CN107529530B - Preparation method and application of special fertilizer for planting astragalus membranaceus - Google Patents
Preparation method and application of special fertilizer for planting astragalus membranaceus Download PDFInfo
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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Abstract
The invention discloses a preparation method and application of a special fertilizer for planting astragalus membranaceus, wherein the preparation method of the special fertilizer for planting astragalus membranaceus comprises the following steps: s1, preparing an elicitor solution, preparing a solution from SNP by using distilled water, S2, fermenting green manure, S3, preparing a nitrogen-fixing bacterial liquid, respectively culturing nitrogen-fixing strains T16 and T21 to prepare a bacterial liquid, and S4, mixing the green manure, the elicitor solution and the nitrogen-fixing bacterial liquid to prepare the solid fertilizer. The stem and leaf of the astragalus are fermented and then mixed with three elicitors and two azotobacter to prepare the solid fertilizer, and after fertilization treatment, the dry weight of the astragalus, the accumulation amount of flavone in the astragalus root, the accumulation amount of saponin in the astragalus root and the accumulation amount of polysaccharide in the astragalus root are all obviously improved. Moreover, the special fertilizer can improve the nutrient content of soil, so that the contents of organic matters, total nitrogen, phosphorus and potassium and effective nitrogen, phosphorus and potassium in the soil are all improved.
Description
Technical Field
The invention relates to the technical field of astragalus cultivation, in particular to a preparation method and application of a special fertilizer for astragalus planting.
Background
Astragalus membranaceus is a plant used as both medicine and food, and the medicinal use of astragalus membranaceus has been in more than 2000 years so far. Astragalus is rich in various bioactive components, of which the most important and studied are flavone, saponin and polysaccharide. The dietetic or medicinal astragalus root can strengthen heart, improve immunity, and has the functions of resisting aging, etc., and is widely applied to cardiovascular diseases. Astragalus membranaceus in Shanxi province formally becomes a Chinese medicinal material in 2004, and the best known astragalus membranaceus is produced from the same muddy source. The muddy source astragalus has high topography, cool climate, rich sunshine, loose soil, rich nutrient elements and unique ecological environment, and provides a good environment for the growth and development of the astragalus, so the muddy source astragalus is not in demand in the medicinal material market. However, the planting mode is backward, more plants depend on the natural culture, the management is incomplete, the planting degradation is caused, the effective components are unstable, the difference is large, the yield is low, and the economic benefit is directly influenced.
The early-stage research result of a laboratory shows that the water content of the soil is 60-65 percent which is most suitable for the growth of the astragalus and the accumulation of the effective components; the synthesis and accumulation of flavonoids under yellow light are increased, saponin substances under blue light are more accumulated, and the polysaccharide substances are more suitable for synthesis under natural light. These tests create a growth adversity for astragalus root, excite some signal molecules in astragalus root, stimulate the secondary metabolism path in astragalus root and promote the synthesis and accumulation of main medicinal components, but these adversity stimulations are only suitable for small-area artificial planting, and have great limitation for astragalus root planted under large-area wild or semi-wild conditions, so we choose to use elicitors as substitutes for these adversity stimulations because elicitors can also excite signal molecules in secondary metabolism path to play the same role. The perennial stem and leaf of the astragalus are prepared into green manure which is mixed with compound fertilizer to effectively promote the growth of the corn; azotobacteria T16 and T21 directly separated from radix astragali rhizosphere have good nitrogen fixation effect.
Based on the test results, the inventor synthesizes the several influence factors in advance to prepare the astragalus fertilizer, and researches the change of the yield, the quality and the soil nutrient of the astragalus under different fertilizer ratios to screen out the optimal fertilizer ratio which ensures the yield and the quality of the astragalus, lays a foundation for perfecting the standardized artificial cultivation technical system of the astragalus mongholicus, and provides reasonable scientific basis.
Disclosure of Invention
The invention solves the technical problems that the existing astragalus planting mode is lagged behind, the management is incomplete, the planting degradation is caused, the effective components are unstable, the difference is large, the yield is low, and the economic benefit is directly influenced, thereby providing the preparation method and the application of the special fertilizer for planting astragalus.
The technical scheme of the invention is as follows:
a preparation method of a fertilizer special for planting astragalus comprises the following steps:
s1, preparing an elicitor solution:
preparing Sodium Nitroprusside (SNP) into a solution with the concentration of 10-25mmol/L by using distilled water for later use;
s2, fermenting green manure:
1) drying fresh stem and leaf of radix astragali at low temperature until water content is lower than 30%, and pulverizing to 0.5-1 cm;
2) pouring the 'leap fruit microbial agent' into warm water at the temperature of 30-35 ℃ according to the weight ratio of 1:50-100, and fully and uniformly mixing to form a microbial liquid for later use;
3) mixing the microbial liquid with the crushed astragalus stem and leaf to control the final water content of the mixture to be 50-65%;
4) fermenting the mixture at 36-40 deg.C for 2-10 days;
s3, preparing nitrogen-fixing bacteria liquid:
1) respectively inoculating the stored nitrogen-fixing strains T16 and T21 into LB (luria-bertani) liquid culture medium, culturing at 28 deg.C and 125r/min for 48h, and allowing them to grow sufficiently to obtain bacterial suspension;
2) putting 150mL of LB liquid culture medium into two 500mL triangular flasks, sterilizing, placing at room temperature for 1-2 d, respectively inoculating 15mL of each bacterial suspension after checking no pollution, and culturing at 28 ℃ and 125r/min for 2-3 d to obtain mother liquor;
3) the mother liquor was diluted to the desired concentration with distilled water: t16 is 105-107Each ml, T21 is 107-109Per ml;
s4, mixing the green manure with an elicitor solution and a nitrogen-fixing bacterium solution to prepare a solid fertilizer:
weighing 1.23-4.9kg of the green manure fermented in the step S2, T1675-225ml of the azotobacteria liquid prepared in the step S3, T2175-225ml of the azotobacteria liquid prepared in the step S1, 150ml of the SA solution prepared in the step S1, 300ml of the SNP solution 120-.
As an improvement, the preparation method of the special fertilizer for planting astragalus comprises the following steps:
s1, preparing an elicitor solution:
preparing Salicylic Acid (SA) into a solution with the concentration of 10-20umol/L by using distilled water, preparing Sodium Nitroprusside (SNP) into a solution with the concentration of 10-25mmol/L by using distilled water, and preparing naphthylacetic acid (NAA) into a solution with the concentration of 1-1.2mg/L by using distilled water for later use;
s2, fermenting green manure:
1) drying fresh stem and leaf of radix astragali at low temperature until water content is lower than 30%, and pulverizing to 0.5-1 cm;
2) pouring the 'leap fruit microbial agent' into warm water at the temperature of 30-35 ℃ according to the weight ratio of 1:50-100, and fully and uniformly mixing to form a microbial liquid for later use;
3) mixing the microbial liquid with the crushed astragalus stem and leaf to control the final water content of the mixture to be 50-65%;
4) fermenting the mixture at 36-40 deg.C for 2-10 days;
s3, preparing nitrogen-fixing bacteria liquid:
1) respectively inoculating the stored nitrogen-fixing strains T16 and T21 into LB (luria-bertani) liquid culture medium, culturing at 28 deg.C and 125r/min for 48h, and allowing them to grow sufficiently to obtain bacterial suspension;
2) putting 150mL of LB liquid culture medium into two 500mL triangular flasks, sterilizing, placing at room temperature for 1-2 d, respectively inoculating 15mL of each bacterial suspension after checking no pollution, and culturing at 28 ℃ and 125r/min for 2-3 d to obtain mother liquor;
3) the mother liquor was diluted to the desired concentration with distilled water: t16 is 105-107Each ml, T21 is 107-109Per ml;
s4, mixing the green manure with an elicitor solution and a nitrogen-fixing bacterium solution to prepare a solid fertilizer:
weighing 1.23-4.9kg of the green manure fermented in the step S2, T1675-225ml of the azotobacteria liquid prepared in the step S3, T2175-225ml of the azotobacteria liquid prepared in the step S1, 150ml of the SA solution prepared in the step S1, 300ml of the SNP solution 120-.
Further, in the above scheme, in the step S3, when the bacterial suspensions are prepared, after the bacterial strains are sufficiently grown, an absorbance value (OD600, optical density) of each bacterial suspension at 600nm is measured by using an ultraviolet spectrophotometer; when the OD value is more than 0.5, the OD values are adjusted to be equal, and then each strain suspension is continuously allowed to grow fully.
Further, in the above embodiment, in the step S3, when the mother solution is prepared, after culturing for 2 to 3 days, the OD value of the fermentation broth is measured, and when the OD value is greater than 0.5 (wavelength 660nm), the mother solution is obtained by injecting the broth into a sterilized glass bottle by a sterilized syringe, and sealing and storing at normal temperature.
As an optimal choice, in the step S3, the mother liquor is diluted to T16 of 106Each ml, T21 is 109Per ml; in the step S4, the dosage of the nitrogen-fixing bacteria liquid T16 is 75ml, and the dosage of T21 is 75 ml.
Further, in the above-mentioned aspect, in the step S4, when the solid fertilizer is prepared, 0.01-0.1% (wt) of the preservative and 2-5% (wt) of the binder are added.
Further, in the above scheme, the preservative is sodium diformate or potassium benzoate.
Further, in the above scheme, the binder is bentonite or clay.
Further, in the scheme, a special device is used in the process of preparing the solid fertilizer, the device comprises a low-temperature drying device, a crushing device, a stirring and mixing device, a three-dimensional layered fermentation device, a dynamic mixing device and a granulating device, when the fertilizer is prepared, fresh astragalus stems and leaves stored in a green manure storage device are conveyed to the low-temperature drying device through a belt type feeding device to be dried at low temperature until the water content is lower than 30%, then the fresh astragalus stems and leaves are lifted to the crushing device through a lifting device to be crushed to 0.5-1cm, the 'leap solid microbial inoculum' is poured into warm water at 30-35 ℃ according to the weight ratio of 1:50-100 to be fully mixed uniformly to form a microbial inoculum, the microbial inoculum is filled into a zymogen liquid storage device, then the crushed astragalus stems and leaves and the microbial inoculum are uniformly mixed in the stirring and mixing device, controlling the final water content of the mixture to 50-65%; feeding the mixture into a three-dimensional layered fermentation device for fermentation, and keeping the fermentation temperature at 36-40 ℃ for 2-10 days by a temperature control device; three-dimensional layering fermenting installation includes casing, layering frame, top cap, base, air-supply line, exhaust pipe, air-blower, exhaust fan, temperature sensor, the layering is put up inside the casing, the top cap is located casing upper portion, the base is located the casing bottom, the air-supply line is located the base, and connect to the air-blower is equipped with a plurality of air intakes on the air-supply line, the air intake accesss to inside the casing, the exhaust pipe is located the top cap, and with the exhaust fan links to each other, is equipped with a plurality of air outlets on the exhaust pipe, inside a plurality of air outlets also accesss to the casing, still is equipped with 3 at least temperature sensor in the layering frame, lies in the upper, middle and lower position of layering frame respectively, temperature sensor is connected to temperature regulating device through the wire. In the fermentation process, on one hand, the temperature can be adjusted through the temperature control device, the temperature control device adjusts according to the temperature information transmitted back by the temperature sensor in real time, on the other hand, the temperature can be adjusted through the air inlet pipeline and the air outlet pipeline, especially, when the temperature is too high, air is directly introduced into the three-dimensional layered fermentation device through the air blower and the air inlet pipeline to cool, and then is discharged through the air outlet pipeline and the air exhaust fan, after the fermentation is finished, cold air can be introduced into the three-dimensional layered fermentation device through the air blower and the air inlet pipeline to cool the fermented green manure, redundant heat is taken away by the air and is discharged through the air outlet pipeline and the air exhaust fan, after the temperature is reduced to a proper temperature, the green manure is conveyed to the dynamic mixing device through the lifting device, and simultaneously the five components are added into the dynamic mixing device according to the proportion through the containers of SA solution, the SNP solution, the NAA solution, the azotobacter solution T16 and, and then the motor drives the dynamic mixing device to work, the dynamic mixing device is uniformly mixed, the mixture is dried by a low-temperature drying device until the water content is lower than 5 percent, and finally the mixture is sent into a granulation tower for granulation to obtain the solid fertilizer.
Furthermore, the 0.01-0.1% (wt) preservative and 2-5% (wt) binder are added into the dynamic mixing device, then the mixture is dried by a low-temperature drying device until the water content is lower than 5%, and finally the mixture is sent into a granulation tower for granulation, so that the solid fertilizer is prepared.
Furthermore, the containers for holding the SA solution, the SNP solution, the NAA solution, the azotobacter solution T16 and the azotobacter solution T21 are a container I, a container II, a container III, a container IV and a container V respectively, each container is connected to a dynamic mixing device through a pipeline, and each pipeline is provided with a mass flowmeter and a distribution valve, so that the dosage of each component can be accurately and quantitatively controlled.
The invention also provides a method for cultivating astragalus by using the special fertilizer for astragalus planting, which comprises the following steps:
s1: soaking radix astragali seed in 98% concentrated sulfuric acid for 3min, washing with running water for 60min until the residual solution of concentrated sulfuric acid is washed, and air drying on filter paper;
s2: after land preparation and ploughing, manually sowing the astragalus seeds treated by the S1, wherein the average plant spacing and the average row spacing of the astragalus are 15cm and 25cm, sowing solid fertilizer, and performing conventional field management until harvesting.
The invention has the beneficial effects that: the stem and leaf of the astragalus are fermented and then mixed with three elicitors and two azotobacter to prepare the solid fertilizer, after fertilization, the dry weight of the astragalus is improved by 50.52 percent, the accumulation amount of flavone in the astragalus root is improved by 74.5 percent, the accumulation amount of saponin in the astragalus root is improved by 63.18 percent, the accumulation amount of polysaccharide in the astragalus root is improved by 30.7 percent, and the yield and the quality of the astragalus are obviously improved. And the nutrient content of the soil can be improved, so that the organic matter in the soil is improved by 7.23%, the total nitrogen phosphorus potassium content is respectively improved by 3.69%, 17.59% and 2.15%, and the effective nitrogen phosphorus potassium content is respectively improved by 8.96%, 33.16% and 10.33%. In addition, the special fertilizer for astragalus disclosed by the invention is simple in preparation method and low in investment cost.
Drawings
FIG. 1 is a schematic view of the construction of an apparatus for preparing the fertilizer of the present invention;
FIG. 2 is a schematic structural view of a three-dimensional layered fermentation apparatus.
Wherein, 1-green manure storage device, 2-belt type feeding device, 3-low temperature drying device, 4-lifting device, 5-crushing device, 6-stirring mixing device, 7-zymophyte liquid storage device, 8-three-dimensional layered fermentation device, 81-layered frame, 82-top cover, 83-base, 84-air inlet pipeline, 841-air inlet, 85-air outlet pipeline, 851-air outlet, 86-blower, 87-air outlet, 88-temperature control device, 89-temperature sensor, 810-lead, 9-dynamic mixing device, 92-motor, 93-container I, 94-container II, 95-container III, 96-container IV, 97-container V, 98-mass flow meter, 99-distribution valve, 10-granulation device.
Detailed Description
The invention will be described in further detail below with reference to the accompanying drawings and specific embodiments.
Example 1:
a preparation method of a fertilizer special for planting astragalus comprises the following steps:
s1, preparing an elicitor solution:
preparing Salicylic Acid (SA) into a solution with the concentration of 10umol/L by using distilled water, preparing Sodium Nitroprusside (SNP) into a solution with the concentration of 10mmol/L by using distilled water, and preparing naphthylacetic acid (NAA) into a solution with the concentration of 1mg/L by using distilled water for later use;
salicylic Acid (SA) -purchased from Tianjin Beichen Fangzheng chemical reagent plant with molecular weight of 138.12, Sodium Nitroprusside (SNP) -purchased from Tianjin Tianli chemical reagent plant with molecular weight of 297.94, naphthylacetic acid (NAA) -purchased from Tianjin Tianda chemical reagent plant with molecular weight of 186.21;
s2, fermenting green manure:
1) collecting fresh stem and leaf of radix astragali from radix astragali of the same turbid radix astragali base, drying at low temperature until the water content is lower than 30%, and pulverizing to 0.5 cm;
2) pouring the bissextile fungus agent into warm water at the temperature of 1:50 by weight ratio, and fully and uniformly mixing to form a microorganism bacterium liquid for later use;
3) mixing the microbial liquid with crushed astragalus stem and leaf to control the final water content of the mixture to be 50%; the judgment standard for whether the water content is proper is to grip a material tightly, and the material is preferably scattered in a scattered manner without dripping when water is in the gaps;
4) fermenting the mixture, keeping the fermentation temperature at 36 ℃ and the fermentation time at 2 days;
s3, preparing nitrogen-fixing bacteria liquid:
1) respectively inoculating the stored nitrogen-fixing strains T16 and T21 into LB (Luria-bertani) liquid culture medium, culturing at 28 deg.C and 125r/min for 48h, and measuring the absorbance value (OD600, optical density) of each strain suspension at 600nm with an ultraviolet spectrophotometer after the strains grow sufficiently; when the OD value is larger than 0.5, adjusting the OD values to be equal, and then continuing to enable each strain suspension to fully grow to obtain a bacterial suspension for later use;
2) putting 150mL LB liquid culture medium into two 500mL triangular flasks, sterilizing, placing at room temperature for 1d, respectively inoculating 15mL of each bacterial suspension after checking no pollution, culturing at 28 ℃ and 125r/min for 2d, measuring the OD value of zymogen liquid, and injecting the liquid into a sterilized glass bottle by using a sterilized injector for normal-temperature sealed storage when the OD value is more than 0.5 (the wavelength is 660nm), thus obtaining the mother solution;
3) the mother liquor was diluted to the desired concentration with distilled water: t16 is 105Each ml, T21 is 107Per ml;
s4, mixing the green manure with an elicitor solution and a nitrogen-fixing bacterium solution to prepare a solid fertilizer:
weighing 1.23kg of the green manure fermented in the step S2, the nitrogen-fixing bacteria liquid T1675ml and T2175ml prepared in the step S3, 150ml of SA solution prepared in the step S1, 120ml of SNP solution and 120ml of NAA solution, mixing, and preparing the solid fertilizer.
The special equipment is used in the process of preparing the solid fertilizer, the special equipment comprises a low-temperature drying device 3, a crushing device 5, a stirring and mixing device 6, a three-dimensional layered fermentation device 8, a dynamic mixing device 9 and a granulating device 10, when preparing the fertilizer, fresh astragalus stems and leaves stored in a green manure storage device 1 are conveyed to a low-temperature drying device 3 through a belt type feeding device 2 to be dried at low temperature until the water content is lower than 30 percent, then lifting to a crushing device 5 through a lifting device 4, crushing to 0.5cm, pouring the intercalated microorganism bacterium agent into warm water at 30 ℃ according to the weight ratio of 1:50, fully and uniformly mixing to form a microorganism bacterium solution, filling the microorganism bacterium solution into a zymocyte liquid storage device 7, then uniformly mixing the crushed stem and leaf of the astragalus with the microbial liquid in a stirring and mixing device 6 to control the final water content of the mixture to be 50 percent; feeding the mixture into a three-dimensional layered fermentation device 8 for fermentation, and keeping the fermentation temperature at 36 ℃ for 2 days through a temperature control device 88; the three-dimensional layered fermentation device 8 comprises a shell, a layered frame 81, a top cover 82, a base 83, an air inlet pipeline 84, an air outlet pipeline 85, a blower 86, an air outlet fan 87 and a temperature sensor 89, the shelf 81 is located inside the housing, the top cover 82 is located on the top of the housing, the base 83 is located on the bottom of the housing, the air inlet duct 84 is located in the base 83, and connected to the blower 86, a plurality of air inlets 841 are provided on the air inlet pipe 84, the air inlets 841 lead to the inside of the housing, the air outlet pipe 85 is positioned in the top cover 82, and is connected with the exhaust fan 87, the exhaust duct 85 is provided with a plurality of exhaust outlets 851, the exhaust outlets 851 are also communicated with the inside of the shell, at least 3 temperature sensors are arranged in the layering rack 81 and are respectively positioned at the upper, middle and lower positions of the layering rack 81, and the temperature sensors 89 are connected to the temperature control device 88 through leads 810. In the fermentation process, on one hand, the temperature can be adjusted through the temperature control device 88, the temperature control device 88 adjusts the temperature in real time according to the temperature information transmitted back by the temperature sensor 89, on the other hand, the temperature can be adjusted through the air inlet pipeline 84 and the air outlet pipeline 85, especially when the temperature is too high, the air is directly introduced into the three-dimensional layered fermentation device 8 through the air blower 86 and the air inlet pipeline 84 to cool, the air is discharged through the air outlet pipeline 85 and the air exhaust fan 87, after the fermentation is finished, the cold air can be introduced into the three-dimensional layered fermentation device 8 through the air blower 86 and the air inlet pipeline 84 to cool the fermented green manure, the redundant heat is taken away by the air and is discharged through the air outlet pipeline 85 and the air exhaust fan 87, after the temperature is reduced to the proper temperature, the green manure is conveyed to the dynamic mixing device 9 through the lifting device 4, the SNP solution, the NAA solution, the azotobacter liquid T16 and the azotobacter liquid T21 are added into the dynamic mixing device 9 according to the proportion, then the dynamic mixing device 9 is driven by a motor 92 to work and is uniformly mixed, then the mixture is dried by the low-temperature drying device 3 until the water content is lower than 5 percent, and finally the mixture is sent into a granulation tower for granulation, so that the solid fertilizer is prepared.
The containers for holding the SA solution, the SNP solution, the NAA solution, the azotobacter solution T16 and the azotobacter solution T21 are a container I93, a container II 94, a container III 95, a container IV 96 and a container V97 respectively, all the containers are connected to the dynamic mixing device 9 through pipelines, and each pipeline is provided with a mass flow meter 98 and a distribution valve 99, so that the dosage of each component can be accurately and quantitatively controlled.
The method for cultivating astragalus membranaceus by using the prepared special fertilizer for planting astragalus membranaceus comprises the following steps:
s1: soaking radix astragali seed in 98% concentrated sulfuric acid for 3min, washing with running water for 60min until the residual solution of concentrated sulfuric acid is washed, and air drying on filter paper;
s2: after land preparation and ploughing, manually sowing the astragalus seeds treated by the S1, wherein the average plant spacing and the average row spacing of the astragalus are 15cm and 25cm, sowing solid fertilizer, and performing conventional field management until harvesting.
Example 2:
a preparation method of a fertilizer special for planting astragalus comprises the following steps:
s1, preparing an elicitor solution:
preparing Salicylic Acid (SA) into a solution with the concentration of 15umol/L by using distilled water, preparing Sodium Nitroprusside (SNP) into a solution with the concentration of 15umol/L by using distilled water, and preparing naphthylacetic acid (NAA) into a solution with the concentration of 1.1mg/L by using distilled water for later use;
salicylic Acid (SA) -purchased from Tianjin Beichen Fangzheng chemical reagent plant with molecular weight of 138.12, Sodium Nitroprusside (SNP) -purchased from Tianjin Tianli chemical reagent plant with molecular weight of 297.94, naphthylacetic acid (NAA) -purchased from Tianjin Tianda chemical reagent plant with molecular weight of 186.21;
s2, fermenting green manure:
1) collecting fresh stem and leaf of radix astragali from radix astragali of the same turbid radix astragali base, drying at low temperature until the water content is lower than 30%, and pulverizing to 0.7 cm;
2) pouring the bissextile fungus agent into warm water at 33 ℃ according to the weight ratio of 1:75, and fully and uniformly mixing to form a microorganism bacterium liquid for later use;
3) mixing the microbial liquid with the crushed astragalus stem and leaf to control the final water content of the mixture to be 58%; the judgment standard for whether the water content is proper is to grip a material tightly, and the material is preferably scattered in a scattered manner without dripping when water is in the gaps;
4) fermenting the mixture, keeping the fermentation temperature at 38 ℃ and the fermentation time at 6 days;
s3, preparing nitrogen-fixing bacteria liquid:
1) respectively inoculating the stored nitrogen-fixing strains T16 and T21 into LB (Luria-bertani) liquid culture medium, culturing at 28 deg.C and 125r/min for 48h, and measuring the absorbance value (OD600, optical density) of each strain suspension at 600nm with an ultraviolet spectrophotometer after the strains grow sufficiently; when the OD value is larger than 0.5, adjusting the OD values to be equal, and then continuing to enable each strain suspension to fully grow to obtain a bacterial suspension for later use;
2) 150mL of LB liquid culture medium is filled into two 500mL triangular flasks, the triangular flasks are placed at room temperature for 1.5d after sterilization, 15mL of each bacterial suspension is respectively inoculated after checking no pollution, the culture is carried out at 28 ℃ and 125r/min for 2.5d, the OD value of the zymogen liquid is measured, and when the OD value is more than 0.5 (the wavelength is 660nm), the zymogen liquid is injected into a sterilized glass bottle by a sterilized injector and is sealed and stored at normal temperature, thus obtaining the mother liquid;
3) the mother liquor was diluted to the desired concentration with distilled water: t16 is 106Each ml, T21 is 108Per ml;
s4, mixing the green manure with an elicitor solution and a nitrogen-fixing bacterium solution to prepare a solid fertilizer:
weighing 2.4kg of the green manure fermented in the step S2, the azotobacter solution T16150ml and T21150ml prepared in the step S3, 225ml of SA solution prepared in the step S1, 210ml of SNP solution and 210ml of NAA solution, mixing, adding 0.05% (wt) of preservative sodium diformate and 3.5% (wt) of binder bentonite, and preparing into a solid fertilizer.
The special equipment is used in the process of preparing the solid fertilizer, the special equipment comprises a low-temperature drying device 3, a crushing device 5, a stirring and mixing device 6, a three-dimensional layered fermentation device 8, a dynamic mixing device 9 and a granulating device 10, when preparing the fertilizer, fresh astragalus stems and leaves stored in a green manure storage device 1 are conveyed to a low-temperature drying device 3 through a belt type feeding device 2 to be dried at low temperature until the water content is lower than 30 percent, then lifting to a crushing device 5 through a lifting device 4, crushing to 0.7cm, pouring the intercalated microorganism bacterium agent into warm water at 33 ℃ according to the weight ratio of 1:75, fully and uniformly mixing to form a microorganism bacterium solution, filling the microorganism bacterium solution into a zymocyte liquid storage device 7, then uniformly mixing the crushed astragalus stem leaves and the microbial liquid in a stirring and mixing device 6 to control the final water content of the mixture to be 58%; feeding the mixture into a three-dimensional layered fermentation device 8 for fermentation, and keeping the fermentation temperature at 38 ℃ for 6 days through a temperature control device 88; the three-dimensional layered fermentation device 8 comprises a shell, a layered frame 81, a top cover 82, a base 83, an air inlet pipeline 84, an air outlet pipeline 85, a blower 86, an air outlet fan 87 and a temperature sensor 89, the shelf 81 is located inside the housing, the top cover 82 is located on the top of the housing, the base 83 is located on the bottom of the housing, the air inlet duct 84 is located in the base 83, and connected to the blower 86, a plurality of air inlets 841 are provided on the air inlet pipe 84, the air inlets 841 lead to the inside of the housing, the air outlet pipe 85 is positioned in the top cover 82, and is connected with the exhaust fan 87, the exhaust duct 85 is provided with a plurality of exhaust outlets 851, the exhaust outlets 851 are also communicated with the inside of the shell, at least 3 temperature sensors are arranged in the layering rack 81 and are respectively positioned at the upper, middle and lower positions of the layering rack 81, and the temperature sensors 89 are connected to the temperature control device 88 through leads 810. In the fermentation process, on one hand, the temperature can be adjusted through the temperature control device 88, the temperature control device 88 adjusts the temperature in real time according to the temperature information transmitted back by the temperature sensor 89, on the other hand, the temperature can be adjusted through the air inlet pipeline 84 and the air outlet pipeline 85, especially when the temperature is too high, the air is directly introduced into the three-dimensional layered fermentation device 8 through the air blower 86 and the air inlet pipeline 84 to cool, the air is discharged through the air outlet pipeline 85 and the air exhaust fan 87, after the fermentation is finished, the cold air can be introduced into the three-dimensional layered fermentation device 8 through the air blower 86 and the air inlet pipeline 84 to cool the fermented green manure, the redundant heat is taken away by the air and is discharged through the air outlet pipeline 85 and the air exhaust fan 87, after the temperature is reduced to the proper temperature, the green manure is conveyed to the dynamic mixing device 9 through the lifting device 4, the SNP solution, the NAA solution, the azotobacteria liquid T16 and the azotobacteria liquid T21 are added into the dynamic mixing device 9 according to the proportion, then 0.06 percent (wt) of sodium diformate and 3.5 percent (wt) of bentonite are added, then the dynamic mixing device 9 is driven to work by a motor 92, the components are uniformly mixed, then the mixture is dried by the low-temperature drying device 3 until the water content is lower than 5 percent, and finally the mixture is sent into a granulation tower for granulation, so that the solid fertilizer is prepared.
The containers for holding the SA solution, the SNP solution, the NAA solution, the azotobacter solution T16 and the azotobacter solution T21 are a container I93, a container II 94, a container III 95, a container IV 96 and a container V97 respectively, all the containers are connected to the dynamic mixing device 9 through pipelines, and each pipeline is provided with a mass flow meter 98 and a distribution valve 99, so that the dosage of each component can be accurately and quantitatively controlled.
The method for cultivating astragalus membranaceus by using the prepared special fertilizer for planting astragalus membranaceus comprises the following steps:
s1: soaking radix astragali seed in 98% concentrated sulfuric acid for 3min, washing with running water for 60min until the residual solution of concentrated sulfuric acid is washed, and air drying on filter paper;
s2: after land preparation and ploughing, manually sowing the astragalus seeds treated by the S1, wherein the average plant spacing and the average row spacing of the astragalus are 15cm and 25cm, sowing solid fertilizer, and performing conventional field management until harvesting.
Example 3:
a preparation method of a fertilizer special for planting astragalus comprises the following steps:
s1, preparing an elicitor solution:
preparing Salicylic Acid (SA) into a solution with the concentration of 20umol/L by using distilled water, preparing Sodium Nitroprusside (SNP) into a solution with the concentration of 25mmol/L by using distilled water, and preparing naphthylacetic acid (NAA) into a solution with the concentration of 1.2mg/L by using distilled water for later use;
salicylic Acid (SA) -purchased from Tianjin Beichen Fangzheng chemical reagent plant with molecular weight of 138.12, Sodium Nitroprusside (SNP) -purchased from Tianjin Tianli chemical reagent plant with molecular weight of 297.94, naphthylacetic acid (NAA) -purchased from Tianjin Tianda chemical reagent plant with molecular weight of 186.21;
s2, fermenting green manure:
1) collecting fresh stem and leaf of radix astragali from radix astragali of the same turbid radix astragali base, drying at low temperature until the water content is lower than 30%, and pulverizing to 1 cm;
2) pouring the bissextile fungus agent into warm water at 35 ℃ according to the weight ratio of 1:100, and fully and uniformly mixing to form a microorganism bacterium liquid for later use;
3) mixing the microbial liquid with crushed astragalus stem and leaf to control the final water content of the mixture to be 65%; the judgment standard for whether the water content is proper is to grip a material tightly, and the material is preferably scattered in a scattered manner without dripping when water is in the gaps;
4) fermenting the mixture, keeping the fermentation temperature at 40 ℃ and the fermentation time at 10 days;
s3, preparing nitrogen-fixing bacteria liquid:
1) respectively inoculating the stored nitrogen-fixing strains T16 and T21 into LB (Luria-bertani) liquid culture medium, culturing at 28 deg.C and 125r/min for 48h, and measuring the absorbance value (OD600, optical density) of each strain suspension at 600nm with an ultraviolet spectrophotometer after the strains grow sufficiently; when the OD value is larger than 0.5, adjusting the OD values to be equal, and then continuing to enable each strain suspension to fully grow to obtain a bacterial suspension for later use;
2) putting 150mL LB liquid culture medium into two 500mL triangular flasks, sterilizing, placing at room temperature for 2d, respectively inoculating 15mL of each bacterial suspension after checking no pollution, culturing at 28 ℃ and 125r/min for 3d, measuring the OD value of zymogen liquid, and injecting the liquid into a sterilized glass bottle by using a sterilized injector for normal-temperature sealed storage when the OD value is more than 0.5 (the wavelength is 660nm), thus obtaining the mother solution;
3) the mother liquor was diluted to the desired concentration with distilled water: t16 is 107Each ml, T21 is 109Per ml;
s4, mixing the green manure with an elicitor solution and a nitrogen-fixing bacterium solution to prepare a solid fertilizer:
weighing 1.23-4.9kg of the green manure fermented in the step S2, 300ml of the azotobacter liquid T16225ml and T21225ml prepared in the step S3, 300ml of the SA solution, 300ml of the SNP solution and 300ml of the NAA solution prepared in the step S1, adding 0.1% (wt) of preservative potassium benzoate and 5% (wt) of binder clay, mixing, and preparing into a solid fertilizer.
The special equipment is used in the process of preparing the solid fertilizer, the special equipment comprises a low-temperature drying device 3, a crushing device 5, a stirring and mixing device 6, a three-dimensional layered fermentation device 8, a dynamic mixing device 9 and a granulating device 10, when preparing the fertilizer, fresh astragalus stems and leaves stored in a green manure storage device 1 are conveyed to a low-temperature drying device 3 through a belt type feeding device 2 to be dried at low temperature until the water content is lower than 30 percent, then lifting to a crushing device 5 through a lifting device 4, crushing to 1cm, pouring the Runfei microbial inoculum into warm water at 35 ℃ according to the weight ratio of 1:100, mixing uniformly to form a microbial inoculum, filling into a zymocyte liquid storage device 7, then uniformly mixing the crushed stem and leaf of the astragalus mongholicus and the microbial solution in a stirring and mixing device 6 to control the final water content of the mixture to be 65%; feeding the mixture into a three-dimensional layered fermentation device 8 for fermentation, and keeping the fermentation temperature at 40 ℃ for 10 days through a temperature control device 88; the three-dimensional layered fermentation device 8 comprises a shell, a layered frame 81, a top cover 82, a base 83, an air inlet pipeline 84, an air outlet pipeline 85, a blower 86, an air outlet fan 87 and a temperature sensor 89, the shelf 81 is located inside the housing, the top cover 82 is located on the top of the housing, the base 83 is located on the bottom of the housing, the air inlet duct 84 is located in the base 83, and connected to the blower 86, a plurality of air inlets 841 are provided on the air inlet pipe 84, the air inlets 841 lead to the inside of the housing, the air outlet pipe 85 is positioned in the top cover 82, and is connected with the exhaust fan 87, the exhaust duct 85 is provided with a plurality of exhaust outlets 851, the exhaust outlets 851 are also communicated with the inside of the shell, at least 3 temperature sensors are arranged in the layering rack 81 and are respectively positioned at the upper, middle and lower positions of the layering rack 81, and the temperature sensors 89 are connected to the temperature control device 88 through leads 810. In the fermentation process, on one hand, the temperature can be adjusted through the temperature control device 88, the temperature control device 88 adjusts the temperature in real time according to the temperature information transmitted back by the temperature sensor 89, on the other hand, the temperature can be adjusted through the air inlet pipeline 84 and the air outlet pipeline 85, especially when the temperature is too high, the air is directly introduced into the three-dimensional layered fermentation device 8 through the air blower 86 and the air inlet pipeline 84 to cool, the air is discharged through the air outlet pipeline 85 and the air exhaust fan 87, after the fermentation is finished, the cold air can be introduced into the three-dimensional layered fermentation device 8 through the air blower 86 and the air inlet pipeline 84 to cool the fermented green manure, the redundant heat is taken away by the air and is discharged through the air outlet pipeline 85 and the air exhaust fan 87, after the temperature is reduced to the proper temperature, the green manure is conveyed to the dynamic mixing device 9 through the lifting device 4, the SNP solution, the NAA solution, the azotobacteria liquid T16 and the azotobacteria liquid T21 are added into the dynamic mixing device 9 according to the proportion, then 0.1 percent (wt) of potassium benzoate and 5 percent (wt) of clay are added, then the dynamic mixing device 9 is driven by a motor 92 to work and uniformly mixed, then the mixture is dried by the low-temperature drying device 3 until the water content is lower than 5 percent, and finally the mixture is sent into a granulation tower for granulation to obtain the solid fertilizer.
The containers for holding the SA solution, the SNP solution, the NAA solution, the azotobacter solution T16 and the azotobacter solution T21 are a container I93, a container II 94, a container III 95, a container IV 96 and a container V97 respectively, all the containers are connected to the dynamic mixing device 9 through pipelines, and each pipeline is provided with a mass flow meter 98 and a distribution valve 99, so that the dosage of each component can be accurately and quantitatively controlled.
The method for cultivating astragalus membranaceus by using the prepared special fertilizer for planting astragalus membranaceus comprises the following steps:
s1: soaking radix astragali seed in 98% concentrated sulfuric acid for 3min, washing with running water for 60min until the residual solution of concentrated sulfuric acid is washed, and air drying on filter paper;
s2: after land preparation and ploughing, manually sowing the astragalus seeds treated by the S1, wherein the average plant spacing and the average row spacing of the astragalus are 15cm and 25cm, sowing solid fertilizer, and performing conventional field management until harvesting.
Example 4:
a preparation method of a fertilizer special for planting astragalus comprises the following steps:
s1, preparing an elicitor solution:
preparing Sodium Nitroprusside (SNP) into a solution with the concentration of 15mmol/L by using distilled water for later use; sodium Nitroprusside (SNP) -purchased from Tianjin Tianli chemical reagent factory with molecular weight of 297.94;
s2, fermenting green manure:
1) collecting fresh stem and leaf of radix astragali from radix astragali of the same turbid radix astragali base, drying at low temperature until the water content is lower than 30%, and pulverizing to 1 cm;
2) pouring the bissextile fungus agent into warm water at the temperature of 1:50 by weight ratio, and fully and uniformly mixing to form a microorganism bacterium liquid for later use;
3) mixing the microbial liquid with crushed astragalus stem and leaf to control the final water content of the mixture to be 50%; the judgment standard for whether the water content is proper is to grip a material tightly, and the material is preferably scattered in a scattered manner without dripping when water is in the gaps;
4) fermenting the mixture, keeping the fermentation temperature at 36 ℃ and the fermentation time at 2 days;
s3, preparing nitrogen-fixing bacteria liquid:
1) respectively inoculating the stored nitrogen-fixing strains T16 and T21 into LB (Luria-bertani) liquid culture medium, culturing at 28 deg.C and 125r/min for 48h, and measuring the absorbance value (OD600, optical density) of each strain suspension at 600nm with an ultraviolet spectrophotometer after the strains grow sufficiently; when the OD value is larger than 0.5, adjusting the OD values to be equal, and then continuing to enable each strain suspension to fully grow to obtain a bacterial suspension for later use;
2) putting 150mL LB liquid culture medium into two 500mL triangular flasks, sterilizing, placing at room temperature for 1d, respectively inoculating 15mL of each bacterial suspension after checking no pollution, culturing at 28 ℃ and 125r/min for 2d, measuring the OD value of zymogen liquid, and injecting the liquid into a sterilized glass bottle by using a sterilized injector for normal-temperature sealed storage when the OD value is more than 0.5 (the wavelength is 660nm), thus obtaining the mother solution;
3) the mother liquor was diluted to the desired concentration with distilled water: t16 is 106Each ml, T21 is 109Per ml;
s4, mixing the green manure with an elicitor solution and a nitrogen-fixing bacterium solution to prepare a solid fertilizer:
weighing 1.23kg of the green manure fermented in the step S2, 200ml of the nitrogen-fixing bacteria liquid T1675ml and T2175ml prepared in the step S3 and the SNP solution prepared in the step S1, mixing, and preparing the solid fertilizer.
The special equipment is used in the process of preparing the solid fertilizer, the special equipment comprises a low-temperature drying device 3, a crushing device 5, a stirring and mixing device 6, a three-dimensional layered fermentation device 8, a dynamic mixing device 9 and a granulating device 10, when preparing the fertilizer, fresh astragalus stems and leaves stored in a green manure storage device 1 are conveyed to a low-temperature drying device 3 through a belt type feeding device 2 to be dried at low temperature until the water content is lower than 30 percent, then lifting to a crushing device 5 through a lifting device 4, crushing to 1cm, pouring the intercalated microorganism bacterium agent into warm water at 30 ℃ according to the weight ratio of 1:50, fully and uniformly mixing to form microorganism bacterium liquid, filling the microorganism bacterium liquid into a zymocyte liquid storage device 7, then uniformly mixing the crushed stem and leaf of the astragalus with the microbial liquid in a stirring and mixing device 6 to control the final water content of the mixture to be 50 percent; feeding the mixture into a three-dimensional layered fermentation device 8 for fermentation, and keeping the fermentation temperature at 36 ℃ for 2 days through a temperature control device 88; the three-dimensional layered fermentation device 8 comprises a shell, a layered frame 81, a top cover 82, a base 83, an air inlet pipeline 84, an air outlet pipeline 85, a blower 86, an air outlet fan 87 and a temperature sensor 89, the shelf 81 is located inside the housing, the top cover 82 is located on the top of the housing, the base 83 is located on the bottom of the housing, the air inlet duct 84 is located in the base 83, and connected to the blower 86, a plurality of air inlets 841 are provided on the air inlet pipe 84, the air inlets 841 lead to the inside of the housing, the air outlet pipe 85 is positioned in the top cover 82, and is connected with the exhaust fan 87, the exhaust duct 85 is provided with a plurality of exhaust outlets 851, the exhaust outlets 851 are also communicated with the inside of the shell, at least 3 temperature sensors are arranged in the layering rack 81 and are respectively positioned at the upper, middle and lower positions of the layering rack 81, and the temperature sensors 89 are connected to the temperature control device 88 through leads 810. In the fermentation process, on one hand, the temperature can be adjusted through the temperature control device 88, the temperature control device 88 adjusts the temperature in real time according to the temperature information transmitted back by the temperature sensor 89, on the other hand, the temperature can be adjusted through the air inlet pipeline 84 and the air outlet pipeline 85, especially when the temperature is too high, air is directly introduced into the three-dimensional layered fermentation device 8 through the air blower 86 and the air inlet pipeline 84 to cool, the air is discharged through the air outlet pipeline 85 and the air exhaust fan 87, after the fermentation is finished, cold air can be introduced into the three-dimensional layered fermentation device 8 through the air blower 86 and the air inlet pipeline 84 to cool the fermented green manure, redundant heat is taken away by the air and is discharged through the air outlet pipeline 85 and the air exhaust fan 87, after the temperature is reduced to a proper temperature, the green manure is conveyed to the dynamic mixing device 9 through the lifting device 4, and the three components are added to the dynamic mixing device 9 according to the ratio through the container in which the SNP solution, the azotobac In the mixing device 9, the motor 92 drives the dynamic mixing device 9 to work, the materials are uniformly mixed, the materials are dried by the low-temperature drying device 3 until the water content is lower than 5%, and finally the materials are sent into a granulation tower for granulation to obtain the solid fertilizer.
The containers of the SNP solution, the azotobacter liquid T16 and the azotobacter liquid T21 are a second container 94, a fourth container 96 and a fifth container 97 respectively, each container is connected to the dynamic mixing device 9 through a pipeline, each pipeline is provided with a mass flow meter 98 and a distribution valve 99, and the dosage of each component can be accurately and quantitatively controlled.
The method for cultivating astragalus membranaceus by using the prepared special fertilizer for planting astragalus membranaceus comprises the following steps:
s1: soaking radix astragali seed in 98% concentrated sulfuric acid for 3min, washing with running water for 60min until the residual solution of concentrated sulfuric acid is washed, and air drying on filter paper;
s2: after land preparation and ploughing, manually sowing the astragalus seeds treated by the S1, wherein the average plant spacing and the average row spacing of the astragalus are 15cm and 25cm, sowing solid fertilizer, and performing conventional field management until harvesting.
And (3) experimental verification:
1. influence of different elicitors on germination and growth of astragalus seeds
The test is carried out in a Shanxi province Changzhongdong scientific and technological park, belongs to the southeast of the Shanxi province, and the west foot of the middle section of the Taihang mountain is positioned at the edge of the southeast of the Changzhong basin, is positioned between 35 degrees 52 'and 36 degrees 9' in the north latitude, is positioned between 112 degrees 58 'and 113 degrees 11' in the east longitude, and has the altitude of more than 1200 meters. The climate in Changzhi county belongs to continental monsoon climate in warm temperature zone, and has clear four seasons, sufficient sunshine, moderate rainfall and small day-night temperature difference. In cold and warm semi-dry areas, the annual average temperature is 9 ℃, the temperature is-6.2 ℃ in one month, the temperature is 22.9 ℃ in July, the annual rainfall is 411mm, the frost period is from the first ten days of October to the middle ten days of April of the next year, and the frost period is 160 d.
In the test, Mongolian milkvetch roots in the same turbid astragalus base are treated by 98% concentrated sulfuric acid, washed clean by tap water and slightly dried for later use. Seed treatment was performed with different elicitors (SA, SNP, NAA) and tap water as a blank control (see table 1). After 7 days (namely 5 months and 8 days in 2015), the seeds are sown in flowerpots, 10 grains are planted in each flowerpot, the soil in each flowerpot is watered thoroughly in advance, and the soil is covered uniformly, so that nutrients required by the growth of the seeds are ensured, and the respiration effect of the seeds cannot be influenced. Watering, weeding and killing insects during the later growth process of the astragalus, and building a sunshade at the seedling stage to prevent the astragalus seedlings from being burnt by the sun. After the seedling stage (2015, 6 months and 20 days), the sunshade net can be gradually removed, so that the photosynthesis of the astragalus membranaceus plants is ensured.
TABLE 1 different elicitor treatments
Sampling in 2015 for 9 months (the dry matter accumulation reaches the highest), selecting 15 Astragalus membranaceus with similar growth vigor under the same treatment condition, dividing into 3 groups, and measuring the biomass and the content of effective components.
The result shows that SA, SNP and NAA with different concentrations obviously influence the growth of astragalus and the accumulation of active ingredients, SA can effectively promote the synthesis of flavone, and the influence degree is 15umol/L, 10umol/L and 20umol/L from large to small; the SNP can promote the synthesis of the saponin, and the influence degrees are 15mmol/L, 10mmol/L, 25mmol/L and 20mmol/L from large to small; NAA is beneficial to polysaccharide accumulation, and the influence degree is 1.0mg/L >0.6mg/L >0.8mg/L >1.2mg/L from large to small.
2. Research on optimal proportion of astragalus fertilizer
The orthogonal design used for this test was performed by SPSS software, the test protocol is shown in table 2, and the factor levels are shown in table 3.
Table 2 test protocol table
TABLE 3 levels of test factors
Turning over the land, preparing the land, partitioning and making marks in 2016, 4, 7 months; fertilizing in 4 months and 8 days, and manually sowing. The plot length is 5m, width is 3m, the average plant spacing of astragalus is 15cm, the average row spacing is 25cm, and each group treats about 60g of astragalus seeds. The indexes of investigation during growth include: plant height, root length, thicker, fresh weight, chlorophyll in leaves, superoxide dismutase (SOD), Peroxidase (POD), Catalase (CAT), Malondialdehyde (MDA), Phenylalanine Ammonia Lyase (PAL), and post-harvest investigation indexes including: astragalus dry matter weight, root flavonoid content, root saponin content, root polysaccharide, astragalus root flavonoid economic yield, saponin economic yield, and polysaccharide economic yield.
The result shows that the treatment effect of the treatment 4 is most obvious, namely the optimal mixture ratio combination is as follows: 15mmol/L SNP, 1.23Kg green manure, 106/ml T16，109/ml T21. Compared with blank control, the dry weight of radix astragali is increased by 50.52%, flavone content in root is increased by 74.5%, polysaccharide accumulation is increased by 30.7%, saponin content is increased by 63.18%, and yield and effective component content of radix astragali are increased.
Table 4 test results of treatment 4 compared to blank control
Note: in the table ". star" indicates that the statistical analysis was very significant; "+" indicates statistically significant differences.
As can be seen from Table 4, the plant height, PAL activity, dry weight, root flavone, saponin and polysaccharide content of the former and the economic content of root flavone, saponin and polysaccharide are significantly higher than those of the latter in the treatment 4 compared with the blank control cultivation method; the root length and root thickness of the astragalus root in the former is obviously higher than those in the latter; the former has higher activities of fresh weight of radix astragali, chlorophyll and POD than the latter; the former astragalus SOD and MDA are lower than the latter, but the difference is not obvious.
3. Influence of different fertilizer ratios on soil nutrients
The soil physical and chemical property measurement is respectively carried out in 2016 (4 months are original values, 6 months, 8 months and 10 months are values after fertilization treatment), each cell collects 0-20cm soil layer soil samples according to an S-shaped 5-point sampling method to prepare a mixed sample, and the depth, soil body and quantity of each sampling part are ensured to be consistent during sampling. Uniformly mixing the soil samples, removing impurities such as dead branches, fallen leaves, stones and the like, taking part of the fresh soil samples, storing the fresh soil samples in a 4-degree refrigerator, air-drying the rest of the soil samples in a clean and dry place, removing the rest, bagging and marking, and measuring the contents of soil organic matters, total nitrogen phosphorus potassium, available nitrogen, available phosphorus and available potassium.
The results show that different fertilizer ratios can improve the nutrient content of the soil and promote the growth of the astragalus membranaceus, wherein the treatment effect of the treatment 4 is the best, the content is stable, compared with a blank control, the treatment 4 improves the organic matter in the soil by 7.23%, the total nitrogen, phosphorus and potassium contents by 3.69%, 17.59% and 2.15%, and the effective nitrogen, phosphorus and potassium contents by 8.96%, 33.16% and 10.33%, respectively.
Claims (2)
1. A preparation method of a fertilizer special for planting astragalus membranaceus is characterized by comprising the following steps:
s1, preparing an elicitor solution:
preparing Salicylic Acid (SA) into a solution with the concentration of 10-20umol/L by using distilled water, preparing Sodium Nitroprusside (SNP) into a solution with the concentration of 10-25mmol/L by using distilled water, and preparing naphthylacetic acid (NAA) into a solution with the concentration of 1-1.2mg/L by using distilled water for later use;
s2, fermenting green manure:
drying fresh stem and leaf of radix astragali at low temperature until water content is lower than 30%, and pulverizing to 0.5-1 cm;
pouring the leap fruit microbial agent into warm water at the temperature of 30-35 ℃ according to the weight ratio of 1:50-100, and fully and uniformly mixing to form a microbial liquid for later use;
mixing the microbial liquid with the crushed astragalus stem and leaf to control the final water content of the mixture to be 50-65%;
fermenting the mixture at 36-40 deg.C for 2-10 days;
s3, preparing nitrogen-fixing bacteria liquid:
1) respectively inoculating the stored nitrogen-fixing strains T16 and T21 into LB (luria-bertani) liquid culture medium, culturing at 28 deg.C and 125r/min for 48h, and allowing them to grow sufficiently to obtain bacterial suspension;
2) putting 150mL of LB liquid culture medium into two 500mL triangular flasks, sterilizing, placing at room temperature for 1-2 d, respectively inoculating 15mL of each bacterial suspension after checking no pollution, and culturing at 28 ℃ and 125r/min for 2-3 d to obtain mother liquor;
3) the mother liquor was diluted to the desired concentration with distilled water: t16 is 105-107Each ml, T21 is 107-109Per ml;
s4, mixing the green manure with an elicitor solution and a nitrogen-fixing bacterium solution to prepare a solid fertilizer:
weighing 1.23-4.9kg of the green manure fermented in the step S2, T1675-225ml of the azotobacteria liquid prepared in the step S3, T2175-225ml of the SNP solution prepared in the step S1 and 300ml of the SNP solution prepared in the step S1, and mixing to prepare a solid fertilizer;
in step S3, when preparing the bacterial suspension, after the bacterial strains grow sufficiently, measuring an absorbance value (OD600, optical density) of each bacterial strain suspension at 600nm by using an ultraviolet spectrophotometer; when the OD value is larger than 0.5, adjusting the OD values to be equal, and then continuing to allow each strain suspension to fully grow;
in the step S3, when the mother solution is prepared, after culturing for 2-3 days, measuring the OD value of the zymogen liquid, when the OD value is greater than 0.5, at this time, the wavelength is 660nm, injecting the bacteria liquid into a sterilized glass bottle by using a sterilized syringe, and sealing and storing at normal temperature to obtain the mother solution;
in the step S3, the mother liquor is diluted to T16 of 106Each ml, T21 is 109Per ml;
in the step S4, the dosage of the nitrogen-fixing bacterium liquid T16 is 75ml, and the dosage of T21 is 75 ml;
in the step S4, 0.01-0.1wt% of preservative and 2-5wt% of binder are added when preparing the solid fertilizer;
the preservative is sodium diformate or potassium benzoate; the binder is bentonite or clay.
2. The cultivation method for preparing astragalus membranaceus by using the preparation method according to claim 1, which is characterized by comprising the following steps of:
s1: soaking radix astragali seed in 98% concentrated sulfuric acid for 3min, washing with running water for 60min until the residual solution of concentrated sulfuric acid is washed, and air drying on filter paper;
s2: after soil preparation, uniformly spreading solid fertilizer, then turning the soil, uniformly mixing the soil and the fertilizer, ditching every other day, manually sowing the astragalus membranaceus seeds treated by the S1, wherein the average plant spacing and the average row spacing of the astragalus membranaceus are 15cm and 25cm respectively, and performing conventional field management until harvesting.
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