Preparation and method for preventing and treating tea tree diseases
By spraying rhamnolipid solution with a concentration of 0.5 mg/mL to 10 mg/mL on tea trees, the problem of unstable environmental pollution and biological control effects in the prevention and control of tea trees is solved, and rapid and effective disease prevention and control and tea trees are achieved.
Patent Information
- Application Number
- CN202510321523.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-18
- Publication Date
- 2025-06-20
AI Technical Summary
The prior art has problems such as environmental pollution and pesticide residues when preventing and controlling fungal diseases in tea tree, and the biological control methods are unstable and the action speed is slow.
Rhamnolipid solution was used, with a concentration of 0.5 mg/mL to 10 mg/mL, and was used to prevent and treat tea ring spots and tea anthrax, and was applied to tea trees by spraying.
Rhamnolipid solution has a significant antibacterial effect on pathogens of tea trenchymal disease and tea anthrax, which can quickly improve the disease resistance of tea trees and reduce or replace the use of chemical pesticides.
Smart Images
Figure SMS_1 
Figure HDA0005317532890000011 
Figure HDA0005317532890000012
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of agricultural planting. Specifically, the present invention relates to a preparation and method for preventing and controlling tea tree diseases. Background Art
[0003] As a perennial plant, tea trees are inevitably threatened by various pests and diseases during their growth process. Among them, fungal diseases in tea tree diseases are one of the important factors affecting the healthy growth of tea trees. There are many types of tea tree fungal diseases, such as tea leaf blight, tea anthracnose, tea wilt disease, etc. These diseases damage the tea tree tissues, hinder their physiological functions, lead to growth retardation or even plant death, and thus greatly reduce the yield and quality of tea leaves. At the same time, the diseases will also change the chemical composition of tea leaves, affect the taste and nutritional value, further weaken their market competitiveness, and affect the entire tea industry chain. Therefore, it is urgent to take effective measures to prevent and control tea tree fungal diseases to ensure the healthy growth of tea trees and further improve the quality of tea in China.
[0004] At present, the main means of preventing and controlling tea tree fungal diseases is chemical control. However, with the increasing attention of people to environmental protection and food safety, the traditional chemical control methods have been controversial due to various problems such as potential environmental pollution and pesticide residues. It may cause various problems such as the degradation of the physical and chemical properties of tea garden soil and the harm to the health of consumers, which greatly restricts the ecological sustainability and high-quality development of the tea industry in different regions. In contrast, the biological control method of using microorganisms or their metabolites to inhibit pathogenic bacteria or induce systemic resistance in tea trees, as an environmentally friendly and sustainable disease control means, is gradually becoming an important strategy for preventing and controlling tea tree fungal diseases. However, the biological control effect is unstable, easily affected by environmental conditions such as temperature and humidity, and the action speed is relatively slow, often taking a long time to take effect. Summary of the Invention
[0005] Based on this, the purpose of the present invention is to provide a preparation and method for quickly preventing and controlling tea tree fungal diseases.
[0006] The specific technical solutions for achieving the above invention purposes are as follows.
[0007] In the first aspect of the present invention, there is provided the application of rhamnolipid solution in preventing and controlling tea tree diseases, wherein the concentration of the rhamnolipid solution is 0.5 mg / mL to 10 mg / mL, and the tea tree diseases are tea leaf blight and / or tea anthracnose.
[0008] In the second aspect of the present invention, there is provided the use of a rhamnolipid solution in inhibiting pathogenic bacteria of tea tree diseases, wherein the concentration of the rhamnolipid solution is 0.5 mg / mL to 10 mg / mL, and the pathogenic bacteria of tea tree diseases are Pseudopestalotiopsis sp., Neopestalotiopsis sp. and / or Colletotrichum sp.
[0009] In the third aspect of the present invention, there is provided a method for controlling tea tree diseases, comprising the following steps: spraying a rhamnolipid solution with a concentration of 0.5 mg / mL to 10 mg / mL on tea trees infected or not infected with tea target spot and / or tea anthracnose.
[0010] In the fourth aspect of the present invention, there is provided a method for improving the disease resistance of tea trees, comprising the following steps: spraying a rhamnolipid solution with a concentration of 0.5 mg / mL to 10 mg / mL on tea trees infected or not infected with tea target spot and / or tea anthracnose.
[0011] The inventors of the present invention found that a rhamnolipid solution with a certain concentration has good antibacterial effects on Pseudopestalotiopsis sp. and Neopestalotiopsis sp. which are the pathogenic bacteria causing tea target spot, and Colletotrichum sp. which is the pathogenic bacteria causing tea anthracnose. Therefore, the rhamnolipid solution provides a new way for the green control of the main tea tree diseases, tea target spot and tea anthracnose, and the control effect is rapid, and it is expected to reduce or replace the use of traditional chemical pesticides and fertilizers in tea gardens.
[0012] In addition, the inventors also found that after treating the healthy tea tree leaves with the rhamnolipid solution, the activities of catalase, peroxidase and superoxide dismutase in the tea tree leaves were significantly increased, and the contents of phenylalanine ammonia-lyase, chitin and proline were significantly increased. Therefore, a rhamnolipid solution with a certain concentration can positively induce the accumulation of metabolites related to the disease resistance of tea trees, thus having the potential to enhance the disease resistance of tea trees. Description of the Drawings
[0013] Figure 1 It shows the induction effect of rhamnolipid solutions with different concentrations on the activity of catalase (CAT) in tea tree leaves in Example 2 of the present invention.
[0014] Figure 2 It shows the induction effect of rhamnolipid solutions with different concentrations on the activity of peroxidase (POD) in tea tree leaves in Example 2 of the present invention.
[0015] Figure 3The induction effect of rhamnolipid solutions with different concentrations in Example 2 of the present invention on the superoxide dismutase (SOD) activity in tea tree leaves.
[0016] Figure 4 The induction effect of rhamnolipid solutions with different concentrations in Example 2 of the present invention on phenylalanine ammonia-lyase in tea tree leaves.
[0017] Figure 5 The induction effect of rhamnolipid solutions with different concentrations in Example 2 of the present invention on chitin in tea tree leaves.
[0018] Figure 6 The induction effect of rhamnolipid solutions with different concentrations in Example 2 of the present invention on proline in tea tree leaves. Detailed implementation manners
[0019] To facilitate the understanding of the present invention, the present invention will be described more comprehensively below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, these embodiments are provided to make the understanding of the disclosed content of the present invention more thorough and comprehensive.
[0020] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those skilled in the technical field to which the present invention belongs. The terms used in the description of the present invention are only for the purpose of describing specific embodiments and are not used to limit the present invention. The term "and / or" used in the present invention includes any and all combinations of one or more of the related listed items.
[0021] If not specifically specified, the embodiments are all carried out under conventional experimental conditions, such as those in the Molecular Cloning Experimental Manual by Sambrook et al. (Sambrook J & Russell DW, Molecular Cloning: a Laboratory Manual, 2013), or under the conditions recommended by the manufacturer's instructions.
[0022] In some embodiments of the present invention, the application of rhamnolipid solutions in preventing and controlling tea tree diseases is disclosed. The concentration of the rhamnolipid solution is 0.5 mg / mL to 10 mg / mL, and the tea tree diseases are tea target spot and / or tea anthracnose.
[0023] In some embodiments, the rhamnolipid solution is an aqueous rhamnolipid solution.
[0024] In some embodiments, the concentration of the rhamnolipid solution is 2 mg / mL to 10 mg / mL.
[0025] In some embodiments, the concentration of the rhamnolipid solution is 2 mg / mL to 6 mg / mL.
[0026] In some of these embodiments, the concentration of the rhamnolipid solution is 2 mg / mL to 5 mg / mL.
[0027] In some of these embodiments, the concentration of the rhamnolipid solution is 2 mg / mL to 3 mg / mL.
[0028] In some of these embodiments, the concentration of the rhamnolipid solution is 2.4 mg / mL to 2.6 mg / mL.
[0029] In some other embodiments of the present invention, the application of the rhamnolipid solution in inhibiting pathogenic bacteria of tea tree diseases is disclosed. The concentration of the rhamnolipid solution is 0.5 mg / mL to 10 mg / mL, and the pathogenic bacteria of tea tree diseases are Pseudopestalotiopsis sp., Neopestalotiopsis sp., and / or Colletotrichum sp.
[0030] In some of these embodiments, the concentration of the rhamnolipid solution is 2 mg / mL to 10 mg / mL.
[0031] In some of these embodiments, the concentration of the rhamnolipid solution is 2 mg / mL to 6 mg / mL.
[0032] In some of these embodiments, the concentration of the rhamnolipid solution is 2 mg / mL to 5 mg / mL.
[0033] In some of these embodiments, the concentration of the rhamnolipid solution is 2 mg / mL to 3 mg / mL.
[0034] In some of these embodiments, the concentration of the rhamnolipid solution is 2.4 mg / mL to 2.6 mg / mL.
[0035] In some other embodiments of the present invention, a method for controlling tea tree diseases is disclosed, including the following steps: spraying a rhamnolipid solution with a concentration of 0.5 mg / mL to 10 mg / mL on tea trees infected or not infected with tea zonate spot and / or tea anthracnose.
[0036] In some of these embodiments, the spraying site is the branches, seedlings, and / or leaves of the tea tree.
[0037] In some of these embodiments, it is sprayed once every 22 h to 26 h, and a total of 3 to 4 times.
[0038] In some other embodiments of the present invention, a method for improving the disease resistance of tea plants is disclosed, which includes the following steps: spraying a rhamnolipid solution with a concentration of 0.5 mg / mL to 10 mg / mL on tea plants infected or not infected with tea zonate leaf spot and / or tea anthracnose.
[0039] In some of these embodiments, the spraying sites are the branches, seedlings, and / or leaves of the tea plants.
[0040] In some of these embodiments, spraying is performed once every 22 h to 26 h, and a total of 3 to 4 times.
[0041] The rhamnolipid (Rhamonlipid, RLs) used in the following embodiments of the present invention has a purity of ≥90% and is purchased from Sigma. The 4 kinds of pathogenic bacteria used in the following embodiments of the present invention were isolated from diseased tea plants by the applicant using conventional methods and verified by molecular biology experiments, and are respectively: strain FHC2-2 (Pseudopestalotiopsis sp., the pathogenic bacterium causing tea zonate leaf spot, GenBank serial number PQ814330), strain XQS5-2 (Neopestalotiopsis sp., the pathogenic bacterium causing tea zonate leaf spot, GenBank serial number PQ814332), strain XQS2-3 (Colletotrichum sp., the pathogenic bacterium causing tea anthracnose, GenBank serial number PQ814331), strain DNS1-2-1 (Fusarium sp., the pathogenic bacterium causing tea wilt disease, GenBank serial number PQ870410). The tea plant variety used in the following embodiments of the present invention is Huangjincha No. 1, which is purchased from Zhaoping County, Hezhou City, Guangxi Zhuang Autonomous Region.
[0042] The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.
[0043] Example 1 Inhibitory effect of rhamnolipid solution on pathogenic bacteria of different tea plant diseases
[0044] This example tested the inhibitory effect of rhamnolipid solutions with different concentrations on pathogenic bacteria of different tea plant diseases, and specifically included the following steps:
[0045] 1. Transfer the tea plant pathogenic fungi (strains FHC2-2, XQS5-2, XQS2-3, DNS1-2-1) isolated, purified and identified in the previous laboratory to new PDA plates respectively, and culture them in an inverted manner in an incubator at 25°C ± 1°C for 3 to 5 days. Use a punch with a diameter of 5 mm to punch out a 5-mm-diameter fungal cake along the edge of the colony for standby.
[0046] 2. Dilute rhamnolipid with sterile water to prepare a stock solution with a concentration of 2.5 g / mL. Filter and sterilize it with a 0.22 μm filter membrane and store it at 4°C for later use.
[0047] 3. Take the rhamnolipid stock solution and add it to the PDA medium that has been sterilized and cooled to 60°C. After mixing well, pour it into petri dishes to prepare PDA media with different concentrations of rhamnolipid. The final concentrations of rhamnolipid are 0.25 mg / mL, 0.5 mg / mL, 1.0 mg / mL, 2.5 mg / mL, 5.0 mg / mL, and 10.0 mg / mL respectively. Inoculate a pathogen disc in the center of each plate. Use the PDA medium without rhamnolipid as the blank control (CK). Repeat each treatment 3 times. All the above operations are carried out on a laminar flow bench.
[0048] 4. After inoculation, place the petri dishes in an incubator at 25°C ± 1°C and incubate them in an inverted position for 5 days. Observe the colony growth situation, measure the colony growth diameter using the cross method, and calculate the growth inhibition rate of different concentrations of rhamnolipid on the tea tree pathogenic fungi.
[0049] Inhibition rate (%) = (control colony diameter - rhamnolipid-treated colony diameter) / control colony diameter × 100%.
[0050] 5. Experimental results
[0051] The antibacterial activities of rhamnolipid solutions with different concentrations against different tea tree disease pathogens are shown in Table 1.
[0052] Table 1
[0053]
[0054] Table 1 results show that rhamnolipid solutions at different concentrations have no obvious inhibitory effect on the tea blight pathogen strain DNS1-2-1 (Fusarium sp.); while they have inhibitory effects on the tea zonate leaf spot pathogen - strain FHC2-2 (Pseudopestalotiopsis sp.), strain XQS5-2 (Neopestalotiopsis sp.), and the tea anthracnose pathogen strain XQS2-3 (Colletotrichum sp.), and the antibacterial effects show similar patterns. Specifically, when the concentration of rhamnolipid exceeds 0.5 mg / mL, it has a certain antibacterial effect. As the concentration increases to 2.5 mg / mL and 5.0 mg / mL, the antibacterial effect is significantly improved, basically exceeding 80%. However, when the concentration continues to increase to 10.0 mg / mL, the antibacterial effect decreases instead. Therefore, rhamnolipid solutions with concentrations of 0.5 mg / mL to 10.0 mg / mL have good effects on inhibiting the tea zonate leaf spot pathogen and the tea anthracnose pathogen. Among them, when the concentration of the rhamnolipid solution is 2.5 mg / mL, the antibacterial effect is the best.
[0055] Example 2: Effects of rhamnolipid solution on substances related to inducing tea tree resistance
[0056] Based on the antibacterial effect experiment in Example 1, this example further studied the effects of rhamnolipid solutions at different concentrations on the accumulation changes of endogenous related resistance substances in tea trees.
[0057] After selecting healthy two-year-old Huangjincha No. 1 seedlings with soil and bringing them back to the laboratory, in a greenhouse at 25°C ± 1°C, different concentrations of rhamnolipid solutions were evenly sprayed on the upper leaves of the tea trees until droplets appeared on the leaf surface. Spraying was carried out once every 24 hours for 3 consecutive times (5 seedlings were taken for each concentration for the experiment, and 3 biological replicates were carried out). On the 4th day, the upper young tea tree leaves were taken for the detection and analysis of metabolites closely related to tea tree disease resistance (including antioxidant enzyme activity, phenylalanine ammonia-lyase, chitin, and proline).
[0058] 1. Antioxidant enzyme activity
[0059] Catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) are the main ROS enzyme scavenging systems in plants. The three work together to effectively scavenge free radicals and peroxides in plants, thereby improving the disease resistance of plants.
[0060] After spraying and treating with rhamnolipid solutions at different concentrations, the activity results of catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) in tea tree leaves are as Figures 1 to 3As shown in the figure. The results show that the treatment with rhamnolipid solutions at concentrations of 0.5 mg / mL to 10.0 mg / mL increased the activities of CAT, POD, and SOD in tea tree leaves to varying degrees, thus having the potential to enhance the disease resistance of tea trees. Among them, the rhamnolipid solution at 2.5 mg / mL had the highest induction effect on the antioxidant enzyme activities of tea trees.
[0061] 2. Phenylalanine ammonia-lyase, chitin, proline
[0062] When plants are attacked by pathogens, they often synthesize some other metabolites closely related to disease resistance to protect themselves to the greatest extent. Among them, phenylalanine ammonia-lyase (PAL) is a key enzyme in secondary metabolism closely related to the disease resistance of plants and is closely related to the disease resistance / tolerance of plants. Chitinase (CHIT) is a glycoside hydrolase that can hydrolyze the natural polymer chitin and is a key enzyme in the biodegradation of chitin. It can degrade the cell walls of most fungi and prevent fungi from infecting, colonizing, and spreading in plants, playing a very important role in the study of plant antifungal diseases. In addition, proline, as an important secondary metabolite involved in plant resistance, plays a very important role in maintaining the integrity of cell membrane structure under stress conditions and eliminating the excessive accumulation of ammonia in plants.
[0063] After spraying and treating with rhamnolipid solutions at different concentrations, the results of the contents of phenylalanine ammonia-lyase, chitin, and proline in tea tree leaves are as Figures 4 to 6 shown. The results show that the treatment with rhamnolipid solutions at 0.5 mg / mL to 10.0 mg / mL can induce the accumulation levels of phenylalanine ammonia-lyase, chitin, and proline in tea tree leaves to varying degrees. Among them, the rhamnolipid solution at 2.5 mg / mL has the best induction effect on phenylalanine ammonia-lyase, and the rhamnolipid solutions at 2.5 mg / mL and 5.0 mg / mL have comparable induction levels on chitin and proline.
[0064] The results of this example show that rhamnolipid solutions at 0.5 mg / mL to 10.0 mg / mL have a positive effect on inducing the levels of metabolites related to the disease resistance of tea trees, thus having the potential to enhance the disease resistance of tea trees.
[0065] The technical features of the above-described embodiments can be combined arbitrarily. For the sake of brevity of description, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered as the scope described in this specification.
[0066] The above-described embodiments merely represent several implementation manners of the present invention. The description thereof is relatively specific and detailed, but it should not be construed as a limitation on the scope of the invention patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all fall within the protection scope of the present invention. Therefore, the protection scope of the present invention patent shall be subject to the appended claims.
Claims
1. The application of rhamnolipid solution in preventing and treating tea tree diseases is characterized in that: The concentration of the rhamnolipid solution is 0.5 mg / mL to 10 mg / mL, and the tea tree disease is tea ring spot and / or tea anthracnose.
2. The use according to claim 1, characterized in that: The rhamnolipid solution is a rhamnolipid aqueous solution; And / or, the concentration of the rhamnolipid solution is 2 mg / mL to 10 mg / mL.
3. The use according to claim 2, characterized in that: The concentration of the rhamnolipid solution is 2 mg / mL to 6 mg / mL.
4. The use according to claim 3, characterized in that: The concentration of the rhamnolipid solution is 2 mg / mL to 5 mg / mL.
5. The use according to claim 4, characterized in that: The concentration of the rhamnolipid solution is 2 mg / mL to 3 mg / mL, more preferably 2.4 mg / mL to 2.6 mg / mL.
6. The application of rhamnolipid solution in inhibiting tea tree disease pathogens, characterized in that: The concentration of the rhamnolipid solution is 0.5 mg / mL to 10 mg / mL, and the tea tree disease pathogens are Pseudopestalotiopsis sp., Neopestalotiopsis sp. and / or Colletotrichum sp.
7. The use according to claim 6, characterized in that: The concentration of the rhamnolipid solution is 2 mg / mL to 10 mg / mL, preferably 2 mg / mL to 6 mg / mL, preferably 2 mg / mL to 5 mg / mL, preferably 2 mg / mL to 3 mg / mL, and more preferably 2.4 mg / mL to 2.6 mg / mL.
8. A method for preventing and controlling tea tree diseases, characterized in that: The following steps are involved: For tea trees infected or not infected with tea ring spot and / or tea anthracnose, spray a rhamnolipid solution with a concentration of 0.5 mg / mL to 10 mg / mL.
9. The method for preventing and controlling tea tree diseases according to claim 8, characterized in that: The concentration of the rhamnolipid solution is 2 mg / mL to 10 mg / mL, preferably 2 mg / mL to 6 mg / mL, preferably 2 mg / mL to 5 mg / mL, preferably 2 mg / mL to 3 mg / mL, and more preferably 2.4 mg / mL to 2.6 mg / mL; And / or, the spraying site is branches, seedlings, and / or leaves of tea trees; And / or, spray once every 22h to 26h, for a total of 3 to 4 times.
10. A method for improving disease resistance of tea plants, characterized in that: The following steps are involved: For tea trees infected or not infected with tea ring spot and / or tea anthracnose, rhamnolipid with a concentration of 0.5 mg / mL to 10 mg / mL is sprayed, and the concentration of the rhamnolipid solution is preferably 2 mg / mL to 10 mg / mL; preferably 2 mg / mL to 6 mg / mL; preferably 2 mg / mL to 5 mg / mL; preferably 2 mg / mL to 3 mg / mL; more preferably 2.4 mg / mL to 2.6 mg / mL.