Exogenous induction method for increasing EGCG content of tea tree

Abstract

The invention belongs to the technical field of tea planting, and particularly relates to an exogenous induction method for increasing the EGCG content of tea trees. The invention provides an exogenous induction method for increasing the EGCG content of a tea tree. An allogenic substance adenosine triphosphate is applied to the tea trees and lasts for a certain time after application, and the purpose of increasing the EGCG content is achieved. According to the method, the EGCG content of tea trees including the Jinxuan tea tree variety can be increased to 0.5%-28%. According to the method, more EGCG can be accumulated in the tea tree in a short time by using a simple means and easily available and safe materials, and the method has important significance on tea processing and deep processing.

Description

Technical Field

[0001] The present invention belongs to the technical field of tea tree planting, and more specifically relates to an exogenous induction method for increasing the EGCG content in tea trees. Background Art

[0002] EGCG (epigallocatechin gallate) is an important secondary metabolite in tea plants, exhibiting significant antioxidant, antibacterial, anti-inflammatory, and antiviral properties. While EGCG can be produced synthetically, tea plants are the primary natural source of EGCG. In recent years, tea breeders have focused on increasing EGCG content through two primary approaches: screening for naturally high-EGCG tea resources and developing exogenous induction techniques to promote EGCG synthesis in tea plants.

[0003] At present, the exogenous induction methods for increasing the EGCG content of tea trees mainly include sucrose induction, light induction, hormone induction, temperature induction and chemical induction. For example, Chinese patent application CN103493694A discloses an exogenous inducer solution containing esters, salts and solubilizers, which can effectively increase the EGCG content of Longjing 43 tea varieties. However, this method has obvious limitations: the methyl ester substances in the inducer are toxic and may have a negative impact on the environment and tea tree growth. Long-term use of these chemicals may lead to soil pollution or a decrease in tea tree resistance. In addition, this method requires continuous spraying for at least 4 days, which is time-consuming.

[0004] In view of these shortcomings of existing methods, developing a safe, rapid and effective exogenous induction method to increase the EGCG content in tea plants has become an urgent need in current research. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to overcome the defects and shortcomings of existing induction reagents for increasing the EGCG content in tea plants, such as certain toxicity and time-consuming use, and to provide an application of the exogenous substance adenosine triphosphate in promoting the biosynthesis of EGCG in tea plants.

[0006] The purpose of the present invention is to provide an exogenous induction method for increasing the EGCG (epigallocatechin gallate) content in tea plants.

[0007] The above-mentioned purpose of the present invention is achieved through the following technical solutions: The present invention protects the application of exogenous substance adenosine triphosphate (ATP) in promoting the biosynthesis of EGCG in tea plants.

[0008] ATP is clinically used as a coenzyme drug, commonly used to treat conditions such as heart failure, myocarditis, and cerebral arteriosclerosis. By improving cellular metabolism and providing energy support, it promotes the repair and regeneration of damaged cells, thereby improving patients' symptoms and quality of life. The inventors have discovered that applying exogenous ATP to tea plants can promote the synthesis of EGCG and increase its content.

[0009] Furthermore, the varieties of the tea trees include one or more of Jinxuan (Taiwan Tea No. 12), Fuding Dabaicha (Huacha No. 1), Tieguanyin (Huacha No. 7), and Yinghong No. 9.

[0010] Preferably, the concentration of ATP is 0.05-10 mM. ATP of different concentrations is prepared by diluting with water.

[0011] More preferably, the concentration of ATP is 0.1-2 mM.

[0012] The present invention also protects an exogenous induction method for increasing the EGCG content of tea plants, comprising the following steps: applying an exogenous substance adenosine triphosphate to the tea plants for a certain period of time to achieve the purpose of increasing the EGCG content.

[0013] The inventors discovered that applying exogenous ATP to tea plants can promote the synthesis of EGCG and increase the EGCG content in tea plants. This method not only avoids the use of toxic chemicals but also shortens the induction time, ensuring environmental friendliness and tea plant health.

[0014] Preferably, the concentration of ATP is 0.05-10 mM.

[0015] More preferably, the concentration of ATP is 0.1-2 mM.

[0016] Preferably, the duration after applying is 1 to 25 hours.

[0017] More preferably, the duration after application is 4 to 24 hours.

[0018] Furthermore, the application method is spraying.

[0019] As a preferred embodiment, when the concentration of ATP is 0.1 mM≤≤1 mM, the duration after application is 12 to 24 hours; when the concentration of ATP is 1 mM<≤2 mM, the duration after application is 6 to 12 hours.

[0020] Furthermore, the time for applying the exogenous substance adenosine triphosphate is when the tea tree has grown at least one bud and two leaves.

[0021] Furthermore, the applied dose is 0.05 to 0.2 liters / m 2 , more preferably 0.08~0.12 L / m 2 .

[0022] Furthermore, the EGCG content is detected by high performance liquid chromatography.

[0023] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides an exogenous induction method for increasing the EGCG content in tea plants, comprising the following steps: applying an exogenous substance, adenosine triphosphate, to the tea plants for a period of time to increase the EGCG content. This method can increase the EGCG content of tea plants, including the Jinxuan tea variety, to 0.5% to 28%. This method, using simple means and readily available, safe materials, allows tea plants to accumulate significant amounts of EGCG in a short period of time, and has significant implications for tea processing and further processing. DETAILED DESCRIPTION

[0024] The present invention is further described below with reference to specific examples, which, however, are not intended to limit the present invention in any way. Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional reagents, methods, and equipment in the art.

[0025] Unless otherwise specified, the reagents and materials used in the following examples were commercially available.

[0026] In the embodiment, the Jinxuan tea tree variety in the tea garden of South China Agricultural University was used as the experimental object, and different concentrations (0.1 mM, 1 mM, 2 mM, 5 mM, 10 mM; specifically, 10 mM ATP was first prepared (dissolved in pure water), and then diluted downward according to the concentration gradient, and the final concentrations of the prepared ATP solutions were 0.1 mM, 1 mM, 2 mM, 5 mM, and 10 mM, respectively), and the spraying dose was 0.1 liter / m 2 The picking time was 6 h, 12 h, and 1 day, respectively. One bud and two leaves were picked and their EGCG content was determined by high performance liquid chromatography.

[0027] The HPLC detection conditions were as follows: C18 column, column temperature 35 °C, injection volume 10 μL, mobile phase A: methanol, mobile phase B: 1% trifluoroacetic acid in water, flow rate 1 mL / min, detection wavelength 278 nm, elution gradient: 0-2 min, 2% A; 2-8 min, A increased to 8%; 8-30 min, A increased to 10%; 30-35 min, A increased to 20%; 35-45 min, A increased to 25%; 45-50 min, A increased to 30% and held for 5 min; 55-60 min, A increased to 35%.

[0028] Example 1 An exogenous induction method for increasing the EGCG content of Jinxuan tea variety Select the Jinxuan tea garden with good growth rate of 10 m 2 One bud and two leaves of Jinxuan tea tree were picked 6 hours after spraying with pure water, freeze-dried and solidified, and the EGCG content was detected by high performance liquid chromatography, which was 79.88 mg / g, and used as the control for exogenous induction treatment.

[0029] 10 m above the Jinxuan Tea Garden 2 In the experiment, one bud and two leaves of Jinxuan tea tree were picked 6 hours after spraying 0.1 mM ATP, and the samples were freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 85.49 mg / g, an increase of 7.02% compared with before exogenous induction treatment.

[0030] Example 2: An exogenous induction method for increasing the EGCG content of Jinxuan tea variety Select the Jinxuan tea garden with good growth rate of 10 m 2 One bud and two leaves of Jinxuan tea tree were picked 6 hours after spraying with pure water, freeze-dried and solidified, and the EGCG content was detected by high performance liquid chromatography, which was 79.88 mg / g, and used as the control for exogenous induction treatment.

[0031] 10 m above the Jinxuan Tea Garden 2 In the experiment, one bud and two leaves of Jinxuan tea tree were picked 6 hours after spraying 1 mM ATP, and the samples were freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 86.20 mg / g, an increase of 7.92% compared with that before exogenous induction treatment.

[0032] Example 3: An exogenous induction method for increasing the EGCG content of Jinxuan tea variety Select the Jinxuan tea garden with good growth rate of 10 m 2 One bud and two leaves of Jinxuan tea tree were picked 6 hours after spraying with pure water, freeze-dried and solidified, and the EGCG content was detected by high performance liquid chromatography, which was 79.88 mg / g, and used as the control for exogenous induction treatment.

[0033] 10m above the Jinxuan Tea Garden2 In the experiment, one bud and two leaves of Jinxuan tea tree were picked 6 hours after spraying 2 mM ATP, freeze-dried and solidified. The EGCG content was detected by high performance liquid chromatography, which was 97.24 mg / g, an increase of 21.73% compared with that before exogenous induction treatment.

[0034] Example 4: An exogenous induction method for increasing the EGCG content of Jinxuan tea variety Select the Jinxuan tea garden with good growth rate of 10 m 2 One bud and two leaves of Jinxuan tea tree were picked 12 hours after spraying with pure water, freeze-dried and solidified, and the EGCG content was detected by high performance liquid chromatography, which was 86.20 mg / g, and used as the control for exogenous induction treatment.

[0035] 10 m above the Jinxuan Tea Garden 2 In the experiment, one bud and two leaves of Jinxuan tea tree were picked 12 hours after spraying 0.1 mM ATP, and the samples were freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 96.18 mg / g, an increase of 11.59% compared with that before exogenous induction treatment.

[0036] Example 5: An exogenous induction method for increasing the EGCG content of Jinxuan tea variety Select the Jinxuan tea garden with good growth rate of 10 m 2 One bud and two leaves of Jinxuan tea tree were picked 12 hours after spraying with pure water, freeze-dried and solidified, and the EGCG content was detected by high performance liquid chromatography, which was 86.20 mg / g, and used as the control for exogenous induction treatment.

[0037] 10m from the above Jinxuan Tea Garden 2 In the experiment, one bud and two leaves of Jinxuan tea tree were picked 12 hours after spraying 1 mM ATP, and the samples were freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 101.762 mg / g, an increase of 18.05% compared with that before exogenous induction treatment.

[0038] Example 6: An exogenous induction method for increasing the EGCG content of Jinxuan tea variety Select the Jinxuan tea garden with good growth rate of 10 m 2 One bud and two leaves of Jinxuan tea tree were picked 12 hours after spraying with pure water, freeze-dried and solidified, and the EGCG content was detected by high performance liquid chromatography, which was 86.20 mg / g, and used as the control for exogenous induction treatment.

[0039] 10 m above the Jinxuan Tea Garden 2 In the mu, one bud and two leaves of Jinxuan tea tree were picked 12 hours after spraying 2 mM ATP, and the solid samples were freeze-dried. The EGCG content was determined by high performance liquid chromatography-mass spectrometry and was 110.17 mg / g, which was 27.81% higher than that before exogenous induction treatment.

[0040] Example 7: An exogenous induction method for increasing the EGCG content of Jinxuan tea variety Select the Jinxuan tea garden with good growth rate of 10 m 2 One bud and two leaves of Jinxuan tea tree were picked one day after spraying with pure water, and the solid sample was freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 106.71 mg / g, and used as the control for exogenous induction treatment.

[0041] 10 m above the Jinxuan Tea Garden 2 In the experiment, one bud and two leaves of Jinxuan tea tree were picked one day after spraying 0.1 mM ATP, and the samples were freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 119.95 mg / g, an increase of 12.41% compared with that before exogenous induction treatment.

[0042] Example 8: An exogenous induction method for increasing the EGCG content of Jinxuan tea variety Select the Jinxuan tea garden with good growth rate of 10 m 2 One bud and two leaves of Jinxuan tea tree were picked one day after spraying with pure water, and the solid sample was freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 106.71 mg / g, and used as the control for exogenous induction treatment.

[0043] 10 m above the Jinxuan Tea Garden 2 In the experiment, one bud and two leaves of Jinxuan tea tree were picked one day after spraying 1 mM ATP, and the samples were freeze-dried. The EGCG content was determined by high performance liquid chromatography (HPLC) and was 122.92 mg / g, which was 15.19% higher than that before exogenous induction treatment.

[0044] Example 9: An exogenous induction method for increasing the EGCG content of Jinxuan tea variety Select the Jinxuan tea garden with good growth rate of 10 m 2 One bud and two leaves of Jinxuan tea tree were picked one day after spraying with pure water, and the solid sample was freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 106.71 mg / g, and used as the control for exogenous induction treatment.

[0045] 10 m above the Jinxuan Tea Garden 2 In the experiment, one bud and two leaves of Jinxuan tea tree were picked one day after spraying 2 mM ATP, and the samples were freeze-dried. The EGCG content was determined by high performance liquid chromatography (HPLC) and was 107.27 mg / g, which was 0.52% higher than that before exogenous induction treatment.

[0046] The test results of Examples 1 to 9 are summarized as shown in Table 1.

[0047] Table 1 Effects of different ATP concentrations and treatment times on EGCG content in Jinxuan tea cultivar

[0048] Note: All samples were harvested from the Jinxuan tea garden at South China Agricultural University. One bud and two leaves were picked from different Jinxuan tea trees for testing. Three biological replicates were obtained, and each sample was subjected to three technical replicates. The average result was taken as the final result.

[0049] The test results are shown in Table 1. ATP spraying at a concentration of 0.1 to 2 mM can increase the EGCG content in tea plants. EGCG content increased 6 h, 12 h, and 1 day after spraying. Specifically, when the ATP spraying concentration was 0.1 mM, EGCG content increased significantly 12 h to 1 day after spraying, by 11% to 13%; when the ATP spraying concentration was 1 mM, EGCG content increased significantly 12 h to 1 day after spraying, by 15% to 19%; and when the ATP spraying concentration was 2 mM, EGCG content increased significantly 6 to 12 h after spraying, by 21% to 28%.

[0050] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered as equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. Application of exogenous substance adenosine triphosphate in promoting EGCG biosynthesis in tea plants.

2. The application according to claim 1, characterized in that The varieties of the tea trees include one or more of Jinxuan, Fuding Dabaicha, Tieguanyin, and Yinghong No.

9.

3. The use according to claim 1, characterized in that The concentration of the adenosine triphosphate is 0.05-10 mM.

4. The use according to claim 3, characterized in that The concentration of ATP is 0.1-2 mM.

5. An exogenous induction method for increasing the EGCG content in tea plants, characterized in that: The method comprises the following steps: applying exogenous substance adenosine triphosphate to tea trees, and applying the substance for a certain period of time to achieve the purpose of increasing the EGCG content.

6. The method according to claim 5, wherein: The concentration of the adenosine triphosphate is 0.05-10 mM.

7. The method according to claim 6, wherein: The concentration of ATP is 0.1-2 mM.

8. The method according to claim 5, wherein: The duration after application is 1 to 25 hours.

9. The method according to claim 8, wherein The duration after application is 4 to 24 hours.

10. The method according to claim 5, characterized in that: The time for applying the exogenous substance adenosine triphosphate is when the tea tree has grown at least one bud and two leaves.

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