Cultivation method for improving effective components of golden camellia

Abstract

The invention provides a cultivation method for improving effective components of camellia nitidissima, and relates to the technical field of camellia nitidissima cultivation. The method comprises the steps that 1, a biological treatment agent A is applied to the roots of camellia nitidissima flower buds in the physiological differentiation period in a watering mode, and the biological treatment agent A is formed by compounding dark partitioned endophytic fungus HS40 fermentation liquor and a bamboo leaf extracting solution according to the volume ratio of (1-3): 1; 2) spraying a biological treatment agent B on leaf surfaces 30-45 days before harvesting, wherein the biological treatment agent B is prepared by performing enzymolysis on camellia nitidissima flower extraction residues; (3) directional pruning is conducted before sprouting in spring, so that the included angle between a main branch and a trunk is kept at 45-60 degrees, and it is guaranteed that the light transmittance of a tree crown is not lower than 40%. According to the method, two biological treatment agents are applied in a specific period and combined with directional pruning, all technical means act synergistically, the content of total saponins and total flavonoids in the golden camellia leaves is effectively increased, and meanwhile resource recycling of agricultural by-products is achieved.

Description

Technical Field

[0001] This invention relates to the field of Camellia chrysantha cultivation technology, specifically to a cultivation method for enhancing the effective components of Camellia chrysantha. Background Technology

[0002] Golden camellia tea As a rare and precious plant of the Camellia family unique to my country, the golden camellia is highly prized for its golden flowers and is known as the "Queen of Teas," possessing both high ornamental and medicinal value. In recent years, with its health benefits becoming increasingly recognized, the market demand for golden camellia has continued to grow, driving the development of the artificial cultivation industry.

[0003] The core value of golden camellia lies primarily in the various active ingredients contained within its leaves. Tests show that golden camellia is rich in tea polysaccharides, tea polyphenols, total saponins, total flavonoids, tea pigments, as well as various vitamins, amino acids, and trace elements (such as selenium, germanium, zinc, and vanadium). These components endow golden camellia with multiple physiological functions, including lowering blood sugar, lowering blood lipids, lowering cholesterol, anti-oxidation, and enhancing immunity. Therefore, the quality of golden camellia largely depends on the content of key active ingredients such as saponins and flavonoids in its leaves.

[0004] However, existing cultivation techniques often have significant limitations in practice. Many cultivation measures focus on promoting vegetative growth and increasing fresh leaf yield, such as optimizing nitrogen, phosphorus, and potassium fertilization or using general-purpose organic fertilizers. While these methods can increase biomass to some extent, they often fail to effectively and specifically increase the content of secondary metabolites such as saponins and flavonoids in the leaves. Sometimes, excessive vegetative growth may even "dilute" the concentration of active ingredients, resulting in the medicinal quality failing to reach the ideal state. Furthermore, Camellia chrysantha itself has high requirements for environmental conditions, preferring a semi-shaded, cool, and humid environment. In artificial cultivation, if the environment (such as light transmittance) is unsuitable, it may affect its growth and the accumulation of active ingredients, thus limiting the full realization of its medicinal potential.

[0005] Therefore, there is an urgent need in this field for a cultivation method that can directly and significantly enrich the core effective components of Camellia chrysantha, in order to break through the quality bottleneck of the current industry development. Summary of the Invention

[0006] The purpose of this invention is to provide a cultivation method that enhances the effective components of Camellia chrysantha. By applying a specific compound biological treatment agent A during the physiological differentiation period of Camellia chrysantha flower buds and applying biological treatment agent B during the critical period before harvesting, the technical problem of the existing cultivation technology being unable to target and enrich key active ingredients is effectively solved.

[0007] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0008] A cultivation method for enhancing the effective components of Camellia chrysantha includes the following steps:

[0009] (1) During the physiological differentiation period of the flower buds of Camellia chrysantha, apply biological treatment agent A once by watering the roots;

[0010] (2) Apply biological treatment agent B by foliar spraying 30-45 days before harvesting the golden flower tea leaves;

[0011] Wherein, step (1) biological treatment agent A contains fermentation broth of dark-colored septate endophytic fungi HS40 and bamboo leaf extract; biological treatment agent B contains enzymatic hydrolysate of extract residue from Camellia chrysantha flowers.

[0012] The aforementioned dark-colored septate endophytic fungus HS40 has the accession number CGMCC NO.14345. This strain was previously disclosed in the applicant's earlier invention patent application, application number 201710833508.3.

[0013] The physiological differentiation period of flower buds in the Guinan region is from mid to late June, and the 30-45 days before harvest in the Guinan region is from early to mid-September.

[0014] In this invention, the biological treatment agent A is further prepared by compounding a fermentation broth of dark-colored septate endophytic fungus HS40 with a bamboo leaf extract at a volume ratio of (1-3):1; the viable count of the HS40 fermentation broth is not less than 1×10⁻⁶. 8 CFU / mL;

[0015] In this invention, the biological treatment agent A is further diluted 50-100 times when applied, and the amount of diluted solution applied per plant is 10-20L.

[0016] In this invention, the bamboo leaf extract is further prepared by the following method: dried bamboo leaves are pulverized, and extracted by reflux at 70-80°C for 2-3 hours with 60-70% ethanol solution as solvent at a material-to-liquid ratio of 1:(15-20). After filtration, the extract is concentrated until there is no alcohol odor.

[0017] In this invention, the biological treatment agent B is further prepared by the following method: the wet residue obtained after extracting Camellia chrysantha flowers is mixed with water at a mass ratio of 1:(10-15), the pH is adjusted to 4.5-5.5, 1-2% of the total mass of the system of compound cellulase is added, and the mixture is enzymatically hydrolyzed at 45-55℃ for 2-4 hours. After enzyme inactivation, the mixture is filtered, and the supernatant is obtained.

[0018] In this invention, the biological treatment agent B is further diluted 200-300 times when applied, and the amount sprayed per plant is 0.5-1.5L of diluted solution.

[0019] Furthermore, in this invention, the method also includes directional pruning of the Camellia chrysantha plant before the spring shoots emerge. The directional pruning specifically includes pulling the main branches to maintain the angle between the main branches and the main trunk at 45°-60°, and removing crossing branches, overlapping branches, and upright overgrown branches in the inner canopy.

[0020] Furthermore, in this invention, after the directional pruning, the light transmittance of the tree canopy is guaranteed to be no less than 40%.

[0021] The present invention also discloses a special composition for enhancing the effective components of Camellia chrysantha, which consists of biostimulant A and biostimulant B;

[0022] Wherein, biostimulant A is a complex compounded by fermentation broth of dark-colored septate endophytic fungus HS40 and bamboo leaf flavonoid extract in a volume ratio of (1-3):1; biostimulant B is an enzymatic hydrolysate obtained by biological enzymatic hydrolysis of extract residue from Camellia chrysantha flowers.

[0023] Compared with the prior art, the present invention has at least the following advantages:

[0024] 1. This invention achieves effective regulation of the secondary metabolism of Camellia chrysantha tea leaves by applying a specific compound treatment agent at a specific growth stage. Experimental data shows that the method of this invention can increase the total saponin and total flavonoid content in Camellia chrysantha tea leaves by more than 40% compared with conventional cultivation methods, effectively solving the technical problem of the difficulty in targeted enrichment of key active ingredients in existing cultivation techniques. Specifically, this invention produces synergistic effects through the combined application of biological treatment agent A, the precise timing of spraying biological treatment agent B, and the physical regulation of targeted pruning. The data shows that the complete technical solution (experimental group) significantly increases the effective ingredient content by exceeding any single technical means such as applying HS40 fermentation liquid alone, applying bamboo leaf extract alone, applying biological treatment agent B alone, or performing targeted pruning alone. It also significantly exceeds the incomplete solutions that change the timing of biological treatment agent B application or omit the pruning step. This proves that the various technical features in the cultivation method provided by this invention are interconnected and work together, forming an inseparable organic whole.

[0025] 2. The treatment agent of this invention includes treatment agent A, which is applied to the roots during the physiological differentiation period of flower buds. This treatment agent is a compound of fermentation broth of dark-colored septate endophytic fungus HS40 and bamboo leaf extract at a specific volume ratio (1-3):1. This specific compounding scheme has a more significant effect on enhancing the effective components compared to using either component alone. The applicant believes that this may be because the active substances in the HS40 fermentation broth and bamboo leaf extract work together on the plant, synergistically activating the secondary metabolic system of Camellia chrysantha through different signaling pathways, promoting the secondary metabolic process of Camellia chrysantha, and thus more effectively increasing the accumulation of saponins and flavonoids.

[0026] 3. The treatment agent of this invention also includes treatment agent B, which is applied 30-45 days before the harvest of Camellia chrysantha tea leaves. Firstly, in terms of raw materials, it creatively selects the residue extracted from Camellia chrysantha flowers themselves, rather than other plant materials or tea dregs. This choice not only achieves high-value recycling of processing by-products, but more importantly, the applicant discovered that the enzymatic hydrolysate derived from the same Camellia chrysantha flowers may have a higher affinity and matching degree with the secondary metabolic pathways of the leaves, thus enabling more precise stimulation of the synthesis of target components. Secondly, in terms of timing, applying it during this specific window before harvest coincides precisely with the peak period of natural accumulation of effective components in Camellia chrysantha. This precise synchronization of "self-signaling substances" and "endogenous metabolic rhythms" achieves a highly efficient linkage between exogenous induction and endogenous enrichment, thereby obtaining the best component enrichment effect that is difficult to achieve with a single method.

[0027] 4. Finally, this invention involves directional pruning of the Camellia chrysantha plants before spring shoot emergence. This is achieved by pulling the branches to maintain the angle between the main branches and the trunk at 45°-60°, and thinning out overly dense branches in the inner canopy to ensure that the canopy light transmittance is not less than 40%. This pruning method improves canopy light and ventilation, creating a favorable physical environment for healthy plant growth and the accumulation of active ingredients. Experimental data demonstrates that this pruning step, combined with the application of biological treatment agents, makes an indispensable contribution to enhancing the final effective ingredient content. Together with other features in the technical solution, it constitutes a complete and efficient cultivation program.

[0028] In summary, this invention effectively increases the content of core medicinal components in Camellia chrysantha by organically combining two biological treatment agents applied at specific times with targeted pruning measures, while simultaneously achieving the recycling of agricultural resources. Detailed Implementation

[0029] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments and comparative examples. It should be understood that the specific embodiments described herein are for illustrative purposes only and are not intended to limit the invention.

[0030] Example 1:

[0031] This embodiment discloses a cultivation method for enhancing the effective components of Camellia chrysantha, comprising the following steps:

[0032] (1) Application of biological treatment agent A: During the physiological differentiation period of flower buds of Camellia chrysantha, biological treatment agent A is applied once by watering the roots. Biological treatment agent A is obtained by mixing fermentation broth of dark-colored septate endophytic fungus HS40 (preservation number CGMCC NO.14345) with bamboo leaf extract at a volume ratio of 1:1. The bamboo leaf extract is prepared by pulverizing dried bamboo leaves, using 60-70% ethanol solution as solvent, and refluxing at 70℃ for 2 hours at a material-to-liquid ratio of 1:15. After filtration, the extract is concentrated until no alcohol odor remains. The viable count of the HS40 fermentation broth is not less than 1×10⁻⁶. 8 CFU / mL; Biological treatment agent A is diluted 50 times when applied, and the amount of diluted solution applied per plant is 10L.

[0033] (2) Application of biological treatment agent B: 30 days before the harvest of Camellia chrysantha tea leaves, apply biological treatment agent B by foliar spraying once. The biological treatment agent B is prepared by the following method: The wet residue obtained after extracting Camellia chrysantha flowers is mixed with water at a mass ratio of 1:10, the pH is adjusted to 4.5, 1% of the total mass of compound cellulase is added, and the mixture is enzymatically hydrolyzed at 45℃ for 2 hours. After enzyme inactivation, the mixture is filtered, and the supernatant is obtained. The biological treatment agent B is diluted 200 times during application, and the spraying amount per plant is 0.5L of diluted solution.

[0034] (3) Pruning: From late February to mid-March (i.e., before spring shoot emergence), perform directional pruning on the Camellia chrysantha plants. Specifically, directional pruning includes pulling the main branches to maintain the angle between the main branches and the main trunk at 45°-60°, and removing crossing branches, overlapping branches, and upright overgrown branches in the inner canopy. After directional pruning, ensure that the light transmittance of the canopy is not less than 40%.

[0035] Example 2:

[0036] This embodiment discloses a cultivation method for enhancing the effective components of Camellia chrysantha, comprising the following steps:

[0037] (1) Application of biological treatment agent A: During the physiological differentiation period of flower buds of Camellia chrysantha, biological treatment agent A is applied once by watering the roots. Biological treatment agent A is obtained by mixing the fermentation broth of dark-colored septate endophytic fungus HS40 (preservation number CGMCC NO.14345) with bamboo leaf extract at a volume ratio of 2:1. The bamboo leaf extract is prepared by pulverizing dried bamboo leaves, using 65% ethanol solution as solvent, and refluxing at 75°C for 2.5 hours at a material-to-liquid ratio of 1:18. After filtration, the extract is concentrated until no alcohol odor remains. The viable count of the HS40 fermentation broth is not less than 1×10⁻⁶. 8 CFU / mL; Biological treatment agent A is diluted 75 times when applied, and the amount of diluted solution applied per plant is 15L.

[0038] (2) Application of biological treatment agent B: 38 days before the harvest of Camellia chrysantha tea leaves, apply biological treatment agent B by foliar spraying once. The biological treatment agent B is prepared by the following method: The wet residue obtained after extracting Camellia chrysantha flowers is mixed with water at a mass ratio of 1:12, the pH is adjusted to 5.0, and 1.5% of a compound cellulase is added to the mixture. The mixture is then enzymatically hydrolyzed at 50°C for 3 hours. After enzyme inactivation, the mixture is filtered, and the supernatant is obtained. The biological treatment agent B is diluted 250 times during application, and the spraying amount per plant is 1.0 L of diluted solution.

[0039] (3) Pruning: From late February to mid-March (i.e., before spring shoot emergence), perform directional pruning on the Camellia chrysantha plants. Specifically, directional pruning includes pulling the main branches to maintain the angle between the main branches and the main trunk at 45°-60°, and removing crossing branches, overlapping branches, and upright overgrown branches in the inner canopy. After directional pruning, ensure that the light transmittance of the canopy is not less than 40%.

[0040] Example 3:

[0041] This embodiment discloses a cultivation method for enhancing the effective components of Camellia chrysantha, comprising the following steps:

[0042] (1) Application of biological treatment agent A: During the physiological differentiation period of flower buds of Camellia chrysantha, biological treatment agent A is applied once by watering the roots. Biological treatment agent A is obtained by mixing fermentation broth of dark-colored septate endophytic fungus HS40 (preservation number CGMCC NO.14345) with bamboo leaf extract at a volume ratio of 3:1. The bamboo leaf extract is prepared by pulverizing dried bamboo leaves, using 70% ethanol solution as solvent, and refluxing at 80℃ for 3 hours at a material-to-liquid ratio of 1:20. After filtration, the extract is concentrated until no alcohol odor remains. The viable count of the HS40 fermentation broth is not less than 1×10⁻⁶. 8 CFU / mL; Biological treatment agent A is diluted 100 times when applied, and the amount of diluted solution applied per plant is 20L.

[0043] (2) Application of biological treatment agent B: 45 days before the harvest of Camellia chrysantha tea leaves, apply biological treatment agent B by foliar spraying once. The biological treatment agent B is prepared as follows: The wet residue obtained after extracting Camellia chrysantha flowers is mixed with water at a mass ratio of 1:15, the pH is adjusted to 5.5, 2% (by mass) of a compound cellulase is added, and the mixture is enzymatically hydrolyzed at 55°C for 4 hours. After enzyme inactivation, the mixture is filtered, and the supernatant is obtained. The biological treatment agent B is diluted 300 times during application, and the amount sprayed per plant is 1.5 L of diluted solution.

[0044] (3) Pruning: From late February to mid-March (i.e., before spring shoot emergence), perform directional pruning on the Camellia chrysantha plants. Specifically, directional pruning includes pulling the main branches to maintain the angle between the main branches and the main trunk at 45°-60°, and removing crossing branches, overlapping branches, and upright overgrown branches in the inner canopy. After directional pruning, ensure that the light transmittance of the canopy is not less than 40%.

[0045] Experimental example:

[0046] To demonstrate the practical value of this application, the applicant conducted a cultivation experiment at the Camellia chrysantha Planting Resource Nursery (Nanning), Institute of Flower Research, Guangxi Academy of Agricultural Sciences. The test material consisted of four-year-old 'Fangcheng No. 2' Camellia chrysantha seedlings with uniform growth. The experiment lasted for one complete growth cycle (February 2023 to October 2023).

[0047] The experimental groups are as follows:

[0048] Control group: Conventional cultivation and management (equal amount of water applied, no directional pruning).

[0049] Comparative Example 1: CK + HS40 fermentation broth applied alone (diluted 75 times, application time and method are the same as in Example 2 of this invention).

[0050] Comparative Example 2: CK + Bamboo leaf extract alone (diluted 75 times, application time and method are the same as in Example 2 of this invention).

[0051] Comparative Example 3: CK + biological treatment agent B applied alone (application timing and method are the same as in Example 2 of this invention).

[0052] Comparative Example 4: Only directional pruning was performed (the pruning method was the same as in Example 2 of this invention), but no biological treatment agent was applied.

[0053] Comparative Example 5: The complete scheme of Example 2 of the present invention was adopted, but the spraying time of biological treatment agent B was advanced to the same period as biological treatment agent A (mid-to-late June).

[0054] Comparative Example 6: The complete scheme of Embodiment 2 of the present invention is adopted, but the spring directional pruning step is omitted.

[0055] Experimental group: The complete scheme of Example 2 of this invention was adopted (i.e., directional pruning + application of compound biological treatment agent A during the physiological differentiation period of flower buds + application of biological treatment agent B 38 days before harvest).

[0056] Each group had 3 replicates, with 10 plants per replicate. Except for the emphasized differences, all other methods were the same for each group. Mature, healthy leaves from the middle section of plants in each treatment group were harvested uniformly on October 20, 2023. After treatment, the total saponin and total flavonoid content (mg / g) in the leaves was determined. The mean ± standard deviation was recorded, and the results are shown in Table 1.

[0057] Table 1 Comparison of effective ingredient content in Golden Flower Tea Leaves of Different Groups

[0058]

[0059] This indicates that the difference compared with the control group was statistically significant (P < 0.05).

[0060] The data in Table 1 clearly demonstrate that the method of this invention can effectively increase the main active ingredients of golden chrysanthemum tea leaves. The total saponin and total flavonoid content in the experimental group of this invention were significantly higher than those in the control group and any single treatment group (Comparative Examples 1-4), indicating that a synergistic effect was produced between the compound components of biological treatment agent A, biological treatment agent B, and the targeted pruning measures. Although the active ingredient content in Comparative Example 5 (incorrect timing) and Comparative Example 6 (no pruning) was better than that of the single treatment, it was significantly lower than that in the experimental group of the complete scheme. This proves that precise application timing and targeted pruning are indispensable key links in the technical solution of this invention, and their absence will lead to a significant decrease in the final effect.

[0061] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A cultivation method for enhancing the effective components of Camellia chrysantha, characterized in that, Includes the following steps: (1) During the physiological differentiation period of the flower buds of Camellia chrysantha, apply biological treatment agent A once by watering the roots; (2) Apply biological treatment agent B by foliar spraying 30-45 days before harvesting the golden flower tea leaves; Wherein, step (1) biological treatment agent A contains fermentation broth of dark-colored septate endophytic fungi HS40 and bamboo leaf extract; biological treatment agent B contains enzymatic hydrolysate of extract residue from Camellia chrysantha flowers.

2. The method according to claim 1, characterized in that, The biological treatment agent A is obtained by compounding the fermentation broth of dark-colored septate endophytic fungi HS40 with bamboo leaf extract at a volume ratio of (1-3):1; the viable count of the HS40 fermentation broth is not less than 1×10⁻⁶. 8 CFU / mL.

3. The method according to claim 1, characterized in that, The biological treatment agent A is diluted 50-100 times when applied, and the amount of diluted solution applied per plant is 10-20L.

4. The method according to claim 1, characterized in that, The bamboo leaf extract is prepared by the following method: dried bamboo leaves are pulverized, and extracted by reflux at 70-80°C for 2-3 hours with 60-70% ethanol solution as solvent at a material-to-liquid ratio of 1:(15-20). After filtration, the extract is concentrated until there is no alcohol taste.

5. The method according to claim 1, characterized in that, The biological treatment agent B is prepared by the following method: the wet residue obtained after extracting Camellia chrysantha flowers is mixed with water at a mass ratio of 1:(10-15), the pH is adjusted to 4.5-5.5, 1-2% of the total mass of the system of compound cellulase is added, and the mixture is enzymatically hydrolyzed at 45-55℃ for 2-4 hours. After enzyme inactivation, the mixture is filtered and the supernatant is obtained.

6. The method according to claim 2, characterized in that, The biological treatment agent B is diluted 200-300 times when applied, and the amount sprayed per plant is 0.5-1.5L of diluted solution.

7. The method according to claim 1, characterized in that, The method also includes directional pruning of the Camellia chrysantha plant before the spring shoots emerge. The directional pruning specifically includes pulling the main branches to maintain the angle between the main branches and the main trunk at 45°-60°, and removing crossing branches, overlapping branches and upright overgrown branches in the inner canopy.

8. The method according to claim 7, characterized in that, After the directional pruning, the light transmittance of the tree canopy is guaranteed to be no less than 40%.

9. A special composition for enhancing the effective components of Camellia chrysantha, characterized in that, It is composed of biostimulant A and biostimulant B; Wherein, biostimulant A is a complex compounded by fermentation broth of dark-colored septate endophytic fungus HS40 and bamboo leaf flavonoid extract in a volume ratio of (1-3):1; biostimulant B is an enzymatic hydrolysate obtained by biological enzymatic hydrolysis of extract residue from Camellia chrysantha flowers.

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