CN110683870A - Preparation method and application of biofertilizer prepared from kelp residues and bacillus - Google Patents

Preparation method and application of biofertilizer prepared from kelp residues and bacillus Download PDF

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Publication number
CN110683870A
CN110683870A CN201911074826.1A CN201911074826A CN110683870A CN 110683870 A CN110683870 A CN 110683870A CN 201911074826 A CN201911074826 A CN 201911074826A CN 110683870 A CN110683870 A CN 110683870A
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bacillus
residue
kelp residue
biofertilizer
microwave treatment
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李义强
袁源
张成省
王晓强
邹平
耿玉亭
周金辉
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Tobacco Research Institute of CAAS
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Priority to AU2020101766A priority patent/AU2020101766A4/en
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    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
    • 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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  • Organic Chemistry (AREA)
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  • Life Sciences & Earth Sciences (AREA)
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  • General Chemical & Material Sciences (AREA)
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  • Molecular Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
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Abstract

The invention provides a preparation method of a biofertilizer taking kelp residue and bacillus as raw materials, belonging to the technical field of biofertilizer preparation and comprising the following steps: 1) mixing the kelp residue with water, performing microwave treatment, centrifuging the obtained microwave treatment product to obtain a first filter residue and a first clear liquid, centrifuging the first clear liquid to obtain a second clear liquid and a second filter residue, and freeze-drying the second clear liquid to obtain a kelp residue degradation product; 2) dissolving the degraded kelp residue obtained in the step 1) in water, inoculating bacillus, and fermenting to obtain the biofertilizer. The invention takes kelp residue and bacillus as raw materials to prepare the biological fertilizer, and the biological fertilizer can promote the growth of plants and can prevent and treat plant diseases.

Description

Preparation method and application of biofertilizer prepared from kelp residues and bacillus
Technical Field
The invention belongs to the technical field of biological fertilizer preparation, and particularly relates to a preparation method and application of a biological fertilizer taking kelp residue and bacillus as raw materials.
Background
China is a traditional big agricultural country, and agriculture occupies a very important position in national production. The modern agricultural technology promotes the rapid development of agricultural production, and the popularization of chemical fertilizers enables the agricultural production to obtain great economic benefits. In recent years, the problems of environmental pollution, soil hardening, soil fertility decline, ecological deterioration and the like caused by using a large amount of fertilizers are increasingly serious, the environment is damaged, the soil fertility is influenced, and the quality of agricultural products is reduced. In order to realize sustainable development of agriculture and achieve the aims of high yield, high quality, ecology and safety, the development of economic and efficient natural fertilizers is of great significance.
Disclosure of Invention
In view of the above, the present invention aims to provide a preparation method and application of a biofertilizer prepared from kelp residue and bacillus.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides a preparation method of a biofertilizer taking kelp residue and bacillus as raw materials, which comprises the following steps:
1) mixing the kelp residue with water, performing microwave treatment, centrifuging the obtained microwave treatment product to obtain a first filter residue and a first clear liquid, centrifuging the first clear liquid to obtain a second clear liquid and a second filter residue, and freeze-drying the second clear liquid to obtain a kelp residue degradation product;
2) dissolving the degraded kelp residue obtained in the step 1) in water, inoculating bacillus, and fermenting to obtain the biofertilizer.
Preferably, the bacillus of step 2) comprises bacillus subtilis and/or bacillus amyloliquefaciens.
Preferably, the volume ratio of the mass of the kelp residue in the step 1) to the volume of water is (1-5) g to (10-50) ml.
Preferably, the microwave treatment conditions in step 1) include: the temperature of the microwave treatment is 150-200 ℃, the power of the microwave treatment is 800-1200W, and the time of the microwave treatment is 10-30 min.
Preferably, the centrifugation conditions of step 1) include: the rotating speed of the centrifugation is 9000-11000 rpm, and the time of the centrifugation is 15-25 min.
Preferably, the bacillus is inoculated by a bacillus liquid, and the OD of the bacillus liquid600The value is 0.1 to 0.5.
Preferably, the volume ratio of the mass of the kelp residue degradation product to the bacillus liquid is (1-5) g to (1-5) ml.
Preferably, the fermentation conditions of step 2) include: the fermentation time is 90-150 h, the fermentation temperature is 25-30 ℃, and the fermentation rotating speed is 150-200 rpm.
The invention also provides application of the biological fertilizer prepared by the preparation method in the technical scheme in promoting plant growth.
The invention also provides application of the biological fertilizer prepared by the preparation method in the technical scheme in plant disease control.
The invention provides a preparation method of a biofertilizer taking kelp residue and bacillus as raw materials, which comprises the following steps: 1) mixing the kelp residue with water, performing microwave treatment, centrifuging the obtained microwave treatment product to obtain a first filter residue and a first clear liquid, centrifuging the first clear liquid to obtain a second clear liquid and a second filter residue, and freeze-drying the second clear liquid to obtain a kelp residue degradation product; 2) dissolving the degraded kelp residue obtained in the step 1) in water, inoculating bacillus, and fermenting to obtain the biofertilizer. The invention takes kelp residue and bacillus as raw materials to prepare the biological fertilizer, and the biological fertilizer can promote the growth of plants and can prevent and treat plant diseases.
Drawings
FIG. 1 shows the growth promotion results of the bio-fertilizer of example 1, wherein LJW + C is the bio-fertilizer prepared from Enteromorpha prolifera and Bacillus amyloliquefaciens, and NTC is the control;
FIG. 2 is the growth promotion result of the bio-fertilizer of example 1, wherein LJW + T is the bio-fertilizer prepared by using Enteromorpha prolifera and Bacillus subtilis as raw materials, and NTC is a control;
FIG. 3 shows the results of preventing plant diseases in the biofertilizer of example 2, wherein LJW + C is a biofertilizer prepared from Enteromorpha and Bacillus amyloliquefaciens as raw materials, and NTC is a control;
fig. 4 shows the results of preventing plant diseases by the biofertilizer of example 2, wherein LJW + T is the biofertilizer prepared from enteromorpha and bacillus subtilis, and NTC is the control.
Detailed Description
The invention provides a preparation method of a biofertilizer taking kelp residue and bacillus as raw materials, which comprises the following steps:
1) mixing the kelp residue with water, performing microwave treatment, centrifuging the obtained microwave treatment product to obtain a first filter residue and a first clear liquid, centrifuging the first clear liquid to obtain a second clear liquid and a second filter residue, and freeze-drying the second clear liquid to obtain a kelp residue degradation product;
2) dissolving the degraded kelp residue obtained in the step 1) in water, inoculating bacillus, and fermenting to obtain the biofertilizer.
Mixing kelp residue with water, performing microwave treatment, centrifuging the obtained microwave treatment product to obtain a first filter residue and a first clear liquid, centrifuging the first clear liquid to obtain a second clear liquid and a second filter residue, and freeze-drying the second clear liquid to obtain a kelp residue degradation product.
In the invention, the kelp residue is preferably residue left after iodine and sodium alginate are extracted from kelp.
In the present invention, the ratio of the mass of the kelp residue to the volume of water is preferably (1 to 5) g to (10 to 50) ml, and more preferably (2 to 4) g to (20 to 40) ml.
In the present invention, the conditions of the microwave treatment preferably include: the temperature of the microwave treatment is preferably 150-200 ℃, and more preferably 160-180 ℃; the power of the microwave treatment is preferably 800-1200W, and more preferably 900-1100W; the time of the microwave treatment is preferably 10-30 min, and more preferably 15-25 min. The invention degrades the kelp residue through microwave treatment.
In the invention, the centrifugation is preferably carried out when the temperature of the microwave treatment object is 50-70 ℃, and the centrifugation conditions preferably comprise: the rotation speed of the centrifugation is preferably 9000-11000 rpm, and more preferably 10000 rpm; the time for centrifugation is preferably 15-25 min, and more preferably 20 min. In the present invention, the centrifugation removes solid matter of the kelp residue.
The time for freeze-drying the second clear liquid is not particularly limited, and the conventional freeze-drying condition is adopted. In the present invention, the obtained degraded kelp residue is preferably stored at-20 ℃.
The invention dissolves the kelp residue degradation product in water, inoculates bacillus and then ferments to obtain the biological fertilizer.
In the present invention, the bacillus preferably includes bacillus subtilis and/or bacillus amyloliquefaciens, and when the bacillus includes bacillus subtilis and bacillus amyloliquefaciens, they are mixed in an arbitrary number ratio. In the invention, the bacillus subtilis is preferably bacillus subtilis Tbp55 with the preservation number of CGMCC No. 2853; the bacillus amyloliquefaciens is preferably bacillus amyloliquefaciens Cas02 with the preservation number of CGMCC NO. 15514. In the present invention, the bacillus ferments the degraded kelp residue.
In the present invention, the volume ratio of the mass of the degraded product of kelp residue to water is preferably (1 to 5) g to (0.1 to 2) L, and more preferably (2 to 3) g to (0.8 to 1.5) L.
In the present invention, the bacillus is preferably inoculated with a bacillus solution, the OD of which600The value is preferably 0.1 to 0.5, more preferably 0.3. In the present invention, the preparation of the bacillus liquid preferably includes: inoculating Bacillus in LB liquid culture medium, culturing, diluting the obtained culture solution, and measuring OD600The value is 0.1 to 0.5. The LB liquid culture medium is not particularly limited, and the conventional LB liquid culture medium can be adopted. In the invention, the culture temperature is preferably 25-30 ℃, and the culture time is preferably 20-30 h.
In the present invention, the conditions of the fermentation preferably include: the fermentation time is preferably 90-150 h, and more preferably 110-130 h; the fermentation temperature is preferably 25-30 ℃; the rotation speed of the fermentation is preferably 150-200 rpm, and more preferably 160-180 rpm.
In the invention, the biological fertilizer is a liquid fertilizer, the content of laminarin in the liquid fertilizer is 1-3 g/L, and the number of bacillus is 1 multiplied by 1010～1011And (2) per liter.
The invention also provides application of the biological fertilizer prepared by the preparation method in the technical scheme in promoting plant growth.
The invention also provides application of the biological fertilizer prepared by the preparation method in the technical scheme in plant disease control.
In the present invention, the plant preferably includes pepper, tobacco, eggplant or cucumber.
In the present invention, the plant disease preferably includes a pepper bacterial wilt.
In the invention, the use method of the biological fertilizer is preferably 15-25 ml, preferably 20ml, of the biological fertilizer used for each plant, and the use method is root irrigation.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
1) Degradation of kelp residues: taking 2g of kelp residue, adding 20mL of water, degrading for 20min by using a microwave reactor (CEM Mars 6) under the conditions of 180 ℃ and 1000W of microwave power, centrifugally separating the extract when the reaction system is cooled to 60 ℃, and centrifugally treating the obtained extracting solution again to obtain the final extracting solution. Vacuum freeze drying the final extractive solution to obtain degraded product of herba Zosterae Marinae residue, and storing in refrigerator at-20 deg.C. Wherein, the residue left after extracting iodine and sodium alginate from the kelp residue and kelp.
2) And (3) microbial fermentation: culturing Bacillus subtilis Tbp55 in LB culture medium for 24h, and diluting to OD600The value is 0.3, and the bacillus subtilis Tbp55 bacterial liquid is obtained. Taking 5g seaAnd dissolving the degraded product with the residues in 1L of water, inoculating 1mL of bacillus subtilis Tbp55 bacterial liquid, and performing shake culture at 28 ℃ and 175rpm for 120h to prepare the kelp residue bacillus subtilis Tbp55 functional liquid fertilizer.
Or
And (3) microbial fermentation: culturing Bacillus amyloliquefaciens Cas02 in LB culture medium for 24h, and diluting to OD600The value is 0.3, and a bacillus amyloliquefaciens Cas02 bacterial liquid is obtained. Dissolving 5g of kelp residue degradation product in 1L of water, adding 1mL of bacillus amyloliquefaciens TCas02 bacterial liquid, and performing shake culture at 28 ℃ and 175rpm for 120h to prepare the kelp residue bacillus amyloliquefaciens Cas02 functional liquid fertilizer.
Example 2
Transplanting pepper seedlings with consistent growth vigor into a sterilized matrix, and slowly filling the liquid fertilizer prepared in the example 1 into pepper rhizosphere 20ml per plant when the seedlings grow to two true leaves. The control group was watered with an equal volume of water. Irrigate every 5 days three times in succession. After the third irrigation for 7 days, the growth promoting effect of the liquid fertilizer on the pepper is measured, and the results are shown in table 1 and figures 1-2.
TABLE 1 growth promoting effect on Capsici fructus
Treatment of Plant height mm Number of blades mm Leaf length mm Leaf width mm
NTC 88.80±11.67 8.40±1.06 70.13±8.44 37.30±4.81
LJW+C 116.53±22.87 10.13±1.13 79.80±9.66 41.13±8.41
LJW+T 88.93±16.02 9.07±1.10 70.53±12.52 38.00±5.10
Note that: NTC-water blank; LJW-degradation products of kelp residue; T-Bacillus subtilis; C-Bacillus amyloliquefaciens
As can be seen from Table 1 and FIGS. 1-2, the Bacillus laminariae residue functional fertilizer has a significant growth promoting effect on capsicum.
Example 3
After the pepper was transplanted, the liquid fertilizer prepared in example 1 was slowly poured around the roots of the pepper, 20ml per plant, and the control group was poured with an equal volume of Hoagland's nutrient solution. Irrigate every 5 days three times in succession. Inoculating Laurella solanacearum around the rhizosphere of Capsici fructus 7 days after the third irrigation (the bacterium is added for allowing Capsici fructus to have bacterial wilt), and inoculating (OD) in the form of bacterial liquid6000.3), 20ml per plant, and the results of statistics of disease onset starting 5d after inoculation are shown in Table 2 and FIGS. 3-4. The classification into 5 grades according to the classification standard of bacterial wilt disease proposed by Kempe (1983) and 0, 1, 2, 3 and 4.
Level 0: no symptoms of bacterial wilt;
level 1: 1-25% of leaves show wilting symptoms;
and 2, stage: the leaf withering symptom appears in 26-50% of leaves;
and 3, level: 51-75% of leaves have a wilting symptom;
4, level: 76-100% of leaves show wilting symptoms.
The disease index and the biocontrol effect are calculated according to the following formulas:
disease index is 100 × Σ (number of diseased plants × number of disease steps)/(total number of plants × number of highest disease steps);
relative prevention and treatment effect (%) (control disease index-treatment disease index)/control disease index × 100.
TABLE 2 statistics of the onset of pepper bacterial wilt
Figure BSA0000194154240000061
Note that: NTC-water blank; LJW-degradation products of kelp residue; T-Bacillus subtilis; C-Bacillus amyloliquefaciens
As can be seen from Table 2 and FIGS. 3 to 4, the functional fertilizer for Bacillus laminariae residues has significant control effect on bacterial wilt of capsicum.
As can be seen from the above examples, the present invention prepares a biofertilizer using kelp residue and bacillus as raw materials, the biofertilizer can promote the growth of plants and can control plant diseases.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims (10)

1. A preparation method of a biofertilizer taking kelp residue and bacillus as raw materials is characterized by comprising the following steps:
1) mixing the kelp residue with water, performing microwave treatment, centrifuging the obtained microwave treatment product to obtain a first filter residue and a first clear liquid, centrifuging the first clear liquid to obtain a second clear liquid and a second filter residue, and freeze-drying the second clear liquid to obtain a kelp residue degradation product;
2) dissolving the degraded kelp residue obtained in the step 1) in water, inoculating bacillus, and fermenting to obtain the biofertilizer.
2. The method according to claim 1, wherein the Bacillus of step 2) comprises Bacillus subtilis and/or Bacillus amyloliquefaciens.
3. The method according to claim 1, wherein the ratio of the mass of the kelp residue in the step 1) to the volume of water is (1 to 5) g to (10 to 50) ml.
4. The method according to claim 1, wherein the microwave treatment conditions of step 1) include: the temperature of the microwave treatment is 150-200 ℃, the power of the microwave treatment is 800-1200W, and the time of the microwave treatment is 10-30 min.
5. The method of claim 1, wherein the conditions for centrifugation in step 1) comprise: the rotating speed of the centrifugation is 9000-11000 rpm, and the time of the centrifugation is 15-25 min.
6. The method according to claim 1, wherein the bacillus is inoculated with a bacillus solution, and the OD of the bacillus solution600The value is 0.1 to 0.5.
7. The method according to claim 1 or 6, wherein the mass ratio of the degraded product of the kelp residue to the volume of the Bacillus subtilis solution is (1-5) g to (1-5) ml.
8. The method according to claim 1, wherein the conditions for the fermentation in step 2) include: the fermentation time is 90-150 h, the fermentation temperature is 25-30 ℃, and the fermentation rotating speed is 150-200 rpm.
9. Use of the biofertilizer prepared by the preparation method according to any one of claims 1 to 8 for promoting plant growth.
10. Use of the biofertilizer prepared by the preparation method according to any one of claims 1 to 8 for controlling plant diseases.
CN201911074826.1A 2019-11-06 2019-11-06 Preparation method and application of biofertilizer prepared from kelp residues and bacillus Pending CN110683870A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111908982A (en) * 2020-08-10 2020-11-10 济南航晨生物科技有限公司 Method for extracting seaweed by electromagnetic radiation
CN112226391A (en) * 2020-11-04 2021-01-15 河南柏裕植物免疫科技有限公司 Microbial agent containing bacillus and preparation method and application thereof

Citations (4)

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Publication number Priority date Publication date Assignee Title
CN103304278A (en) * 2012-11-06 2013-09-18 青岛明月海藻集团有限公司 Seaweed fertilizer comprising seaweed leach liquor and preparation method thereof
CN104774094A (en) * 2015-04-01 2015-07-15 大连理工大学 Preparation method of compound microbial seaweed fertilizer
KR101579485B1 (en) * 2015-06-12 2015-12-22 농업회사법인 이수농산 주식회사 Organic fertilizers and the preparation thereof
CN109796279A (en) * 2019-04-03 2019-05-24 成都云图控股股份有限公司 A kind of multi-element biological organic fertilizer and preparation method thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103304278A (en) * 2012-11-06 2013-09-18 青岛明月海藻集团有限公司 Seaweed fertilizer comprising seaweed leach liquor and preparation method thereof
CN104774094A (en) * 2015-04-01 2015-07-15 大连理工大学 Preparation method of compound microbial seaweed fertilizer
KR101579485B1 (en) * 2015-06-12 2015-12-22 농업회사법인 이수농산 주식회사 Organic fertilizers and the preparation thereof
CN109796279A (en) * 2019-04-03 2019-05-24 成都云图控股股份有限公司 A kind of multi-element biological organic fertilizer and preparation method thereof

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111908982A (en) * 2020-08-10 2020-11-10 济南航晨生物科技有限公司 Method for extracting seaweed by electromagnetic radiation
CN112226391A (en) * 2020-11-04 2021-01-15 河南柏裕植物免疫科技有限公司 Microbial agent containing bacillus and preparation method and application thereof

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Application publication date: 20200114