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

[0002] Jinxuan, also known as Taiwan Tea No. 12, was selected and bred by the Taiwan Tea Improvement Station in China and was recognized as a provincial tea variety in 1981. This variety is a shrub type, medium-leaf type, with strong fertility and neat buds and leaves, suitable for making black tea, green tea, and oolong tea. At present, the Jinxuan tea variety has the largest cultivation area in Taiwan, Fujian, Guangdong, and Guangxi, and its adaptability and resistance are relatively strong. The red, green, and oolong teas made from the Jinxuan tea variety have good quality characteristics and are deeply loved by consumers in the market. The black tea made from it has the following characteristics: the color of the dry tea is dark and moist with golden hair, the soup color is bright red, the floral fragrance is long-lasting, and the taste is sweet and refreshing. The green tea made from it has the following characteristics: the color is green and moist with obvious hair, the aroma is long and high, the soup color is clear and bright green, the taste is fresh and smooth, and the bottom of the leaves is bright green. The finished oolong tea has the following characteristics: the dry tea is dark green and glossy, the grains are compact and heavy, the aroma is rich, the variety is fragrant, with elegant milk or fresh flower fragrance, mellow taste, bright honey yellow soup color, and soft leaves. In addition, the Jinxuan tea tree variety is also the backbone parent for the breeding of black tea, green tea and oolong tea varieties in my country, and an excellent resource for breeding high-quality and special new varieties.

[0003] EGCG (epigallocatechin gallate) is an important secondary metabolite in tea trees, with significant biological functions such as antioxidant, antibacterial, anti-inflammatory and antiviral. Although EGCG can also be prepared by artificial synthesis, under natural conditions, tea trees are the main biological source of EGCG. In recent years, tea tree breeders have been committed to increasing the EGCG content through two main approaches: one is to screen natural high-EGCG tea tree resources, and the other is to develop exogenous induction technology to promote tea trees to synthesize more EGCG.

[0004] 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 have certain toxicity 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 decreased resistance of tea trees. In addition, this method requires continuous spraying for at least 4 days, which is time-consuming.

[0005] In view of these defects 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

[0006] The technical problem to be solved by the present invention is to overcome the defects and shortcomings of the existing induction reagents for increasing the EGCG content of tea trees, such as certain toxicity and time-consuming use, and to provide an application of an exogenous substance adenosine triphosphate in promoting the biosynthesis of EGCG in tea trees.

[0007] The invention aims to provide an exogenous induction method for increasing the content of EGCG (epigallocatechin gallate) in tea plants.

[0008] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0009] The invention protects the application of exogenous substance adenosine triphosphate (ATP) in promoting the biosynthesis of EGCG in tea plants.

[0010] ATP is used as a coenzyme drug in clinical practice, and is often used to treat heart failure, myocarditis, cerebral arteriosclerosis and other diseases. It can improve the patient's symptoms and quality of life by improving cell metabolism and providing energy support, promoting the repair and regeneration of damaged cells. The inventor creatively discovered that applying exogenous ATP to tea trees can promote the synthesis of EGCG in tea trees and increase the content of EGCG in tea trees.

[0011] Furthermore, the varieties of the tea trees include one or more of Jinxuan (Taiwan Tea No. 12), Fuding Dabaicha (Hua Tea No. 1), Tieguanyin (Hua Tea No. 7), and Yinghong No. 9.

[0012] Preferably, the concentration of ATP is 0.05-10 mM. ATP of different concentrations is prepared by diluting with water.

[0013] More preferably, the concentration of ATP is 0.1-2 mM.

[0014] The present invention also protects an exogenous induction method for increasing the EGCG content of tea trees, comprising the following steps: applying an exogenous substance adenosine triphosphate to the tea trees for a certain period of time to achieve the purpose of increasing the EGCG content.

[0015] The inventors found that applying exogenous ATP to tea trees can promote the synthesis of EGCG in tea trees and increase the content of EGCG in tea trees. This method can not only avoid the use of toxic chemicals, but also shorten the induction time to ensure environmental friendliness and tea tree health.

[0016] Preferably, the concentration of ATP is 0.05-10 mM.

[0017] More preferably, the concentration of ATP is 0.1-2 mM.

[0018] Preferably, the duration after applying is 1 to 25 hours.

[0019] More preferably, the duration after applying is 4 to 24 hours.

[0020] Furthermore, the application method is spraying.

[0021] 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.

[0022] Furthermore, the exogenous substance adenosine triphosphate is applied when the tea tree has grown at least one bud and two leaves.

[0023] Furthermore, the applied dosage is 0.05 to 0.2 liters / m 2 , more preferably 0.08 to 0.12 liters / m 2 .

[0024] Furthermore, the EGCG content is detected by high performance liquid chromatography.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The present invention provides an exogenous induction method for increasing the EGCG content of tea trees, which specifically comprises the following steps: applying exogenous substance adenosine triphosphate to tea trees, and continuing the application for a certain period of time to achieve the purpose of increasing the EGCG content. The present invention can increase the EGCG content of tea trees including the Jinxuan tea tree variety to 0.5% to 28%. The method can use simple means and easily available safe materials to enable tea trees to accumulate more EGCG in a short period of time, which is of great significance to the processing and deep processing of tea leaves. DETAILED DESCRIPTION

[0027] The present invention is further described below with reference to specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0028] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0029] 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 (0.1mM, 1mM, 2mM, 5mM, 10mM; specifically, 10mM ATP was first prepared (dissolved in pure water), and then diluted downward according to the concentration gradient, and the final concentrations of ATP solutions were 0.1mM, 1mM, 2mM, 5mM, and 10mM, respectively) were prepared using a spray pot, and the spraying dose was 0.1L / m 2 The picking time was 6h, 12h and 1d (day) respectively. One bud and two leaves were picked for high performance liquid chromatography to detect the EGCG content.

[0030] The HPLC detection conditions were as follows: C18 column, column temperature of 35°C, injection volume of 10 μL, mobile phase A was methanol, mobile phase B was 1% trifluoroacetic acid aqueous solution, flow rate of 1 mL / min, detection wavelength of 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 maintained for 5 min; 55-60 min, A increased to 35%.

[0031] Example 1 An exogenous induction method for increasing the EGCG content of Jinxuan tea variety

[0032] Select the Jinxuan tea garden with good growth 10m 2 One bud and two leaves of Jinxuan tea tree were picked 6 hours after spraying with pure water, and the solid sample was freeze-dried. The EGCG content was 79.88 mg / g by high performance liquid chromatography detection, which was used as the control for exogenous induction treatment.

[0033] 10m from the above Jinxuan Tea Garden 2 In the experiment, one bud and two leaves of Jinxuan tea tree were picked 6 hours after spraying 0.1mM ATP, and the solid samples were freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 85.49mg / g, an increase of 7.02% compared with before exogenous induction treatment.

[0034] Example 2 An exogenous induction method for increasing the EGCG content of Jinxuan tea variety

[0035] Select the Jinxuan tea garden with good growth 10m 2 One bud and two leaves of Jinxuan tea tree were picked 6 hours after spraying with pure water, and the solid sample was freeze-dried. The EGCG content was 79.88 mg / g by high performance liquid chromatography detection, which was used as the control for exogenous induction treatment.

[0036] 10m from the above Jinxuan Tea Garden 2In the experiment, one bud and two leaves of Jinxuan tea tree were picked 6 hours after spraying 1mM ATP, and the solid samples were freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 86.20mg / g, an increase of 7.92% compared with that before exogenous induction treatment.

[0037] Example 3: An exogenous induction method for increasing the EGCG content of Jinxuan tea variety

[0038] Select the Jinxuan tea garden with good growth 10m 2 One bud and two leaves of Jinxuan tea tree were picked 6 hours after spraying with pure water, and the solid sample was freeze-dried. The EGCG content was 79.88 mg / g by high performance liquid chromatography detection, which was used as the control for exogenous induction treatment.

[0039] 10m above the Jinxuan Tea Garden 2 In the experiment, one bud and two leaves of Jinxuan tea tree were picked 6 hours after spraying 2mM ATP, and the solid samples were freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 97.24mg / g, an increase of 21.73% compared with that before exogenous induction treatment.

[0040] Example 4: An exogenous induction method for increasing the EGCG content of Jinxuan tea variety

[0041] Select the Jinxuan tea garden with good growth 10m 2 One bud and two leaves of Jinxuan tea tree were picked 12 hours after spraying pure water, and the solid sample was freeze-dried. The EGCG content was 86.20 mg / g by high performance liquid chromatography detection, which was used as the control for exogenous induction treatment.

[0042] 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 0.1mM ATP, and the solid samples were freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 96.18mg / g, an increase of 11.59% compared with that before exogenous induction treatment.

[0043] Example 5: An exogenous induction method for increasing the EGCG content of Jinxuan tea variety

[0044] Select the Jinxuan tea garden with good growth 10m 2 One bud and two leaves of Jinxuan tea tree were picked 12 hours after spraying pure water, and the solid sample was freeze-dried. The EGCG content was 86.20 mg / g by high performance liquid chromatography detection, which was used as the control for exogenous induction treatment.

[0045] 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 1mM ATP, and the solid samples were freeze-dried. The EGCG content was 101.762mg / g by HPLC, which was 18.05% higher than that before exogenous induction treatment.

[0046] Example 6 An exogenous induction method for increasing the EGCG content of Jinxuan tea variety

[0047] Select the Jinxuan tea garden with good growth 10m 2 One bud and two leaves of Jinxuan tea tree were picked 12 hours after spraying pure water, and the solid sample was freeze-dried. The EGCG content was 86.20 mg / g by high performance liquid chromatography detection, which was used as the control for exogenous induction treatment.

[0048] 10m from the above Jinxuan Tea Garden 2 In one mu, fresh leaves with one bud and two leaves from Jinxuan tea tree were picked 12 hours after spraying 2mM ATP, and the solid samples were freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 110.17 mg / g, an increase of 27.81% compared with before exogenous induction treatment.

[0049] Example 7 An exogenous induction method for increasing the EGCG content of Jinxuan tea variety

[0050] Select the Jinxuan tea garden with good growth 10m 2 One bud and two leaves of Jinxuan tea tree were picked one day after spraying pure water, and the solid sample was freeze-dried. The EGCG content was 106.71 mg / g by high performance liquid chromatography detection, which was used as the control for exogenous induction treatment.

[0051] 10m from the above Jinxuan Tea Garden 2 In the experiment, one bud and two leaves of Jinxuan tea tree were picked one day after spraying 0.1mM ATP, and the solid samples were freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 119.95mg / g, an increase of 12.41% compared with that before exogenous induction treatment.

[0052] Example 8 An exogenous induction method for increasing the EGCG content of Jinxuan tea variety

[0053] Select the Jinxuan tea garden with good growth 10m 2 One bud and two leaves of Jinxuan tea tree were picked one day after spraying pure water, and the solid sample was freeze-dried. The EGCG content was 106.71 mg / g by high performance liquid chromatography detection, which was used as the control for exogenous induction treatment.

[0054] 10m from the above Jinxuan Tea Garden 2 In the experiment, one bud and two leaves of Jinxuan tea tree were picked one day after spraying 1mM ATP, and the solid samples were freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 122.92 mg / g, an increase of 15.19% compared with that before exogenous induction treatment.

[0055] Example 9: An exogenous induction method for increasing the EGCG content of Jinxuan tea variety

[0056] Select the Jinxuan tea garden with good growth 10m2 One bud and two leaves of Jinxuan tea tree were picked one day after spraying pure water, and the solid sample was freeze-dried. The EGCG content was 106.71 mg / g by high performance liquid chromatography detection, which was used as the control for exogenous induction treatment.

[0057] 10m from the above Jinxuan Tea Garden 2 In the experiment, one bud and two leaves of Jinxuan tea tree were picked one day after spraying 2mM ATP, and the solid samples were freeze-dried. The EGCG content was detected by high performance liquid chromatography, which was 107.27 mg / g, an increase of 0.52% compared with that before exogenous induction treatment.

[0058] The test results of Examples 1 to 9 are summarized as shown in Table 1.

[0059] Table 1 Effects of different ATP concentrations and different treatment times on the EGCG content of Jinxuan tea variety

[0060]

[0061] Note: All samples were picked from the Jinxuan variety of the tea garden of South China Agricultural University. One bud and two leaves were picked from different Jinxuan tea trees for testing, with a total of 3 biological replicates, and each sample was technically replicated 3 times, and the average value was taken as the test result.

[0062] The test results are shown in Table 1. The ATP spraying concentration of 0.1-2mM can increase the EGCG content of tea plants, and the EGCG content increased 6h, 12h, and 1d after spraying. Specifically, when the ATP spraying concentration is 0.1mM, the EGCG content increased significantly after 12h-1d of spraying, with an increase of 11%-13%; when the ATP spraying concentration is 1mM, the EGCG content increased significantly after 12h-1d of spraying, with an increase of 15%-19%; when the ATP spraying concentration is 2mM, the EGCG content increased significantly after 6-12h of spraying, with an increase of 21%-28%.

[0063] 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 equivalent replacement methods and are included in the protection scope 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 application 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 adenosine triphosphate 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 continuing the application for a certain period of time to achieve the purpose of increasing the EGCG content.

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

7. The method according to claim 6, characterized in that: The concentration of adenosine triphosphate is 0.1-2 mM.

8. The method according to claim 5, characterized in that: The duration after applying is 1 to 25 hours.

9. The method according to claim 8, characterized in that: 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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