Nothing Special   »   [go: up one dir, main page]

CN116083297A - Microbial agent for preventing and treating strawberry root rot and preparation method thereof - Google Patents

Microbial agent for preventing and treating strawberry root rot and preparation method thereof Download PDF

Info

Publication number
CN116083297A
CN116083297A CN202211532735.XA CN202211532735A CN116083297A CN 116083297 A CN116083297 A CN 116083297A CN 202211532735 A CN202211532735 A CN 202211532735A CN 116083297 A CN116083297 A CN 116083297A
Authority
CN
China
Prior art keywords
bacillus
root rot
paenibacillus polymyxa
pseudomycoides
paenibacillus
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202211532735.XA
Other languages
Chinese (zh)
Inventor
杨瑞先
刘萍
李娟�
卢明媚
孙高峰
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Luoyang Institute of Science and Technology
Original Assignee
Luoyang Institute of Science and Technology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Luoyang Institute of Science and Technology filed Critical Luoyang Institute of Science and Technology
Priority to CN202211532735.XA priority Critical patent/CN116083297A/en
Publication of CN116083297A publication Critical patent/CN116083297A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION 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
    • A01N63/00Biocides, 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
    • A01N63/20Bacteria; Substances produced thereby or obtained therefrom
    • A01N63/22Bacillus
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION 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
    • A01N63/00Biocides, 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
    • A01N63/20Bacteria; Substances produced thereby or obtained therefrom
    • A01N63/25Paenibacillus
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01PBIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P3/00Fungicides
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12RINDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00Microorganisms ; Processes using microorganisms
    • C12R2001/01Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07Bacillus
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12RINDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00Microorganisms ; Processes using microorganisms
    • C12R2001/01Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07Bacillus
    • C12R2001/12Bacillus polymyxa ; Paenibacillus polymyxa

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Environmental Sciences (AREA)
  • Plant Pathology (AREA)
  • Virology (AREA)
  • Pest Control & Pesticides (AREA)
  • Agronomy & Crop Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Dentistry (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Mycology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

A microbial agent for preventing and treating strawberry root rot is prepared from Paenibacillus polymyxa Paenibacillus polymyxa and Bacillus pseudolaricis Bacillus pseudomycoides by compounding, wherein the preservation number of Paenibacillus polymyxa Paenibacillus polymyxa is CGMCC No.25858, and the preservation number of Bacillus pseudolaricis Bacillus pseudomycoides is CGMCC No.25859. The preparation method comprises the following steps: after the two bacteria are fermented and cultured, the ratio of the two bacteria is 1: the compound of the proportion 1 is wettable powder, and the application mode is root irrigation. The microbial agent prepared by the invention can effectively prevent and treat strawberry root rot, promote strawberry plant growth, and can not cause pathogenic bacteria drug resistance generation, pesticide residues and environmental pollution after use.

Description

Microbial agent for preventing and treating strawberry root rot and preparation method thereof
Technical Field
The invention relates to the technical field of agricultural microorganisms, in particular to a microbial agent for preventing and treating strawberry root rot and a preparation method thereof.
Background
Strawberry is an important commercial crop, and in recent years, strawberry is changed from a single open field cultivation mode to a facility cultivation mode with an increase in planting area and an adjustment in planting structure. However, due to the characteristics of low temperature, high humidity, sealing, continuous cropping planting and annual production in the strawberry facility cultivation process, the physical and chemical properties of the planting soil are deteriorated, the number of pathogenic bacteria is increased in an accumulated manner, so that diseases are serious, the quality and the yield of strawberries are seriously influenced, and the sustainable development of the strawberry industry is restricted.
Strawberry root rot is a soil-borne disease with serious harm and wide influence range, and is a main factor causing the reduction of the yield and quality of strawberries. The strawberry root rot field disease symptoms are complex, pathogenic bacteria causing disease are more, disease can be caused in the whole growth period of the strawberry, the strawberry root rot field disease has the characteristics of sudden and destructive disease, once the disease happens, the disease is difficult to control, and huge loss is brought to strawberry production. At present, the strawberry variety with high root rot resistance is lacking, so that the control of the strawberry root rot is mainly chemical control, and the green control technology with biological control as a core is also lacking in efficient microbial inoculum products and application technology thereof.
In recent years, the problem of pesticide residues generated by preventing crop diseases by chemical agents and the related food safety problem caused by the problem are very prominent, and the biocontrol microbial inoculum is increasingly applied to preventing and controlling crop diseases due to the characteristics of broad-spectrum disease prevention effect, multiple disease resistance mechanisms and environmental friendliness, so that the biocontrol microbial inoculum becomes an important resource for reducing or replacing pesticides. Therefore, development of a high-efficiency biocontrol microbial agent is needed, and the biocontrol microbial agent is applied to green prevention and control of strawberry root rot, so that sustainable development of the strawberry industry is strongly promoted.
Disclosure of Invention
The invention aims to provide a microbial agent for preventing and treating strawberry root rot and a preparation method thereof, and the microbial agent can be used for effectively preventing and treating strawberry root rot so as to promote the growth of strawberry plants; in addition, the microbial agent has the characteristics of environmental protection and no pesticide residue, and can prevent pathogenic bacteria from generating drug resistance after use.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
a microbial agent for preventing and treating strawberry root rot is prepared from Paenibacillus polymyxa Paenibacillus polymyxa and Bacillus pseudolaris Bacillus pseudomycoides through compounding, and is prepared by separating and preserving in the experiment. The Paenibacillus polymyxa Paenibacillus polymyxa is preserved in China general microbiological culture Collection center (CGMCC) at a preservation number of CGMCC No.25858 and at a preservation address of North Chen Xili No.1 and 3 in the Korean region of Beijing in 2022, 09 and 30 days; the bacillus pseudomycosis Bacillus pseudomycoides is preserved in China general microbiological culture Collection center (CGMCC) at 30-09-2022, and has a preservation number of CGMCC No.25859 and a preservation address of North Chenxi Lu No. 1-3 in the Korean region of Beijing city.
Preferably, the mass ratio of the paenibacillus polymyxa bacterial agent to the pseudofungus bacillus polymyxa bacterial agent is 1:1.
further, the effective viable count of the Paenibacillus polymyxa Paenibacillus polymyxa and the Paenibacillus pseudomycoides Bacillus pseudomycoides is not less than 1.0X10 10 CFU/g。
The invention also provides a preparation method of the microbial agent for preventing and treating strawberry root rot, which comprises the following steps:
s1 strain screening and identification
Inoculating pathogenic bacteria of strawberry root rot, namely Mucor pseudodisc and Fusarium oxysporum, to the center of a potato dextrose agar culture medium, inoculating strains to be screened to the edge of the culture medium, culturing for 5 days in a constant temperature incubator at 28 ℃, observing the antibacterial condition of antagonistic bacteria on the pathogenic bacteria of strawberry root rot, and screening and identifying according to experimental results to obtain a Paenibacillus polymyxa Paenibacillus polymyxa strain and a Paenibacillus pseudomycoides Bacillus pseudomycoides with obvious antibacterial effect;
s2 Strain fermentation
(1) Culturing strains: inoculating Bacillus polymyxa Paenibacillus polymyxa and Bacillus pseudomycoides Bacillus pseudomycoides to NA solid medium, culturing to obtain single colony, inoculating to NB medium, shake culturing at 30deg.C and 180r/min for 18-24 hr to obtain seed solution with concentration not lower than 1.0X10 8 CFU/mL;
(2) Fermentation culture: inoculating Paenibacillus polymyxa seed solution and pseudofungus bacillus polymyxa seed solution obtained in the step (1) into a fermentation tank according to an inoculum size of 5%, respectively, culturing for 36h under the conditions of 30 ℃ and 180r/min at a rotating speed and 20% -22% of dissolved oxygen, and completing fermentation, thereby ensuring that the concentration of thalli in the fermentation liquid is 1.0x10% 10 CFU/mL, the spore yield reaches more than 90%;
(3) Preparing a bacillus microbial inoculum: respectively adding 5% of soluble starch into the Paenibacillus polymyxa fermentation liquor and the Paenibacillus pseudomycoides fermentation liquor obtained in the step (2), uniformly stirring, and directly performing spray drying to obtain Paenibacillus polymyxa microbial inoculum and Paenibacillus pseudomycoides microbial inoculum;
(4) Preparation of microbial agent: the paenibacillus polymyxa bacterial agent and the pseudofungus bacillus polymyxa bacterial agent obtained in the step (3) are mixed according to the mass ratio of 1:1, mixing to obtain the microbial agent for preventing and treating strawberry root rot.
Preferably, in step (2), the medium formulation (g/L) of the fermentation culture: beef extract powder 5, peptone 10, yeast extract powder 5, sodium chloride 5, glucose 10 and pH7.0.
Compared with the prior art, the invention has the following beneficial effects:
according to the invention, the paenibacillus polymyxa bacterial agent and the pseudo-fungus bacillus cereus bacterial agent are compounded to be used as microbial bacterial agents, so that the incidence rate of strawberry root rot is remarkably reduced, the effect of the paenibacillus polymyxa and the pseudo-fungus bacillus cereus compounded bacterial agent is better than that of independently inoculating the paenibacillus polymyxa and the pseudo-fungus bacillus cereus bacterial agent, and the paenibacillus polymyxa bacterial agent and the pseudo-fungus bacillus cereus bacterial agent have synergistic effects in the aspect of preventing and treating strawberry root rot, and can be effectively and stably applied to green prevention and control of strawberry root rot. The invention can effectively prevent and treat the strawberry root rot, thereby promoting the growth of strawberry plants; in addition, the microbial agent has the characteristics of environmental protection and no pesticide residue, and can avoid pathogenic bacteria from generating drug resistance after use.
Drawings
FIG. 1 antagonism of Paenibacillus polymyxa Paenibacillus polymyxa against the root rot pathogen Paecilomyces polytrichosporus (a) and Fusarium oxysporum (b);
FIG. 2 antagonism of Bacillus pseudomycoides Bacillus pseudomycoides against the root rot strawberry root rot pathogen Mucor pseudodisci (a) and Fusarium oxysporum (b);
FIG. 3 is a graph comparing growth conditions of strawberries before and after Paenibacillus polymyxa Paenibacillus polymyxa microbial inoculum treatment has been inoculated with a strawberry root rot pathogen pot; (a) Inoculating a root rot pathogen and a Paenibacillus polymyxa Paenibacillus polymyxa microbial inoculum; (b) inoculating root rot pathogen;
FIG. 4 is a graph comparing growth conditions of strawberries before and after the treatment of the pseudo-bacillus mycoides Bacillus pseudomycoides microbial inoculum has been inoculated with a strawberry root rot pathogen pot; (a) Inoculating root rot pathogen + pseudofungus bacillus cereus Bacillus pseudomycoides; (b) inoculating root rot pathogen;
FIG. 5 is a graph showing the comparison of the growth of strawberries before and after the treatment with the microbial agent of the present invention and after the potting of the strawberry root rot pathogen; (a) Inoculating root rot pathogen, paenibacillus polymyxa Paenibacillus polymyxa inoculant and pseudofungus bacillus sphaeroides Bacillus pseudomycoides; (b) inoculating root rot pathogen.
Detailed Description
The Paenibacillus polymyxa Paenibacillus polymyxa and the pseudofungus bacillus polymyxa Bacillus pseudomycoides provided by the invention are preserved in China general microbiological culture Collection center (CGMCC) No.25858 and CGMCC No.25859 respectively in 2022 and 09 months and have a preservation address of North-West-Lu No.1 and 3 in the Chaoyang area of Beijing city.
The following specific examples are set forth to further illustrate the beneficial effects of the present invention, and are specifically described to aid in a better understanding of the present invention and are in no way limiting to the scope of the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The reagents or apparatus used in the examples below were conventional products available for purchase by a regular distributor without the manufacturer's knowledge.
The strawberry root rot pathogen in the examples is separated from the root tissue of a strawberry planting base in the Menjin area of the Luoyang city of Henan province, and the microorganism strain in the examples is separated from the root soil and the root tissue of a healthy strawberry in the strawberry planting base.
Example 1
A preparation method of a microbial agent for preventing and treating strawberry root rot comprises the following steps:
s1 strain screening and identification
(1) Screening of biocontrol Strain
Activating bacteria separated from healthy strawberry rhizosphere soil and root tissues, determining antagonism of bacterial strains to strawberry root rot pathogen trichoderma pseudodisc and fusarium oxysporum by a plate counter-surface culture method, specifically implementing the method by grafting bacterial strain blocks of strawberry root rot pathogen bacteria with the diameter of 5mm at the center of an improved PDA culture medium plate with the diameter of 9cm, symmetrically grafting bacterial strains at a position 9mm away from the edge of a culture dish, grafting pathogenic bacteria mycelium blocks in a control group, culturing for 5 days at the constant temperature of 28 ℃, measuring the width of a bacteriostatic zone, and calculating the bacteriostatic rate.
Figure SMS_1
The test results are shown in Table 1, FIG. 1 and FIG. 2, wherein the width of the antibacterial zone of the strain CMS5 on the strawberry root rot pathogen, the trichoderma pseudodisc and the fusarium oxysporum is 8.2mm and 2.5mm respectively, and the width of the antibacterial zone of the strain CMR12 on the strawberry root rot pathogen, the trichoderma pseudodisc and the fusarium oxysporum is 7.6mm and 3.6mm respectively; the bacteriostasis rates of the strain CMS5 on the strawberry root rot pathogen, namely the trichoderma pseudodisc and the fusarium oxysporum are 57.1 percent and 38.4 percent respectively, and the bacteriostasis rates of the strain CMR12 on the strawberry root rot pathogen, namely the trichoderma pseudodisc and the fusarium oxysporum are 24.4 percent and 30.0 percent respectively.
TABLE 1 Flat plate antagonism of CMS5 and CMR12 strains against strawberry root rot pathogen
Figure SMS_2
In this embodiment, the potato dextrose medium (PDA) consists of 200g potato, 20g dextrose, 17g agar and 1000mL distilled water, with a natural pH.
(2) Identification of biocontrol strains CMS5 and CMR12
The implementation of the invention mainly adopts a method combining the observation of the bacterial body shape, the measurement of the physiological and biochemical characteristics and the molecular biology means to identify the strains CMS5 and CMR 12. As shown in FIG. 2, the strains CMS5 and CMR12 grew well on NA medium, and colonies thereof were spread, dried, opaque, white, and positive after gram staining, and the thallus was rod-shaped to produce spores. The PCR amplification result of the 16S rRNA shows that the amplification products of the strain CMS5 and the strain CMR12 are 1426bp and 1436bp respectively, sequencing is carried out by the Shanghai biological engineering company, the complete sequences of the genes of the strain CMS5 and the strain CMR1216S rRNA are obtained, the BLAST software in GenBank is utilized to analyze the sequences of the genes of the strain 16S rRNA, the 16S rRNA sequence of the strain CMS5 has higher homology with the Paenibacillus polymyxa, the 16S rRNA sequence of the strain CMR12 has higher homology with the pseudomycoides, and the similarity is up to 99%. Strains CMS5 and CMR12 were identified as Paenibacillus polymyxa Paenibacillus polymyxa and Bacillus pseudomycoides Bacillus pseudomycoides, respectively, by combining morphological, physiological and biochemical characteristics with 16S rRNA gene sequence analysis.
The 16S rDNA gene sequences of the strains CMS5 and CMR12 are shown in sequence tables SEQ ID NO.1 and SEQ ID NO. 2.
In this embodiment, the beef extract peptone medium (NA) is composed of beef extract 3g, peptone 5g, glucose 2.5g, agar 17g and 1000mL distilled water, pH7.2.
S2 Strain fermentation
(1) Culturing strains: inoculating Bacillus polymyxa Paenibacillus polymyxa and Bacillus pseudomycoides Bacillus pseudomycoides to NA solid medium, culturing to obtain single colony, inoculating to NB medium, shake culturing at 30deg.C and 180r/min for 18-24 hr to obtain seed solution with concentration not lower than 1.0X10 8 CFU/mL;
(2) Fermentation culture: inoculating the bacillus seed liquid obtained in the step (1) into a fermentation tank according to 5% of inoculation amount, culturing for 36h under the conditions of 30 ℃ and 180r/min of rotating speed and 20% -22% of dissolved oxygen, and finishing fermentation, thereby ensuring that the concentration of thalli in the fermentation liquid is 1.0x10 10 CFU/mL, the spore yield reaches more than 90%;
(3) Preparing a bacillus microbial inoculum: respectively adding 5% of soluble starch into the fermentation liquor of the paenibacillus polymyxa and the pseudo-fungus bacillus, uniformly stirring, and directly performing spray drying to obtain the paenibacillus polymyxa and the pseudo-fungus bacillus;
(4) And (3) preparing a microbial agent, namely mixing the two bacillus agents obtained in the step (3) according to the mass ratio of the paenibacillus polymyxa to the pseudomycoid bacillus agent of 1:1 to obtain the microbial agent.
The culture medium used for the strain fermentation culture in this example is BPY medium, which consists of beef extract powder 5g, peptone 10g, yeast extract powder 5g, sodium chloride 5g, glucose 10g, pH7.0.
Example 2
Application of microbial agent in preventing and treating strawberry root rot
(1) Test materials
The variety of the strawberries to be tested is red in color, and the transplanted seedlings are provided by a strawberry base in Shi Li Vanilla in Monjin area.
The microbial preparation to be tested was the microbial agent prepared in this example 1.
(2) Test design
Healthy strawberry seedlings (red skin) with good growth vigor and consistent size are selected, transplanted into pot bodies, the diameter of the pot mouth is 20cm, all soil is matrix soil, sterilization is carried out for 2 hours at 160 ℃, each pot body contains 1 plant, and conventional management is carried out.
The test had 4 treatments: t1 (strawberry seedling+root rot pathogen); t2 (strawberry seedling+root rot pathogen+paenibacillus polymyxa Paenibacillus polymyxa inoculant); t3 (strawberry seedling + root rot pathogen + pseudomonas Bacillus pseudomycoides inoculant); t4 (strawberry seedling+root rot pathogen+microbial inoculum of the invention).
(3) Test method
After the strawberry is transplanted and the seedling is recovered for 1 month, the strawberry seedling is subjected to root injury treatment, 2 to 3 of the adventitious roots of the strawberry are cut off by a blade after the strawberry is disinfected by 75 percent of ethanol, the root irrigation treatment is carried out by pathogenic bacteria spore suspension, and 50mL of the seedling root is respectively inoculated with the concentration of 1 multiplied by 10 7 Is prepared from the spore suspension of Mucor pseudodisci and Fusarium oxysporum. After pathogenic bacteria are inoculated for 5 days, a root irrigation method is adopted to inoculated with a biocontrol microbial inoculum, the microbial inoculum is diluted by 100 times of clear water, 100mL of each plant is irrigated, 3 repetitions are arranged for each treatment of 5 pots, potting experiments are finished after 2 months, and the morbidity is counted and the microbial inoculum control effect is calculated.
(4) Test results and statistics
The calculation formula of the potted plant morbidity and the microbial inoculum control effect is as follows:
potted morbidity (%) = number of diseased potted plants/total number of potted plants investigated x 100
The disease index is calculated according to the following formula:
disease index = Σ (number of seedlings at each stage×each stage)/(number of total plants investigated×highest disease stage) ×100
TABLE 2 disease classification for strawberry root rot potting experiments
Disease level Description of the disease
Level 0 No disease occurrence
Level 1 The number of the diseased potted plants accounts for 1 to 5 percent of the total number of the potted plants for investigation
Level 2 The number of the diseased potted plants accounts for 5 to 15 percent of the total number of the potted plants for investigation
3 grade The number of the diseased potted plants accounts for 15 to 25 percent of the total number of the cultivated plants for investigation
Grade 4 The number of the diseased potted plants accounts for 25 to 50 percent of the total number of the potted plants for investigation
Grade 5 The number of the diseased potted plants accounts for 50-100% of the total number of the cultivated plants for investigation
Control effect (%) = (control disease index-treatment disease index)/control disease index×100
(5) Test results
As shown in fig. 3-5, after 2 months of treatment of potted strawberry plants with different bactericides, the control group T1 and the treatment groups (T2, T3 and T4) have different degrees of disease symptoms, the disease symptoms of the potted strawberry in the control group T1 without the bactericides are that the leaf margin of the strawberry leaf is dry and curled, the strawberry plants have wilting and lodging phenomena, the disease symptoms of the potted strawberry plants in the inoculation bactericides treatment groups (T2, T3 and T4) are relatively light, and leaf disease spots are not spread or basically not spread. Wherein the incidence rate of the control group T1 is 83.3 percent, and the disease index is 83.3; secondly, the morbidity of the treatment of the Paenibacillus polymyxa Paenibacillus polymyxa microbial agent T2 and the treatment of the Paenibacillus pseudomycoides Bacillus pseudomycoides microbial agent T3 are 38.9 percent and 33.3 percent respectively, and the disease indexes are 28.9 and 26.7 respectively; the incidence rate of the root rot of the potted strawberries after the treatment of the microbial agent T4 is 16.7 percent, and the disease index is 10.0. According to the disease index calculation, the relative control effect of different microbial agents is calculated, wherein the relative control effect of the paenibacillus polymyxa Paenibacillus polymyxa microbial agent is 65.3 percent, the relative control effect of the pseudofungus bacillus sphaericus Bacillus pseudomycoides microbial agent is 68.0 percent, the control effect of the microbial agent is 88.0 percent, and the control effect of the microbial agent is obviously better than that of the single microbial agent.
TABLE 3 Effect of different treatments on the incidence of strawberry root rot
Treatment of Incidence of disease Index of disease condition Relative control effect
T1 83.3% 83.3 /
T2 38.9% 28.9 65.3%
T3 33.3% 26.7 68.0%
T4 16.7% 10.0 88.0%
Meanwhile, compared with a control group, the potted strawberry with the microbial agent is found to be vigorous in plant growth, green in leaves and thick in plant root system by comparing the growth conditions of different treated strawberries, and the microbial agent is obviously superior to the control group in clear water treatment, so that the microbial agent has a certain growth promoting effect on strawberries.
Therefore, the microbial agent suitable for the strawberry root rot prepared by the invention has high efficiency, stability, strong antagonism to pathogenic bacteria and remarkable synergistic effect, is applied to the control process of the strawberry root rot, can be used for stable field planting, effectively reduces the incidence of the strawberry root rot, and effectively improves the growth condition of strawberries.
While only certain embodiments of the present invention have been described, it will be apparent to those skilled in the art that other modifications and improvements can be made without departing from the inventive concept of the present invention.

Claims (5)

1. The microbial agent for preventing and treating the strawberry root rot is characterized by being formed by compounding paenibacillus polymyxa Paenibacillus polymyxa and pseudofungus bacillus thuringiensis Bacillus pseudomycoides, wherein paenibacillus polymyxa Bacillus pseudomycoides is preserved in China general microbiological culture collection center (CGMCC) No.25858 in the year 09 and 30 of 2022; the bacillus pseudomycosis Bacillus pseudomycoides is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.25859 in the year 09 and 30 of 2022.
2. The microbial agent for preventing and treating strawberry root rot according to claim 1, wherein the mass ratio of the paenibacillus polymyxa agent to the pseudomycoides agent is 1:1.
3. the microbial agent for preventing and treating strawberry root rot according to claim 1 or 2, wherein the effective viable count of the Paenibacillus polymyxa Paenibacillus polymyxa and the Paenibacillus pseudomycoides Bacillus pseudomycoides is not less than 1.0X10 10 CFU/g。
4. A method for preparing a microbial agent for controlling strawberry root rot according to any one of claims 1 to 3, comprising the steps of:
s1 strain screening and identification
Inoculating pathogenic bacteria of strawberry root rot, namely Mucor pseudodisc and Fusarium oxysporum, to the center of a potato dextrose agar culture medium, inoculating strains to be screened to the edge of the culture medium, culturing for 5 days in a constant temperature incubator at 28 ℃, observing the antibacterial condition of antagonistic bacteria on the pathogenic bacteria of strawberry root rot, and screening and identifying according to experimental results to obtain a Paenibacillus polymyxa Paenibacillus polymyxa strain and a Paenibacillus pseudomycoides Bacillus pseudomycoides with obvious antibacterial effect;
s2 Strain fermentation
(1) Culturing strains: inoculating Bacillus polymyxa Paenibacillus polymyxa and Bacillus pseudomycoides Bacillus pseudomycoides to NA solid medium, culturing to obtain single colony, inoculating to NB medium, shake culturing at 30deg.C and 180r/min for 18-24 hr to obtain seed solution with concentration not lower than 1.0X10 8 CFU/mL;
(2) Fermentation culture: inoculating Paenibacillus polymyxa seed solution and pseudofungus bacillus polymyxa seed solution obtained in the step (1) into a fermentation tank according to an inoculum size of 5%, respectively, culturing for 36h under the conditions of 30 ℃ and 180r/min at a rotating speed and 20% -22% of dissolved oxygen, and completing fermentation, thereby ensuring that the concentration of thalli in the fermentation liquid is 1.0x10% 10 CFU/mL, the spore yield reaches more than 90%;
(3) Preparing a bacillus microbial inoculum: respectively adding 5% of soluble starch into the Paenibacillus polymyxa fermentation liquor and the Paenibacillus pseudomycoides fermentation liquor obtained in the step (2), uniformly stirring, and directly performing spray drying to obtain Paenibacillus polymyxa microbial inoculum and Paenibacillus pseudomycoides microbial inoculum;
(4) Preparation of microbial agent: the paenibacillus polymyxa bacterial agent and the pseudofungus bacillus polymyxa bacterial agent obtained in the step (3) are mixed according to the mass ratio of 1:1, mixing to obtain the microbial agent for preventing and treating strawberry root rot.
5. The method for preparing a microbial agent for preventing and treating strawberry root rot of claim 4, wherein in the step (2), the formula (g/L) of the culture medium for fermentation culture is: beef extract 5, peptone 10, yeast extract 5, sodium chloride 5, glucose 10g, pH7.0.
CN202211532735.XA 2022-12-02 2022-12-02 Microbial agent for preventing and treating strawberry root rot and preparation method thereof Pending CN116083297A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202211532735.XA CN116083297A (en) 2022-12-02 2022-12-02 Microbial agent for preventing and treating strawberry root rot and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202211532735.XA CN116083297A (en) 2022-12-02 2022-12-02 Microbial agent for preventing and treating strawberry root rot and preparation method thereof

Publications (1)

Publication Number Publication Date
CN116083297A true CN116083297A (en) 2023-05-09

Family

ID=86211078

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202211532735.XA Pending CN116083297A (en) 2022-12-02 2022-12-02 Microbial agent for preventing and treating strawberry root rot and preparation method thereof

Country Status (1)

Country Link
CN (1) CN116083297A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114703094A (en) * 2022-03-18 2022-07-05 山东野田生物科技有限公司 Bacillus pseudomycoides and application thereof in preparation of plant pathogenic bacteria bacteriostat

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114703094A (en) * 2022-03-18 2022-07-05 山东野田生物科技有限公司 Bacillus pseudomycoides and application thereof in preparation of plant pathogenic bacteria bacteriostat
CN114703094B (en) * 2022-03-18 2023-09-05 山东野田生物科技有限公司 Bacillus pseudomycosis and application thereof in preparation of plant pathogenic bacteria bacteriostat

Similar Documents

Publication Publication Date Title
CN107779420B (en) Two-strain endogenous Bacillus belgii for antagonizing tobacco bacterial wilt and application thereof
CN106754557B (en) Bacillus subtilis YBM-4 and application thereof in preventing and treating tobacco black shank and promoting growth
CN110452832B (en) Acid-resistant bacillus amyloliquefaciens Kc-5 and application thereof
CN111944716B (en) Special compound microbial agent for tobacco seedling culture and preparation method and application thereof
CN115786218B (en) Bacillus bailii and application thereof
CN112342173B (en) Bacillus belgii and application thereof
CN113444651B (en) Saffron endophytic fungus and application thereof in preventing and treating bulb rot
CN108641989B (en) Bacillus methylotrophicus and application thereof
CN110317747A (en) A kind of bacillus amyloliquefaciens JT68 and its application in prevention and treatment tea anthracnose
CN113549578A (en) Siamese bacillus BsNlG13 for inhibiting rice blast germs and promoting seed germination and application thereof
CN111748498A (en) Bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot as well as preparation method and application thereof
CN109971656B (en) Ginger endogenetic trichoderma viride and application thereof
CN116083297A (en) Microbial agent for preventing and treating strawberry root rot and preparation method thereof
CN106635926B (en) Bacillus safensis YJC-4 and application thereof in prevention and treatment of tobacco black shank and growth promotion
CN115927051B (en) Biocontrol bacterium, biocontrol compound microbial inoculum and application thereof
CN113832071B (en) Brevibacillus halotolerans strain and application thereof in preparation of biocontrol microbial inoculum
CN116179430A (en) Bacillus bailii B31 and application thereof
CN115725445A (en) Paenibacillus polymyxa and suspension and application thereof
CN116004419A (en) Bacillus atrophaeus CY-2, microbial inoculum, preparation method and application thereof
CN114456973A (en) Streptomyces rochei strain in tobacco and application thereof in prevention and control of tobacco diseases
CN107629970B (en) Microorganism for preventing and treating ginseng erythroderma and application
CN111979151A (en) Biocontrol strain and application thereof
CN116042461B (en) Streptomyces rohder and application thereof in disease prevention and control of medlar
CN116769607B (en) Application of endophytic fungus A21-1-1 in preventing and controlling rice blast
CN118207129B (en) Bacillus infantis HN01 and application thereof in preventing and treating plant root knot nematode disease

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination