Fermentation method of overground part of tobacco for preventing and treating fusarium oxysporum and application of fermentation method

Abstract

The invention discloses a fermentation method of a tobacco overground part for preventing and treating fusarium oxysporum and application thereof, a tobacco overground part fermentation broth is used for preventing and treating diseases caused by fusarium oxysporum, the fermentation method is as follows: the tobacco overground part fermentation broth adopts the tobacco overground part for fermentation, and during fermentation, the ratio of sugar to the tobacco overground part to water is 1: 3: 10; the fermentation liquor disclosed by the invention not only can prevent and treat diseases caused by fusarium oxysporum, but also can be used as an organic fertilizer in the prevention and treatment process, can reduce the use of chemical fertilizers, is low-carbon and environment-friendly, and reduces soil pollution.

Description

Technical Field

[0001] The present invention belongs to the field of microbial pesticides, and particularly relates to a fermentation method for the aerial parts of tobacco used for controlling Fusarium oxysporum and its application. Background Art

[0002] In recent years, continuous cropping of plants, and long-term random use and abuse of pesticides and chemical fertilizers have caused soil acid-base imbalance, serious hardening, and serious plant root rot diseases. Root rot diseases are characterized by very strong infectivity and destructiveness. Often, due to the failure to timely control local diseases, large areas of dead seedlings are caused. Currently, chemical pesticides are mainly relied on for control. From the perspectives of ecological balance, social safety, and control effects, biological agents are the most ideal method for controlling Fusarium root rot. However, their high costs and instability limit the widespread application of biological agents. Summary of the Invention

[0003] The purpose of the present invention is to provide a fermentation method for the aerial parts of tobacco used for controlling Fusarium oxysporum and its application, and to provide an environmentally friendly and effective control method for diseases caused by Fusarium oxysporum, which can both inhibit Fusarium oxysporum and repair the soil.

[0004] The present invention adopts the following technical solutions:

[0005] Using the fermentation broth of the aerial parts of tobacco to control diseases caused by Fusarium oxysporum.

[0006] Using the fermentation broth of the aerial parts of tobacco to control root rot diseases caused by Fusarium oxysporum.

[0007] A fermentation method for the aerial parts of tobacco used for controlling Fusarium oxysporum. The fermentation broth of the aerial parts of tobacco is fermented using the aerial parts of tobacco, and the sugar: aerial parts of tobacco: water used during fermentation = 1:3:10.

[0008] Further, the fermentation conditions are: 25°C, 6 months. After fermentation is completed, the fermentation broth is obtained by filtration.

[0009] Further, the obtained fermentation broth contains Bacillus subtilis and Flavobacterium humidisoli.

[0010] Further, the fermentation method during fermentation of the fermentation broth is:

[0011] On the 1st day of fermentation: Put sugar, the aerial parts of tobacco, and water into the fermentation device according to the ratio;

[0012] On the 2nd - 7th days of fermentation: Vent once a day. If the raw materials float to the water surface, stir the fermentation broth to make the raw materials immerse in the sugar water;

[0013] Days 8 - 14 of fermentation: Vent the gas once on days 10 and 13;

[0014] Days 15 - 21 of fermentation: Vent the gas once on day 21;

[0015] Days 22 - 27 of fermentation: Vent the gas once on day 27;

[0016] Among them, when venting the gas on day 7, add the previously fermented yeast mother liquor to the fermentation device, and the addition amount of the yeast mother liquor is 5 - 10% of the total fermentation broth in the fermentation device;

[0017] Days 28 - 180 of fermentation: No need to vent the gas, place it in a cool place at 25°C for anaerobic fermentation, and add the previously fermented yeast mother liquor to the fermentation device on any day between days 30 - 35, and the addition amount of the yeast mother liquor is 5 - 10% of the total fermentation broth in the fermentation device.

[0018] Furthermore, the fermentation device used for fermenting the fermentation broth includes:

[0019] Fermentation support, with two layers;

[0020] Fermentation barrel, located at the lower layer of the fermentation support; it contains a mixture of sugar, the above - ground part of tobacco, and water in a predetermined ratio; a fermentation barrel cover is provided on the fermentation barrel, and a ventilation and liquid - guiding hole is provided on the fermentation barrel cover;

[0021] Mother liquor barrel, located at the upper layer of the fermentation support; it contains the fermentation broth of the above - ground part of tobacco that has been previously fermented;

[0022] Liquid - guiding pipe, its upper end is connected to the mother liquor barrel, its lower end passes through the upper - layer plate of the fermentation support, and is connected to the fermentation barrel through the ventilation and liquid - guiding hole of the fermentation barrel; the liquid - guiding pipe is integrally connected and composed of a tubular upper - half connecting pipe, a spherical bulging part made of flexible material, and a tubular lower - half connecting pipe from top to bottom in sequence; the bulging part is located on the upper side of the upper - layer plate of the fermentation support, and the liquid - guiding pipe is used to introduce the yeast mother liquor in the mother liquor barrel into the fermentation barrel at a specific time, and the bulging part is used to reduce the pressure in the fermentation barrel during anaerobic fermentation of the fermentation barrel, and enable the gas generated by fermentation to be collected and buffered at the bulging part.

[0023] Furthermore, the fermentation device also includes: a support mechanism;

[0024] The support mechanism includes:

[0025] A plurality of elastic members, arranged vertically, with their upper ends abutting against the lower side of the bulging part and their lower ends abutting against the upper - layer plate of the fermentation support, for propping up the bulging part upward and keeping the liquid - guiding pipe smooth and preventing the bulging part from sinking into the ventilation and liquid - guiding hole in the contracted state.

[0026] Furthermore, the upper and lower ends of each elastic member are fixedly connected with an upper arc-shaped plate and a lower support plate respectively. The upper arc-shaped plate is used to support the bulged part, and the lower support plate is fixed on the upper layer plate of the fermentation bracket.

[0027] Furthermore, the fermentation method of the fermentation liquid during fermentation is as follows:

[0028] Put sugar, the above-ground part of tobacco, and water into the fermentation device in proportion to start fermentation;

[0029] During the fermentation process on the 1st - 27th day of fermentation, continuously release gas through the upper end of the liquid guide pipe at specified times, so that the gas generated by fermentation accumulated in the bulged part is discharged. After the gas release is completed on the 7th day, add the previously fermented fermentation mother liquor to the fermentation device. The addition amount of the fermentation mother liquor is 5 - 10% of the total fermentation liquid in the fermentation device;

[0030] During the fermentation process on the 28th - 180th day, do not release gas, and on any day between the 30th - 35th day, add the fermented fermentation liquid of the above-ground part of tobacco in the mother liquor barrel into the fermentation device through the liquid guide pipe. The addition amount of the fermentation mother liquor is 5 - 10% of the total fermentation liquid in the fermentation device; during the fermentation process, the gas generated by fermentation continuously accumulates in the bulged part until fermentation is completed.

[0031] The beneficial effects of the present invention are as follows:

[0032] The fermentation liquid of the present invention can not only prevent and control the diseases caused by Fusarium oxysporum, but also be used as an organic fertilizer during the prevention and control process, which can reduce the use of chemical fertilizers, is low-carbon and environmentally friendly, and reduces soil pollution;

[0033] The fermentation method of the present invention can obtain a fermentation liquid with a relatively high bacterial content. Add 5 - 10% of the previously fermented fermentation mother liquor of the total fermentation liquid on any day between the 7th day and the 30th - 35th day, so that the fermentation liquid in the fermentation barrel can ferment quickly and supplement new and active bacterial groups into the fermentation barrel; the fermentation liquid of the present invention contains Bacillus subtilis and Flavobacterium humi, which further verifies the prevention and control mechanism of the diseases caused by Fusarium oxysporum;

[0034] The fermentation device of the present invention can not only supplement the mother liquor into the fermentation barrel through the liquid guide pipe; it can also release gas outward through the liquid guide pipe; and it can also collect the gas generated by fermentation through the bulged part on the liquid guide pipe, reduce the pressure of the fermentation barrel, and make the gas generated by fermentation accumulate and buffer at the bulged part, thereby increasing the capacity of the fermentation liquid in the fermentation barrel;

[0035] The fermentation device of the present invention is provided with a support mechanism to support the bulged part and maintain the stability of the bulged part, avoiding the left - right shaking of the bulged part, which affects the exhaust, liquid guiding and collection of fermentation gas of the fermentation barrel. Description of the Drawings

[0036] Figure 1 Schematic diagram of the effect of the fermentation broth of the present invention on the stem length of Dendrobium officinale after treatment

[0037] Figure 2 Schematic diagram of the effect of the fermentation broth of the present invention on the number of stem nodes of Dendrobium officinale after treatment

[0038] Figure 3 Schematic diagram of the effect of the fermentation broth of the present invention on the stem diameter of Dendrobium officinale after treatment

[0039] Figure 4 State diagram of the mycelium of the blank control in the embodiment of the present invention under the microscope

[0040] Figure 5 State diagram of the mycelium under the microscope after treatment with 25 mg / L fermentation broth in the embodiment of the present invention

[0041] Figure 6 State diagram of the mycelium under the microscope after treatment with 50 mg / L fermentation broth in the embodiment of the present invention

[0042] Figure 7 Schematic diagram of the structure of the fermentation device of the present invention

[0043] Figure 8 Schematic diagram of the structure of the fermentation device of the present invention

[0044] Wherein: 10, fermentation support; 11, fermentation barrel; 12, mother liquor barrel; 13, liquid guide pipe; 14, bulging part; 15, elastic part; 16, upper arc plate; 17, lower support plate. Detailed implementation manners

[0045] The present invention will be described in detail below with reference to the accompanying drawings and specific implementation manners.

[0046] The present invention discloses the use of the fermentation broth of the above-ground part of tobacco to prevent and control diseases caused by Fusarium oxysporum.

[0047] The present invention also discloses the use of the fermentation broth of the above-ground part of tobacco to prevent and control root rot caused by Fusarium oxysporum.

[0048] The present invention also discloses a fermentation method for the above-ground part of tobacco used to prevent and control Fusarium oxysporum. The fermentation broth of the above-ground part of tobacco is fermented using the above-ground part of tobacco, and the sugar: above-ground part of tobacco: water used during fermentation = 1:3:10.

[0049] The fermentation conditions are: 25°C, 6 months, and after fermentation, the fermentation broth is obtained by filtration.

[0050] The obtained fermentation broth contains Bacillus subtilis and Flavobacterium humidisoli.

[0051] The fermentation method of the fermentation broth during fermentation is as follows:

[0052] On the 1st day of fermentation: Put sugar, the above-ground part of tobacco, and water into the fermentation device in proportion.

[0053] On the 2nd - 7th days of fermentation: Vent once a day. If the raw materials float to the water surface, stir the fermentation broth to immerse the raw materials in the sugar water.

[0054] On the 8th - 14th days of fermentation: Vent on the 10th and 13th days.

[0055] On the 15th - 21st days of fermentation: Vent on the 21st day.

[0056] On the 22nd - 27th days of fermentation: Vent on the 27th day.

[0057] Among them, when venting on the 7th day, add the previously fermented fermentation mother liquor to the fermentation device, and the addition amount of the fermentation mother liquor is 5 - 10% of the total fermentation broth in the fermentation device.

[0058] On the 28th - 180th days of fermentation: No need to vent, place it in a cool place at 25°C for anaerobic fermentation, and add the previously fermented fermentation mother liquor to the fermentation device on any day between the 30th and 35th days. The addition amount of the fermentation mother liquor is 5 - 10% of the total fermentation broth in the fermentation device.

[0059] The fermentation device used for the fermentation of the fermentation broth includes: a fermentation support 10, a fermentation barrel 11, a mother liquor barrel 12, and a liquid guide pipe 13.

[0060] The fermentation support 10 has two layers.

[0061] The fermentation barrel 11 is located on the lower layer of the fermentation support 10; the fermentation barrel 11 contains a mixture of sugar, the above-ground part of tobacco, and water in a predetermined proportion; a fermentation barrel cover is provided on the fermentation barrel 11, and an air vent and liquid guide hole is provided on the fermentation barrel cover.

[0062] The mother liquor barrel 12 is located on the upper layer of the fermentation support 10; the mother liquor barrel 12 contains the previously fermented fermentation broth of the above-ground part of tobacco.

[0063] The upper end of the liquid guide pipe 13 is connected to the mother liquid barrel 12, and the lower end of the liquid guide pipe 13 passes through the upper layer plate of the fermentation support 10 and is connected to the fermentation barrel 11 through the ventilation and liquid guide hole of the fermentation barrel 11; the liquid guide pipe 13 is integrally connected and composed of a tubular upper half connecting pipe, a spherical bulging part 14 made of flexible material, and a tubular lower half connecting pipe in sequence from top to bottom; the bulging part 14 is located on the upper side of the upper layer plate of the fermentation support 10, and the liquid guide pipe 13 is used to introduce the fermentation mother liquid in the mother liquid barrel 12 into the fermentation barrel 11 at a specific time, and the bulging part 14 is used to reduce the pressure in the fermentation barrel 11 during the anaerobic fermentation of the fermentation barrel 11 and enable the gas generated by fermentation to be collected and buffered at the bulging part 14.

[0064] The fermentation device further includes: a support mechanism; the support mechanism includes: a plurality of elastic members 15, the plurality of elastic members 15 are arranged vertically, the upper ends of the plurality of elastic members 15 abut against the lower side of the bulging part 14, the lower ends of the plurality of elastic members 15 abut against the upper layer plate of the fermentation support 10, and the plurality of elastic members 15 are used to support the bulging part 14 upward and keep the liquid guide pipe 13 smooth and prevent the bulging part 14 from sinking into the ventilation and liquid guide hole in the contracted state.

[0065] Upper arc-shaped plates 16 and lower support plates 17 are respectively fixedly connected to the upper and lower ends of each elastic member 15. The upper arc-shaped plate 16 is used to support the bulging part 14, and the lower support plate 17 is fixed on the upper layer plate of the fermentation support 10.

[0066] The fermentation method during fermentation of the fermentation broth can also be:

[0067] Put sugar, the above-ground part of tobacco, and water into the fermentation device in proportion and start fermentation;

[0068] During the fermentation process on the 1st - 27th day of fermentation, continuously release gas through the upper end of the liquid guide pipe 13 at a specified time, so that the gas generated by fermentation accumulated in the bulging part 14 is discharged. After the gas release is completed on the 7th day, add the previously fermented fermentation mother liquid to the fermentation device, and the addition amount of the fermentation mother liquid is 5 - 10% of the total fermentation broth in the fermentation device;

[0069] Do not release gas during the fermentation process on the 28th - 180th day, and add the fermented fermentation broth of the above-ground part of tobacco in the mother liquid barrel 12 to the fermentation device through the liquid guide pipe 13 on any day between the 30th - 35th day, and the addition amount of the fermentation mother liquid is 5 - 10% of the total fermentation broth in the fermentation device; during the fermentation process, the gas generated by fermentation continuously accumulates in the bulging part 14 until the fermentation is completed.

[0070] The upper end of the liquid guide tube 13 of the present invention can also be provided with a three-way valve. The first passage communicates with the mother liquor barrel 12, the second passage communicates with the outside, and the third passage communicates with the fermentation barrel 11. In this way, when gas needs to be discharged, the second passage connecting the fermentation barrel 11 and the outside is conducted. When fermentation mother liquor needs to be added, the first passage connecting the fermentation barrel 11 and the mother liquor barrel 12 is conducted. When anaerobic fermentation is required, the third passage is closed to maintain anaerobic fermentation, and the gas generated by fermentation is collected at the bulging part 14.

[0071] For the setting of the mother liquor barrel 12, it can be set as a syringe-type mother liquor barrel 12. When it is necessary to introduce the fermentation mother liquor in the mother liquor barrel 12 into the fermentation barrel 11, through pressurization, the mother liquor in the mother liquor barrel 12 can be introduced into the fermentation barrel 11. When it is not necessary to introduce, the outlet valve can be closed.

[0072] For the setting of the mother liquor barrel 12, it can also be set as a pump-pressurized barrel body. Through pressurization and weighing, the fermentation mother liquor in the mother liquor barrel 12 can be introduced into the fermentation barrel 11.

[0073] Example

[0074] The tested Fusarium oxysporum is a strain preserved in the laboratory of the Scientific and Technological Innovation Team for the Protection and Utilization of Orchid Dendrobium Plant Resources in Yunnan Forestry Vocational and Technical College, Yunnan Province.

[0075] In this example, the mycelial growth rate method was used to determine the antibacterial activity of the fermentation broth against the tested pathogenic bacteria. First, the strain was cultured on the PDA medium plane to form a colony for inoculation. Different concentrations of the fermentation broth were mixed with the sterilized and melted medium to make a medicated medium plane. A bacterial cake taken from the cultured colony was inoculated on the medicated medium plane, and the diameter of the colony growth (the average value was taken by the cross method) was measured respectively. The inhibition rate was calculated according to the colony diameter size of each treatment, and the EC 50 、EC 90 values of each fermentation broth against the pathogenic bacteria were obtained, and the virulence of each fermentation broth against each tested pathogenic bacteria was compared.

[0076] A mixed solution was prepared in the ratio of sugar: above-ground part of tobacco: water = 1:3:10 and fermented at 25°C for 6 months, and then filtered to obtain the tobacco fermentation broth.

[0077] Take 200 g of washed and peeled potatoes, cut them into small pieces, boil them in 1000 mL of distilled water for half an hour, filter out the potatoes with a gauze, add 17 g of agar and 15 g of sucrose, heat to melt the agar, then divide and fill it into 250 mL Erlenmeyer flasks, accurately inject 75 mL into each Erlenmeyer flask, add cotton plugs and sterilize under high pressure for 30 min (121°C), and set aside.

[0078] Under aseptic conditions, when the culture medium cooled to about 50 °C, 7.5 mL, 3.75 mL, 1.875 mL, 0.937 mL, and 0.468 mL of the fermentation broth were added to 5 Erlenmeyer flasks containing 75 mL of the culture medium respectively to prepare medicated culture media with contents of 100 mg / L, 50 mg / L, 25 mg / L, 12.5 mg / L, and 6.25 mg / L; only the culture medium was placed in 1 Erlenmeyer flask containing the culture medium as a blank control. After shaking well, it was poured into 9-cm petri dishes to prepare medicated and blank control plates for standby.

[0079] Under aseptic conditions, the pre-activated bacterial source was punched with a puncher with a diameter of 0.5 cm, and the bacterial cake was inoculated into the plate with sterile forceps, 1 bacterial cake was inoculated per dish, and it was repeated 5 times (the operation method of the control was the same as this). After inoculation, it was placed in an incubator at a constant temperature of 26 °C for cultivation.

[0080] Result observation: After inoculation, observe the growth of colonies in each treatment and measure the diameter of the colonies. The colony diameter was measured by the vertical cross method, and its average value was taken (the unit was expressed in cm). For each treatment, the average value of the colony diameters of 5 repetitions was used to calculate the growth inhibition rate.

[0081] The growth inhibition rate of the fermentation broth against the pathogenic bacteria was calculated by the following formula:

[0082]

[0083] After obtaining the growth inhibition rates of the medicaments in each treatment, the growth inhibition rates and concentrations of the medicaments were subjected to probit-logarithmic transformation, the virulence regression equation of each treatment was obtained, and EC 50 and its 95% confidence limit were calculated. SPSS 17.0 was used to analyze the experimental data.

[0084] As can be seen from Table 1 and Table 2, the inhibitory effect of the fermentation broth on the mycelial growth of Fusarium oxysporum increased with the increase of the treatment dose. The inhibition rate of the 100-mg / L fermentation broth against Fusarium oxysporum could reach 100%; the inhibition rate of the 50-mg / L fermentation broth against Fusarium oxysporum could also reach 100% after 1 day of treatment, but the inhibition rate could reach 90% after 2 - 4 days of treatment, and on the 6th day of treatment, its inhibition rate could also reach 82%.

[0085] Table 1 Average colony diameters (cm) of the fermentation broth treatment and the blank control

[0086]

[0087] Table 2 Inhibition rates (%) of the fermentation broth on the mycelial growth of Fusarium oxysporum

[0088]

[0089]

[0090] Note: Different letters in the same row indicate significant differences (P < 0.05).

[0091] Table 3 Virulence of the fermentation broth in inhibiting the mycelial growth of Fusarium oxysporum

[0092] Fusarium oxysporum Toxicity regression equation Correlation coefficient R EC50 (mg / L) 95% confidence limit (mg / L) 1d y = 5.4161x - 5.8104 0.961 11.825 4.449-6.384 2d y = 3.401x - 3.854 0.985 13.590 2.785-4.016 3d y = 2.473x - 2.353 0.952 8.935 1.931-3.016 4d y = 2.722x - 2.739 0.953 10.15 2.161-3.282 5d y = 2.255x - 2.120 0.993 8.706 1.741-2.770 6d y = 2.033x - 1.858 0.997 8.200 1.541-2.526

[0093] As can be seen from Table 3, the EC of the fermentation broth against Fusarium oxysporum at 1 d 50 was 11.825 mg / L, and the EC at 6 d 50 was 8.2 mg / L.

[0094] After culturing each treatment and the control colonies of Fusarium oxysporum at 26 °C for 7 d, mycelia at the edge of the colonies were picked to prepare slide specimens, and the effects of the fermentation broth at different concentrations on the mycelial morphology of Fusarium oxysporum were observed under a biological microscope.

[0095] From Figure 4 it can be seen that the inclusions in normal mycelia are evenly distributed, and the inclusions such as protoplasm and nuclei are evenly distributed; from Figure 5 it can be seen that when the concentration of the fermentation broth is 25 mg / L, the mycelia are uneven in thickness and shrunken; from Figure 6 it can be seen that when the concentration of the fermentation broth is 50 mg / L, the inclusions flow out and empty tubes appear.

[0096] Taking the precious Yunnan medicinal material Dendrobium officinale as the research object and substituting it for foliar fertilizer, the experimental data show that the fermentation broth has a growth-promoting effect and can replace chemical fertilizers. After detection, the contents in the tobacco fermentation broth are shown in Table 4.

[0097] Table 4 Contents of various elements in the tobacco fermentation broth

[0098]

[0099]

[0100] Table 5 Contents of various elements in the commercially available conventional chemical fertilizers

[0101] Sample name Hua Wuque Diluted 1000 times (ppm) Total nitrogen (N) 20.0％ 200 <![CDATA[Water-soluble phosphorus (P2O5)]]> 20.0％ 200 <![CDATA[Water-soluble potassium (K2O)]]> 20.0％ 200 EDTA-Copper (Cu) 0.05 0.5 EDTA-Iron (Fe) 0.1 1.0 EDTA-Manganese (Mn) 0.1 1.0 EDTA-Zinc (Zn) 0.1 1.0 Boron (B) 0.15 1.5

[0102] Table 5 is the table of the contents of various elements of the 20-20-20+TE general balanced fertilizer produced by Shanghai Yongtong Ecological Engineering Co., Ltd. As can be seen from Table 4 and Table 5, the nutrient content of the fermentation broth is lower than that of the chemical fertilizer. However, when it is formulated into a solution and sprayed on the above-ground part of Dendrobium officinale by foliar application, from the data of the growth phenotypes (stem length, number of stem nodes, stem diameter) of Dendrobium officinale (see Figure 1 - 3 ), the fermentation broth has a significant growth-promoting effect compared with spraying with clear water, but there is no significant difference between the fermentation broth and the chemical fertilizer treatment, and the fermentation broth can almost replace the water-soluble fertilizer.

[0103] Through market research, it is found that for growing Chinese herbal medicine plants in greenhouses, usually at least 2 packs of water-soluble fertilizers, 1 pack of high-N fertilizer, 1 pack of balanced fertilizer are required per mu of land per year. Each pack costs 260 yuan and weighs 15 kg, totaling 520 yuan per mu. And producing one ton of chemical fertilizer generates approximately 2.6 - 4.2 tons of carbon dioxide emissions. Therefore, if the fermentation broth prepared in this embodiment is used, the carbon dioxide emissions per mu of land can be reduced by 0.08 - 0.13 tons.

[0104] After using the fermentation broth prepared in this embodiment as the foliar fertilizer for Dendrobium officinale, the heavy metal content in the Dendrobium officinale plants was detected, as shown in Table 6.

[0105] Table 6 Analysis of heavy metal content in Dendrobium officinale plants under different treatments

[0106]

[0107] Water (one year) means using water as a control and spraying it on the leaves of Dendrobium officinale; Fermentation broth (one year) means spraying the fermentation broth prepared in the embodiment on the leaves of Dendrobium officinale; Chemical fertilizer (one year) means spraying Huawuque on the leaves of Dendrobium officinale.

[0108] As can be seen from Table 6, the heavy metal content (Cu, Cd, Hg) in the Dendrobium officinale plants sprayed with the fermentation broth as the foliar fertilizer for one year decreased significantly.

[0109] After using the fermentation broth prepared in this embodiment as the foliar fertilizer for Dendrobium officinale, the content of medicinal components in the Dendrobium officinale plants was detected, as shown in Table 7.

[0110] Table 7 Analysis of medicinal component content in Dendrobium officinale plants under different treatments

[0111]

[0112]

[0113] Water (one year) means using water as a control and spraying it on the leaves of Dendrobium officinale; Fermentation broth (one year) means spraying the fermentation broth prepared in the embodiment on the leaves of Dendrobium officinale; Chemical fertilizer (one year) means spraying Huawuque on the leaves of Dendrobium officinale.

[0114] As can be seen from Table 7, the contents of soluble sugar and mannose in the Dendrobium officinale plants sprayed with the fermentation broth as the foliar fertilizer for one year increased significantly. This shows that the fermentation broth contains rich macroelements and trace elements, which can be directly absorbed and utilized by plants, and can promote the growth of new shoots, leaves, increase the chlorophyll content, single fruit weight and yield of plants. While promoting plant growth, the medicinal components and their contents in the plant body also change.

[0115] DNA extraction was performed on the strain samples in the tobacco fermentation broth prepared in this example. Amplification sequencing was carried out using universal bacterial 16S primers, and the sequencing results were used for comparison in the NCBI database to preliminarily identify the strains in the tobacco fermentation broth.

[0116]

[0117] It was detected that the tobacco fermentation broth also contained Bacillus subtilis. Bacillus subtilis belongs to the domain Bacteria → phylum Bacillati → phylum Bacillota → class Bacilli → order Bacillales → family Bacillaceae → genus Bacillus; its sequence is:

[0118] CTCCTAAAGGTTACCTCACCGACTTCGGGTGTTACAAACTCTCGTGGTGTG

[0119] ACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATC

[0120] CGCGATTACTAGCGATTCCAGCTTCACGCAGTCGAGTTGCAGACTGCGATC

[0121] CGAACTGAGAACAGATTTGTGGGATTGGCTTAACCTCGCGGTTTCGCTGCC

[0122] CTTTGTTCTGTCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGA

[0123] TGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCACCTTA

[0124] GAGTGCCCAACTGAATGCTGGCAACTAAGATCAAGGGTTGCGCTCGTTGCG

[0125] GGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACC

[0126] TGTCACTCTGCCCCCGAAGGGGACGTCCTATCTCTAGGATTGTCAGAGGAT

[0127] GTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCC

[0128] ACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAGTCTTGCGACCGT

[0129] ACTCCCCAGGCGGAGTGCTTAATGCGTTAGCTGCAGCACTAAGGGGCGGAA

[0130] ACCCCCTAACACTTAGCACTCATCGTTTACGGCGTGGACTACCAGGGTATCT

[0131] AATCCTGTTCGCTCCCCACGCTTTCGCTCCTCAGCGTCAGTTACAGACCAG

[0132] AGAGTCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCACCGCT

[0133] ACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGTTCCCCAGTTTCCAAT

[0134] GACCCTCCCCGGTTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCC

[0135] TGCGAGCCCTTTACGCCCAATAATTCCGGACAACGCTTGCCACCTACGTATT

[0136] ACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTC

[0137] AAGGTACCGCCCTATTCGAACGGTACTTGTTCTTCCCTAACAACAGAGCTTT

[0138] ACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGACTTTCG

[0139] TCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGT

[0140] CTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTCGC

[0141] CTTGGTGAGCCGTTACCTCACCAACTAGCTAATGCGCCGCGGGTCCATCTGT

[0142] AAGTGGTAGCCGAAGCCACCTTTTATGTTTGAACCATGCGGTTCAAACAAC

[0143] CATCCGGTATTAGCCCCGGTTTCCCGGAGTTATCCCAGTCTTACAGGCAGGT

[0144] TACCCACGTGTTACTCACCCGTCCGCCGCTAACATCAGGGAGCAAGCTCCC

[0145] ATCTGTCCGCTCG

[0146] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. Using the fermentation broth of the above-ground part of tobacco to control the diseases caused by Fusarium oxysporum.

2. Using the fermentation broth of the above-ground part of tobacco to control the root rot caused by Fusarium oxysporum.

3. A fermentation method for the above-ground part of tobacco used to control Fusarium oxysporum, characterized in that, The fermentation broth of the above-ground part of tobacco is prepared by fermenting the above-ground part of tobacco, and the ratio of sugar: the above-ground part of tobacco: water used in fermentation is 1:3:

10.

4. The fermentation method of the above-ground part of tobacco used for preventing and controlling Fusarium oxysporum according to claim 3, characterized in that, The fermentation conditions are: 25 °C, 6 months. After fermentation, the fermentation broth is obtained by filtration.

5. A fermentation method for the above-ground part of tobacco used for preventing and controlling Fusarium oxysporum, characterized in that, The obtained fermentation broth contains Bacillus subtilis and Flavobacterium humi disoli.

6. A fermentation method for the above-ground part of tobacco used to control Fusarium oxysporum, characterized in that, The fermentation method of the fermentation broth during fermentation is as follows: On the 1st day of fermentation: Put sugar, the above-ground part of tobacco, and water into the fermentation device according to the ratio. On the 2nd - 7th days of fermentation: Vent once a day. If the raw materials float to the water surface, stir the fermentation broth to make the raw materials immerse in the sugar water. On the 8th - 14th days of fermentation: Vent on the 10th and 13th days. On the 15th - 21st days of fermentation: Vent on the 21st day. On the 22nd - 27th days of fermentation: Vent on the 27th day. Among them, when venting on the 7th day, add the previously fermented fermentation mother liquor to the fermentation device, and the addition amount of the fermentation mother liquor is 5 - 10% of the total fermentation broth in the fermentation device. On the 28th - 180th days of fermentation: No need to vent, place it in a cool place at 25 °C for anaerobic fermentation, and add the previously fermented fermentation mother liquor to the fermentation device on any day between the 30th and 35th days, and the addition amount of the fermentation mother liquor is 5 - 10% of the total fermentation broth in the fermentation device.

7. A fermentation method for the above-ground part of tobacco used for preventing and controlling Fusarium oxysporum, characterized in that, The fermentation device used for the fermentation of the fermentation broth includes: A fermentation support (10) with two layers. A fermentation barrel (11) located on the lower layer of the fermentation support (10); it contains a mixture of sugar, the above-ground part of tobacco, and water in a predetermined ratio; a fermentation barrel cover is provided on the fermentation barrel (11), and a ventilation and liquid conduction hole is provided on the fermentation barrel cover. A mother liquor barrel (12) located on the upper layer of the fermentation support (10); it contains the fermentation broth of the above-ground part of tobacco that has been fermented previously. A liquid conduction pipe (13), the upper end of which is connected to the mother liquor barrel (12), and the lower end passes through the upper plate of the fermentation support (10) and is connected to the fermentation barrel (11) through the ventilation and liquid conduction hole of the fermentation barrel (11); the liquid conduction pipe (13) is integrally connected by a tubular upper half connection pipe, a spherical bulging part (14) made of flexible material, and a tubular lower half connection pipe from top to bottom in sequence; the bulging part (14) is located on the upper side of the upper plate of the fermentation support (10), and the liquid conduction pipe (13) is used to introduce the fermentation mother liquor in the mother liquor barrel (12) into the fermentation barrel (11) at a specific time, and the bulging part (14) is used to reduce the pressure in the fermentation barrel (11) during anaerobic fermentation of the fermentation barrel (11) and collect and buffer the gas generated by fermentation at the bulging part (14).

8. A fermentation method for the above-ground part of tobacco used for preventing and controlling Fusarium oxysporum, characterized in that, The fermentation device also includes: a support mechanism; The support mechanism includes: A plurality of elastic members (15), vertically arranged, with their upper ends abutted against the lower side of the bulging part (14) and their lower ends abutted against the upper layer plate of the fermentation bracket (10), are used to support the bulging part (14) upward and keep the liquid guide pipe (13) smooth and prevent the bulging part (14) from sinking into the ventilation liquid guide hole in the contracted state.

9. A fermentation method for the above-ground part of tobacco used to control Fusarium oxysporum, characterized in that, Upper arc-shaped plates (16) and lower support plates (17) are respectively fixedly connected to the upper and lower ends of each elastic member (15). The upper arc-shaped plates (16) are used to support the bulging part (14), and the lower support plates (17) are fixed on the upper layer plate of the fermentation bracket (10).

10. A fermentation method for the above-ground part of tobacco used to control Fusarium oxysporum, characterized in that, The fermentation method of the fermentation liquid during fermentation is as follows: Put sugar, the above-ground part of tobacco, and water into the fermentation device in proportion to start fermentation; During the fermentation process on the 1st - 27th day of fermentation, continuously release gas through the upper end of the liquid guide pipe (13) at specified times, so that the gas generated by fermentation accumulated in the bulging part (14) is discharged. After the gas release is completed on the 7th day, add the previously fermented fermentation mother liquor to the fermentation device. The addition amount of the fermentation mother liquor is 5 - 10% of the total fermentation liquid in the fermentation device; Do not release gas during the fermentation process on the 28th - 180th day, and add the fermented fermentation liquid of the above-ground part of tobacco in the mother liquor barrel (12) to the fermentation device through the liquid guide pipe (13) on any day between the 30th - 35th day. The addition amount of the fermentation mother liquor is 5 - 10% of the total fermentation liquid in the fermentation device; during the fermentation process, the gas generated by fermentation continuously accumulates in the bulging part (14) until fermentation is completed.

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