CN117551596A - Method for relieving bacteriostatic action of soil on biocontrol fungi - Google Patents
Method for relieving bacteriostatic action of soil on biocontrol fungi Download PDFInfo
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N3/00—Spore forming or isolating processes
-
- 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
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P5/00—Nematocides
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/79—Paecilomyces
Abstract
The invention discloses a method for relieving the bacteriostatic action of soil on biocontrol fungi, which relates to the technical field of microbial pesticides, and improves the spore germination rate of biocontrol fungi under the inhibition of soil by activating Adenosine 5' -monophosphate-activated protein kinase (AMPK protein kinase for short) of fungi by an activator; the AMPK activator is any one of acadine or metformin. The AMPK activator can be used as a functional auxiliary agent of fungus biopesticide, improves the spore germination rate of the fungus biopesticide under the inhibition of soil, and can be used for biological control of soil-borne diseases.
Description
Technical Field
The invention relates to the technical field of microbial pesticides, in particular to a method for relieving the bacteriostatic action of soil on biocontrol fungi.
Background
Soil-borne diseases caused by pathogenic fungi, oomycetes, parasitic nematodes and the like cause a large amount of economic losses each year (Lopes EA, ellen JC, gomes VA, bruno SV, parreira DF, neves WS. 2022. Anaerobic soil disinfestation for the management of soilborne pathogens: A review. Applied Soil Ecology, 174,104408.). Pathogenic fungi of the genus Fusarium (Fusarium), rhizoctonia (Rhizoctonia), and the genus Phytophthora (Phytophthora), pythum (Pythum), and the like often cause rot, cataplexy (Rhizoctonia), and other soil-borne diseases of cereal and vegetable crops (Stefanie L, barbara T, thomas O, johanna M, jacques GF, franco W, florian MF, christian HA 2020, harnessing the Microbiomes of Suppressive Composts for Plant Protection: from Metagenomes to Beneficial Microorganisms and Reliable diagnostics Frontiers in Microbiology, 11, 1810.). Root knot nematode disease caused by Root knot nematode infestation of plant roots is another major class of soil-borne diseases (Moens M, perry RN, starr JL. 2009. Meloidoyne species-a diverse group of novel and important plant antigens: in: perry RN, moens M, starr JL, eds. Root-knot nematodes. London, UK: CAB International, 1-13.), the most harmful 4 species of the genus Meloidogyne: meloidogyne incognita (m. Incognita), meloidogyne arachidis hypogaeae (m. Arenicia), meloidogyne northern (m. Hapla) and meloidogyne javanica (m. Javanica).
Pathogenic microorganisms causing soil-borne diseases can survive in soil for many years through the formation of a variety of propagules such as sclerotium, chlamydospores, oospores, eggs and cysts, and are difficult to control. Chemical pesticides have long played an absolute leading role in the control of crop soil-borne diseases. However, the problems of agricultural products and ecological pollution are very prominent due to the high toxicity and long-term excessive use of chemical pesticides. Therefore, the use of chemical pesticides is reduced. Biological control is an important prevention and control means for replacing chemical pesticides and guaranteeing stable and high yield of crops due to the advantages of no pesticide residue, environmental protection, difficult generation of drug resistance and the like.
Trichoderma fungi such as Trichoderma harzianum and Trichoderma viride (Ferriera FV, musumeci MA. 2021 Trichoderma as biological control agent: scope and prospects to improve efaciens World J Microbiol Biotechnol, 37 (5), 90.) are commonly used for controlling soil-borne pathogenic microorganisms such as Fusarium, phytophthora, botrytis and Rhizoctonia. The biological control Fungi such as the sarcomere oligospora (Arthrobotrys oligospora), the Pochonia chlamydosporia (Pochonia chlamydosporia) and the Paecilomyces lilacinus (Purpureocillium lilacinum) are mainly used for preventing and controlling root-knot nematodes (Zhang Y, li S, li H, wang R, zhang KQ, U.S. J.2020. Fungi-Nematode Interactions: diversity, ecology, and Biocontrol Prospects in agricultural, journal of Fungi (Basel), 6 (4), 206). However, most microbial agents have well known common problems: the field control effect is low and the control effect is unstable (Ruan Yingying, liu Feng, 2020. The action mechanism and the application research progress of trichoderma biological control. Zhejiang agricultural science, 61 (11), 2290-2294).
The precondition of biocontrol fungi for preventing and controlling soil-borne diseases is that the fungus agent can germinate and grow well in soil so as to establish enough population density. However, germination and growth of biocontrol agents in soil can be severely inhibited by "soil bacteriostasis" (bonomi G, gaglione SA, incerti G, zoina a.2013. Biochemical quality of organic amendments affects soil furtistasis Applied Soil Ecology, 72, 135-142.). Inhibition strength of hundreds of agricultural soils for various vegetation against various biocontrol fungi was evaluated: the germination rate of most fungi in soil is lower than 50%; the inhibition rate of all soil samples on the germination of three trichoderma biocontrol fungi (Trichoderma harzianum, trichoderma viride, trichoderma hamatum) and paecilomyces lilacinus (P. Lilacinum) spores is above 95%; the average inhibition rate of all soil samples on spore germination of Pochonia chlamydosporia (P. Chlamydosporia) was more than 70%. Therefore, the inhibition of the soil on the biocontrol fungi is relieved, so that the spore germination rate of the biocontrol fungi spores in the soil is improved, and the method is very important for improving and stabilizing the field control effect of the biocontrol fungi.
The results of the last 70 years of research indicate that: lack of soil nutrition; abiotic factors related to soil physicochemical properties such as soil pH, heavy metal ions and the like; and bacteriostatic factors derived from soil microorganisms mediate the formation of soil bacteriostasis (Garbeva P, hol WHG, termorsumizen AJ, kowalchuk GA, boer WD. 2011. Fungistasis and general soil biostasis-A new systems Soil Biology and Biochemistry, 43 (3), 469-477.). For the elimination of SF of the bacteriostatic action of the soil, studies have found that the addition of nutrients required by microorganisms can temporarily eliminate SF (Lockwood JL. 1977. Fungistasis in solids Biological Reviews, 52 (1), 1-43); SF (Wu M, zhang H, li X, zhang Y, su Z, zhang C.2008. Soil fungistasis and its relations to soil microbial composition and diversity: a case study of a series of soils with different fungstasis. Journal of Environmental Sciences, 20 (7), 871-877.) can also be relieved by heat sterilization and chemical fumigation of the soil to reduce the microbial species and numbers thereof. However, these disarming methods do not work well in field practice for reasons including: the added nutrient substances are rapidly consumed by indigenous microorganisms, and the action time is too short to enable the biocontrol fungi to establish enough population density; the soil sterilization modes such as chemical fumigation and the like have great ecological damage to soil microorganisms, and are easy to cause rampant outbreaks of soil-borne diseases. At present, a method for effectively relieving the inhibition effect of soil on biocontrol fungi is lacking.
AMPK protein kinase plays an important role in physiological activity and signal transduction in eukaryotes (Alao JP, legon L, dabrowska a, tricoloci a M, kumar J, rallis c.2023. Interplays of AMPK and TOR in Autophagy Regulation in yeast, cells, 12 (4), 519.), academine (Drew BG, kingwell ba.2008, academine, an adenosine-regulating agent with the potential for widespread indications, expert Opin Pharmacother, 9 (12), 2137-2144) and Metformin (Ma T, tian X, zhang B, li M), wang Y, yang C, wu J, wei X, qu Q, yu Y, long S, feng JW, li C, zhang C, xie C, wu Y, xu Z, chen J, yu Y, huang X, he Y, yao L, zhang L, zhu M, wang W, wang ZC, zhang M, bao Y, jia W, linSY, ye Z, piao HL, deng X, zhang CS, lin SC. 2022, low-dose Metformin targets the lysosomal AMPK pathway through PEN. Nature, 603 (7899), 159-165 and Foretz M, guide B, bertrand L, pollak M, viollet B.2014. Metfor: from mechanisms of action to therapies, cell metanolism, 20 (6), 953-966) are activators of AMPK protein kinase, a method for relieving the inhibition of soil to fungal spore germination by using an AMPK activator and application of the method in soil-borne disease control are not reported.
Disclosure of Invention
The invention aims to provide a method for relieving the inhibition effect of soil on biocontrol fungi, which helps to improve the colonization abundance of biocontrol fungi in soil.
In order to achieve the purposes of the application, the technical scheme adopted by the application is as follows: the invention discloses a method for relieving the bacteriostatic action of soil on biocontrol fungi, which discovers that the AMPK activator of 2-4 mM can improve the spore germination rate of biocontrol fungi under the inhibition of soil, and can relieve the inhibition of soil on the spore germination of biocontrol fungi and improve the colonization abundance of biocontrol fungi in soil by directly spraying an aqueous solution of the AMPK activator into the soil in a spraying manner.
The invention discloses a method for relieving the bacteriostatic action of soil on biocontrol fungi, which is characterized by comprising the following steps: the spore germination rate of the biocontrol fungi under the inhibition of soil is improved by directly adding an AMPK activator into the soil to activate the AMPK protein kinase of the fungi, wherein the AMPK activator is any one of 2-4 mM/LAcadesine (alcalidin, 5-aminoimidazole-4-formamide-1-B-D-ribofuranoside) or 2-4 mM/L metformin.
The AMPK activator comprises acadine or metformin, can be used as a functional auxiliary agent of fungus biopesticide, and can improve the colonization abundance of biocontrol fungi under the inhibition of soil.
Further, the biocontrol fungi suitable for the AMPK activator comprise one or a combination of more of nematode biocontrol fungi paecilomyces lilacinus, podoconcha chlamydosporium or sarcophagospora oligospora.
The beneficial effects are that: the invention can effectively relieve the inhibition of the soil on spore germination of fungus biopesticide, improve the colonization abundance of biocontrol fungi in the soil, and can be used for biological control of root-knot nematodes.
Compared with the prior art, the invention has the following advantages:
(1) The invention has simple application and long duration of the release effect on the soil bacteriostasis;
(2) The invention can not destroy the microbial ecology of the soil.
Detailed Description
In order to make the technical problems, technical schemes and beneficial effects to be solved by the present application more clear, the present application is further described in detail below with reference to the embodiments. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the present application.
The invention discloses a method for relieving the bacteriostatic action of soil on biocontrol fungi, which discovers that the AMPK activator of 2-4 mM can improve the spore germination rate of biocontrol fungi under the inhibition of soil, and can relieve the inhibition of soil on the spore germination of biocontrol fungi and improve the colonization abundance of biocontrol fungi in soil by directly spraying an aqueous solution of the AMPK activator into the soil in a spraying manner.
The invention discloses a method for relieving the bacteriostatic action of soil on biocontrol fungi, which is characterized by comprising the following steps: the spore germination rate of the biocontrol fungi under the inhibition of soil is improved by directly adding an AMPK activator to the soil to activate the AMPK protein kinase of the fungi, wherein the AMPK activator is any one of 2-4 mM/L academe (acardite, 5-aminoimidazole-4-formamide-1-B-D-ribofuranoside) or 2-4 mM/L metformin.
The AMPK activator comprises 5-aminoimidazole-4-formamide-1-B-D-ribofuranoside acadine or metformin, can be used as a functional auxiliary agent of fungus biopesticide, and can improve the colonization abundance of biocontrol fungi under the soil inhibition.
The biocontrol fungi suitable for the AMPK activator comprise nematode biocontrol fungi paecilomyces lilacinus (Purpureocillium lilacinum, girardi NS, sosa AL, etheverry MG, pasone MA.. 2022. In vitro characterization bioassays of the nematophagous fungus Purpureocillium lilacinum: evaluation on growth, extracellular enzymes, 823-mat. Fungal Biol, 126 (4), 300-307), pochonia chlamydosporia (Pochonia chlamydosporia, bo T, kong C, zouS, mo M, liu Y, 2022. Bacillus nematocida B16 Enhanced the Rhizosphere Colonization of Pochonia chlamydosporia ZK 7-Knot Nematode Meloidogyne incognita, micro-organs, 10 (2), 218.) and oligospora (Arthrobotrys oligospora, yang J, wang L, ji X, feng Y, li X, zou C, xu J, ren Y, mi Q, wu J, liu S, liu Y, huang X, wah, niu X, li J, liang L, luo Y, yu K, Y, yu W, 92, Z.35, Z.37, Z.20119) or a combination of two species of the fungi.
Example 1
AMPK activator is directly added into soil suspension
According to the method for relieving the bacteriostatic action of the soil on the biocontrol fungi, disclosed by the invention, the spore germination rate of the biocontrol fungi under the inhibition of the soil is improved by activating the Adenosine 5' -monophosphate-activated protein kinase (AMPK protein kinase for short) of the fungi by the AMPK activator; the AMPK activator includes acadine or metformin.
The AMPK activator acadine or metformin is used as a functional auxiliary agent of fungus biopesticide, so that the spore germination rate of the biocontrol fungus under the inhibition of soil is improved.
The biocontrol fungus strain is a paecilomyces lilacinus strain, a Pochonia chlamydosporia strain and a Nostoc oligosporus strain,
collecting conidium: inoculating three strains of bacteria on PDA culture medium contained in 250ml triangular flask, culturing at 28deg.C for 7-14 days, adding sterilized water and glass beads, slightly shaking and vibrating to make conidium fall off, collecting liquid, centrifuging to obtain conidium, adding appropriate amount of water, and counting by blood cell counting plate to adjust spore concentration to 1-2×10 5 Individual spores/mL.
Preparing soil suspension: collecting soil with 20cm surface layer from the farmland, weighing 1 kg of soil, placing in a beaker, adding 1000ml of sterile water, stirring for 30 minutes, standing for 24 hours, and sucking the upper liquid to obtain soil suspension for testing the antibacterial strength of the soil.
Spore germination rate test. Control group: 1ml of the spore suspension was put into a protein dialysis bag, then the spore suspension was placed into 200ml of the prepared soil suspension, and after 24 hours at 28 degrees, the spore suspension was sucked from the dialysis bag, and the spore germination rate was observed and counted by a microscope. Experimental group: the AMPK activator, academine or metformin, was added to 200ml of the above soil suspension to achieve a final concentration of 5-aminoimidazole-4-carboxamide-1-B-D-ribofuranoside, academine, of 2mM/L or metformin, of 4mM/L, and then added to a dialysis bag containing 1ml of spore suspension as in the control group, and after standing at 28℃for 24 hours, the spore germination rate was observed and counted by microscopy.
Example 2
Example 2 differs from example 1 in that: the spore germination rate of the biocontrol fungi under the inhibition of soil is improved by directly adding an AMPK activator into the soil to activate the AMPK protein kinase of the fungi, wherein the AMPK activator is any one of 3 mM/L5-aminoimidazole-4-formamide-1-B-D-ribofuranoside or 2mM/L metformin.
Example 3
Example 3 differs from example 1 in that: the spore germination rate of the biocontrol fungi under the inhibition of soil is improved by directly adding an AMPK activator into the soil to activate the AMPK protein kinase of the fungi, wherein the AMPK activator is 4 mM/L5-aminoimidazole-4-formamide-1-B-D-ribofuranoside.
Example 4
Example 4 differs from example 1 in that: the AMPK activator is any one of 3mM/L metformin.
Test example 1
Experiment of bacteriostasis of AMPK activator academine and metformin on biocontrol fungi by soil release
Soil sampling site: the Yunnan university is under the control.
Test strain: purpureocillium lilacinum (accession number: YMF 1.00943), arthrobotrys oligospora (accession number: YMF 1.00051), pochonia chlamydosporia (accession number: YMF 1.06480), conidia of the three strains were collected for testing.
The experimental method comprises the following steps: three treatments were designed altogether.
Process 1
Soil suspension 200ml, without addition of acadine or metformin, as a control, conidium of three biocontrol fungi (number 1-2×10 5 And respectively placing the obtained materials in dialysis bags, immersing the dialysis bags in the soil suspension, sampling after 24 hours to detect spore germination rate, and sampling 3 parts of conidia of each biocontrol fungus.
Process 2
200 to ml of soil suspension, adding 2 to mM/L of acadine, and 1 to 2X 10 conidia of three biocontrol fungi 5 And respectively placing the obtained materials in dialysis bags, immersing the dialysis bags in the soil suspension, sampling after 24 hours to detect spore germination rate, and sampling 3 parts of conidia of each biocontrol fungus.
Process 3
200 to ml to which metformin with a final concentration of 4 to mM/L was added, and conidia of three biocontrol fungi (number 1 to 2X 10 5 And respectively placing the obtained materials in dialysis bags, immersing the dialysis bags in the soil suspension, sampling after 24 hours to detect spore germination rate, and sampling 3 parts of conidia of each biocontrol fungus.
Experimental results of the agadine and metformin enhancement of spore germination rate of conidia in soil suspension are shown in table 1:
TABLE 1
| Strain | Process 1 | Process 2 | Process 3 |
| Arthrobotrys oligospora | 42.7% | 75.3% | 69.3% |
| Purpureocillium lilacinum | 12.5% | 25.6% | 21.3% |
| Pochonia chlamydosporia | 16.3% | 30.1% | 26.8% |
As shown in table 1, compared with the control treatment 1, both academine and metformin can significantly improve the spore germination rate of the conidia of the three biocontrol fungi in the soil suspension, namely, the academine and the metformin have a significant relieving effect on the bacteriostasis of the soil on the three biocontrol fungi.
Test example 2
Soil sampling site: county of eyebrow mountain is Sichuan province.
Test strain: purpureocillium lilacinum (accession number: YMF 1.00943), arthrobotrys oligospora (accession number: YMF 1.00051), pochonia chlamydosporia (accession number: YMF 1.06480), conidia of the three strains were collected for testing.
The experimental method comprises the following steps: three treatments were designed altogether.
Process 4
Soil suspension 200ml, without addition of acadine or metformin, as a control, conidium of three biocontrol fungi (number 1-2×10 5 And respectively placing the obtained materials in dialysis bags, immersing the dialysis bags in the soil suspension, sampling after 24 hours to detect spore germination rate, and sampling 3 parts of conidia of each biocontrol fungus.
Process 5
Soil suspension200ml, adding 2mM/L of academine, and 1-2×10 conidia of three biocontrol fungi 5 And respectively placing the obtained materials in dialysis bags, immersing the dialysis bags in the soil suspension, sampling after 24 hours to detect spore germination rate, and sampling 3 parts of conidia of each biocontrol fungus.
Process 6
200 to ml to which metformin with a final concentration of 4 to mM/L was added, and conidia of three biocontrol fungi (number 1 to 2X 10 5 And respectively placing the obtained materials in dialysis bags, immersing the dialysis bags in the soil suspension, sampling after 24 hours to detect spore germination rate, and sampling 3 parts of conidia of each biocontrol fungus.
Experimental results of the agadine and metformin enhancement of spore germination rate of conidia in soil suspension are shown in table 2:
TABLE 2
| Strain | Process 4 | Process 5 | Process 6 |
| Arthrobotrys oligospora | 35.4% | 64.1% | 58.8% |
| Purpureocillium lilacinum | 8.6% | 18.4% | 15.2% |
| Pochonia chlamydosporia | 12.3% | 22.6% | 20.4% |
As shown in table 2, compared with the control group treatment 4, both academine and metformin can significantly improve the spore germination rate of the conidia of the three biocontrol fungi in the soil suspension, namely, the academine and the metformin have a significant relieving effect on the bacteriostasis of the soil on the three biocontrol fungi.
Test example 3
Spraying the AMPK activator aqueous solution into the soil
Example 1:
the soil from the Guanyan university is split into round flowerpots with the diameter of 20 cm/25 cm, and each flowerpot is split into 3 kg of soil, and the treatment is divided into 4 treatments:
treatment 1: sterile water 500 and ml is poured into the soil of the flowerpot.
Treatment 2: 1g of freshly collected oligospore is stirred and mixed evenly from conidium and soil in a flowerpot, and then 500ml of water is uniformly sprayed on the soil.
Treatment 3: 1g of freshly collected oligospore was stirred and mixed uniformly from the conidium and the soil in the pot, then 400ml of water was uniformly applied to the soil, and 100ml of metformin at a concentration of 4mM/L was applied.
Treatment 4: 1g of freshly collected oligospore was stirred and mixed uniformly from the conidium and the soil in the pot, then 400ml of water was uniformly applied to the soil, and 100ml of 2mM/L of academine was applied.
3 replicates of each treatment were placed in a greenhouse, periodically watered, kept at 30-40% soil humidity, after 50 days of placement 20g of soil samples were taken from the soil in the middle of each pot, 3 replicates of each treatment were mixed into one sample, the samples were stored in a 4 degree refrigerator, and then sent to the Shanghai Meiji biological medicine technologies Co., ltd for sequencing of the soil microbiome, 5 sequencing replicates were set for each sample, after sequencing, the relative abundance of fungi in the soil was analyzed at the genus level, and the results are shown in Table 3. The results show that the relative abundance of the Arthrobotrys fungus to which the S.oligospora belongs after metformin (treatment 3) application is 8.1% significantly higher than that of the control group (treatment 2) (77.1% improvement). After the application of acadine (treatment 4), the relative abundance of the fungi of the genus Arthrobotrys was 5.81%, which was 26.9% higher than that of the control group (treatment 2). The effect of metformin or academine on Arthrobotrys oligospora soil colonisation abundance is shown in table 3:
TABLE 3 Table 3
As shown in table 3: treatment 1, control soil without any material or bacterial liquid added. Treatment 2, soil to which Arthrobotrys oligospora conidia were applied. Treatment 3, soil to which metformin and Arthrobotrys oligospora conidia were applied. Treatment 4, soil to which acadine and Arthrobotrys oligospora conidia were applied.
The foregoing has shown and described the basic principles, principal features and advantages of the invention. It will be understood by those skilled in the art that the foregoing test examples and descriptions are merely illustrative of the principles of the present invention and that various changes and modifications may be made therein without departing from the spirit and scope of the invention, the scope of which is defined in the appended claims, specification and their equivalents.
Claims (2)
1. A method for relieving the bacteriostatic action of soil on biocontrol fungi is characterized by comprising the following steps: the spore germination rate of the biocontrol fungi under the inhibition of soil is improved by directly adding an AMPK activator into the soil to activate the AMPK protein kinase of the fungi, wherein the AMPK activator is any one of 2-4 mM/L5-aminoimidazole-4-formamide-1-B-D-ribofuranoside or 2-4 mM/L metformin.
2. The method for relieving the bacteriostatic action of soil on biocontrol fungi according to claim 1, which is characterized by comprising the following steps: the biocontrol fungi comprise one or a combination of a plurality of paecilomyces lilacinus, pochonia chlamydosporia or Arthropoda oligospora.
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