Chinese herbal medicine cortex phellodendri preparation for inhibiting tomato basal stem rot and use method of Chinese herbal medicine cortex phellodendri preparation

Abstract

The invention discloses a Chinese herbal medicine cortex phellodendri preparation for inhibiting tomato stem rot and a use method thereof.The Chinese herbal medicine preparation is a cortex phellodendri preparation, and the preparation method comprises the steps that a proper amount of cortex phellodendri is weighed and put into a glass container, water with the volume 10 times that of Chinese herbal medicine is added, and the Chinese herbal medicine is soaked for 1 hour; heating for 30 minutes, filtering and collecting liquid medicine, adding the same amount of water, and heating for 30 minutes; filtering and collecting the medicine liquid, combining the medicine liquid obtained twice, and diluting to the required concentration; the application method comprises the following steps: after tomato plants are attacked, tomato seedlings are treated in a root irrigation manner once every three days, and treatment is carried out for three times in total. The cortex phellodendri preparation is a novel pollution-free biostimulant, the resistance of tomatoes to stem rot is improved, the symptoms of the tomato stem rot can be remarkably relieved, plant growth is promoted, the photosynthetic capacity of leaves is enhanced, and the activity of antioxidant enzyme is improved. Compared with a traditional chemical pesticide, the preparation is obvious in treatment effect, environment-friendly and safer.

Description

Technical Field

[0001] The invention relates to the technical field of plant biostimulants, in particular to a Chinese herbal medicine phellodendron preparation for inhibiting tomato stem base rot and a use method thereof. Background Art

[0002] Tomato is one of the most widely grown nightshade vegetables in the world. It is deeply loved by consumers for its good taste, wide variety and rich nutrition. Stem rot caused by Fusarium solani is one of the most common soil-borne diseases in greenhouse tomato growth in northern China in recent years, causing great losses to the tomato industry.

[0003] At present, chemical control is the most commonly used means to prevent and control tomato stem rot. However, long-term use of chemical pesticides will cause a series of environmental problems. Biostimulants have excellent effects on weight loss, efficiency, and resistance improvement and are widely used in agricultural production. Biostimulants mainly include humic acid, protein hydrolyzate and other nitrogen-containing compounds, seaweed extracts, chitosan and other biopolymers, inorganic compounds, microbial agents, etc. When applied to plant leaves or rhizosphere, they can stimulate plant physiological responses, thereby promoting plant absorption and utilization of nutrients, improving the crop rhizosphere ecological environment, and improving the ability of crops to resist biological stress. In recent years, biostimulants have the effects of fertilizer enhancement and soil improvement, which have attracted the attention and choice of more and more fertilizer companies and farmer consumers.

[0004] my country is rich in Chinese herbal medicine resources. Natural Chinese herbal medicines have the characteristics of high efficiency, antibacterial, safety and no residue that ordinary chemical agents do not have. They can solve problems such as pathogen resistance, environmental pollution and pesticide residues. With the development of green agriculture, Chinese herbal medicines are increasingly widely used in biological control. As a traditional Chinese herbal medicine in my country, Phellodendron chinense belongs to the Rutaceae family. Its dried bark is used as medicine with a wide range of pharmacological activities, including anti-inflammatory, antifungal, antioxidant and other pharmacological effects. It is cheap and rich in resources. It is an excellent resource for the development of new biostimulants. Therefore, we select moderately priced Chinese herbal medicines, prepare them into environmentally friendly Chinese herbal medicine extracts, and apply them in the field of crop disease resistance, opening up new ways for the research and development of natural, green and pollution-free biostimulants.

[0005] Therefore, we propose a Chinese herbal medicine Phellodendron amurense preparation for inhibiting tomato stem base rot and a method for using the same. Summary of the invention

[0006] The invention aims to provide a Chinese herbal medicine phellodendron preparation for inhibiting tomato stem base rot and a use method thereof.

[0007] In order to achieve the above object, the present invention adopts the following technical scheme: In a first aspect, the present invention provides a Chinese herbal medicine preparation for inhibiting tomato stem base rot, wherein a component of the Chinese herbal medicine preparation is a Phellodendron chinense preparation.

[0008] In a second aspect, the present invention also provides a method for preparing a Chinese herbal medicine preparation for inhibiting tomato stem rot, comprising the following steps: (1) Weigh an appropriate amount of Phellodendron chinense into a glass container, add 10 times the volume of water of the Chinese herbal medicine, and soak for 1 hour; (2) Heat in a water bath at 100°C for 30 minutes, filter and collect the solution using a funnel and filter paper, add an equal amount of water to the residue, and heat in a water bath at 100°C for 30 minutes; (3) Use a funnel and filter paper to filter and collect the drug solution, combine the two solutions, and dilute to the required concentrations, which are 4, 8, and 12 mg / mL respectively.

[0009] Preferably, the water used in step 1 and step 2 is distilled water.

[0010] In a third aspect, the present invention also provides a method for inoculating tomato stem rot, comprising the following steps: (1) When tomatoes have three leaves and one heart, use a sterilized cotton swab to pick up Fusarium solani ( Fusarium solani ) bacterial liquid, scratch the base of the stem; (2) using a hole puncher to punch holes in the PDA culture medium infested with Fusarium solani; (3) Place the mycelium block directly over the abrasion site and wrap the infected site with sterilized cotton soaked in the Fusarium solani liquid; (4) Place a transparent cover on the tray, spray water on the cover to provide moist conditions for disease, and place in a low-light location for cultivation.

[0011] In a fourth aspect, the present invention also provides a method for using a Chinese herbal medicine preparation for inhibiting tomato stem base rot: the target crop is tomato, and the target site is the base of the stem; after the tomato plants become diseased, the tomato seedlings are treated by root irrigation, once every three days, for a total of three times.

[0012] The beneficial effects of the Chinese herbal medicine Phellodendron amurense preparation for inhibiting tomato stem base rot and the use method thereof of the present invention are specifically as follows: 1. The present invention uses a natural Chinese herbal medicine Phellodendron chinense preparation as a biostimulant, and treats tomato plants by root irrigation, which significantly reduces the symptoms of tomato stem base rot, promotes plant growth, enhances leaf photosynthetic capacity, and improves antioxidant enzyme activity. Therefore, the Phellodendron chinense preparation, as a natural, green, pollution-free new biostimulant, can not only improve the resistance of tomatoes to stem base rot, but also does not cause heavy metal and pesticide residues in tomato plants, and has higher safety; 2. The formula of the present invention is a pure Chinese medicine formula, which is a natural substance with little toxic and side effects, mild efficacy, and no drug resistance. It can not only be effectively absorbed by organisms, but also is not easy to cause soil environmental pollution. It is safe, pollution-free, residue-free, green, healthy and environmentally friendly, and can effectively improve the benefits of tomato planting. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0014] Figure 1 Schematic diagram of the effect of different concentrations of the Cortex Phellodendri preparations provided by the present invention on the aboveground phenotype of tomato after grafting; wherein, Figure A is a diagram of the overall growth phenotype of the plant; Figure B is a diagram of the lesions at the base of the stem; Figure C is the area of ​​the lesions at the base of the stem; Figure D is the plant height; Figure E is the stem diameter; sterile water indicates that the plant has no grafted stem base rot and is treated with sterile water; Figure 2 Schematic diagram of the effect of different concentrations of the Cortex Phellodendri preparations provided by the present invention on the root size of tomato after grafting; wherein, Figure A is the root phenotype of tomato; Figure B is the total length of the root system; C is the root surface area; D is the root volume; E is the average root diameter; F is the number of root tips; G is the fractal dimension; sterile water indicates that the plant has no grafting stem base rot and is treated with sterile water; Figure 3 Schematic diagram of the effect of different concentrations of the Cortex Phellodendri preparations provided by the present invention on the biomass of tomato plants after grafting; wherein, Figure A is the fresh weight of the aboveground part of the plant; Figure B is the fresh weight of the underground part; Figure C is the dry weight of the aboveground part; Figure D is the dry weight of the underground part; Figure E is the fresh root-crown ratio; Figure F is the dry root-crown ratio; sterile water means that the plant has no grafted stem base rot and is treated with sterile water; Figure 4 Schematic diagram of the effect of different concentrations of the phellodendron amurense preparation provided by the present invention on the photosynthetic parameters of tomato leaves; wherein, Figure A is the net photosynthetic rate; Figure B is the intercellular carbon dioxide concentration; Figure C is the stomatal conductance; Figure D is the transpiration rate; Figure E is the actual photochemical efficiency of photosystem II; Figure F is the maximum photochemical efficiency of photosystem II; sterile water indicates that the plant has no grafted stem rot; Figure 5 Schematic diagram of the effect of different concentrations of the Cortex Phellodendri preparations provided by the present invention on the photosynthetic pigments of tomato leaves after grafting; wherein, Figure A is the chlorophyll a content of tomato leaves; Figure B is the chlorophyll b content; Figure C is the total chlorophyll content; Figure D is the carotenoid content; sterile water indicates that the plant has no grafted stem base rot and is treated with sterile water; Figure 6Schematic diagram of the effect of different concentrations of Phellodendron amurense preparations provided by the present invention on the antioxidant enzyme activity and malondialdehyde content of tomato leaves after grafting disease; Figure A is the superoxide dismutase activity of the leaves; Figure B is the peroxidase activity; Figure C is the catalase activity; Figure D is the malondialdehyde content; sterile water indicates that the plants do not have grafted stem base rot and are treated with sterile water. DETAILED DESCRIPTION

[0015] In order to make the technical means, creative features, objectives and effects of the present invention easy to understand, the present invention is further described below in conjunction with specific implementation methods. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. Example

[0016] like Figures 1 to 6 As shown, the present invention provides a Chinese herbal medicine Phellodendron chinense preparation for inhibiting tomato stem base rot, and the component of the Chinese herbal medicine preparation is Phellodendron chinense preparation.

[0017] In this embodiment, the Chinese herbal medicine Cortex Phellodendri preparation is obtained by the following steps: (1) Weigh an appropriate amount of Phellodendron chinense into a glass container, add 10 times the volume of water of the Chinese herbal medicine, and soak for 1 hour; (2) Heat in a water bath at 100°C for 30 minutes, filter and collect the solution using a funnel and filter paper, add an equal amount of water to the residue, and heat in a water bath at 100°C for 30 minutes; (3) Use a funnel and filter paper to filter and collect the drug solution, combine the two solutions, and dilute to the required concentrations, which are 4, 8, and 12 mg / mL respectively.

[0018] The tomato stem base rot inoculation method in this embodiment is as follows: (1) When tomatoes have three leaves and one heart, use a sterilized cotton swab to soak in the fungus solution of Fusarium solani and scratch the base of the stem; (2) using a hole puncher to punch holes in the PDA culture medium infested with Fusarium solani; (3) Place the mycelium block directly over the abrasion site and wrap the infected site with sterilized cotton soaked in the Fusarium solani liquid; (4) Place a transparent cover on the tray, spray water on the cover to provide moist conditions for disease, and place in a low-light location for cultivation.

[0019] The method of using the Chinese herbal medicine preparation in this embodiment is as follows: After the tomato plants become sick, treat the tomato seedlings by root irrigation, once every three days, for a total of three times.

[0020] Among them, the targeted crop is tomato and the targeted part is the base of the stem.

[0021] Test example: The tomato variety used for the test was "Heinz 1706".

[0022] Experimental grouping: The experimental material of this study is the tomato cultivar "Heinz 1706". The seeds were first washed with 75% anhydrous ethanol in a clean bench for 1 minute, then soaked with 25% sodium hypochlorite, shaken in a shaker at 28 ℃ for 20 minutes, washed 10 times with sterile water, and the seed coat was washed until it turned slightly white before being sown in MS medium. After culturing in the dark at 28 °C for 3 days, the seedlings were transferred to light for culturing. When the first true leaf grew out, the seedlings were transplanted into a mixed matrix of vermiculite, perlite, and soil in a ratio of 1:1:2. The seedlings were watered thoroughly and kept in a light cycle of 16 h (light) and 8 h (dark), a day-night temperature cycle of 25 °C / 20 °C, and a light intensity of 150 μmol·m −2 ·s −1 and grown under conditions of 50% relative humidity; After the disease occurred at the base of the stem, the cotton was removed and the base of the stem was irrigated with 0, 4, 8, and 12 mg / mL concentrations of Phellodendron chinense preparations, respectively. Sterile water indicated that the plants were not infected with stem base rot bacteria and were treated with sterile water. Each tomato seedling was irrigated with 30 ml of the Chinese herbal medicine preparation, and the treatment was performed once every 3 days for a total of 3 times.

[0023] Experimental method: Completely randomized block design with 6 biological replicates.

[0024] Experimental results: Refer to the attached Figure 1 There were significant differences in the lesion area between different concentrations at the statistical level, and with the increase of treatment concentration, it showed a trend of first decreasing and then increasing. Among them, the 8 mg / mL treatment had the most obvious inhibitory effect on the lesion area. Compared with the 0 mg / mL treatment group, the plant height of the infected tomato treated with 8 mg / mL increased by 40.44% and the stem diameter increased by 11.27%; Refer to the attached Figure 2 After being treated with different concentrations of Phellodendron amurense preparations, the root length, root surface area, root volume, root average diameter, and root tip number of tomato roots increased significantly, and with the increase of treatment concentration, they first increased and then decreased, and a peak value appeared at the treatment concentration of 8 mg / mL. Refer to the attached Figure 3Compared with the control group, only the treatment concentration of 8 mg / mL significantly increased the dry weight of the aboveground and underground parts, and the promotion effect of the other treatments was not significant. There was no significant difference in the fresh root-to-crown ratio among the treatments, but the dry root-to-crown ratio of the 8 mg / mL treatment increased significantly by 22.60% compared with the 0 mg / mL treatment; the fresh root-to-crown ratio was the largest at the 4 mg / mL treatment concentration, which was significantly increased by 12.89% compared with the 0 mg / mL treatment; See attached Figure 4 , the stomatal conductance and transpiration rate of infected tomatoes in the 8 mg / mL treatment concentration also increased significantly, by 67.95% and 39.50% respectively compared with the 0 mg / mL treatment, while the intercellular carbon dioxide concentration decreased by 13.55% under the 8 mg / mL treatment. ΦPSⅡ (actual photochemical efficiency of photosystem Ⅱ) and Fv / Fm (maximum photochemical efficiency of photosystem Ⅱ) in the 4 mg / mL and 8 mg / mL treatment concentrations were significantly higher than those in the two control groups, and the 12 mg / mL treatment concentration did not change significantly; See attached Figure 5 , the contents of chlorophyll a and chlorophyll b at 4 mg / mL concentration were 0.4458 mg / g and 0.1270 mg / g, respectively, which were 44.18% and 37.74% higher than those at 0 mg / mL. There was no significant difference in carotenoid content, but it also reached the maximum at 4 mg / mL, which was 14.43% higher than that at 0 mg / mL. See attached Figure 6 Compared with the uninoculated group, the antioxidant enzyme activities of the infected tomato leaves treated with Phellodendron chinense preparation were significantly higher than those of the untreated group. The activities of SOD (superoxide dismutase activity), POD (peroxidase activity), and CAT (catalase activity) in the infected tomato leaves showed a trend of first increasing and then decreasing under different concentration treatments, and reached the maximum value at 8 mg / mL. The malondialdehyde content in the infected tomato leaves was significantly higher than that in the uninoculated control group, and showed a trend of first decreasing and then increasing under different concentration treatments. The MDA (malondialdehyde content) content was the lowest at the 8 mg / mL treatment concentration, which was reduced by 67% compared with the 0 mg / mL treatment.

[0025] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. It should be understood by those skilled in the art that the present invention is not limited to the above embodiments. The above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention to be protected. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. The application of Chinese herbal medicine preparation in inhibiting tomato stem rot, characterized in that: The component of the Chinese herbal medicine preparation is a Phellodendron chinense preparation; and the preparation method of the Chinese herbal medicine Phellodendron chinense preparation comprises the following steps: (1) Weigh an appropriate amount of Phellodendron chinense into a glass container, add 10 times the volume of water of the Chinese herbal medicine, and soak for 1 hour; (2) Heat in a 100°C water bath for 30 minutes, filter and collect the solution using a funnel and filter paper, add an equal amount of water to the residue, and heat in a 100°C water bath for 30 minutes; (3) Use a funnel and filter paper to filter and collect the drug solution, combine the two solutions, and dilute to the required concentrations, which are 4, 8, and 12 mg / mL respectively.

2. The method for preparing a Chinese herbal medicine preparation for inhibiting tomato stem rot according to claim 1, characterized in that: The water used in step (1) and step (2) is distilled water.

3. An application of the Chinese herbal medicine Phellodendron chinense preparation as claimed in claim 1 in inhibiting tomato stem rot, characterized in that: The following steps are involved: (1) When tomatoes have three leaves and one heart, treat the tomato seedlings with stem base rot; (2) After the tomato plants become diseased, the tomato seedlings are treated with the Chinese herbal medicine Phellodendron amurense preparation once every three days for a total of three times.

4. The use of the Chinese herbal medicine Phellodendron chinense preparation in inhibiting tomato stem rot according to claim 3, characterized in that: In the step (1), the inoculation treatment of tomato against stem base rot comprises the following steps: (1) When tomatoes have three leaves and one heart, use a sterilized cotton swab to pick up the fungus Fusarium solani ( Fusarium solani ) bacterial liquid, scratch the base of the stem; (2) using a hole puncher to punch holes in the PDA culture medium infested with Fusarium solani; (3) Place the mycelium block directly over the abrasion site and wrap the infected site with sterilized cotton soaked in the Fusarium solani liquid; (4) Place a transparent cover on the tray and spray water on the cover to provide moist conditions for the disease to occur. Place the tray in a low-light location and wait for the tomato plants to become diseased.

5. The use of the Chinese herbal medicine Phellodendron chinense preparation in inhibiting tomato stem rot according to claim 3, characterized in that: In step (2), the part of the tomato seedlings that is treated is the base of the stem, and the treatment method is root irrigation.

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