CN110655424A - Preparation method and application of biofertilizer taking enteromorpha and burkholderia pyrrocinia as raw materials - Google Patents

Preparation method and application of biofertilizer taking enteromorpha and burkholderia pyrrocinia as raw materials Download PDF

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Publication number
CN110655424A
CN110655424A CN201911074553.0A CN201911074553A CN110655424A CN 110655424 A CN110655424 A CN 110655424A CN 201911074553 A CN201911074553 A CN 201911074553A CN 110655424 A CN110655424 A CN 110655424A
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enteromorpha
preparation
burkholderia pyrrocinia
microwave treatment
clear liquid
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袁源
王晓强
李义强
张成省
邹平
耿玉亭
周金辉
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Tobacco Research Institute of CAAS
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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
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/03—Algae
    • 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
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
    • C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
    • 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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  • Chemical & Material Sciences (AREA)
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  • Pest Control & Pesticides (AREA)
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Abstract

The invention provides a preparation method of a biological fertilizer taking enteromorpha and burkholderia pyrrocinia as raw materials, belonging to the technical field of biological fertilizer preparation and comprising the following steps: 1) mixing enteromorpha with water, carrying out microwave treatment, centrifuging the obtained microwave treatment substance 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 an enteromorpha degradation product; 2) dissolving the enteromorpha degradation product obtained in the step 1) in water, inoculating Burkholderia pyrrocinia, and fermenting to obtain the biological fertilizer. According to the invention, the enteromorpha prolifera and the burkholderia pyrrocinia are used as raw materials to prepare the biological fertilizer, and the biological fertilizer can promote the growth of plants and can prevent and control plant diseases.

Description

Preparation method and application of biofertilizer taking enteromorpha and burkholderia pyrrocinia as raw materials
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 prepared from enteromorpha and Burkholderia pyrrocinia.
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 invention aims to provide a preparation method and application of a biofertilizer taking enteromorpha and burkholderia pyrrocinia as raw materials.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides a preparation method of a biological fertilizer taking enteromorpha and burkholderia pyrrocinia as raw materials, which comprises the following steps:
1) mixing enteromorpha with water, carrying out microwave treatment, centrifuging the obtained microwave treatment substance 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 an enteromorpha degradation product;
2) dissolving the enteromorpha degradation product obtained in the step 1) in water, inoculating Burkholderia pyrrocinia, and fermenting to obtain the biological fertilizer.
Preferably, the mass ratio of the enteromorpha degradation product in the step 2) to the volume ratio of water is (5-10) g to (0.1-2) L.
Preferably, the ratio of the mass of the enteromorpha prolifera 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 Burkholderia pyrrocinia is inoculated with a Burkholderia pyrrocinia bacterial liquid, and OD of the Burkholderia pyrrocinia bacterial liquid600The value is 0.1 to 0.5.
Preferably, the volume ratio of the mass of the enteromorpha degradation product to the Burkholderia pyrrocinia bacterial liquid is (5-10) g to (1-2) ml.
Preferably, the fermentation conditions of step 2) include: the fermentation time is 100-140 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 biological fertilizer taking enteromorpha and burkholderia pyrrocinia as raw materials, which comprises the following steps: 1) mixing enteromorpha with water, carrying out microwave treatment, centrifuging the obtained microwave treatment substance 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 an enteromorpha degradation product; 2) dissolving the enteromorpha degradation product obtained in the step 1) in water, inoculating Burkholderia pyrrocinia, and fermenting to obtain the biological fertilizer. According to the invention, the enteromorpha prolifera and the burkholderia pyrrocinia are used as raw materials to prepare the biological fertilizer, and the biological fertilizer can promote the growth of plants and can prevent and control plant diseases.
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FIG. 1 is a result of growth promotion by the bio-fertilizer of example 1, in which PE + L is a bio-fertilizer prepared from Enteromorpha and Burkholderia pyrrocinia as raw materials, and NTC is a control;
fig. 2 shows the results of preventing plant diseases by the biofertilizer of example 1, wherein PE + L is a biofertilizer prepared from enteromorpha and burkholderia pyrrocinia as raw materials, and NTC is a control.
Detailed Description
The invention provides a preparation method of a biological fertilizer taking enteromorpha and burkholderia pyrrocinia as raw materials, which comprises the following steps:
1) mixing enteromorpha with water, carrying out microwave treatment, centrifuging the obtained microwave treatment substance 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 an enteromorpha degradation product;
2) dissolving the enteromorpha degradation product obtained in the step 1) in water, inoculating Burkholderia pyrrocinia, and fermenting to obtain the biological fertilizer.
According to the invention, enteromorpha prolifera and water are mixed and then subjected to microwave treatment, the obtained microwave treatment object is centrifuged to obtain a first filter residue and a first clear liquid, the first clear liquid is centrifuged to obtain a second clear liquid and a second filter residue, and the second clear liquid is freeze-dried to obtain an enteromorpha prolifera degradation product.
In the invention, the volume ratio of the mass of the enteromorpha prolifera to the water is preferably (1-5) g to (10-50) ml, and more preferably (2-4) g to (20-40) 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 enteromorpha is degraded 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 invention, the enteromorpha residue is removed by centrifugation.
The time for freeze-drying the second clear liquid is not particularly limited, and the conventional freeze-drying condition is adopted. The enteromorpha degradation product is preferably stored at the temperature of minus 20 ℃.
According to the invention, the enteromorpha degradation product is dissolved in water, and fermentation is carried out after Burkholderia pyrrocinia is inoculated to obtain the biological fertilizer.
In the present invention, the Burkholderia pyrrocinia preferably employs the strain Lyc2 of Burkholderia pyrrocinia disclosed in the literature of Cloning and Analysis of Genes Controlling Activities of Burkholderia pyrrocinia strain Lyc2.Xiaoqiang Wang et al Current Microbiology. In the invention, the Burkholderia pyrrocinia ferments enteromorpha degradation products.
In the invention, the volume ratio of the mass of the enteromorpha degradation product to water is preferably (5-10) g to (0.1-2) L, more preferably (6-9) g to (0.8-1.5) L, and most preferably (7-8) g to (1-1.2) L.
In the present invention, the Burkholderia pyrrocinia is preferably inoculated with a Burkholderia pyrrocinia solution, and OD of the Burkholderia pyrrocinia solution600The value is preferably 0.1 to 0.5, more preferably 0.3. In the invention, the preparation of the burkholderia pyrrocinia bacterial liquid preferably comprises the following steps: inoculating Burkholderia pyrrocinia to LB liquid culture medium for culture, 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 100-140 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 enteromorpha polysaccharide in the liquid fertilizer is 2-3 g/L, and the number of Burkholderia bigeri 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 crops such as pepper, tobacco, eggplant and the like.
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) Degrading enteromorpha prolifera: adding 20mL of water into 2g of enteromorpha, 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 system is cooled to 60 ℃ after the reaction is finished, and centrifugally treating the obtained extracting solution again to obtain the final extracting solution. And (4) carrying out vacuum freeze drying on the obtained final extracting solution to obtain an enteromorpha degradation product, and storing the enteromorpha degradation product in a refrigerator at the temperature of-20 ℃.
2) And (3) microbial fermentation: culturing Burkholderia pyrrocinia Lyc2 in LB culture medium for 24h, and diluting to OD600The value was 0.3, and a bacterial solution of Burkholderia pyrrocinia Lyc2 was obtained. Dissolving 5g of enteromorpha degradation product in 1L of water, inoculating 1mL of Burkholderia pyrrocinia Lyc2 bacterial liquid, and performing shake culture at 28 ℃ and 175rpm for 120h to prepare the enteromorpha Burkholderia pyrrocinia Lyc2 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 watering for 7d, the growth promoting effect of the liquid fertilizer on the pepper was measured, and the results are shown in table 1 and fig. 1.
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
PE+L 113.33±26.50 10.93±1.03 90.73±9.79 46.27±4.64
Note that: NTC-water blank; PE-enteromorpha degradation product; L-Burkholderia
From the table 1 and the figure 1, it can be obtained that the Enteromorpha prolifera Burkholderia Lyc2 functional fertilizer has a remarkable growth promoting effect on the peppers.
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 water. 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 strain, and the results of statistics of disease onset starting 5d after inoculation are shown in Table 2 and FIG. 2. 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 BSA0000194154090000061
Note that: NTC-water blank; PE-enteromorpha degradation product; L-Burkholderia
As can be seen from the table 2 and the figure 2, the Enteromorpha prolifera Burkholderia pyrrocinia Lyc2 functional fertilizer has a remarkable prevention and treatment effect on the pepper bacterial wilt.
According to the embodiments, the enteromorpha prolifera and the burkholderia pyrrocinia Lyc2 are used as raw materials to prepare the biological fertilizer, and the biological fertilizer can promote the growth of plants and can prevent and 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 enteromorpha and Burkholderia pyrrocinia as raw materials is characterized by comprising the following steps:
1) mixing enteromorpha with water, carrying out microwave treatment, centrifuging the obtained microwave treatment substance 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 an enteromorpha degradation product;
2) dissolving the enteromorpha degradation product obtained in the step 1) in water, inoculating Burkholderia pyrrocinia, and fermenting to obtain the biological fertilizer.
2. The preparation method according to claim 1, wherein the ratio of the mass of the enteromorpha degradation product in the step 2) to the volume of water is (5-10) g to (0.1-2) L.
3. The preparation method according to claim 1, wherein the ratio of the mass of the enteromorpha prolifera in the step 1) to the volume of the water is (1-5) g to (10-50) 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 Burkholderia pyrrocinia is inoculated with a Burkholderia pyrrocinia solution, and the OD of the Burkholderia pyrrocinia solution600The value is 0.1 to 0.5.
7. The preparation method according to claim 1 or 6, wherein the volume ratio of the mass of the enteromorpha degradation product to the Burkholderia pyrrocinia bacterial liquid is (5-10) g to (1-2) ml.
8. The method according to claim 1, wherein the conditions for the fermentation in step 2) include: the fermentation time is 100-140 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.
CN201911074553.0A 2019-11-06 2019-11-06 Preparation method and application of biofertilizer taking enteromorpha and burkholderia pyrrocinia as raw materials Pending CN110655424A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11807585B1 (en) 2023-02-28 2023-11-07 King Faisal University Fertilizer composition

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CN104262008A (en) * 2014-09-30 2015-01-07 青岛嘉瑞生物技术有限公司 Process for preparing liquid seaweed fertilizer through enzymolysis and fermentation
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CN104341181A (en) * 2013-07-23 2015-02-11 山东恩宝生物科技有限公司 Multifunctional fermented kelp liquid and manufacturing method thereof
CN103820354A (en) * 2013-11-21 2014-05-28 上海海洋大学 Burkholderia FH4 bacterial strain, and screening method and application thereof
CN104262008A (en) * 2014-09-30 2015-01-07 青岛嘉瑞生物技术有限公司 Process for preparing liquid seaweed fertilizer through enzymolysis and fermentation
CN105602995A (en) * 2016-01-21 2016-05-25 青岛海洋生物医药研究院股份有限公司 Method for preparing Enteromorpha bio-fertilizer through liquid deep quick fermentation
CN108409372A (en) * 2018-05-11 2018-08-17 天津市辰圣海洋生物技术有限公司 It is a kind of using Enteromorpha as the alga fertilizer preparation method of raw material

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11807585B1 (en) 2023-02-28 2023-11-07 King Faisal University Fertilizer composition

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Application publication date: 20200107