CN116762825A - Chrysanthemum straw fermentation product of bacillus amyloliquefaciens and application of chrysanthemum straw fermentation product in prevention and control of soil-borne bacterial wilt - Google Patents
Chrysanthemum straw fermentation product of bacillus amyloliquefaciens and application of chrysanthemum straw fermentation product in prevention and control of soil-borne bacterial wilt Download PDFInfo
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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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
- C12N1/20—Bacteria; Culture media therefor
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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
- A01N37/00—Biocides, 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
- A01N37/36—Biocides, 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 at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N37/00—Biocides, 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
- A01N37/36—Biocides, 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 at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids
- A01N37/38—Biocides, 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 at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids having at least one oxygen or sulfur atom attached to an aromatic ring system
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- 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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- A01N63/22—Bacillus
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Abstract
The invention discloses a chrysanthemum straw fermentation product of bacillus amyloliquefaciens T-5, which is prepared by taking bacillus amyloliquefaciens T-5 as a functional microorganism and taking chrysanthemum straw as a substrate through solid fermentation and leaching; the preservation number of the bacillus amyloliquefaciens T-5 is CGMCC No.8547. The chrysanthemum straw is fermented by T-5 to obtain a fermentation product, a small amount of leaching liquor can remarkably inhibit the growth of bacterial wilt, and the order of magnitude of T-5 after the fermentation is finishedAt 10 9 ‑10 10 Between them. The metabonomics and the verification experiment result show that the substances such as partial phenolic acid after T-5 fermentation are key to exerting bacteriostasis, and the secondary metabolites such as p-hydroxy phenylpropionic acid and 2-hydroxy-3-phenylpropionic acid have remarkable inhibition effects on the growth of the bacterial wilt. The invention also discloses application of the chrysanthemum straw fermentation product of the bacillus amyloliquefaciens T-5 in preventing and controlling soil-borne bacterial wilt.
Description
Technical Field
The invention belongs to the field of microorganisms, and relates to a chrysanthemum straw fermentation product of bacillus amyloliquefaciens and application thereof in preventing and controlling soil-borne bacterial wilt.
Background
The microbial fermentation technology is one of important means for synthesizing a certain or a certain class of compounds by utilizing a substrate by functional microorganisms, and has wide application in various fields of medicine, food, agriculture and the like. In the past, most of the compounds rely on chemical synthesis and plant extraction, and although meeting the requirements to a great extent, serious environmental and ecological damages are unavoidable. With the advancement of society and urgent demands for high quality of life, microbial fermentation technology is beginning to gradually replace the two substance acquisition means, and is becoming a research hotspot.
Soil-borne bacterial wilt is a soil-borne bacterial disease caused by Lawsonia solanaceae (Ralstonia solanacearum) and causing irreversible damage to solanaceous crops (such as tomatoes, potatoes, peppers and the like). Although conventional chemical control means (such as chemical pesticide spraying) alleviate the spread of diseases and the attack on crops to some extent, pathogenic bacteria gradually evolve resistance to pesticides over time. Moreover, the application of a large amount of pesticides brings about food safety problems, soil environmental problems, pesticide residue problems and the like.
Disclosure of Invention
The inventor takes a plurality of different straws as substrates and bacillus amyloliquefaciens T-5 as functional microorganisms, explores the inhibition effect of different straw fermentation products on bacterial wilt, and analyzes functional substances which exert key antibacterial effects. The inventors found that: only the leaching liquor of the chrysanthemum straw after being fermented by the bacillus amyloliquefaciens T-5 can remarkably inhibit the growth of the bacterial wilt; further combining with metabonomics, the chrysanthemum straw is fermented by bacillus amyloliquefaciens T-5, and then the enrichment of secondary metabolites such as alkaloids, flavonoids, phenolic acids and the like occurs.
The invention aims at realizing the following technical scheme:
the chrysanthemum straw fermentation product of the bacillus amyloliquefaciens T-5 is prepared by taking the bacillus amyloliquefaciens T-5 as a functional microorganism and taking chrysanthemum straw as a substrate through solid fermentation and leaching.
The chrysanthemum straw fermentation product contains parahydroxybenzoic acid and 2-hydroxy-3-phenylpropionic acid.
Bacillus amyloliquefaciens T-5, which is classified as Bacillus amyloliquefaciens (Bacillus amyloliquefaciens), is preserved in China general microbiological culture Collection center (CGMCC) at 12-month 06 in 2013, and has an address of China national institute of microbiology, china academy of sciences, CGMCC No.8547. Bacillus amyloliquefaciens T-5 is a known microorganism, and the separation and identification and physiological and biochemical characteristics of Bacillus amyloliquefaciens T-5 are described in example 2 of Chinese patent application CN 108835127A.
The invention also provides a preparation method of the chrysanthemum straw fermentation product of the bacillus amyloliquefaciens T-5, which comprises the following steps: the chrysanthemum straw is dried, ground, sieved and sterilized, the water content of the chrysanthemum straw powder is regulated to 65-75% by adopting sterile water, bacillus amyloliquefaciens T-5 bacterial liquid is inoculated according to the inoculum size of 1-2%, solid fermentation is carried out, and then sterile water is added according to the solid-liquid ratio (mass ratio) of 1:5-1:10 for extraction, thus obtaining the fermentation product.
Preferably, the chrysanthemum straw is ground into powder and then is sieved by a 20-mesh sieve.
Preferably, the sterilization is carried out at 115 ℃ for 30min.
Preferably, the concentration of the bacillus amyloliquefaciens T-5 bacterial liquid is OD 600 ＝0.5～0.6。
The bacillus amyloliquefaciens T-5 bacterial liquid is prepared by the following method: selecting a single colony of T-5, inoculating the single colony to NA liquid culture medium, culturing at a rotating speed of 160-200 rpm and a temperature of 25-35 ℃ overnight, absorbing bacterial liquid into a sterile centrifuge tube, centrifuging, discarding supernatant, adopting sterile water to wash the culture medium, adopting sterile water to adjust OD of bacillus amyloliquefaciens T-5 bacterial liquid 600 0.5 to 0.6.
Preferably, the solid fermentation is carried out at an ambient temperature of 26-35 ℃, an air relative humidity of 70-80% and a solid fermentation time of 7-10 days.
Preferably, the leaching conditions are as follows: leaching for 0.5-2 h at normal temperature with the rotating speed of 160-200 rpm.
The invention also aims to provide an application of the chrysanthemum straw fermentation product of the bacillus amyloliquefaciens T-5 in preventing and controlling soil-borne bacterial wilt.
A method for preventing and controlling soil-borne bacterial wilt, comprising: after transplanting tomato seedlings in the trefoil period for 2 weeks, diluting the chrysanthemum straw fermentation product by 10-15 times, and applying 20-50 mL of diluted chrysanthemum straw fermentation product to each tomato.
Preferably, the chrysanthemum straw fermentation product is diluted by 10-15 times by adding water.
The invention also aims to provide an application of the chrysanthemum straw fermentation product of the bacillus amyloliquefaciens T-5 in preparing a pesticide preparation or a fertilizer for preventing and controlling soil-borne bacterial wilt.
The invention also aims to provide the application of the p-hydroxy-phenylpropionic acid or the 2-hydroxy-3-phenylpropionic acid in preparing pesticide preparations or fertilizers for preventing and controlling soil-borne bacterial wilt.
The invention has the beneficial effects that:
the chrysanthemum straw is fermented by T-5 to obtain a fermentation product, a small amount of leaching liquor can remarkably inhibit the growth of bacterial wilt, and the order of magnitude of T-5 after the fermentation is finished is 10 9 -10 10 Between them. The metabonomics and the verification experiment result show that substances such as partial phenolic acid after T-5 fermentation are key to exerting bacteriostasis, and secondary metabolites such as parahydroxy phenylpropionic acid and 2-hydroxy-3-phenylpropionic acid have remarkable inhibitory effects on the growth of the bacterial wilt, and the growth of the bacterial wilt can be remarkably inhibited when the concentration of the parahydroxy phenylpropionic acid or the 2-hydroxy-3-phenylpropionic acid is 0.2 mM.
Drawings
FIG. 1 shows the effect of chrysanthemum straw extract on bacterial wilt biomass.
FIG. 2 is the effect of chrysanthemum straw on T-5 biomass.
FIG. 3 is a comparison of the abundance of different secondary metabolites in post-fermentation straw.
FIG. 4 shows the effect of different concentrations of the secondary metabolite 3- (4-hydroxyphenolyl) -propionic acid on bacterial growth.
FIG. 5 shows the effect of different concentrations of the secondary metabolite 2-Hydroxy-3-phenylpropanoic acid (2-Hydroxy-3-phenylpropionic acid) on growth of ralstonia solanacearum.
Figure 6 shows the effect of different treatments of the extract on the bacterial wilt index of tomatoes.
Biological material preservation information
T-5, its classification name is Bacillus amyloliquefaciens (Bacillus amyloliquefaciens), which is preserved in China general microbiological culture Collection center (CGMCC) at 12 and 06 days in 2013, and has an address of Beijing, chaoyang, north Xiyun No. 1, china academy of sciences microbiological culture Collection center (CGMCC) No.8547.
Detailed Description
The technical scheme of the invention is further described below with reference to the specific embodiments.
Preparing a T-5 bacterial liquid:
streaking the T-5 fungus liquid glycerol tube stored at-80deg.C on NA solid culture medium, standing at 30deg.CPlaced until single colonies are grown. Inoculating single bacteria to NA liquid culture medium, culturing overnight at 170rpm and 30 ℃, then sucking 2mL of bacterial liquid, centrifuging at 6000rpm in a sterile centrifuge tube for 4 min, discarding supernatant, then re-suspending with sterile water, repeating the above operation for 2-3 times to fully wash off the culture medium, measuring absorbance with a microplate reader to obtain absorbance of 600 bacterial liquid, and diluting or concentrating to OD according to absorbance 600 0.5.
Preparation of a green orange fungus (RS) fungus solution:
the laboratory separates and obtains bacterial wilt QL-Rs1115 with strong pathogenicity from tomato plants with onset in kylin town in Nanjing, and the bacterial wilt QL-Rs are called RS (references: wei, Z., yang, X.M., YIn, S.X., shen, Q.R., ran, W).&Xu, y.c. Efficacy of Bacillus-fortified organic fertiliser in controlling bacterial wilt of tomato in the field. Appl. Oil ecol.,2011,48,152-159.). And (3) streaking the RS bacterial liquid glycerol pipe stored at the temperature of minus 80 ℃ on the NA solid culture medium, and standing at the temperature of 30 ℃ until single colonies grow. Inoculating single bacteria to NA liquid culture medium, culturing overnight at 170rpm and 30deg.C, sucking 2mL of bacterial liquid into sterile centrifuge tube, centrifuging at 6000rpm for 4 min, discarding supernatant, re-suspending with sterile water, repeating the above steps for 2-3 times to completely wash off the culture medium to obtain OD 600 0.5 RS bacterial liquid.
Example 1
Inhibition effect of different straw leaching solutions on bacterial wilt
Straw: chrysanthemum straw, corn straw, wheat straw, soybean straw and chilli straw
And (5) taking straws, removing roots and leaves, drying, grinding into powder and sieving with a 20-mesh sieve. Weighing 0.80g of straw powder into a 50mL centrifuge tube, sealing by using a sealing film, and sterilizing for 30min at 115 ℃; setting a straw-T-5 fermentation group and a straw-unfermented group (without inoculation); straw-T-5 fermentation group: regulating water content (mass fraction) to 70% with sterile water in ultra clean bench, and inoculating T-5 bacteria solution (OD) at a ratio of 1% 600 =0.5), and standing and fermenting at 30 ℃ with 80% relative humidity of air for 7d; after fermentation, taking out the centrifuge tube, adding sterile water according to the solid-to-liquid ratio (i.e. mass ratio) of 1:6, andleaching for 1.5h at the normal temperature with the rotating speed of 170 rpm. Chrysanthemum straw-unfermented group: the T-5 bacterial liquid is not connected, and other conditions are controlled to be consistent with the chrysanthemum straw-T-5 fermentation group.
After the completion of the leaching, the mixture was allowed to stand for half an hour, and the supernatant was collected and sterilized by filtration (0.22 μm filter head) to obtain a sterile leaching solution. In 96-well plates according to 10% NA medium (i.e., 10-fold dilution of NA medium), the extract of the straw-T-5 fermentation group and RS broth (OD 600 =0.5) was mixed to a total volume of 200 μl at a volume ratio=178:20:2, and simultaneously incubated with equal volumes of sterile water (CK) and the extract of the chrysanthemum straw-unfermented group as controls at 30 ℃ for 48h at 170rpm, respectively, to determine the OD of the bacterial wilt under different treatments 600 The effect of the leaching solution prepared by fermenting the straw with T-5 on the growth of the ralstonia solanacearum is analyzed, and the results are shown in figure 1 and table 1. After the chrysanthemum straw is fermented by bacillus amyloliquefaciens T-5, the leaching solution can remarkably inhibit the growth of bacterial wilt.
TABLE 1 influence of different treatments on bacterial wilt biomass
Example 2
Determination of T-5 biomass
Straw: chrysanthemum straw, corn straw, wheat straw, soybean straw and chilli straw
And (5) taking straws, removing roots and leaves, drying, grinding into powder and sieving with a 20-mesh sieve. Weighing 0.80g of straw powder into a 50mL centrifuge tube, sealing by using a sealing film, and sterilizing for 30min at 115 ℃; then the water content is adjusted to 70% by sterile water in an ultra clean bench, and the T-5 bacterial liquid (OD) is inoculated according to the proportion of 1% 600 =0.5), and standing and fermenting at 30 ℃ with 80% relative humidity of air for 7d; and after fermentation, taking out the centrifuge tube, adding sterile water according to a solid-liquid mass ratio of 1:6, and leaching for 1h at normal temperature under the rotation speed of 170 rpm. Immediately after the leaching, 1mL of leaching solution is taken for density gradient dilution, and the density gradient dilution is carried out on the leaching solution in V8 selection medium (V8 medium: 340mL of V8 fruit and vegetable juice, 33g of NaCl,0.8g of glucose and deionized water to 1L,pH 5.2) were subjected to coating counting. The results are shown in FIG. 2 and Table 2.
As shown by the results, when the chrysanthemum straw is used as a substrate for fermentation, the effective viable count of T-5 can reach 3.58+/-0.95 (10) 9 CFU/g straw).
TABLE 2 influence of different straw on T-5 biomass
| Straw variety | T-5 effective viable count (10) 9 CFU/g straw |
| Soybean straw | 3.75±0.66 |
| Corn stalk | 7.37±2.34 |
| Wheat straw | 2.19±0.32 |
| Chili straw | 2.51±0.49 |
| Chrysanthemum straw | 3.58±0.95 |
Example 3
Metabonomics analysis of different straw before and after fermentation
Different straws are taken, roots and leaves are removed, and the straws are dried, ground into powder and sieved by a 20-mesh sieve. Weighing 0.80g of straw powder into a 50mL centrifuge tube, sealing by using a sealing film, and sterilizing for 30min at 115 ℃; sterile water conditioning in ultra clean benchWater content is reduced to 70%, and the T-5 bacterial liquid (OD) is inoculated according to the proportion of 1% 600 =0.5), and standing, fermenting and culturing for 7d under the conditions of 80% of air relative humidity and 30 ℃ to obtain different straw fermentation samples, and performing metabonomics measurement while taking unfermented straw as a control.
The sample metabonomics analysis procedure is as follows:
sample extraction flow:
(1) Placing the biological sample in a freeze dryer (Scientz-100F) for vacuum freeze drying;
(2) Grinding (30 Hz,1.5 minutes) to powder by a grinding instrument (MM 400, retsch);
(3) Weighing 100mg of powder, and dissolving in 1.2mL of 70% methanol extract;
(4) Vortex once every 30 minutes for 30 seconds, vortex 6 times altogether, place the sample in a refrigerator at 4 ℃ overnight;
(5) After centrifugation (speed 12000rpm,10 min), the supernatant was aspirated, and the sample was filtered through a microfiltration membrane (0.22 μm pore size) and stored in a sample bottle for UPLC-MS/MS analysis.
Chromatographic mass spectrometry acquisition conditions:
the data acquisition instrument system mainly comprises ultra-high performance liquid chromatography (Ultra Performance Liquid Chromatography, UPLC, SHIMADZU Nexera X2, https:// www.shimadzu.com.cn /) and tandem mass spectrometry (Tandem mass spectrometry, MS/MS, applied Biosystems 4500QTRAP, http:// www.appliedbiosystems.com.cn /).
By combining metabonomics, firstly analyzing secondary metabolites which are remarkably enriched after chrysanthemum straw fermentation, comparing with unfermented chrysanthemum straw metabonomics information, and screening out the secondary metabolites with highest relative abundance in a chrysanthemum straw fermented sample; and then compared with other fermented straw metabonomics transversely, the screening range of key antibacterial substances is further narrowed.
Results: after the chrysanthemum straw is fermented, 45 secondary metabolites are obviously increased, and as can be seen from fig. 4, the total relative abundance of 8 substances is highest, the ratio of the 8 substances is more than 10%, and phenolic acid substances are mainly used.
The inventor adopts an indoor bacteriostasis experiment to verify that: standard stock solutions with the concentration of 100mM are prepared by taking DMSO as a solvent, the standard stock solutions with the final concentrations of 20 mM, 40 mM, 60 mM, 80 mM and 100mM are respectively obtained by gradient dilution with DMSO, and filtration sterilization is carried out by using a 0.22 μm organic phase filter membrane. 178. Mu.L of 10% NA medium, 18. Mu.L of sterile water, 2. Mu.L of standard solutions of the above concentrations and 2. Mu.L of Qingzhujun fungus solution (OD) were added to a 96-well plate 600 =0.5), and culturing at 30deg.C and 170rpm for 24 hr with 2 μl DMSO instead of standard solution as control to determine biomass of Citrus reticulata
The results of indoor bacteriostasis experiments are shown in fig. 4 and 5, and show that the parahydroxybenzoic acid and the 2-hydroxy-3-phenylpropionic acid have remarkable inhibitory effect on the growth of the bacterial wilt, and the growth of the bacterial wilt can be remarkably inhibited when the concentration of the two substances is 0.2 mM.
Example 4
Bacterial wilt prevention and control effect of chrysanthemum straw fermentation leaching liquor
Transplanting tomato seedlings which grow to the trefoil period and have consistent growth vigor into 6-hole seedling trays with 100g of matrix in each hole, culturing in a greenhouse, and controlling the environmental temperature to be 30-35 ℃ and the air humidity to be 60-80%; inoculating bacterial wilt after two weeks, controlling the concentration of bacterial wilt to 10 6 CFU/g matrix, after 3d inoculation and stable colonization with ralstonia solanacearum, three treatment groups were set: RSF1 group: example 1 leaching solution prepared by chrysanthemum straw-T-5 fermentation group was diluted 10 times with water, and 20mL of diluted chrysanthemum straw fermentation leaching solution was applied to each seedling by root irrigation, RSNF1 group: example 1 leaching solution prepared from chrysanthemum straw-unfermented group was diluted 10 times with water, and 20mL of diluted leaching solution of chrysanthemum straw-unfermented group, CK: 20mL of clear water is added to each seedling in a root irrigation mode. And then recording disease prevention conditions of tomatoes every day, and calculating disease indexes at different times according to disease grades, wherein the plant disease grade is classified into 5 grades: 0=healthy plants, 1=1% -25% of the plant parts show symptoms, 2=26% -50% of the plant parts show symptoms, 3=51% -75% of the plant parts show symptoms, 4=plant is completely ill or is illPlants had died (ref: biological control of bacterial wilt of potatoes caused by Pseudomonas solanacearum).
Plant disease index (%) = [ Σ (number of individual stages of disease × number of corresponding stages)/(total number of investigation × highest-stage value) ] ×100 (ref: biological control of bacterial wilt of potatoes: attempts to induce resistance by treating tubes with bacteria).
As shown in fig. 6, the incidence trend of tomato bacterial wilt is remarkably reduced after the leaching solution of the chrysanthemum straw-T-5 fermentation group is added.
Claims (10)
1. The chrysanthemum straw fermentation product of the bacillus amyloliquefaciens T-5 is characterized in that: the bacillus amyloliquefaciens T-5 is used as a functional microorganism, chrysanthemum straw is used as a substrate, and the bacillus amyloliquefaciens is prepared through solid fermentation and leaching;
bacillus amyloliquefaciens T-5, which is classified as Bacillus amyloliquefaciens Bacillus amyloliquefaciens, is preserved in China general microbiological culture Collection center (CGMCC) with a strain preservation number of CGMCC No.8547 in 12-06 of 2013.
2. The chrysanthemum straw fermentation product of bacillus amyloliquefaciens T-5 according to claim 1, which is characterized in that: the chrysanthemum straw fermentation product contains parahydroxy phenylpropionic acid and 2-hydroxy-3-phenylpropionic acid.
3. A method for preparing a chrysanthemum straw fermentation product of bacillus amyloliquefaciens T-5 according to claim 1, which is characterized in that: comprising the following steps: the chrysanthemum straw is dried, ground, sieved and sterilized, the water content of the chrysanthemum straw powder is regulated to 65-75% by adopting sterile water, bacillus amyloliquefaciens T-5 bacterial liquid is inoculated according to the inoculum size of 1-2%, solid fermentation is carried out, and then sterile water is added according to the solid-liquid ratio of 1:5-1:10 for extraction, thus obtaining the fermentation product.
4. The method for preparing the chrysanthemum straw fermentation product of the bacillus amyloliquefaciens T-5, which is characterized in that: grinding the chrysanthemum straw, and sieving with a 20-mesh sieve; the sterilization is carried out for 30min at 115 ℃.
5. The method for preparing the chrysanthemum straw fermentation product of the bacillus amyloliquefaciens T-5, which is characterized in that: the concentration of the bacillus amyloliquefaciens T-5 bacterial liquid is OD 600 ＝0.5～0.6。
6. The method for preparing the chrysanthemum straw fermentation product of the bacillus amyloliquefaciens T-5, which is characterized in that: the ambient temperature of the solid fermentation is 26-35 ℃, the relative humidity of air is 70-80%, and the solid fermentation time is 7-10 days.
7. The method for preparing the chrysanthemum straw fermentation product of the bacillus amyloliquefaciens T-5, which is characterized in that: the leaching conditions are as follows: leaching for 0.5-2 h at normal temperature with the rotating speed of 160-200 rpm.
8. The use of the chrysanthemum straw fermentation product of bacillus amyloliquefaciens T-5 in preventing and controlling soil-borne bacterial wilt.
9. A method for preventing and controlling soil-borne bacterial wilt is characterized by comprising the following steps: comprising the following steps: after transplanting tomato seedlings in the trefoil period for 2 weeks, diluting the chrysanthemum straw fermentation product in the method of 10-15 times, and applying 20-50 mL of diluted chrysanthemum straw fermentation product to each tomato in a root irrigation mode.
10. The application of the parahydroxybenzoic acid or the 2-hydroxy-3-phenylpropionic acid in preparing pesticide preparations or fertilizers for preventing and controlling soil-borne bacterial wilt.
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