CN107523508B

CN107523508B - Aspergillus niger JXJ A01 with activities of decomposing potassium and phosphorus and degrading organophosphorus pesticides and application thereof

Info

Publication number: CN107523508B
Application number: CN201710843897.8A
Authority: CN
Inventors: 张炳火; 李汉全
Original assignee: Jiujiang University
Current assignee: Jiujiang University
Priority date: 2017-09-15
Filing date: 2017-09-15
Publication date: 2020-09-08
Anticipated expiration: 2037-09-15
Also published as: CN107523508A

Abstract

The invention relates to the technical field of microbial application, in particular to aspergillus niger JXJ A01 with activities of dissolving potassium and phosphorus and degrading organophosphorus pesticides, and also relates to application of aspergillus niger JXJ A01 in dissolving insoluble potassium, organophosphorus and inorganic phosphorus and degrading organophosphorus pesticides, and application of aspergillus niger JXJ A01 in microbial fertilizer aspect, wherein the aspergillus niger JXJ A01 is preserved in China general microbiological culture Collection center (CGMCC No. 13775). The Aspergillus niger JXJ A01 has a strong dissolving effect on insoluble phosphorus and potassium ore powder, so that the Aspergillus niger JXJ A01 is converted into soluble phosphorus and potassium elements which can be absorbed by plants, and can degrade organophosphorus pesticide omethoate to form nontoxic soluble phosphate radicals, reduce the concentration of pesticides and increase the soluble phosphorus elements in soil.

Description

Aspergillus niger JXJ A01 with activities of decomposing potassium and phosphorus and degrading organophosphorus pesticides and application thereof

Technical Field

The invention relates to the technical field of microbial application, in particular to application of Aspergillus niger JXJ A01 in microbial fertilizer aspect, which can convert insoluble potassium, organic phosphorus and inorganic phosphorus into soluble potassium and phosphorus, and application of Aspergillus niger JXJ A01 in reducing organophosphorus pesticide residue in farmland and improving and repairing farmland soil.

Background

Phosphorus is an essential element essential for the normal growth and development of plants. More than 95 percent of phosphorus elements in soil are insoluble phosphorus which is difficult to be directly absorbed and utilized by plants, about 74 percent of farmland cultivated land in China is lack of phosphorus in soil, and the application of organic and chemical phosphate fertilizers is a main source of soil phosphorus. However, soil minerals have strong reactivity to phosphorus elements and are easy to form insoluble ineffective phosphate, so that the current utilization rate of applied phosphate fertilizers is only 5% -25%. Phosphate fertilizer mainly comes from ground phosphate rock, but ground phosphate rock is the non-renewable resource, will consume in 50 ~ 100 years, consequently, how to increase soluble phosphorus element content in the soil, improve the phosphorus fertilizer utilization efficiency, is the urgent problem that needs to solve of agricultural.

Potassium is one of the major elements essential for plant growth and development. Under general production conditions, the potassium demand of crops is equivalent to that of nitrogen. Although the soil contains a large amount of potassium element (all mineral nutrient elements of the crust are in the 4 th position), more than 90 percent of the soil is ineffective potassium which cannot be absorbed and utilized by plants. Although the insoluble potassium ore resources are rich in China, the soluble potassium ore resources are relatively deficient, and 50-70% of the dosage depends on import. The shortage of effective potassium in farmland has become one of the limiting factors for the sustainable development of agricultural production in China. Therefore, how to convert insoluble invalid potassium in soil into soluble valid potassium has important significance in relieving the shortage of soluble potassium ore resources in China and promoting the sustainable development of agriculture. Many microorganisms can dissolve potassium ore to change insoluble ineffective potassium into soluble effective potassium, most of the current researches are potassium-decomposing bacteria, and filamentous fungi are rarely reported. The mold can utilize various agricultural wastes for solid fermentation due to the developed hypha, has rapid growth and high spore yield, and is suitable for rapidly producing the microbial solid fertilizer.

In the past decades, agriculture in China is characterized by using a large amount of chemical fertilizers and organophosphorus pesticides, so that soil pollution is serious, particularly, pesticide residue is high, and the quality of crops and the development of modern ecological agriculture are seriously influenced. The microorganism has strong passivation and degradation effects on a plurality of toxic substances, so that harmful heavy metals such as soluble cadmium, lead and the like are passivated into insoluble cadmium and lead, the absorption of crops is reduced, and toxic substances such as pesticides and the like are degraded into nontoxic substances and even into nutrient components beneficial to the growth of the plants. Therefore, the application of related microorganisms in the soil can effectively reduce heavy metal pollution, the residual quantity and the residual period of pesticides in the soil, improve the soil quality, improve the quality of crops and promote the development of ecological agriculture.

Disclosure of Invention

The invention aims to provide an Aspergillus niger JXJ A01 capable of efficiently degrading insoluble potassium, organic phosphorus and inorganic phosphorus into soluble potassium and phosphorus aiming at the sustainable development requirement of modern ecological agriculture, wherein the Aspergillus niger JXJ A01 can degrade organic phosphorus pesticides into nontoxic orthophosphate, eliminate residues of organic phosphorus pesticides in soil and provide absorbable effective phosphorus for plants.

The purpose of the invention is realized by the following technical scheme:

an Aspergillus niger JXJ A01 with potassium-decomposing, phosphorus-decomposing and organophosphorus pesticide degrading activities is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.13775 and the preservation date of 2017, 04 and 20 days. The address of the depository institution: the microbial research institute of

western road

1, 3, national academy of sciences, north-south, morning-yang, Beijing, zip code: 100101, telephone: 86-10-64807355.

The aspergillus JXJ A01 generates brown black radial conidial heads, conidial peduncles are grown on spherical apical sacs, and clustered black spherical conidia are grown on the conidial peduncles and are consistent with the typical morphological characteristics of the aspergillus niger. The strain JXJ A01 grows well on various agricultural waste culture media and produces a large amount of black spores; the spore can germinate and grow at the temperature of 15-47 ℃, and the optimal germination and growth temperature is 28-37 ℃; the spores can still germinate after being treated for 30min under the damp and hot condition of 65 ℃; the spore can germinate and grow under the condition of pH2-12, and the optimum germination growth pH value is about 4-8; both spores and mycelia have certain tolerance and degradation effects on organophosphorus pesticides (such as omethoate). Through comparison and analysis with ITS sequences and 18S rRNA gene sequences in a GenBank database, the similarity of the ITS sequence fragment (591bp) of the strain to Aspergillus niger reaches 100%, the similarity of the 18S rRNA gene sequence (1709bp) to Aspergillus niger is 99%, the strain JXJ A01 is identified as Aspergillus niger (Aspergillus niger) by combining the morphological characteristics, the strain is named as Aspergillus niger JXJA01, the accession numbers of the 18S rRNA gene sequence and the ITS sequence of the strain in GenBank are MF145518 and MF543023 respectively, and specific please refer to SEQ ID NO:1 and SEQ ID NO: 2.

The Aspergillus niger JXJ A01 with the activities of dissolving potassium, dissolving phosphorus and degrading organophosphorus pesticides provided by the invention is detected by adopting a potassium tetraphenylborate gravimetric method in the national standard GB/T8574-2010 of the people's republic of China, and under the condition of a laboratory, after the spores and mycelia of the bacteria are inoculated and cultured for 10 days, the amounts of soluble potassium released from insoluble potassium mineral powder are respectively 158.08 mg.L higher than that of a blank control group^-1And 261.16 mg. L^-1。

Therefore, the invention also relates to the application of the Aspergillus niger JXJ A01 with the activities of dissolving potassium, dissolving phosphorus and degrading organophosphorus pesticides in the soil to convert the potassium which is difficult to dissolve in the soil into the soluble potassium which can be absorbed by plants.

The Aspergillus niger JXJ A01 with the activities of dissolving potassium, dissolving phosphorus and degrading organophosphorus pesticide provided by the invention is detected according to the method of China Ministry of agriculture industry NY/T1847-2010, after spores are inoculated and cultured for 4-10 days, calcium phosphate (20g/L), magnesium phosphate (20g/L), aluminum phosphate (10g/L), iron phosphate (10g/L), calcium phytate (20g/L), lecithin (2 g/L) and phosphorite powder (10g/L, P₂O₅Content is about 25%), the highest content of soluble phosphorus in the culture solution of the culture medium is respectively higher than that of a control group without inoculation by 3784mg/L, 4698mg/L, 813mg/L, 168mg/L, 3082mg/L, 81mg/L and 509mg/L, and the degradation rates of the 7 phosphorus sources are respectively 94.66%, 100%, 32.72%, 15.42%, 59.68%, 100% and 46.61%.

Therefore, the invention also relates to the application of the Aspergillus niger JXJ A01 with the activities of dissolving potassium and phosphorus and degrading organophosphorus pesticides in the soil to convert insoluble organophosphorus and inorganic phosphorus in the soil into soluble phosphorus which can be absorbed by plants.

The spores and the mycelium of the Aspergillus niger JXJ A01 with the activities of decomposing potassium and phosphorus and degrading organophosphorus pesticide provided by the invention are inoculated in a 40% omethoate missible oil phosphorus dissolving culture medium with the volume concentration of 0.1%, and the concentration of soluble phosphate in the culture solution after 5d is respectively 11.77 mg.L higher than that of a control group^-1And 6.79 mg. L^-1。

Therefore, the invention also relates to the application of the Aspergillus niger JXJ A01 with the activities of decomposing potassium and phosphorus and degrading organophosphorus pesticides in soil elimination and improvement and remediation of organophosphorus pesticide contaminated soil.

The Aspergillus niger JXJ A01 with the activities of decomposing potassium and phosphorus and degrading organophosphorus pesticides, provided by the invention, is prepared by inoculating a strain JXJ A01 on sterile wheat bran, culturing, adding a feed after a large number of black spores are formed, and continuously feeding Kunming mice for 4 weeks, wherein the mice grow normally and do not die, so that the bacterium and metabolites thereof have no pathogenicity or toxicity to animals such as mice and the like.

The invention also provides two preparation methods of the Aspergillus niger JXJ A01 solid multifunctional bacterial fertilizer and the soil improvement repairing agent.

The method comprises the following steps: inoculating Aspergillus niger JXJ A01 spores in sterile PDA liquid culture medium or other sterile liquid culture medium (including waste liquid with high BOD value generated in food processing factory, etc.), culturing at 28 deg.C and 160rpm for 1-2 days, spraying and inoculating to mixture of various agricultural wastes (or waste edible fungus chaff, etc.), fowl and livestock feces, phosphate rock powder and potassium ore powder, fermenting at high temperature (or high-temperature steam sterilization), culturing at 30 deg.C and 85% -90% relative humidity for 3-6 days, growing large amount of black spores, drying by natural ventilation drying or hot air drying, adding appropriate amount of trace elements, mixing, and granulating to obtain granules containing large amount of soluble phosphorus and potassium mineral organic matter elements, rich N and N, The high-density aspergillus niger live spore is high-efficiency and environment-friendly multifunctional aspergillus niger bacterial fertilizer.

The second method comprises the following steps: inoculating solid mixture (mass ratio of 60:20:10:10) of various agricultural wastes (or waste edible fungus chaff and the like), livestock and poultry manure, phosphate rock powder, potassium ore powder and the like into Aspergillus niger JXJ A01 liquid strain, stacking and fermenting at high temperature for 5-10 days, and then drying in the sun or naturally drying in the air or drying in hot air for later use. Inoculating aspergillus niger JXJ A01 spores to a sterile PDA (potato dextrose agar) liquid culture medium or other sterile liquid culture media (including waste liquid with a high BOD value and the like generated by a food processing factory and the like), culturing for 2d at 28 ℃ and 160rpm, spraying and inoculating to a sterile wheat bran and straw mixed (the mass ratio is 1:1) solid culture medium, culturing for 3-6d, after a large number of black spores are grown, uniformly mixing with a dry solid mixture stacked and fermented at high temperature according to a certain proportion, and granulating the mixture by using a granulator to obtain the efficient and environment-friendly multifunctional aspergillus niger fertilizer containing a large number of mineral elements such as soluble phosphorus, potassium and the like, rich N and organic matters and high-density aspergillus niger live spores.

The invention has the technical effects that: the Aspergillus niger JXJ A01 is nontoxic, has the effect of converting insoluble potassium, organic phosphorus and inorganic phosphorus into soluble potassium and phosphorus, can reduce organophosphorus pesticide residues in farmlands, improves and repairs farmland soil, can be provided with Aspergillus niger JXJ A01 solid multifunctional bacterial fertilizer and soil improvement and repair agent, and is used as fertilizer and soil repair agent.

Drawings

FIG. 1 is a phosphate solubilizing ring formed by strain JXJ A01 on calcium phosphate and calcium phytate plates.

FIG. 2 shows the degradation capability of the strain JXJ A01 on the insoluble potassium-dissolving ore.

FIG. 3 change of soluble phosphorus content in omethoate culture medium of organophosphorus pesticide after inoculation of JXJ A01 mycelium. Wherein a is the content of the omethoate missible oil with 40 percent and is 0.1 percent (v/v); b is that the content of the omethoate missible oil with 40 percent is 0.2 percent (v/v); c is that the content of the omethoate missible oil with 40 percent is 0.3 percent (v/v); d is that the content of the omethoate missible oil with 40 percent is 0.4 percent (v/v); e is that the content of the omethoate missible oil with 40 percent is 0.5 percent (v/v).

FIG. 4 change of soluble phosphorus content in omethoate medium of organophosphorus pesticide after inoculation of spores of strain JXJ A01. Wherein a is that the content of the omethoate missible oil with 40 percent is 0.1 percent (v/v); b is that the content of the omethoate missible oil with 40 percent is 0.2 percent (v/v).

Detailed Description

1 selection of Aspergillus Phosphonolyticus

Solid plate screening: diluting forest soil collected from Jiujiang City of Jiangxi province with sterile water in a gradient manner, taking 0.2mL of diluent with a proper gradient, and coating the diluent on a phosphate-solubilizing solid culture medium (10g of glucose, (NH) 1847-containing 2010 in the industry standard NY/T1847-₄)₂SO₄0.5g，NaCl 0.3g，MgSO₄·7H₂O 0.3g，KCl 0.3g，FeSO₄·4H₂O 0.036g，MnSO₄·4H₂0.03g of O, 10g of calcium phosphate or calcium phytate, 18g of agar, 1000mL of distilled water and pH7.0), culturing in an incubator at 28 ℃ for 1-7 days, observing the growth condition of bacteria and whether phosphate-solubilizing rings appear around each bacterial colony, and selecting a strain JXJ A01 (figure 1) with obvious phosphate-solubilizing rings for subsequent study.

2 identification of Aspergillus niger JXJ A01

According to the methods of culture characteristics, morphological characteristics, 18S rRNA gene sequence and ITS sequence analysis and the like, the mould JXJ A01 with strong phosphate solubilizing activity is identified. The strain has developed hyphae and many branches, is gray white at the initial stage and then becomes brownish black, and generates brownish black radial conidial heads, conidial peduncles are grown on spherical apical sacs, and clustered black spherical conidia are grown on the conidial peduncles, which belong to the typical morphological characteristics of aspergillus niger. Through comparison and analysis with ITS sequences and 18S rRNA gene sequences in a GenBank database, the similarity of the ITS sequence fragment (591bp) of the strain to Aspergillus niger reaches 100%, the similarity of the 18S rRNA gene sequence (1709bp) to Aspergillus niger reaches 99%, the strain JXJ A01 is identified as Aspergillus (Aspergillus sp.) and named as Aspergillus niger JXJ A01 by integrating the analysis results of morphological characteristics, the ITS sequences and the 18S rRNA gene sequences, the registration numbers of the 18S rRNA gene sequences and the ITS sequences of the strain in the GenBank database are MF145518 and MF543023 respectively, and the specific application is shown in SEQ ID NO:1 and SEQ ID NO: 2. the strain is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.13775 and the preservation date of 2017, 04 months and 20 days. The address of the depository institution: the microbial research institute of

western road

1, 3, China institute of sciences, Japan, in Beijing, Chaoyang, region: 100101, telephone: 86-10-64807355.

3 Potassium-solubilizing Activity of Aspergillus niger JXJ A01

Inoculating strain JXJ A01 spore suspension or mycelium into liquid potassium-decomposing medium (10g glucose, 0.3g NaCl, 1g NaNO)₃，0.5g Na₂HPO₄，0.5g MgSO₄·7H₂O，0.01g FeSO₄·7H₂O, potassium ore10g of powder, 1000mL of distilled water, pH7.4-7.6, sterilization at 121 ℃ for 30min), culturing at 28 ℃ and 160rpm, sampling once every 2d, centrifuging the sample at 4500rpm, and measuring the content of soluble potassium element in the supernatant by adopting a potassium tetraphenylborate gravimetric method in national standard GB/T8574-. With the extension of culture time, potassium mineral powder in the culture medium gradually disappears, soluble potassium element content in the culture solution rapidly increases, and soluble potassium element content in the culture solution inoculated with spore and mycelium after 10d is respectively higher than that in the control group (158.08 + -12.43) mg.L^-1And (261.16. + -. 20.53) mg. L^-1(FIG. 2), the pH of the culture medium was lowered to about 2.4, and therefore, the bacterium had a strong potassium-solubilizing activity, and the production of organic acids was one of the mechanisms for potassium-solubilizing.

4 phosphorus solubilizing Activity of Aspergillus niger JXJ A01

Inoculating Aspergillus niger JXJ A01 spore suspension into liquid phosphate dissolving culture medium (glucose 10g, (NH)₄)₂SO₄0.5g，NaCl0.3g，MgSO₄·7H₂O 0.3g，KCl 0.3g，FeSO₄·4H₂O 0.036g，MnSO₄·4H₂0.03g of O and 1000mL of distilled water, adjusting the pH value to 7.0, and adding a phosphorus source, wherein the iron phosphate, the aluminum phosphate and the powdered rock phosphate are all 10g/L, the lecithin is 2g, or (the glucose is 20g, and the NH content is 20 g)₄)₂SO₄1g，NaCl 0.6g，MgSO₄·7H₂O 0.6g，KCl 0.6g，FeSO₄·4H₂O 0.072g，MnSO₄·4H₂0.06g of O and 1000mL of distilled water, adjusting the pH to 7.0, adding a phosphorus source, wherein the calcium phosphate, the calcium phytate and the magnesium phosphate are all 20g/L) to ensure that the final concentration of the spores is 10⁵～10⁶CFU·mL^-1Culturing at 28 ℃ and 160rpm, sampling once every 24h, centrifuging the sample at 4500rpm, detecting the concentration of soluble effective phosphorus element in the supernatant according to the method of the Chinese Ministry of agriculture industry Standard NY/T1847-2010, adjusting zero by taking deionized water as reference, and taking a blank culture medium without inoculation as a control group. As a result, it was found that the highest content of soluble phosphorus in the culture solution of calcium phosphate, magnesium phosphate, aluminum phosphate, iron phosphate, calcium phytate, lecithin and ground phosphate rock was higher than that in the culture solution of non-cultured calcium phosphate, magnesium phosphate, aluminum phosphate, iron phosphate, calcium phytate, lecithin and ground phosphate rock after 4-10 days of cultureThe control groups of the bacteria have the high contents of 3784mg/L, 4683.2mg/L, 813mg/L, 168mg/L, 3082mg/L, 94mg/L and 512mg/L (Table 1), the degradation rates of the 7 phosphorus sources are respectively 94.66%, 100%, 32.72%, 15.42%, 59.68%, 100% and 47.23%, and the pH of the culture solution is reduced (Table 1), which indicates that the organic acid is generated as one of the phosphate solubilizing mechanisms of the bacteria.

TABLE 1 content (mg. L) of soluble phosphorus in the culture solution after inoculating Aspergillus niger spore culture^-1) And pH change

5 tolerance and degradation effect of Aspergillus niger JXJ A01 on organophosphorus pesticide

Removing phosphorus source in the formula of the phosphate-solubilizing medium of the industry standard NY/T1847-2010 of the Ministry of agriculture, sterilizing, adding 0.1-0.5% (v/v) of 40% omethoate missible oil, inoculating mycelium or spore, culturing at the condition of 28 ℃ and 160rpm, sampling once every 24h, observing the growth condition and the germination condition of the mycelium, centrifuging the sample at 4500rpm, detecting the concentration of soluble effective phosphorus element of the supernatant according to the method of the industry standard NY/T1847-2010 of the Ministry of agriculture, adjusting zero by taking a blank medium without inoculation as a reference, and finding that the mycelium of Aspergillus niger JXJ A01 can survive in the phosphate-solubilizing medium containing 0.3% (v/v) of 40% omethoate missible oil, although the mycelium of the Aspergillus niger is dead in the phosphate-solubilizing medium containing 0.4-0.5% (v/v) of 40% omethoate missible oil, however, the inoculated mycelium still can degrade the omethoate pesticide into soluble phosphorus (figure 3), which shows that the Aspergillus niger JXJ A01 mycelium contains an enzyme for degrading omethoate; spores of Aspergillus niger JXJ A01 could not germinate in a phosphate-solubilizing medium containing 0.3% (v/v) of 40% omethoate emulsifiable concentrate, but when the volume concentration of 40% omethoate emulsifiable concentrate is reduced to 0.2%, the 4 th d of the spores of the bacteria can germinate, when the volume concentration of 40% omethoate emulsifiable concentrate is reduced to 0.1%, the germination inhibition effect on the spores is small, after 1d, the spores germinate and form a large amount of mycelia, and the spores begin to play a role in degrading omethoate immediately after germination, so that the content of soluble phosphorus in the omethoate medium is rapidly increased (figure 4), and the mycelia formed after germination of the spores have stronger degradation effect on omethoate.

6 toxicity of Aspergillus niger JXJ A01 to Kunming mice

The Aspergillus niger JXJ A01 culture solution is inoculated to sterile wheat bran and cultured, after hyphae grow to form a large amount of black spores, the black spores are mixed into feed according to the mass ratio of 1:2, the mixture is continuously fed to Kunming mice for 4 weeks, and the mice grow normally, which indicates that the hyphae, the spores and metabolites thereof of the fungus have no toxicity to animals such as the mice.

Production of 7 Aspergillus niger JXJ A01 bacterial manure

The method comprises the following steps: mixing various dried agricultural wastes (or waste edible fungus chaff and the like), livestock and poultry manure, phosphate rock powder, potassium ore powder and other solids according to the mass ratio of 60:20:10:10, and performing stacking high-temperature fermentation treatment for 5-10 days or high-temperature steam sterilization treatment after uniform mixing for later use. Inoculating aspergillus niger JXJ A01 spores to a sterile PDA (potato dextrose agar) liquid culture medium or other sterile liquid culture media (including waste liquid with a high BOD value and the like generated by a food processing factory and the like), culturing for 2d under the conditions of 28 ℃ and 160rpm, spraying and inoculating to the treated solid mixed culture medium (the thickness of the solid culture medium is about 10-15 cm), culturing for 3-6d under the conditions of about 30 ℃ and 85% -90% of relative humidity, growing a large amount of black spores, drying by natural ventilation or hot air and the like, adding a proper amount of trace elements such as boron, selenium and the like according to the growth requirements of plants, uniformly mixing, and preparing into particles by a granulator to obtain the efficient and environment-friendly multifunctional aspergillus niger bacterial fertilizer containing a large amount of mineral elements such as phosphorus soluble potassium and the like, rich N and organic matters and high-density aspergillus niger live spores.

The second method comprises the following steps: inoculating solid mixture (mass ratio of 60:20:10:10) of various agricultural wastes (or waste edible fungus chaff and the like), livestock and poultry manure, phosphate rock powder, potassium ore powder and the like into Aspergillus niger JXJ A01 liquid strain, stacking and fermenting at high temperature for 5-10 days, and then drying in the sun or naturally drying in the air or drying in hot air until the water content is about 10% -15% for later use. Inoculating Aspergillus niger JXJ A01 spores to a sterile PDA (potato dextrose agar) liquid culture medium or other sterile liquid culture medium (including waste liquid with a high BOD value generated by a food processing factory and the like), culturing at 28 ℃ and 160rpm for 2d, spraying and inoculating the liquid culture medium on a sterile wheat bran and straw mixed solid culture medium (the mass ratio is 1:1), culturing for 3-6d, after a large number of black spores are grown, mixing the liquid culture medium with fermented and dried agricultural wastes (or waste edible fungus chaff and the like), livestock excrement, phosphate rock powder and solid accumulated fermentation products of potassium ore powder according to a certain proportion, adjusting the number of Aspergillus niger live spores in the mixture to be more than 0.2/g, and granulating the mixture by using a granulator to obtain the high-efficiency Aspergillus niger live spores with high density, abundant N and organic matters and abundant mineral matter elements such as soluble phosphorus and potassium, An environment-friendly multifunctional aspergillus niger bacterial fertilizer.

Sequence listing

<110> propylene fire

<120> Aspergillus niger JXJ A01 with activities of potassium and phosphorus dissolving and organophosphorus pesticide degradation and application thereof

<160>2

<170>SIPOSequenceListing 1.0

<210>1

<211>1709

<212>DNA

<213> Aspergillus niger (Aspergillus niger)

<400>1

agaaagcatg tctagtataa gcactttata ctgtgaaact gcgaatgggc tcattaaatc 60

agttatcgtt tatttgatag taccttacta catggatacc tgtggtaatt ctagagctaa 120

tacatgctga aaacctcgac ttcggaaggg gtgtatttat tagataaaaa accaatgccc 180

ttcggggctc cttggtgaat cataataact taacgaatcg catggccttg cgccggcgat 240

ggttcattca aatttctgcc ctatcaactt tcgatggtag gatagtggcc taccatggtg 300

gcaacgggta acggggaatt agggttcgat tccggagagg gagcctgaga aacggctacc 360

acatccaagg aaggcagcag gcgcgcaaat tacccaatcc cgacacgggg aggtagtgac 420

aataaatact gatacggggc tcttttgggt ctcgtaattg gaatgagtac aatctaaatc 480

ccttaacgag gaacaattgg agggcaagtc tggtgccagc agccgcggta attccagctc 540

caatagcgta tattaaagtt gttgcagtta aaaagctcgt agttgaacct tgggtctggc 600

tggccggtcc gcctcaccgc gagtactggt ccggctggac ctttccttct ggggaatctc 660

atggccttca ctggctgtgg ggggaaccag gacttttact gtgaaaaaat tagagtgttc 720

aaagcaggcc tttgctcgaa tacattagca tggaataata gaataggacg tgcggttcta 780

ttttgttggt ttctaggacc gccgtaatga ttaataggga tagtcggggg cgtcagtatt 840

cagctgtcag aggtgaaatt cttggatttg ctgaagacta actactgcga aagcattcgc 900

caaggatgtt ttcattaatc agggaacgaa agttagggga tcgaagacga tcagataccg 960

tcgtagtctt aaccataaac tatgccgact agggatcgga cggtgtttct attatgaccc 1020

gttcggcacc ttacgagaaa tcaaagtttt tgggttctgg ggggagtatg gtcgcaaggc 1080

tgaaacttaa agaaattgac ggaagggcac caccaggcgt ggagcctgcg gcttaatttg 1140

actcaacacg gggaaactca ccaggtccag acaaaataag gattgacaga ttgagagctc 1200

tttcttgatc ttttggatgg tggtgcatgg ccgttcttag ttggtggagt gatttgtctg 1260

cttaattgcg ataacgaacg agacctcggc ccttaaatag cccggtccgc atttgcgggc 1320

cgctggcttc ttagggggac tatcggctca agccgatgga agtgcgcggc aataacaggt 1380

ctgtgatgcc cttagatgtt ctgggccgca cgcgcgctac actgacaggg ccagcgagta 1440

catcaccttg gccgagaggt ctgggtaatc ttgttaaacc ctgtcgtgct ggggatagag 1500

cattgcaatt attgctcttc aacgaggaat gcctagtagg cacgagtcat cagctcgtgc 1560

cgattacgtc cctgcccttt gtacacaccg cccgtcgcta ctaccgattg aatggctcgg 1620

tgaggccttc ggactggctc aggagggttg gcaacgaccc cccagagccg gaaagttggt 1680

caaacccggt cattagagaa gtaagttga 1709

<210>2

<211>591

<212>DNA

<213> Aspergillus niger (Aspergillus niger)

<400>2

tgaacctgcg gaaggatcat taccgagtgc gggtcctttg ggcccaacct cccatccgtg 60

tctattatac cctgttgctt cggcgggccc gccgcttgtc ggccgccggg ggggcgcctt 120

tgccccccgg gcccgtgccc gccggagacc ccaacacgaa cactgtctga aagcgtgcag 180

tctgagttga ttgaatgcaa tcagttaaaa ctttcaacaa tggatctctt ggttccggca 240

tcgatgaaga acgcagcgaa atgcgataac taatgtgaat tgcagaattc agtgaatcat 300

cgagtctttg aacgcacatt gcgccccctg gtattccggg gggcatgcct gtccgagcgt 360

cattgctgcc ctcaagcccg gcttgtgtgt tgggtcgccg tccccctctc cggggggacg 420

ggcccgaaag gcagcggcgg caccgcgtcc gatcctcgag cgtatggggc tttgtcacat 480

gctctgtagg attggccggc gcctgccgac gttttccaac cattttttcc aggttgacct 540

cggatcaggt agggataccc gctgaactta agcatatcaa taagcggagg a 591

Claims

1. The Aspergillus niger JXJA01 with the activities of dissolving potassium and phosphorus and degrading organophosphorus pesticides is characterized in that the Aspergillus niger JXJA01 is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No.13775 and the preservation date of 2017, 04 and 20 days.

2. The use of aspergillus niger JXJA01 having activities of potassium solubilizing, phosphorus solubilizing and organophosphorus pesticide degrading according to claim 1, wherein: is used for converting insoluble potassium into soluble potassium and promoting the absorption of potassium element by plants.

3. The use of aspergillus niger JXJA01 having activities of potassium solubilizing, phosphorus solubilizing and organophosphorus pesticide degrading according to claim 1, wherein: the organic phosphorus fertilizer is used for converting insoluble organic phosphorus into soluble phosphorus and promoting the absorption of phosphorus elements by plants.

4. The application of aspergillus niger JXJA01 with activities of dissolving potassium and phosphorus and degrading organophosphorus pesticides as claimed in claim 1, which is used for converting various insoluble inorganic phosphorus into soluble phosphorus and promoting phosphorus element absorption of plants.

5. The use of aspergillus niger JXJA01 having activities of potassium solubilizing, phosphorus solubilizing and organophosphorus pesticide degrading according to claim 1, wherein: the organic phosphorus degrading agent is used for degrading organic phosphorus pesticide into nontoxic soluble phosphate, reducing pesticide residue in soil, and improving and restoring pesticide pollution of soil.

6. A preparation method of an Aspergillus niger JXJA01 solid multifunctional bacterial fertilizer and a soil improvement repairing agent is characterized in that: inoculating the spores of Aspergillus niger JXJA01 in a sterile PDA liquid culture medium or other sterile liquid culture medium, culturing at 28 ℃ and 160rpm for 1-2 days, spraying and inoculating a mixture of various agricultural wastes, livestock and poultry manure, phosphate rock powder and potassium ore powder subjected to high-temperature stacking fermentation treatment or high-temperature steam sterilization treatment, culturing at 30 ℃ and 85% -90% of relative humidity for 3-6 days, growing a large amount of black spores, drying, adding a proper amount of trace elements according to plant growth requirements, uniformly mixing, and granulating by using a granulator to obtain the multifunctional Aspergillus niger bacterial fertilizer containing a large amount of soluble potassium elements, phosphorus elements and Aspergillus niger live spores.

7. A preparation method of an Aspergillus niger JXJA01 solid multifunctional bacterial fertilizer and a soil improvement repairing agent is characterized in that: inoculating solid mixture of various agricultural wastes, livestock and poultry feces, ground phosphate rock and potassium ore powder into liquid strain containing Aspergillus niger JXJA01 of claim 1, stacking for high temperature fermentation for 5-10 days, and sun drying or naturally air drying or hot air drying for later use; inoculating the spores of Aspergillus niger JXJA01 in a sterile PDA liquid culture medium or other sterile liquid culture medium, culturing at 28 ℃ and 160rpm for 2 days, spraying and inoculating the spores onto a sterile wheat bran and straw mixed solid culture medium, culturing for 3-6 days, after a large number of black spores are grown, uniformly mixing the black spores with a dry solid mixture subjected to high-temperature fermentation treatment according to a certain proportion, and granulating the mixture by using a granulator to obtain the multifunctional Aspergillus niger bacterial fertilizer containing a large number of soluble potassium elements, phosphorus elements and Aspergillus niger live spores.

8. The preparation method of the aspergillus niger JXJA01 solid multifunctional bacterial manure and the soil improvement repairing agent according to claim 7, wherein the preparation method comprises the following steps: the mass ratio of the agricultural wastes to the livestock and poultry manure to the phosphate rock powder to the potassium ore powder is 60:20:10: 10.

9. The preparation method of the aspergillus niger JXJA01 solid multifunctional bacterial manure and the soil improvement repairing agent according to claim 7, wherein the preparation method comprises the following steps: the mass ratio of wheat bran to straws in the wheat bran and straw mixed solid culture medium is 1: 1.