CN113475531B - Actinomycete complex microbial inoculum and preparation method and application thereof - Google Patents

Actinomycete complex microbial inoculum and preparation method and application thereof Download PDF

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CN113475531B
CN113475531B CN202110736778.9A CN202110736778A CN113475531B CN 113475531 B CN113475531 B CN 113475531B CN 202110736778 A CN202110736778 A CN 202110736778A CN 113475531 B CN113475531 B CN 113475531B
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actinomycete
microbial inoculum
algin
chitosan
jasmonic acid
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解晓燕
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Hainan Jinyufeng Biological Engineering Co ltd
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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
    • 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
    • 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
    • A01N37/00Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/42Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing within the same carbon skeleton a carboxylic group or a thio analogue, or a derivative thereof, and a carbon atom having only two bonds to hetero atoms with at the most one bond to halogen, e.g. keto-carboxylic acids
    • 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
    • A01N43/00Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/02Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
    • A01N43/04Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
    • A01N43/14Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
    • A01N43/16Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
    • 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
    • A01N65/00Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/03Algae

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  • Plant Pathology (AREA)
  • Dentistry (AREA)
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Abstract

The invention provides an actinomycete composite microbial inoculum which consists of actinomycete, algin, jasmonic acid, chitosan and a carrier, wherein the actinomycete is streptospora of actinomycetales. The actinomycete composite microbial inoculum can prevent and treat plant diseases such as areca-nut yellowing disease, Hami melon powdery mildew, wilt, premature senility, root rot and the like.

Description

Actinomycete complex microbial inoculum and preparation method and application thereof
Technical Field
The invention relates to the field of plant bactericides, and in particular relates to an actinomycete composite microbial inoculum, and a preparation method and application thereof.
Background
Actinomycetes are a group of gram-positive bacteria with a high (G + C) mol% content (> 55%) and are known for their actinoid-like colonies. It is a prokaryote group, widely distributed in nature, mainly propagated by spores, then propagated by disruptions, like common bacteria, mostly saprophytic and parasitic in a few.
The actinomycetes are widely distributed in nature, mainly exist in soil, air and water in a spore or hypha state, particularly exist in the most quantity in the soil with low water content, rich organic matters and neutral or slightly alkaline state, and are mainly caused by the specific mud fishy smell of the soil and the metabolite of the actinomycetes.
Neurospora belongs to actinomycetes, is widely distributed in natural soil and on gramineous plants, and is particularly easy to occur on corncobs and cottonseed hulls. Spore germination and hypha growth speed of the streptomyces are extremely high, aerial hypha (also called spore hypha) is tenacious, and the streptomyces can penetrate out of a sealing material of a strain to break a strain bag to form a large number of conidium clusters, and has the characteristics of germination on the same day and high-speed spore propagation on every other day. The whole test tube can grow on the slant culture medium within the temperature range of 20-30 ℃ all day and night, and the culture medium of the sawdust and the cotton seed hulls has rapid spread and strong transmission.
Alternaria alternata is mainly characterized in that cysts and cystospores can be formed, and spiral spore silks can be formed sometimes, and the sporocysts are divided into conidia after maturation. The nutrient thalli of the Neurospora is many in branches and rare in transverse septal distance, the diameter is 0.5-1.2 microns, and aerial mycelia form clusters, scattered or concentric ring arrangement and can generate broad-spectrum antibiotics.
Areca catechu L of Areca of Palmaceae contains various essential nutrients and beneficial substances, and the areca nut contains phenols 31.1%, polysaccharide 18.7%, fat 14.0%, crude fiber 10.8%, water 9.9%, ash 3.0% and alkaloid 0.5%.
Betel nuts are the first of four major south medicines, have been planted for more than two thousand years in China, are the main economic crops in Hainan province at present, have the largest planting area and the highest yield value, and the planting area reaches 150 ten thousand mu, so that the betel nuts are the main chewable food for masses in Hainan, Taiwan, Hunan and other places in China and have the habit of consuming the betel nuts from ancient times, and the total scale of the betel nut industry in 2020 exceeds 700 million yuan. In recent years, the areca-nut yellowing disease is more and more serious, and is called as the cancer of the areca-nut by local growers, so that the growers feel yellow and lose gall, and cut down when the areca-nut is yellow, and the prevention and treatment of the areca-nut yellowing disease are very important.
The Hami melon is one of the fruits purchased by residents for daily consumption, and the powdery mildew of the Hami melon is a common disease of the Hami melon in a greenhouse, so that the Hami melon generally has more diseases after the Hami melon is planted in the greenhouse and the yield and the quality of the Hami melon are seriously affected.
Patent CN201510827830.6 discloses actinomycete strain JSX-1 for preventing and treating soybean phytophthora and microbial inoculum thereof, wherein the strain is streptomycete-sp named actinomycete JSX-1. The single colony of the actinomycete strain is round and white, and the surface is dry and opaque; the hyphae are fine, the growth is slow, the colony texture is compact, and the colony has mud fishy smell. The gram staining of the actinomycete strain is purple and positive. The prevention effect of the JSX-1 microbial inoculum on the phytophthora sojae is 55 to 60 percent. The invention solves the control of phytophthora sojae, and does not relate to the control of areca-nut yellowing and Hami melon powdery mildew.
Patent CN201610761037.5 discloses a compound preparation for preventing and treating betel nut yellowing disease and a preparation method thereof, wherein each liter of the compound preparation contains 1.5-43.0 g of inorganic nutrient substances; 3.0-30.0 g of organic nutrient substances; 0.0005 to 0.1 g of plant active substances; 0.01-10.0 g of microbial secondary metabolite; 0.01-5.0 g of bactericide; 0.01-3.0 g of pesticide; the balance of emulsifier and water. The pesticides adopted in the patent comprise organophosphorus pesticides and the like, and the organophosphorus pesticides mostly belong to highly toxic medicaments, are easy to cause acute poisoning of people and livestock, and most varieties are easy to decompose and lose efficacy.
Disclosure of Invention
Therefore, the invention aims to provide a compound microbial inoculum capable of preventing and treating plant diseases, in particular to betel palm yellowing and Hami melon powdery mildew.
An actinomycete composite microbial inoculum comprises actinomycete, algin, jasmonic acid, chitosan and a carrier.
Further, the actinomycetes is an Streptosporangium of Actinomycetales.
Further, the carrier is one or more of an emulsifier, an excipient, a thickening agent, a solubilizer, a dispersant and an antifreeze.
An actinomycete composite bacterial agent is prepared by the following method:
step 1: inoculating the activated actinomycete slant culture in a culture medium for culture;
step 2: adding the algin, the jasmonic acid and the chitosan into the actinomycete preparation obtained in the step 1 according to a certain proportion, and uniformly mixing;
and step 3: and (3) adding the mixture obtained in the step (2) into a carrier, and uniformly mixing.
Further, in the step 1, the culture temperature is 30-35 ℃, and the inoculation amount is 25%.
Furthermore, in the step 2, the viable count of the actinomycete agent is 15-20 × 108 cfu/ml.
Further, the weight ratio of the actinomycete agent to the algin to the jasmonic acid to the chitosan is as follows: 10:4-6:3-5:1-3.
Further, the weight ratio of the actinomyces, the algin, the jasmonic acid and the chitosan is as follows: 10:4:5:1.
Compared with the prior art, the invention has the beneficial effects that:
1) the composite microbial inoculum disclosed by the application has small environmental pollution through biological control;
2) the compound microbial inoculum is wide in application, and can prevent and treat plant diseases such as areca yellowing disease, powdery mildew, wilt, premature senility, root rot and the like of Hami melon.
Detailed Description
For clear and complete description of the technical solutions in the present invention, it is obvious that the inventor combines the embodiments to describe, but the following embodiments describe only some embodiments of the present invention, not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1: yellow transformation of betel nut
In the test land, Hainan province, Qionghai city, in the betel nut test field, the land between the betel nuts is deeply ploughed, actinomycetes complex microbial inoculum is uniformly applied, the Control (CK) is blank control, the betel nut test field is infected by the betel nuts which have already developed the yellowing disease, and the incidence rate of the yellowing disease of the betel nuts is calculated.
Percent inhibition is (number of etiolation diseases of control group-number of etiolation diseases of experimental group)/number of etiolation diseases of control group is 100%
Inoculating activated slant culture of Neurospora at 30-32 deg.C under the inoculation amount of 25% into culture medium, and culturing until viable count reaches 18.9 × 10 8 cfu/ml, and then compounding with the algin, the jasmonic acid, the chitosan and the carrier.
The inhibition rate is calculated according to a Colby method (1966) formula and compared with the actually measured inhibition rate, and the combined effect of the mixture is simply, conveniently and effectively evaluated. The calculation formula is as follows:
E=X1×X2…×Xn/100(n-1)
e is the theoretical inhibition rate of the mixture; n is the number of mixed medicaments; x1 represents the inhibition rate after administration of agent 1; x2 represents the inhibition rate after administration of the 2 nd agent; xn represents the inhibition rate after administration of the nth agent. When the actual inhibition rate of the mixture is greater than the theoretical inhibition rate, synergism is represented; antagonism is indicated when the actual inhibition of the combination is less than the theoretical inhibition.
The results are shown in table 1:
table 1:
Figure BDA0003140278110000041
the actinomycetes can inhibit the areca-nut yellowing disease, wherein the streptospora inhibition effect is obviously better than that of streptomyces griseus; the effect of the streptosporangium matched with the algin, the jasmonic acid and the chitosan is better than that of the streptosporangium used alone, and the effect is synergistic compared with that of the streptosporangium used alone.
Example 2: powdery mildew of Hami melon
The test land is Hainan Ledong city, selecting a test field which frequently generates the powdery mildew of Hami melons, deeply ploughing the land before planting the Hami melons, uniformly applying a No. 1-8 actinomycete compound microbial inoculum, and calculating the powdery mildew incidence rate of the Hami melons by taking a Control (CK) as a blank control.
Inhibition%
The inhibition rate is calculated according to a Colby method (1966) formula and compared with the actually measured inhibition rate, and the combined effect of the mixture is simply, conveniently and effectively evaluated. The calculation formula is as follows:
E=X1×X2…×Xn/100(n-1)
e is the theoretical inhibition rate of the mixture; n is the number of mixed medicaments; x1 represents the inhibition rate after administration of agent 1; x2 represents the inhibition rate following administration of the 2 nd agent; xn represents the inhibition rate after administration of the nth agent. When the actual inhibition rate of the mixture is greater than the theoretical inhibition rate, synergism is represented; antagonism is indicated when the actual inhibition of the combination is less than the theoretical inhibition.
The results are shown in table 2:
table 2:
Figure BDA0003140278110000051
it can be seen that the actinomycetes has an inhibiting effect on powdery mildew of Hami melon, wherein the inhibiting effect of the streptospora is slightly superior to that of streptomyces griseus.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Claims (5)

1. An actinomycete composite microbial inoculum is characterized in that the microbial inoculum consists of actinomycetes, algin, jasmonic acid, chitosan and a carrier, wherein the actinomycetes is streptospora of actinomycetes;
the microbial inoculum is prepared by the following method:
step 1: inoculating the activated actinomycete slant culture in a culture medium for culture;
step 2: adding the algin, the jasmonic acid and the chitosan into the actinomycete preparation obtained in the step 1 according to a certain proportion, and uniformly mixing; the viable count of the actinomycete agent is 15-20 x 10 8 cfu/ml, the weight ratio of actinomycete agent, algin, jasmonic acid and chitosan is as follows: 10:4-6:3-5:1-3,
and step 3: and (3) adding the mixture obtained in the step (2) into a carrier, and uniformly mixing.
2. The actinomycete composite inoculant according to claim 1, wherein the carrier is one or more of an emulsifier, an excipient, a thickener, a solubilizer, a dispersant and an antifreeze.
3. The actinomycete composite microbial inoculum according to claim 1, wherein the weight ratio of the actinomycete, the algin, the jasmonic acid and the chitosan is as follows: 10:4:5:1.
4. The application of the actinomycete composite bacterial agent as claimed in claim 1 in preventing and treating areca-nut yellowing disease.
5. The application of the actinomycete composite bacterial agent as claimed in claim 1 in preventing and treating powdery mildew, wilt, premature senility and root rot of Hami melon.
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