CN114946885A - 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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CN114946885A
CN114946885A CN202210713311.7A CN202210713311A CN114946885A CN 114946885 A CN114946885 A CN 114946885A CN 202210713311 A CN202210713311 A CN 202210713311A CN 114946885 A CN114946885 A CN 114946885A
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chaetomium
leaf spot
aureofaciens
polygonatum odoratum
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CN114946885B (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
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    • C12R2001/645—Fungi ; Processes using fungi
    • 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
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Abstract

The invention relates to the technical field of microbial biology, in particular to a microbial pesticide of Chaetomium aureobasidium for resisting southern blight of polygonatum odoratum, wherein the preservation number of Chaetomium aureobasidium is CGMCC NO. 40190. The invention discovers that fusarium is one of important pathogenic bacteria of polygonatum leaf spot through pathogenic bacteria separation, verifies the antagonistic action of chaetomium aureofaciens on fusarium by adopting a plate confronting method, and the antagonistic action is 1 multiplied by 10 7 Chaetomium aureum at application concentration of cfu/mL has better control effect on polygonatum odoratum leaf spot caused by fusarium, and shows that Chaetomium aureum can be used as a biological pesticide for controlling polygonatum odoratum leaf spot. The invention provides a new idea and a related preparation for the biological control of the polygonatum odoratum leaf spot.

Description

Microbial pesticide for resisting leaf spot disease of polygonatum odoratum
Technical Field
The invention relates to the technical field of microbial biology, in particular to a Chaetomium aureofaciens microbial pesticide preparation for resisting polygonatum odoratum leaf spot.
Background
Polygonatum odoratum, named rhizoma polygonati and radix polygonati officinalis, is one of traditional Chinese medicinal materials in China, is commonly used for treating yin injury of lung and stomach, dryness-heat cough, dry throat and thirst, internal heat diabetes, mild action and no greasy feeling and astringing evil, so the Polygonatum odoratum is widely applied to medicaments for reducing blood sugar, resisting tumors, improving immunity, resisting oxidation, delaying senescence and resisting fatigue. In recent years, the planting scale of polygonatum odoratum in China is getting larger and larger, the planting region is also getting wider day by day, however, the polygonatum odoratum leaf spot disease occurring in the planting of polygonatum odoratum brings serious influence on the production of polygonatum odoratum. The early stage of the disease spots of the polygonatum odoratum leaf spot disease are brown spots, the edges of the disease spots are yellow, the disease spots are gradually expanded to be nearly round, gray in the interior and black brown at the edges, black granular matters appear on the disease spots under the humid condition, and the whole leaf withering can be caused when the disease spots are serious. There are many pathogenic bacteria of leaf spot, and fusarium is also one of the important pathogenic bacteria causing leaf spot.
Chaetomium aureum (Chaetomium aureum) has strong antagonistic effect and biocontrol potential. As early as 1975, scientists began research on the nitrogen fixation of Chaetomium aureum. The literature reports that the chaetomium aureum separated from walnut leaves has strong inhibition effect on the growth of the hypha of phytophthora capsici and can prevent and treat phytophthora capsici; the chaetomium aureofaciens also has better prevention and treatment effects on rice blast. Chaetomium aureofaciens, Lecanicillium angustifolium and Rhizopus arrhizus can be used for biological control of Aphis tabacis.
However, the polygonatum odoratum leaf spot pathogen is mostly controlled by chemical reagents, such as 50% iprodione, 65% zineb, 70% thiophanate-methyl and the like. However, the long-term use of chemical pesticides is liable to cause environmental pollution and ecological system interference, and is highly susceptible to the development of drug resistance. And the microbial biological control has the characteristics of high efficiency, no toxicity, no harm, no pollution and the like, can effectively solve the problem of 3R (Resistance, origin) of chemical pesticides, and promotes the sustainable development of agriculture.
Disclosure of Invention
Therefore, the inventor discovers that fusarium infection can also cause polygonatum leaf spot disease through long-term screening and experiments, and chaetomium aureum or spores thereof have the capacity of antagonizing fusarium, and can play a good effect in preventing and treating polygonatum leaf spot disease. Therefore, the invention provides the following technical scheme:
the invention separates the chaetomium aureum strain from the tuber of polygonatum odoratum, and the strain is identified to belong to the chaetomium aureum which is preserved in the China general microbiological culture Collection center of China general microbiological culture Collection center with the preservation date of 2022 years, 05 months and 07 days, and the preservation number is CGMCC NO. 40190.
In one aspect of the invention, the invention discloses application of chaetomium aureum or spores thereof in preparing a microbial pesticide for resisting southern leaf blight of polygonatum odoratum, wherein the microbial pesticide contains the chaetomium aureum or the spores thereof, and the preservation number of the chaetomium aureum is CGMCC NO. 40190.
In one aspect of the invention, the invention discloses a microbial pesticide for resisting southern blight of polygonatum odoratum, which comprises chaetomium aureofaciens or spores thereof, wherein the preservation number of the chaetomium aureofaciens is CGMCC NO. 40190. Preferably, the chaetomium aureum or the spores thereof in the microbial pesticide is in the form of powder; more preferably, the chaetomium aureum or its spores in the microbial pesticide are present 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 using formulations. The powder can be prepared by freeze-drying and vacuum freeze-drying to maintain the activity of the microorganism to the maximum extent. During the preparation of microbial pesticide, the microbes are separated, purified and identified, and fermented and cultured in large scale to obtain great amount of microbes. The temperature of the fermentation is controlled at 25-28 deg.C, and the pH of the fermentation medium is selected between 4-12, preferably 6-9.
In one aspect of the invention, the invention discloses a microbial pesticide for resisting southern blight, which comprises chaetomium aureofaciens or spores thereof, and the microbial pesticide also comprises a pharmaceutically acceptable auxiliary agent. 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 plays roles of emulsification, wetting, solubilization, defoaming, foaming, stabilization and the like in the microbial pesticide. The protective agent mainly plays a role in protecting microbial cells from being damaged or reducing damage in the preparation and storage processes. The nutrition auxiliary agent mainly provides nutrient substances for microbial thalli and improves the reproductive capacity of the microbes.
In one aspect of the invention, the invention provides application of chaetomium aureum or spores thereof in preparing a preparation for resisting polygonatum odoratum leaf spot.
In one aspect of the invention, the invention provides application of Chaetomium aureum or spores thereof in resisting southern blight of polygonatum, wherein the preservation number of Chaetomium aureum is CGMCC NO. 40190.
In one aspect of the invention, the microbial pesticide or preparation provided by the invention can be combined with other microorganisms resisting polygonatum leaf spot or chemical agents resisting leaf spot.
Advantageous effects
According to the invention, experiments show and prove that Chaetomium aureum can antagonize fusarium, so as to play a role in resisting polygonatum odoratum leaf spot, provide a new idea for biological control of polygonatum odoratum leaf spot, and play a good social and economic effect for control of polygonatum odoratum germs.
Drawings
FIG. 1 shows the antagonism of Chaetomium aureofaciens against Fusarium by plate confrontation.
Detailed Description
The invention is further described with reference to specific examples. The technical solutions of the present invention are conventional in the art unless otherwise specified, and the reagents or materials, which are conventional if not specified, are commercially available.
Golden hair shell contrast bacteria HTC: from the gakerxiang topic group of Shandong university of agriculture.
Example 1 antagonistic action of Chaetomium aureofaciens on Fusarium graminearum
The chaetomium aureofaciens strain is obtained by separating the tuber of the polygonatum odoratum according to a conventional method, is identified as the chaetomium aureofaciens and is preserved, the chaetomium aureofaciens is preserved in the common microorganism center of China general microbiological culture Collection management Committee, the preservation date is 2022 years, 05 months and 07 days, and the preservation number is CGMCC NO. 40190.
The chaetomium aureofaciens culture medium is cultured in a PDB culture medium (potato sucrose culture medium), and the PDB culture medium comprises 200g of potatoes, 20g of cane sugar, 1000 mL of distilled water and pH7.0. Pathogenic bacteria are separated from polygonatum odoratum leaf spots, fusarium is selected as a pathogen which is morphologically identified and separated according to a common bacteria identification manual, and the fusarium is cultured on a PDA (potato dextrose agar) culture medium for later use. PDA culture medium composition: 200g of potato, 20g of glucose, 20g of agar and 1000 mL of distilled water, and the pH value is 7.0.
The plate confrontation method verifies the antagonism of chaetomium aureofaciens on fusarium graminearum. The preparation method comprises preparing each bacterium into cake with a punch with diameter of 5mm, inoculating Fusarium (pathogenic bacteria) in the middle of PDA plate, and placing Chaetomium aureum (antagonistic bacteria) around the plate. The distance of the antagonistic bacteria from the periphery of the culture dish was about 10mm, the distance of inoculation of both bacteria was 30mm, and each treatment was repeated 3 times. The culture was carried out at 28 ℃ for 3 days. Meanwhile, a plate inoculated with only the pathogenic bacteria was used as a control group, and each treatment was repeated 3 times. When the control group pathogenic bacteria colony grows over the culture dish, the colony diameter is measured by a millimeter measuring ruler. And judging whether the biocontrol bacteria have antagonistic action on pathogenic bacteria according to the growth speeds of the 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 and atrophic, and calculating the growth inhibition rate of the chaetomium aureum on fusarium based on the measurement data of the experimental group and the control group. Referring to fig. 1, chaetomium aureum exerts a significant inhibitory effect on the growth of fusarium. Through measurement and calculation, the growth inhibition rate of the chaetomium aureofaciens on fusarium reaches 62.8 percent, and the chaetomium aureofaciens has a relatively obvious inhibition effect.
Example 2 Effect of Chaetomium aureofaciens on the resistance to leaf spot disease of Polygonatum odoratum
Inoculating Chaetomium aureofaciens into a PDB culture medium, carrying out shaking culture at 28 ℃ for 10 days, filtering the fermentation liquor by using gauze, storing in a triangular flask, sealing, and storing in a refrigerator at 4 ℃ for later use.
Healthy polygonatum seedlings with 3-4 leaves are selected and placed in a greenhouse for cultivation two weeks before the test. Separately prepare 1.5X 10 5 CFU/mL、1.5×10 6 CFU/mL、1.5×10 7 CFU/mL、1.5×10 8 CFU/mL fusarium fermentation broth. Sterilizing the polygonatum odoratum leaves, then coating fusarium fermentation liquids with different concentrations on the polygonatum odoratum leaves by using a cotton swab, and coating 5 plants in each group by using sterile water as a control. Then bagging and moisturizing, culturing in an artificial climate box, and observing the pathogenicity of fusarium. Observed at 1.5X 10 7 CFU/mL can reach pathogenic concentrations.
And similarly, selecting healthy polygonatum seedlings with 3-4 leaves, and placing the seedlings in a greenhouse for culture two weeks before the test. Preparation of 1.5X 10 7 And (3) inoculating the CFU/mL fusarium suspension to the polygonatum seedlings leaves by using an inoculating needle, bagging and preserving moisture for 48 hours. Then, at about 1.8X 10 8 cfu/mL, about 1.8X 10 7 cfu/mL, about 1.8X 10 6 cfu/mL, about 1.8X 10 5 cfu/mL Chaetomium aureofaciens fermentation broth, sterile water, 50% iprodione 500-fold diluent, Durio vitamin-Pythium oligandrum (Czech biological preparation International GmbH), and Chaetomium aureofaciens HTC about 1.8 × 10 7 cfu/mL, respectively spraying to the polygonatum seedlings, covering 15 plants in each group, moisturizing, placing in an artificial climate box for 10 days, observing the formation of leaf spots, measuring the sizes of the diseased spots by a cross method, and calculating the antagonistic effect, wherein the calculation formula is as follows:
Figure 558614DEST_PATH_IMAGE001
group of Chaetomium aureofaciens 1.8 10 8 cfu/mL Chaetomium aureofaciens 1.8 10 7 cfu/mL Chaetomium aureofaciens 1.8 10 6 cfu/mL Chaetomium aureofaciens 1.8 10 5 cfu/mL 500 times of 50% iprodione Dilution of Multi-vitamine-oligomate Pythium species Golden hair shell contrast Bacterium HTC Is sterile Water (W)
Inhibition rate 66.1% 65.3% 55.8% 41.4% 73.2% 67.2% 45.8% /
TABLE 1 inhibitory Effect of Chaetomium aureofaciens fermentation broth on leaf spot of Polygonatum odoratum
As can be seen from Table 1, Chaetomium aureofaciens induced fragrant solomonseal rhizome leaves by FusariumThe spot disease has relatively good antagonistic inhibition effect at 1.8 × 10 7 Under the concentration of cfu/mL, the effect of inhibiting the chaetomium aureofaciens is close to that of using a diluent of which the concentration is 500 times that of using 50 percent iprodione, the higher drug concentration does not greatly improve the antibacterial effect, and the use economy is considered, 10 7 The concentration of cfu/mL is preferably used. The chaetomium aureofaciens can be used as a biological control pesticide for the polygonatum odoratum leaf spot caused by fusarium.
While the invention has been described in further detail in connection with specific embodiments thereof, it will be understood that the invention is not limited thereto, and that various other modifications and substitutions may be made by those skilled in the art without departing from the spirit of the invention, which should be considered to be within the scope of the invention as defined by the appended claims.

Claims (8)

1. The microbial pesticide is characterized by comprising chaetomium aureum or spores thereof, wherein the preservation number of the chaetomium aureum is CGMCC NO. 40190.
2. The microbial pesticide of claim 1, wherein the chaetomium aureofaciens is present in powder form.
3. The microbial pesticide of claim 1, wherein the chaetomium aureofaciens is present in the form of granules.
4. The microbial pesticide of any one of claims 1 to 3, wherein the microbial pesticide further comprises a pharmaceutically acceptable adjuvant.
5. The microbial pesticide of any one of claims 1 to 3, wherein the microbial pesticide further comprises other microorganisms or anti-leaf spot chemical agents that are resistant to Yuzhu leaf spot.
6. The microbial pesticide of claim 4, wherein the adjuvant comprises one or more of a carrier, a surfactant, a protective agent, and a nutritional adjuvant.
7. The chaetomium aureofaciens or the spores thereof are applied to the preparation of the preparation for resisting the polygonatum odoratum leaf spot disease, and the preservation number of the chaetomium aureofaciens is CGMCC NO. 40190.
8. The application of Chaetomium aureofaciens or spores thereof in resisting polygonatum odoratum leaf spot disease, wherein the preservation number of the Chaetomium aureofaciens is CGMCC NO. 40190.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116376720A (en) * 2023-04-27 2023-07-04 湖南省烟草公司衡阳市公司 Chaetomium aureum and application thereof in preventing and treating tobacco leaf spot

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Publication number Priority date Publication date Assignee Title
CN116376720A (en) * 2023-04-27 2023-07-04 湖南省烟草公司衡阳市公司 Chaetomium aureum and application thereof in preventing and treating tobacco leaf spot
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