CN114946885B - Microbial pesticide for resisting leaf spot disease of polygonatum odoratum - Google Patents

Microbial pesticide for resisting leaf spot disease of polygonatum odoratum Download PDF

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CN114946885B
CN114946885B CN202210713311.7A CN202210713311A CN114946885B CN 114946885 B CN114946885 B CN 114946885B CN 202210713311 A CN202210713311 A CN 202210713311A CN 114946885 B CN114946885 B CN 114946885B
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leaf spot
chaetomium
microbial pesticide
aureum
spot disease
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CN114946885A (en
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徐瑞
宋荣
周佳民
官迪
谢进
朱校奇
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Hunan Institute Of Agro-Environment And Ecology
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    • 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
    • A01P3/00—Fungicides
    • C—CHEMISTRY; METALLURGY
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    • 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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    • C12R2001/00—Microorganisms ; Processes using microorganisms
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    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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Abstract

The invention relates to the technical field of microorganism, in particular to a microbial pesticide for preventing and treating fragrant solomonseal rhizome leaf spot disease and chaetomium aureum, wherein the preservation number of the chaetomium aureum is CGMCC NO.40190. The invention discovers that fusarium is one of important bacteria of fragrant solomonseal rhizome leaf spot disease through pathogen separation, adopts a flat plate counter method to verify antagonism of golden chaetomium to fusarium, and is 1 multiplied by 10 7 The chaetomium aureum has better control effect on the fragrant solomonseal rhizome leaf spot caused by fusarium at the application concentration of cfu/mL, and shows that the chaetomium aureum can be used as a biopesticide for controlling the fragrant solomonseal rhizome leaf spot. The invention provides a new thought and related preparation for biological control of fragrant solomonseal rhizome leaf spot disease.

Description

Microbial pesticide for resisting leaf spot disease of polygonatum odoratum
Technical Field
The invention relates to the technical field of microorganism, in particular to a chaetomium aureum microbial pesticide preparation for resisting fragrant solomonseal rhizome leaf spot disease.
Background
The fragrant solomonseal rhizome is also called as a fragrant solomonseal rhizome and the tail ginseng is one of traditional Chinese medicinal materials in China, is commonly used for treating lung and stomach yin injury, dryness-heat cough, dry throat and thirst, internal heat and thirst quenching, has mild effect and does not nourish greasy and astringe evil, so that the fragrant solomonseal rhizome has wide application in medicaments for reducing blood sugar, resisting tumors, improving immunity, resisting oxidation, delaying aging and resisting fatigue. In recent years, the scale of planting polygonatum odoratum in China is larger and larger, and the planting region is wider and wider, however, the leaf spot disease of polygonatum odoratum in the planting process of polygonatum odoratum brings serious influence to the production of polygonatum odoratum. The initial stage of the disease spots of the leaf spot of the polygonatum odoratum is brown spots, the edges are yellow, the disease spots are nearly round after being gradually expanded, the inner gray is formed, the edges are black brown, black granular matters appear on the disease spots under the moist condition, and the whole leaf can die when serious. There are a variety of pathogenic bacteria of leaf spot, and fusarium is also one of the important pathogenic bacteria causing leaf spot.
The chaetomium aureum (Chaetomium aureum) has strong antagonism effect and biocontrol potential. As early as 1975, scientists began to study nitrogen fixation of Chaetomium aureum. The literature reports that the chaetomium aureum obtained by separating from walnut leaves has strong inhibition effect on the hypha growth of phytophthora capsici and can prevent and treat the occurrence of pepper epidemic disease; the chaetomium aureum also has better control effect on rice blast. The chaetomium aureum, the lecanii and the rhizopus arrhizus can be used for biological control of the myzus persicae.
However, at present, the fragrant solomonseal rhizome leaf spot disease is mostly prevented and treated by chemical agents, for example, 50% iprodione, 65% zineb, 70% thiophanate methyl and the like are used. However, the use of long-term chemical pesticides is prone to environmental pollution and ecological system disturbances, and is extremely prone to drug resistance. The microbial biological control is efficient, nontoxic, harmless, pollution-free and the like, can effectively solve the problem of 3R (Resistance ) of chemical pesticides, and promotes agricultural sustainable development.
Disclosure of Invention
How to effectively prevent and treat the leaf spot disease of the polygonatum odoratum with low toxicity becomes a common problem for a polygonatum odoratum grower, and the inventor discovers that the long-term screening and test show that the leaf spot disease of the polygonatum odoratum is also caused by fusarium infection, and the golden chaetomium or spores thereof have the capability of antagonizing the fusarium and can play a good role in preventing and treating the leaf spot disease of the polygonatum odoratum. Therefore, the invention provides the following technical scheme:
the invention separates and obtains the golden chaetomium strain from the polygonatum tuber, which belongs to golden chaetomium through identification, the golden chaetomium has been preserved in China general microbiological culture Collection center with the preservation date of 2022, 05 and 07, and the preservation number is CGMCC NO.40190.
The invention discloses an application of chaetomium aureum or spores thereof in preparing a microbial pesticide for resisting fragrant solomonseal leaf spot disease, wherein the microbial pesticide contains chaetomium aureum or spores thereof, and the preservation number of the chaetomium aureum is CGMCC NO.40190.
The invention discloses a microbial pesticide for resisting leaf spot disease of polygonatum, which comprises chaetomium aureum or spores thereof, wherein the collection number of the chaetomium aureum is CGMCC NO.40190. Preferentially, the chaetomium aureum or spores thereof in the microbial pesticide exist in the form of powder; more preferably, chaetomium aureum or spores thereof in the microbial pesticide are in the form of granules. Of course, the preparation of the invention can also be granules, suspending agents, wettable powder and the like, and is suitable for dosage forms used. The powder can be prepared by freeze-drying and vacuum freeze-drying to maintain the activity of the microorganism to the maximum extent. In the process of preparing microbial pesticide, the microorganisms should be separated, purified and identified, and large-scale fermentation culture is performed to obtain a large amount of microorganisms. The fermentation temperature is controlled at 25-28deg.C, and pH of fermentation medium is selected to be 4-12, preferably 6-9.
In one aspect, the invention discloses a microbial pesticide for resisting leaf spot disease of polygonatum odoratum, wherein the microbial pesticide comprises chaetomium aureum or spores thereof, and the microbial pesticide also comprises pharmaceutically acceptable auxiliary agents. The auxiliary agent comprises one or more of a carrier, a surfactant, a protective agent and a nutrition auxiliary agent. The carrier comprises one or more of inorganic carrier and organic carrier, and the surfactant has the functions of emulsifying, wetting, solubilizing, defoaming, foaming, stabilizing and the like in the microbial pesticide. The protective agent has the main function of protecting microbial cells from damage or reducing damage in the preparation and storage processes. The nutrition auxiliary agent mainly provides nutrition for microorganism thallus and improves the reproductive capacity of microorganism.
In one aspect, the invention provides application of chaetomium aureum or spores thereof in preparing a preparation for resisting polygonatum leaf spot disease.
The invention provides an application of chaetomium aureum or spores thereof in resisting polygonatum leaf spot disease, wherein the collection number of chaetomium aureum is CGMCC No.40190.
In one aspect of the invention, the microbial pesticide or formulation provided by the invention can also be combined with other anti-leaf spot microbes or anti-leaf spot chemical agents.
Advantageous effects
The experiment shows that the chaetomium aureum can antagonize fusarium and further play a role in resisting the leaf spot disease of the polygonatum odoratum, a new thought is provided for biological control of the leaf spot disease of the polygonatum odoratum, and a good social and economic effect can be achieved for control of the germ of the polygonatum odoratum.
Drawings
FIG. 1 shows the antagonism of Fusarium with chaetomium aureum by plate confrontation.
Detailed Description
The invention is further described below in connection with specific embodiments. The technical scheme of the invention is a conventional mode in the field unless specifically stated, and the reagent or the material is a conventional reagent and is derived from commercial channels unless specifically stated.
Comparison bacteria HTC of golden hair shell: from the Gao Kexiang subject group of Shandong university of agriculture.
EXAMPLE 1 antagonism of Chaetomium aureum against Fusarium species of Polygonatum odoratum leaf spot
The golden chaetomium strain is obtained by separating the rhizoma polygonati tubers according to a conventional method, and is identified as golden chaetomium and preserved, wherein the golden chaetomium is preserved in China general microbiological culture Collection center with a preservation date of 2022, 05 months and 07 days, and the preservation number is CGMCC NO.40190.
The chaetomium aureum culture medium is cultured in a PDB culture medium (potato sucrose culture medium), wherein the PDB culture medium comprises 200g of potato, 20g of sucrose and 1000 mL of distilled water, and the pH is 7.0. Separating pathogenic bacteria from rhizoma Polygonati Odorati leaf spot, identifying the separated bacteria as Fusarium by morphology with reference to common bacteria identification handbook, and culturing Fusarium on PDA culture medium (potato dextrose agar culture medium) for use. PDA medium composition: 200g of potato, 20g of glucose, 20g of agar and 1000 mL of distilled water, and pH7.0.
The antagonism of the chaetomium aureum on fusarium of polygonatum leaf spot is verified by a plate counter method. Preparing bacterial cakes by using a puncher with the diameter of 5mm, inoculating fusarium (pathogenic bacteria) in the middle of a PDA plate, and placing chaetomium aureum (antagonistic bacteria) around the fusarium. The antagonistic bacteria were about 10mm from the periphery of the dish, the inoculation distance of both bacteria was 30mm, and each treatment was repeated 3 times. Incubated at 28℃for 3 days. Meanwhile, plates inoculated with pathogenic bacteria alone were used as control groups, and each treatment was repeated 3 times. When the control group pathogenic bacteria colony grows up on the culture dish, the colony diameter is measured by a millimeter measuring ruler. Judging whether the biocontrol bacteria have antagonism to pathogenic bacteria according to the growth speed of two bacterial colonies of the experimental group, whether inhibition zones exist between the bacterial colonies and whether hyphae at the edges of the bacterial colonies are sparse or not, and calculating the growth inhibition rate of the chaetomium aureum to fusarium based on the measurement data of the experimental group and the control group. Referring to FIG. 1, chaetomium aureum has a remarkable inhibition effect on growth of Fusarium. Through measurement and calculation, the growth inhibition rate of the chaetomium aureum on fusarium reaches 62.8%, and the chaetomium aureum has obvious inhibition effect.
EXAMPLE 2 Effect of Chaetomium aureum on resistance to leaf spot disease of Polygonatum odoratum
Inoculating Chaetomium aureum into PDB culture medium, shake culturing at 28deg.C for 10 days, filtering the fermentation broth with gauze, storing in triangular flask, sealing, and storing in refrigerator at 4deg.C for use.
Healthy polygonatum seedlings with 3-4 leaves are selected and placed in a greenhouse for cultivation two weeks before the test. Respectively prepare 1.5 multiplied by 10 5 CFU/mL、1.5×10 6 CFU/mL、1.5×10 7 CFU/mL、1.5×10 8 Fusarium fermentation broth of CFU/mL. Sterilizing rhizoma Polygonati Odorati leaf, and smearing fusarium fermentation liquid with different concentrations on rhizoma Polygonati Odorati leaf with cotton swab, wherein each group comprises 5 plants, and the smeared sterile water is used as control. Then bagging for moisturizing, culturing in a climatic chamber, and observing the pathogenicity of fusarium. Observed at 1.5X10 7 CFU/mL can be usedReaching the pathogenic concentration.
Healthy polygonatum seedlings with 3-4 leaves are selected as well and are placed in a greenhouse for cultivation two weeks before the test. Formulation 1.5X10 7 The CFU/mL fusarium suspension is inoculated on the leaves of the polygonatum seedlings by an inoculating needle, and the leaves are covered with bags for moisturizing for 48 hours. Then, at about 1.8X10 8 cfu/mL, about 1.8X10 7 cfu/mL, about 1.8X10 6 cfu/mL, about 1.8X10 5 cfu/mL Chaetomium aureum fermentation broth, sterile water, 50% iprodione 500-fold dilution, vitamin-Mucor dorsalis (Chlamydomonas zech. Co., ltd.), and Chaetomium aureum reference strain HTC about 1.8X10 7 cfu/mL are sprayed onto polygonatum seedlings respectively, 15 plants of each group are put into a climatic chamber for culturing for 10 days after bagging and moisturizing, leaf spot formation is observed, the size of the leaf spot is measured by a crisscross method, and the antagonism effect is calculated by the following calculation formula:
Figure 558614DEST_PATH_IMAGE001
group of Other kinds of things Chaetomium aureum 1.8×10 8 cfu/ mL Chaetomium aureum 1.8×10 7 cfu/ mL Chaetomium aureum 1.8×10 6 cfu/ mL Chaetomium aureum 1.8×10 5 cfu/ mL 50% iprodione 500 times dilution Multi-function vitamin Raw-oligo male Pythium species Golden hair shell Contrast bacteria HTC Without any means for Bacteria (fungus) Water and its preparation method
Inhibiting Manufacturing process Rate of 66.1% 65.3% 55.8% 41.4% 73.2% 67.2% 45.8% /
TABLE 1 inhibition effect of Chaetomium aureum fermentation broth on leaf spot of Polygonatum odoratum
As can be seen from Table 1, chaetomium aureum has good antagonistic and inhibitory effects on leaf spot of Polygonatum odoratum caused by Fusarium, and is 1.8X10 7 At the concentration of cfu/mL, the inhibition effect of chaetomium aureum is close to that of 50% iprodione 500-time diluent, the higher drug concentration has little improvement on the antibacterial effect, and the economy of use is considered, 10 7 The cfu/mL use concentration is a desirable concentration. The chaetomium aureum can be used as a biological control pesticide for preventing and treating the fragrant solomonseal rhizome leaf spot caused by fusarium.
The foregoing is a further elaboration of the present invention in connection with the detailed description, and it is not intended that the invention be limited to the specific embodiments shown, but rather that a number of simple deductions or substitutions be made by one of ordinary skill in the art without departing from the spirit of the invention, should be considered as falling within the scope of the invention as defined in the appended claims.

Claims (8)

1. The microbial pesticide for resisting the leaf spot disease of the polygonatum odoratum is characterized by comprising chaetomium aureum or spores thereof, wherein the collection number of the chaetomium aureum is CGMCC NO.40190.
2. The microbial pesticide according to claim 1, wherein chaetomium aureofaciens in the microbial pesticide is present in the form of a powder.
3. The microbial pesticide according to claim 1, wherein chaetomium aureofaciens in the microbial pesticide is present in the form of granules.
4. A microbial pesticide according to any one of claims 1 to 3, wherein the microbial pesticide further comprises a pharmaceutically acceptable adjuvant.
5. A microbial pesticide according to any one of claims 1 to 3, wherein the microbial pesticide further comprises other anti-leaf spot microorganisms or anti-leaf spot chemical agents of polygonatum odoratum.
6. The microbial pesticide according to claim 4, the adjuvant comprising one or more of a carrier, a surfactant, a protectant, a nutritional adjuvant.
7. The chaetomium aureum or spores thereof are applied to preparing the preparation for resisting the leaf spot disease of polygonatum, and the preservation number of the chaetomium aureum is CGMCC NO.40190.
8. The application of Chaetomium aureum or spores thereof in resisting leaf spot disease of rhizoma polygonati officinalis is disclosed, wherein the preservation number of Chaetomium aureum is CGMCC NO.40190.
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KR20190023689A (en) * 2017-08-30 2019-03-08 영남대학교 산학협력단 Chaetomium aureum TX1 strain producing taxol and uses thereof

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