Sodium orthomolybdate (Ma
2moO
42H
2o) 0.05mg, ironic citrate (FeC
6h
5o
2) 50mg, nitric acid is received (NaNO
3) 60mg, agar 5-10g, water 1000mL, pH7.0.
(2) pea root nodule microbial inoculum, cowpea rhizobium agent, Astragalus Nodules microbial inoculum are accounted for by aforementioned proportion, Slow-growing soybean rhizobia agent mixing, obtain mixed solution;
(3) described mixed solution carries out three grades of enlarged culturing; Described three grades of enlarged culturing comprise shaking table liquid culture, seeding tank liquid culture, fermented liq tank enlarged culturing; Described three grades of inoculum sizes are 15%; Bacterium number reaches the high density finished product of 4,500,000,000/ml.
Described fermentation is aerobic fermentation, and in whole fermenting process, ventilation and stirring are the important method improving aerobic microbial bacteria number.So sterile air filters and plenum system is key prepared by high density nitragin, preferred fermentation condition: the clean air air flow 0.6L (m of sterilizing
3min); Stirring velocity 160r/min; Envrionment temperature 26 DEG C degree; Cultivate 80 hours, compound concentrates finished product microbial inoculum qualified product.
(4) microscopy 5,000,000,000/ml; The compound of obtained enlarged culturing concentrates finished product microbial inoculum.
The present embodiment also provides a kind of symbiotic nitrogen fixation high density compound root nodule solid fungicide, prepares (KG): described compound concentrates microbial inoculum 3, peat composed of rotten mosses solids 1 by following component.
The preparation method of described symbiotic nitrogen fixation high density compound root nodule solid fungicide, comprises the following steps: mixed with peat composed of rotten mosses solids by described liquid concentrated formula, and absorption, makes solid articles; Described peat composed of rotten mosses water content is less than 10%; Described peat composed of rotten mosses fineness is greater than 80 orders; The described peat composed of rotten mosses and microbial inoculum part by weight are 3: 1; After mixing, rapid stirring is even, described solid articles water content 30%.
In order to improve the effect of increasing production of root nodule bacterium, preferably add ammonium molybdate 0.05KG.
The present embodiment nitragin fertilizer obtains raising with bio-feritlizer mixing application effect.Method be by bio-feritlizer with account for gross weight 20% high-concentration bacterial liquid or solid bacterium powder mixes, after mixing with bio-feritlizer, for as seed manure, row replacement, to spread manuer in holes as base fertilizer and seed manure, the percentage of germination of seed can be made to improve about 30%.Also can promote the increment that farm crop root growth doubles, can crop yield 15-25% be impelled.The bio-feritlizer quality used should on be, the organic content of bio-feritlizer should more than 50% compounding application better.Organic content is lower, and application effect is poor.In bio-feritlizer, chemical element fertilizer content accounts for about 8% of total amount and is advisable, and chemical element content is higher, impacts nitragin activity, reduces the useful effect of microbial inoculum.
Claims (9)
1. a symbiotic nitrogen fixation high density compound nitragin is the microbial inoculum complex liquid be prepared from by following less than four parts by weight of component bacterial classification microbial inoculums:
Pea root nodule microbial inoculum 10-30; Cowpea rhizobium agent 10-30; Astragalus Nodules microbial inoculum 40-50; Slow-growing soybean rhizobia agent 10-30;
Described pea root nodule microbial inoculum is Rhizobiumleguminosarum (Frank1879) Frank1889 rhizobium leguminosarum ACCC-16104 ← the Chinese Academy of Agricultural Sciences clay fertilizer institute Liu Hui qin, Ningguo praises and adopted knurl separation from herding in 1986, former numbering CP8636, with pea, broad bean, sweet potato symbiotic nitrogen fixation; 28 ~ 30 DEG C, cultivate and obtain;
Described cowpea rhizobium agent is separated, with peanut symbiotic nitrogen fixation for Bradyrhizobiumvigna cowpea rhizobium ACCC14010 ← Chinese Academy of Agricultural Sciences's clay fertilizer from mung bean root nodule; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains;
Described Astragalus Nodules microbial inoculum is Rhizobiumastragula Rhizobium of Milk Vetch ACCC13004 ← China Agriculture Academe Fertilizer Institute ← Institute of Micro-biology of Zhejiang academy of agricultural sciences; Original number: 38D; With Herba Astragali Melilotoidis (Herba Astragali Sinici) symbiotic nitrogen fixation nitrogenous fertilizer; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains;
Described Slow-growing soybean rhizobia is Bradyrhizobiumjaponicum (Kirchner) Jordan1982 Slow-growing soybean rhizobia ACCC15005 ← Chinese Academy of Agricultural Sciences clay fertilizer 2005 ← U.S. root nodule bacterium company (NitrogenCompany-Inc) 3I1b39; Culture temperature: 28 ~ 30 DEG C, cultivates and obtains.
2. symbiotic nitrogen fixation high density compound nitragin according to claim 1, described four bacterial classification microbial inoculums are following parts by weight of component: pea root nodule microbial inoculum 20; Cowpea rhizobium agent 20; Astragalus Nodules microbial inoculum 30; Slow-growing soybean rhizobia agent 20.
Comprise the following steps:
(1) respectively with nutrition dross medium preparing pea root nodule microbial inoculum, cowpea rhizobium agent, Astragalus Nodules microbial inoculum account for, Slow-growing soybean rhizobia agent;
(2) pea root nodule microbial inoculum, cowpea rhizobium agent, Astragalus Nodules microbial inoculum, Slow-growing soybean rhizobia agent are mixed by aforementioned proportion, obtain mixed solution;
(3) described mixed solution carries out three grades of enlarged culturing; Described three grades of enlarged culturing comprise shaking table liquid culture, seeding tank liquid culture, fermented liq tank enlarged culturing; Bacterium number reaches the high density finished product of 40-50 hundred million/ml.
3. the preparation method of symbiotic nitrogen fixation high density compound nitragin according to claim 2, described nutrition dross substratum is: potassium primary phosphate (KH
2pO
4) 0.22mg, Repone K (KCl) 140mg, magnesium sulfate (MgSO
47H
2o) 70mg, zinc sulfate (ZnSO
47H
2o) 0.25mg, copper sulfate (CuSO
45H
2o) 0.25mg, boric acid (H
3bO
5) solution 0.25mg, Sodium orthomolybdate (Ma
2moO
42H
2o) 0.05mg, ironic citrate (FeC
6h
5o
2) 50mg, nitric acid is received (NaNO
3) 60mg, agar 5-10g, water 1000mL, pH7.0.
4. the preparation method of symbiotic nitrogen fixation high density compound nitragin according to claim 2, described fermentation is aerobic fermentation, fermentation condition: the clean air air flow 0.4-0.8L (m of sterilizing
3min); Stirring velocity 120-200r/min; Envrionment temperature 28-30 DEG C of degree; Cultivate 72-96 hour, obtain compound and concentrate finished product microbial inoculum.
5. according to the symbiotic nitrogen fixation preparation method of high density compound nitragin of claim 1 or 4, also comprise microscopy 40-50 hundred million/more than ml, the compound of obtained enlarged culturing concentrates finished product microbial inoculum.
6. a symbiotic nitrogen fixation high density compound root nodule solid fungicide of one of claim 1-2, comprises the composite solid preparation that following parts by weight of component is prepared from: described compound concentrates microbial inoculum 2-4, peat composed of rotten mosses solids 0.5-1.5.
7. symbiotic nitrogen fixation high density compound root nodule solid fungicide according to claim 6, described ingredients weight parts is: described compound concentrates microbial inoculum 3, peat composed of rotten mosses solids 1.
8. the symbiotic nitrogen fixation preparation method of high density compound root nodule solid fungicide of claim 6 or 7, comprises the following steps: mixed with peat composed of rotten mosses solids by described liquid concentrated formula, and absorption, makes solid articles; Described peat composed of rotten mosses water content is less than 10%; Described peat composed of rotten mosses fineness is greater than 80 orders; The described peat composed of rotten mosses and microbial inoculum parts by weight of component are (2-4): 1, and after mixing, rapid stirring is even, described solid articles water content 30%-35%.
9. the preparation method stating component symbiotic nitrogen fixation high density compound root nodule solid fungicide according to claim 8, adds ammonium molybdate weight part 0.025-0.075.
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| CN106754484A (en) * | 2016-11-28 | 2017-05-31 | 中国农业科学院农业资源与农业区划研究所 | One plant of rhizobium leguminosarum and its fermentation culture method and application |
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| CN110283751A (en) * | 2016-11-28 | 2019-09-27 | 中国农业科学院农业资源与农业区划研究所 | The microbial inoculum and its bacterial strain uses therefor that promote leguminous plant to grow and application |
| CN110283751B (en) * | 2016-11-28 | 2020-08-25 | 中国农业科学院农业资源与农业区划研究所 | Microbial inoculum for promoting growth of leguminous plants and strain and application thereof |
| CN107164261A (en) * | 2017-05-09 | 2017-09-15 | 中国农业科学院农业资源与农业区划研究所 | Rhizobium and its application that one plant of promotion villose vetch increases |
| CN107164261B (en) * | 2017-05-09 | 2019-06-11 | 中国农业科学院农业资源与农业区划研究所 | One plant of rhizobium for promoting villose vetch to increase and its application |
| CN108782092A (en) * | 2018-05-22 | 2018-11-13 | 云南农业大学 | A kind of intercropping culture method of control broad bean continuous cropping obstacle |
| CN108795813A (en) * | 2018-06-25 | 2018-11-13 | 河北省农林科学院农业资源环境研究所 | A kind of composition and preparation method thereof promoting legume nodulation fixed nitrogen |
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