CN111387212A - Complex microbial inoculant for preventing and treating root-knot nematodes by using biocontrol bacteria and preparation method thereof - Google Patents

Complex microbial inoculant for preventing and treating root-knot nematodes by using biocontrol bacteria and preparation method thereof Download PDF

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Publication number
CN111387212A
CN111387212A CN202010155905.1A CN202010155905A CN111387212A CN 111387212 A CN111387212 A CN 111387212A CN 202010155905 A CN202010155905 A CN 202010155905A CN 111387212 A CN111387212 A CN 111387212A
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China
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complex microbial
yeast
verticillium
porous carrier
root
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CN202010155905.1A
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Chinese (zh)
Inventor
彭勇
彭瑞芳
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Wobot Biotechnology Co ltd
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Wobot Biotechnology Co ltd
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Priority to CN202010155905.1A priority Critical patent/CN111387212A/en
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    • 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
    • A01N25/00Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
    • A01N25/08Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing solids as carriers or diluents

Abstract

The invention discloses a compound microbial inoculum for preventing and treating root-knot nematode by using biocontrol bacteria and a preparation method thereof. The compound microbial inoculum for preventing and treating root-knot nematode by using biocontrol bacteria comprises yeast and verticillium dahliae. The compound microbial inoculum provided by the invention is used as a base fertilizer for tomato planting, and can obviously inhibit the attack of root-knot nematodes.

Description

Complex microbial inoculant for preventing and treating root-knot nematodes by using biocontrol bacteria and preparation method thereof
Technical Field
The invention relates to the field of fertilizers, in particular to a compound microbial inoculum for preventing and treating root-knot nematodes by using biocontrol bacteria and a preparation method thereof.
Background
Root-knot nematodes are obligate parasites of more than 30 common vegetables, widely distributed in the tropics and subtropics. Root-knot nematodes affect vegetable growth and reduce vegetable yield.
Verticillium pachyrhizus is a microbial preparation which is widely used at present and used for inhibiting root-knot nematodes. The verticillium dahliae has a great problem of low and unstable control effect in soil environment.
Disclosure of Invention
The invention aims to provide a compound microbial inoculum for biocontrol bacteria to prevent and control root-knot nematodes. The composite microbial inoculum comprises verticillium pachycorum and candida utilis. The complex microbial inoculum has high efficiency and good effect of inhibiting root-knot nematodes.
In the research, the compound microbial inoculum containing candida utilis and verticillium pachycorum is found to have obviously improved inhibition effect on the root-knot nematode in soil compared with the single verticillium pachycorum. The present application is based on this finding.
The invention provides a composite microbial inoculum for preventing and treating root-knot nematodes by using biocontrol bacteria, which comprises yeast and verticillium dahliae.
In some embodiments of the invention, the yeast is one or more of candida utilis and candida tropicalis.
In some embodiments of the invention, the ratio of yeast to Verticillium pachyrhizi is (5-15): (80-110), preferably 10: 100.
In some embodiments of the present invention, the yeast and verticillium dahliae in the complex microbial inoculum are bacteria obtained by filtration or freeze-drying separation.
In some embodiments of the invention, the complex microbial inoculum further comprises a porous carrier;
preferably, the porous carrier is selected from one or more of activated carbon, diatomaceous earth and polymeric microspheres.
In some embodiments of the present invention, the weight ratio of the thalli of the yeast and the verticillium pachyrhizi to the porous carrier in the composite microbial inoculum is not less than 2:8, preferably not less than 3: 7.
The second aspect of the present invention provides a method for preparing the complex microbial inoculum of the first aspect, comprising the following steps:
s01, mixing the separated cells obtained by filtration or freeze-drying of the yeast and Verticillium pachyrhizi.
The third aspect of the present invention provides another method for preparing a complex microbial inoculum according to the first aspect, comprising the following steps:
s11, mixing fermentation liquor and/or thallus of yeast and verticillium pachyrhizi with a porous carrier respectively and/or after mixing;
preferably, the porous carrier is selected from one or more of activated carbon, diatomaceous earth and polymeric microspheres.
In some embodiments of the invention, the method comprises the following steps:
s21, mixing the fermentation liquor of the yeast and/or verticillium pachyrhizi with the porous carrier in batches.
S22, drying;
s23, repeating S21-22 to the target weight ratio of the thallus of the yeast and the verticillium dahliae to the porous carrier;
in some embodiments of the invention, the temperature of the drying is 25-30 ℃;
in some embodiments of the invention, the weight ratio of the fermentation broth to the porous carrier for each mixing is (0.5-3): 10.
the fourth aspect of the invention is to provide an application of the complex microbial inoculum of the first aspect in the prevention and treatment of root-knot nematodes.
The invention has the beneficial effects that:
the composite microbial inoculum for preventing and treating root-knot nematode by using the compound biocontrol bacteria comprises yeast and verticillium dahliae. The composite microbial inoculum provided by the invention is used as a base fertilizer for tomato planting, has an effect of inhibiting root-knot nematodes which is obviously superior to that of single verticillium pachysarum, and can obviously inhibit the attack of the root-knot nematodes.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention.
When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.
Example 1
Complex microbial inoculant for preventing and treating root-knot nematode by using biocontrol bacteria
And respectively mixing 1g of Candida utilis and 10g of freeze-dried thalli of Verticillium pachyrhizi to obtain the composite microbial inoculum.
Example 2
Complex microbial inoculant for preventing and treating root-knot nematode by using biocontrol bacteria
S01, taking 10ml fermentation liquor containing Candida utilis (1.5 g of Candida utilis thallus), mixing with 10g of diatomite for 5 times and 2ml each time, drying at 30 ℃ for 10min, and repeatedly adding the fermentation liquor until the end.
S02, taking fermentation liquor containing 10g of verticillium dahliae, mixing 2ml of fermentation liquor each time with the diatomite loaded with the candida, drying for 10min at 30 ℃, and repeatedly adding the fermentation liquor until the completion to obtain the composite microbial inoculum.
Example 3
Complex microbial inoculant for preventing and treating root-knot nematode by using biocontrol bacteria
And respectively mixing 1g of candida tropicalis and 10g of freeze-dried thalli of verticillium pachydenum to obtain the composite microbial inoculum.
Comparative example 1
Verticillium pachyrhizi bacterial agent
A freeze-dried fungus body of Verticillium pachyrhizus.
Comparative example 2
Candida utilis inoculant
A lyophilized microorganism of Candida utilis.
Experimental example 1
Action of microbial inoculum in tomato on root-knot nematode
The tomato is of a variety of No. 9 medium-impurity, and is planted in a greenhouse after hole-plate seedling cultivation, wherein the row spacing is 65cm, and the plant spacing is 40 cm.
Tomatoes are planted in the greenhouse for three years, and the soil contains root-knot nematodes.
According to the ratio of 20kg of thallus/hm2The inocula of the examples and comparative examples of the present invention were applied in holes, respectively. The examples and comparative examples were each a treatment group, and a blank control group was additionally provided without applying any complex microbial inoculum.
And (4) carrying out greenhouse management according to the same conventional cultivation mode, and inspecting the condition of the root-knot nematodes when the tomatoes pull seedlings.
The grading is carried out according to the grading standards specified in the method of GB/T17980.38-2000. Recording the number of diseased plants at each level, sampling at 5 points on a diagonal line, investigating 3 plants each time, and calculating disease index and prevention effect.
Disease index (number of disease plants at each stage × relative to number of stages)/(total number of investigated plants × 7) × 100.
The prevention effect is (blank page disease index-treatment zone disease index)/(blank page disease index) × 100.
The specific grading method comprises the following steps:
grade 0, no insect gall in root system;
level 1, small galls exist in the root system;
grade 3, two thirds of root systems are full of insect galls;
grade 5, the root system is full of small galls and secondary galls;
and 7, the root system forms a fibrous root group.
The results are shown in Table 1.
TABLE 1 root-knot nematode inhibition by Complex microbial Agents
Index of disease/%) Preventive effect/%)
Example 1 22.69 50.3
Example 2 25.04 45.1
Example 3 26.76 41.4
Comparative example 1 35.62 21.9
Comparative example 2 45.07 1.2
Blank control 45.63 0.0
Researches show that verticillium pachyrhizi has certain effect of inhibiting root-knot nematodes but has poor effect, such as comparative example 1 in table 1; candida hirsutum had little root knot nematode inhibitory effect, such as comparative example 2 of Table 1; candida utilis and Candida tropicalis can promote the effect of Verticillium pachycosum on inhibiting root-knot nematodes, for example, the effect of example 1-3 on inhibiting root-knot nematodes is superior to that of comparative example 1 and comparative example 2.
While the preferred embodiments and examples of the present invention have been described in detail, the present invention is not limited to the embodiments and examples, and various changes can be made without departing from the spirit of the present invention within the knowledge of those skilled in the art.

Claims (10)

1. A composite bacterial preparation for preventing and treating root-knot nematode by using biocontrol bacteria comprises yeast and verticillium dahliae.
2. The complex microbial inoculant according to claim 1, wherein the yeast is one or more of candida utilis and candida tropicalis.
3. The complex microbial inoculum according to claim 1 or 2, characterized in that the ratio of yeast to verticillium pachycorum is (5-15): (80-110), preferably 10: 100.
4. The complex microbial inoculant according to any one of claims 1 to 3, wherein the yeast and Verticillium pachyrhizi are thalli obtained by filtration or freeze-drying separation.
5. The complex microbial inoculant according to any one of claims 1 to 4, wherein the complex microbial inoculant further comprises a porous carrier;
preferably, the porous carrier is selected from one or more of activated carbon, diatomaceous earth and polymeric microspheres.
6. The complex microbial inoculant according to any one of claims 1 to 5, wherein the weight ratio of the thalli of the yeast and the Verticillium pachyrhizi to the porous carrier in the complex microbial inoculant is not less than 2:8, preferably not less than 3: 7.
7. A method for preparing a complex microbial inoculum according to any one of claims 1 to 6, comprising the following steps:
s01, mixing the separated cells obtained by filtration or freeze-drying of the yeast and Verticillium pachyrhizi.
8. A method for preparing a complex microbial inoculum according to any one of claims 1 to 6, comprising the following steps:
s11, mixing fermentation liquor and/or thallus of yeast and verticillium pachyrhizi with a porous carrier respectively and/or after mixing;
preferably, the porous carrier is selected from one or more of activated carbon, diatomaceous earth and polymeric microspheres.
9. The method of claim 8, comprising the steps of:
s21, mixing the fermentation liquor of the yeast and/or verticillium dahliae with the porous carrier in batches;
s22, drying;
s23, repeating S21-22 to the target weight ratio of the thallus of the yeast and the verticillium dahliae to the porous carrier;
preferably, the temperature of the drying is 25-30 ℃;
preferably, when the fermentation liquid and the porous carrier are mixed in batches, the weight ratio of the fermentation liquid to the porous carrier in each mixing is (0.5-3): 10.
10. the application of the complex microbial inoculum according to any one of claims 1 to 6 in the control of root-knot nematodes.
CN202010155905.1A 2020-03-09 2020-03-09 Complex microbial inoculant for preventing and treating root-knot nematodes by using biocontrol bacteria and preparation method thereof Pending CN111387212A (en)

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