CN113598201B - Botanical biocontrol bacterium preparation for preventing and treating rubber anthrax - Google Patents
Botanical biocontrol bacterium preparation for preventing and treating rubber anthrax Download PDFInfo
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- CN113598201B CN113598201B CN202110882925.3A CN202110882925A CN113598201B CN 113598201 B CN113598201 B CN 113598201B CN 202110882925 A CN202110882925 A CN 202110882925A CN 113598201 B CN113598201 B CN 113598201B
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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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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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
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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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/36—Rutaceae [Rue family], e.g. lime, orange, lemon, corktree or pricklyash
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Abstract
The invention provides a waterproofThe botanical biocontrol bacterium preparation for treating rubber anthrax comprises EM bacterium fermentation liquor, radix zanthoxyli aqueous extract and citrus peel ethanol extract, wherein the content of the radix zanthoxyli aqueous extract is 0.1-0.3wt%, the content of the citrus peel ethanol extract is 0.1-0.3wt%, and the total viable count of EM bacteria is more than or equal to 5 multiplied by 10 7 cfu/ml. The active ingredients of the botanical biocontrol bacteria preparation disclosed by the invention consist of EM (effective microorganisms) and fermentation metabolites thereof, zanthoxylum nitidum aqueous extracts and citrus peel ethanol extracts, belong to active ingredients with different action mechanisms, have a better inhibition effect on rubber anthracnose under the synergistic effect, can effectively prevent and treat the rubber anthracnose, and belong to green, environment-friendly and non-toxic active ingredients with double biological regulation functions of pesticide effect and fertilizer effect.
Description
Technical Field
The invention relates to a botanical biocontrol bacterium preparation, in particular to a botanical biocontrol bacterium preparation for preventing and treating rubber anthrax.
Background
The anthrax caused by the infection of various plants by the anthrax bacteria is widely distributed and seriously harmful, and is one of the key research objects of plant pathologists, and the rubber tree anthracnose is one of the most serious. Rubber tree anthracnose is caused by colletotrichum gloeosporioides infection, and causes the falling of numerous rubber tree leaves and even the death of the whole plant in the Hainan and Yunnan areas, thereby causing serious loss to the development of local rubber industry.
Anthrax bacteria overwinter in diseased branches mainly in the form of mycelia, produce conidia as a primary infection source under suitable conditions. The rubber tree seedlings, young trees and mature tapping trees can be infected by the anthrax to cause diseases. The germs can infect tender leaves, leaf stalks, tender tips, fruits and other parts to cause the detachment of the tender leaves, the return withering of the tender tips and the rotting of the fruits, even form stiff fruits to be hung on the trees, can cause the repeated leaf dropping and the return withering of the tender tips of the rubber trees in serious cases, and delay the cutting time. The disease is considered to be one of the main causes of the reduction of the yield of the natural rubber trees. At present, the control of rubber anthracnose still mainly uses chemical bactericides, however, the problems of drug resistance, toxicity and environmental pollution of pathogenic bacteria caused by chemical pesticides are increasingly prominent, the problems become main obstacles restricting the sustainable development of agriculture, and biocontrol microorganisms have the advantages of good environmental compatibility, no toxicity and no harm, and are ideal substitutes of the chemical pesticides. Therefore, the biocontrol microorganism is used for preventing and controlling the anthracnose of the rubber trees, and has important significance for realizing sustainable development of agriculture.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provide a botanical biocontrol bacterium preparation for preventing and treating rubber anthrax.
The invention provides a botanical biocontrol bacterium preparation for preventing and treating rubber anthrax, which consists of EM (effective microorganisms) fermentation liquor, radix zanthoxyli aqueous extract and citrus peel ethanol extract, wherein the content of the radix zanthoxyli aqueous extract is 0.1-0.3wt%, the content of the citrus peel ethanol extract is 0.1-0.3wt%, and the total viable count of EM is more than or equal to 5 multiplied by 10 7 cfu/ml。
Preferably, the EM bacteria fermentation liquid is obtained by mixing crushed corncobs, bean dregs and Chinese medicine dregs with water to prepare a fermentation substrate, inoculating EM bacteria for fermentation for 9-12 days, and filtering.
Preferably, the Chinese medicine residue consists of phellodendron bark residue, angelica dahurica residue and reed rhizome residue.
Still more preferably, the water content of the fermentation substrate is 60-70%, and the fermentation substrate comprises the following components in parts by weight: 40-50 parts of crushed corncobs, 40-50 parts of bean dregs, 5-10 parts of phellodendron bark dregs, 5-10 parts of angelica dahurica dregs, 5-10 parts of reed rhizome dregs and a proper amount of water.
In a second aspect of the present invention, there is provided a method for preparing a plant-derived biocontrol bacterial preparation as described in the first aspect of the present invention, comprising the steps of: (1) Extracting radix Zanthoxyli with water to obtain radix Zanthoxyli water extract; (2) Preparing citrus peel ethanol extract from citrus peel by an ethanol extraction method; (3) Adding water into the crushed corncobs, bean dregs and Chinese medicine dregs, mixing to prepare a fermentation substrate, inoculating EM (effective microorganisms) to ferment for 9-12 days, and filtering to obtain EM fermentation liquor; (4) Mixing EM bacteria fermentation liquid, radix Zanthoxyli water extract and mandarin orange peel ethanol extract uniformly to obtain the final product.
The third aspect of the invention provides the application of the plant-derived biocontrol agent in the first aspect of the invention in controlling rubber anthrax.
The fourth aspect of the invention provides the application of the plant-derived biocontrol bacterial preparation in the control of rubber anthracnose.
The active ingredients of the botanical biocontrol bacteria preparation disclosed by the invention consist of EM (effective microorganisms) and fermentation metabolites thereof, zanthoxylum nitidum aqueous extracts and citrus peel ethanol extracts, belong to active ingredients with different action mechanisms, have a better inhibition effect on rubber anthracnose under the synergistic effect, can effectively prevent and treat the rubber anthracnose, and belong to green, environment-friendly and non-toxic active ingredients with double biological regulation functions of pesticide effect and fertilizer effect.
Detailed Description
The invention will be better understood by reference to the following examples. Those skilled in the art will recognize that the specific techniques or conditions, not specified in the examples, are according to the techniques or conditions described in the literature of the art or according to the product specification. The reagents or instruments used are conventional products which are commercially available, and are not indicated by manufacturers.
Example 1
Preparing radix Zanthoxyli water extract from radix Zanthoxyli by water extraction; the citrus peel is taken to prepare the citrus peel ethanol extract according to an alcohol extraction method.
According to the weight portion, 40 portions of crushed corncobs, 50 portions of bean dregs, 10 portions of phellodendron bark dregs, 10 portions of angelica dahurica dregs, 10 portions of reed rhizome dregs and a proper amount of water are taken to prepare a fermentation substrate, the water content of the fermentation substrate is controlled to be 60-70%, and then EM bacteria are inoculated for fermentation for 12 days and filtered to obtain EM bacteria fermentation liquid.
Mixing EM bacteria fermentation liquid, radix Zanthoxyli water extract and pericarpium Citri Tangerinae ethanol extract uniformly to make the content of radix Zanthoxyli water extract 0.1wt%, the content of pericarpium Citri Tangerinae ethanol extract 0.3wt%, and the total viable count of EM bacteria 5 × 10 7 cfu/ml, and obtaining.
Comparative example 1 (corresponding to example 1)
According to the weight portion, 40 portions of crushed corncobs, 50 portions of bean dregs and a proper amount of water are taken to prepare a fermentation substrate, the water content of the fermentation substrate is controlled to be 60-70%, then EM bacteria are inoculated for fermentation for 12 days, and filtration is carried out to obtain EM bacteria fermentation liquor, wherein the total viable count of the EM bacteria is 5 multiplied by 10 7 cfu/ml。
Example 2
Extracting radix Zanthoxyli with water to obtain radix Zanthoxyli water extract; the citrus peel is taken to prepare the citrus peel ethanol extract according to an ethanol extraction method.
According to the weight portion, 50 portions of crushed corncobs, 40 portions of bean dregs, 5 portions of phellodendron bark dregs, 10 portions of angelica dahurica dregs, 5 portions of reed rhizome dregs and a proper amount of water are taken to prepare a fermentation substrate, the water content of the fermentation substrate is controlled to be 60-70%, and then EM bacteria are inoculated for fermentation for 10 days and filtered to obtain EM bacteria fermentation liquid.
Mixing EM bacteria fermentation liquid, radix Zanthoxyli water extract and pericarpium Citri Tangerinae ethanol extract uniformly to make the content of radix Zanthoxyli water extract 0.3wt%, the content of pericarpium Citri Tangerinae ethanol extract 0.1wt%, and the total viable count of EM bacteria 8 × 10 7 cfu/ml, and obtaining.
Comparative example 2 (corresponding to example 2)
According to the weight portion, 50 portions of crushed corncobs, 40 portions of bean dregs and a proper amount of water are taken to prepare a fermentation substrate, the water content of the fermentation substrate is controlled to be 60-70%, then EM bacteria are inoculated for fermentation for 10 days, and then the EM bacteria fermentation liquid is obtained through filtration.
Example 3
Extracting radix Zanthoxyli with water to obtain radix Zanthoxyli water extract; the citrus peel is taken to prepare the citrus peel ethanol extract according to an ethanol extraction method.
According to the weight portion, 40 portions of crushed corncobs, 50 portions of bean dregs, 10 portions of phellodendron bark dregs, 5 portions of angelica dahurica dregs, 5 portions of reed rhizome dregs and a proper amount of water are taken to prepare a fermentation substrate, the water content of the fermentation substrate is controlled to be 60-70%, and then EM bacteria are inoculated for fermentation for 9 days and filtered to obtain EM bacteria fermentation liquid.
Mixing EM bacteria fermentation liquid, radix Zanthoxyli water extract and pericarpium Citri Tangerinae ethanol extract uniformly to make the content of radix Zanthoxyli water extract 0.12wt%, the content of pericarpium Citri Tangerinae ethanol extract 0.2wt%, and the total viable count of EM bacteria 6 × 10 7 cfu/ml, and obtaining.
Comparative example 3 (corresponding to example 3)
According to the weight portion, 40 portions of crushed corncobs, 50 portions of bean dregs and a proper amount of water are taken to prepare a fermentation substrate, the water content of the fermentation substrate is controlled to be 60-70%, and then EM (effective microorganisms) is inoculated for fermentation for 9 days and filtered to obtain EM fermentation liquid.
Example 4
Extracting radix Zanthoxyli with water to obtain radix Zanthoxyli water extract; the citrus peel is taken to prepare the citrus peel ethanol extract according to an alcohol extraction method.
According to the weight parts, 50 parts of crushed corncobs, 40 parts of bean dregs, 8 parts of phellodendron bark dregs, 7 parts of angelica dahurica dregs, 9 parts of reed rhizome dregs and a proper amount of water are taken to prepare a fermentation substrate, the water content of the fermentation substrate is controlled to be 60-70%, and then EM bacteria are inoculated for fermentation for 11 days and filtered to obtain EM bacteria fermentation liquid.
Mixing EM bacteria fermentation liquid, radix Zanthoxyli water extract and pericarpium Citri Tangerinae ethanol extract uniformly to make the content of radix Zanthoxyli water extract 0.2wt%, the content of pericarpium Citri Tangerinae ethanol extract 0.25wt%, and the total viable count of EM bacteria 9 × 10 7 cfu/ml, and obtaining.
Comparative example 4 (corresponding to example 4)
According to the weight portion, 50 portions of crushed corncobs, 40 portions of bean dregs and a proper amount of water are taken to prepare a fermentation substrate, the water content of the fermentation substrate is controlled to be 60-70%, and then EM (effective microorganisms) are inoculated for fermentation for 11 days and filtered to obtain EM fermentation liquid.
Example 5: experiment for inhibiting bacteria
Preparing a PDA culture medium: weighing peeled potato 200g, cutting, boiling for 30min, filtering with gauze to obtain juice, adding glucose 20.0g and agar 20.0g, dissolving, adding water to 1000mL, packaging into 10 triangular bottles of 150mL, and sterilizing at 121 deg.C for 30 min.
Under aseptic conditions, 20mL of sterilized PDA culture medium cooled to about 45 ℃ is added into a culture dish to prepare a PDA plate. After standing for 24h, inoculating pathogenic bacteria rubber anthrax mycelium in the center of the plate, and culturing at 37 deg.C for 24h. In a well grown rubber anthrax bacteria plate, a round colony block is cut at the edge of a colony by a puncher with the diameter of 1cm and is reserved.
The botanical biocontrol bacteria agents of examples 1-4 were separately taken and the fermentation broth was diluted with sterile water to a total viable count of 5X 10 5 cfu/ml dilution 100. Mu.L of the diluted solution of the plant-derived biocontrol bacteria preparation was added to the PDA plate and spread with an applicator, and the treatment was repeated 5 times each. Inoculating the cut rubber anthrax bacteria block in the center of the coated plate, and inoculating the side with hyphaContacting the flat plate. Culturing at 37 deg.C for 24h, measuring colony diameter (in cm) of rubber anthrax bacteria by ruler cross method, and averaging the results. And centrifuging the mixed fermentation liquor of the comparison example corresponding to each example at 4000r/min for 30min, filtering the supernatant by a 0.22 mu L membrane, and filtering thalli to obtain sterile fermentation supernatant serving as a control. The bacteriostasis rate is calculated according to the following formula:
bacteriostasis rate = (D) Control of -D Treatment of )/D Control X 100%. The results are shown in Table 1.
TABLE 1
| Rate of inhibition of bacteria | |
| Example 1 | 99.12％ |
| Example 2 | 99.01％ |
| Example 3 | 98.39％ |
| Example 4 | 98.76％ |
As can be seen from the table, the invention has good inhibition effect on rubber anthrax.
The embodiments of the present invention have been described in detail, but the embodiments are merely examples, and the present invention is not limited to the embodiments described above. Any equivalent modifications and substitutions for the present invention are within the scope of the present invention for those skilled in the art. Accordingly, equivalent alterations and modifications are intended to be included within the scope of the present invention, without departing from the spirit and scope of the invention.
Claims (4)
1. The application of the botanical biocontrol bacteria preparation for preventing and treating the rubber anthrax is characterized in that the botanical biocontrol bacteria preparation consists of EM (effective microorganisms) fermentation liquor, radix zanthoxyli aqueous extract and citrus peel ethanol extract, wherein the content of the radix zanthoxyli aqueous extract is 0.1-0.3wt%, the content of the citrus peel ethanol extract is 0.1-0.3wt%, and the total viable count of the EM is more than or equal to 5 multiplied by 10 7 cfu/ml; the EM bacteria fermentation liquid is prepared by mixing crushed corncobs, bean dregs and traditional Chinese medicine residues with water to prepare a fermentation substrate, inoculating EM bacteria for fermentation for 9 to 12d, and filtering; the Chinese medicinal residue comprises cortex Phellodendri residue, radix Angelicae Dahuricae residue, and rhizoma Phragmitis residue; the water content of the fermentation substrate is 60-70%, and the fermentation substrate comprises the following components in parts by weight: 40-50 parts of crushed corncobs, 40-50 parts of bean dregs, 5-10 parts of phellodendron bark dregs, 5-10 parts of angelica dahurica dregs, 5-10 parts of reed rhizome dregs and a proper amount of water.
2. The use according to claim 1, wherein the preparation method of the plant-derived biocontrol bacteria preparation comprises the following steps:
(1) Extracting radix Zanthoxyli with water to obtain radix Zanthoxyli water extract;
(2) Preparing citrus peel ethanol extract from citrus peel by an ethanol extraction method;
(3) Adding water into the crushed corncobs, bean dregs and Chinese medicine residues, mixing to prepare a fermentation substrate, inoculating EM (effective microorganisms) to ferment for 9 to 12d, and filtering to obtain EM fermentation liquor;
(4) Mixing EM bacteria fermentation liquid, radix Zanthoxyli water extract and mandarin orange peel ethanol extract uniformly to obtain the final product.
3. The application of the botanical biocontrol agent for preventing and treating the rubber anthracnose is characterized in that the botanical biocontrol agent is prepared from EM (effective microorganisms) fermentation liquor, radix zanthoxyli aqueous extract and citrus peel ethanolThe extract comprises radix Zanthoxyli water extract 0.1-0.3wt%, citrus peel ethanol extract 0.1-0.3wt%, and EM bacteria total viable count not less than 5 × 10 7 cfu/ml; the EM bacteria fermentation liquid is prepared by mixing crushed corncobs, bean dregs and traditional Chinese medicine residues with water to prepare a fermentation substrate, inoculating EM bacteria for fermentation for 9 to 12d, and filtering; the Chinese medicinal residue comprises cortex Phellodendri residue, radix Angelicae Dahuricae residue, and rhizoma Phragmitis residue; the water content of the fermentation substrate is 60 to 70 percent, and the fermentation substrate comprises the following components in parts by weight: 40-50 parts of crushed corncobs, 40-50 parts of bean dregs, 5-10 parts of phellodendron bark dregs, 5-10 parts of angelica dahurica dregs, 5-10 parts of reed rhizome dregs and a proper amount of water.
4. The use according to claim 3, wherein the preparation method of the plant-derived biocontrol bacteria preparation comprises the following steps:
(1) Extracting radix Zanthoxyli with water to obtain radix Zanthoxyli water extract;
(2) Preparing citrus peel ethanol extract from citrus peel by an ethanol extraction method;
(3) Adding water into the crushed corncobs, bean dregs and Chinese medicine residues, mixing to prepare a fermentation substrate, inoculating EM (effective microorganisms) to ferment for 9 to 12d, and filtering to obtain EM fermentation liquor;
(4) Mixing EM bacteria fermentation liquid, radix Zanthoxyli water extract and mandarin orange peel ethanol extract uniformly to obtain the final product.
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| CN1613332A (en) * | 2004-09-15 | 2005-05-11 | 中国农业大学 | Chinese yam preserving agent |
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